Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx 1

Contents lists available at ScienceDirect

European Journal of Pharmaceutical Sciences journal homepage: www.elsevier.com/locate/ejps

2

Review

6 4 7

Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force

5 8

Q1

9 10 11 12 13 14 15

a

Drug Delivery Research Group, University Institute of Pharmaceutical Sciences, Panjab University, 160 014 Chandigarh, India Department of Pharmaceutical Sciences, Dr. H.S. Gour University, Sagar 470 003, MP, India Department of Biotechnology, TRS College, Rewa 486001, MP, India d Department of Periodontics, College of Dental Medicine, Georgia Regents University, Augusta, GA, USA e Department of Biotechnology, IMS Engineering College, Ghaziabad, UP Technical University, UP, India f National Institute of Allergy and Infectious Diseases, National Institutes of Health, TW3/3W15, 12735 Twinbrook Pkwy, Rockville, MD, USA b c

Q2

16 17

a r t i c l e

1 4 9 0 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39

Neeraj K. Garg a,b,⇑,1, Priya Dwivedi c,1, Ashay Jain a,b, Shikha Tyagid d, Rupali Suhag e, Tejram Sahu f, Rajeev K. Tyagi d,⇑

i n f o

Article history: Received 9 December 2013 Received in revised form 5 April 2014 Accepted 28 May 2014 Available online xxxx

Q3

Keywords: Tuberculosis T Cell B Cell Cytokines Liposomes Microparticle Nanoparticles ISCOMS Mucosal vaccine PLGA Chitosan CAF01 DOTAP

a b s t r a c t Despite worldwide availability of the vaccines against most of the infectious diseases, BCG and various programs such as Directly Observed Treatment Short course (DOTS) to prevent tuberculosis still remains one of the most deadly forms of the disease affecting millions of people globally. The evolution of multi drug resistant strains (MDR) has increased the complexity further. Although currently available marketed BCG vaccine has shown sufficient protection against childhood tuberculosis, it has failed to prevent the most common form of disease i.e., pulmonary tuberculosis in adults. However, various vaccine candidates have already entered phase I clinical trials and have shown promising outcomes. The most prominent amongst them is the heterologous prime-boost approach, which shows a great promise towards designing and development of a new efficacious tuberculosis vaccine. It has also been shown that the use of various viral and non-viral vectors as carriers for the potential vaccine candidates will further boost their effect on subsequent immunization. In this review, we briefly summarize the potential of a few novel nano-carriers for developing effective vaccination strategies against tuberculosis. Ó 2014 Published by Elsevier B.V.

41 42 43 44 45 46 47 48 49 50 51 52 53 54

55 56 57

1. Background: epidemiology and control of tuberculosis

58

Tuberculosis (TB), a contagious infectious disease caused by aerosol infection with Mycobacterium tuberculosis (Mtb), is one of the major health issues worldwide that kills millions of people globally every year. Recently, World Health Organization (WHO) published global tuberculosis report 2012, and in 2011 there were

59 60 61 62

⇑ Corresponding authors. Address: Drug Delivery Research Group, University Institute of Pharmaceutical Sciences, Panjab University, 160 014 Chandigarh, India. Tel.: +91 9329615027 (N.K. Garg). Address: Department of Periodontics, College of Dental Medicine, Georgia Regents University, 1120 15th Street, CB 2716, Augusta, GA 30912, USA. Tel.: +1 706 721 8622; fax: +1 706 723 0215 (R.K. Tyagi). E-mail addresses: [email protected] (N.K. Garg), [email protected] (R.K. Tyagi). 1 These authors contributed equally to this work.

8.7 million new cases of Mtb infection (13% co-infected with HIV) and 1.4 million people died of Mtb, accompanying almost one million deaths among HIV-negative individuals and 430,000 among people who were HIV-positive. To date, the only prophylactic available against Mtb is the Bacilli–Calmette–Guerin (BCG) vaccine, an attenuated Mycobacterium bovis (M. bovis) strain that confers protection against several childhood forms of tuberculosis, but fails to prevent pulmonary tuberculosis (PTB) in adults (Meerak et al., 2013). PTB, the most contagious form of TB, requires development of a new and more efficient TB vaccine(s) as BCG vaccine is not efficient in controlling the disease. Beyond vaccines such as BCG, which are administered before tuberculosis infection, one potential strategy to eliminate or control latent tuberculosis and prevent reactivation consists of post-exposure vaccines (Andersen et al., 2007). In both cases, research efforts are directed at conferring broad protection against disease and infection, especially by

http://dx.doi.org/10.1016/j.ejps.2014.05.028 0928-0987/Ó 2014 Published by Elsevier B.V.

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 2

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

Fig. 1. Binding of Mtb by monocytes/macrophages (antigen presenting cell) through one or combination of these mechanisms. (1) Complement receptors (CR1, CR2, CR3 and CR4), (2) Mannose receptors (MR) and (3) Antibody mediated uptake of Mtb by Macrophages.

79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114

provoking cellular immune responses involving CD41 and CD81 T Q4 cells without adverse health consequences (Titball, 2008). The indepth knowledge of pharmaceutical technology and improved understanding of the immunopathology of tuberculosis led to develop candidate subunit vaccines. These vaccines are preferred because of their safety in both normal and immune compromised patients, although their inherent lack of immunogenicity requires the use of adjuvants capable of inducing a protective T-cell response (Schijns, 2003). In order to be protective against Mtb, a candidate vaccine must elicit a specific cell-mediated response, both in immune competent and in immune compromised individuals who are considered a high-risk population for tuberculosis. Selecting the appropriate adjuvant to specific antigenic components will not only enhance the level of immune response but will also determine the type of immune response induced. Although substantial progress has been made in identification and production of antigens, the most widely used adjuvants are still aluminum based compounds (generically called alum). A squalene based oil emulsion (MF59) and virosomes for use in influenza vaccines are approved for human use in Europe (Kallenius et al., 2007). Alum generally promotes weak humoral Th2 type immune responses to protein subunits, and elicits insufficient cell-mediated Th1 immune responses. The failure of alum to stimulate a cell mediated Th1 immune response, required for protective immunity against many cancers and infectious diseases has led to the search for more potent vaccine adjuvants that not only enhance the level of immune activation but also selectively induce cell-mediated immune responses. Therefore there is pre-requisite need of TB vaccine development so that above-mentioned problems such as MDR-TB and inadequate immunity in adults after BCG vaccination might be addressed objectively. Consequently, the development of adjuvants to improve tuberculosis vaccines for human use remains a challenge and is equally important to subunit vaccine formulation as antigen discovery (Hoff et al., 2007; Hoft, 2008)). Here, we review the current state of adjuvant development and its impact on tuberculosis vaccine progress.

115

2. Life cycle of M. tuberculosis (Mtb)

116

Mtb Infection follows cascade of events that have been established through human TB, as well as observations on animal models (Flynn and Chan, 2005; Russell, 2007a,b). The bacilli are

117 118

inhaled as droplet nuclei that have been exhaled into the environment. These droplets are miniature in size and adequately remain air borne for long time even for several hours (Russell, 2007a,b; Russell et al., 2010). The initial stage of infection in the lung is primarily through inference; it is believed that the bacteria are attached through different pathway (Fig. 1) and are phagocytosed by alveolar macrophages, and invades subtending epithelial layer. This causes a localized inflammatory response that leads to recruitment of mononuclear cells from neighboring blood vessels, providing fresh host cells for propagation of bacterial population. Cell-mediated immunity (CMI) is developed within 2–3 week of infection (Fig. 2), and there is an influx of lymphocytes and activated macrophages into the lesion resulting in granuloma formation, which is the defining pathologic feature of this disease (Russell et al., 2009). Primarily the granuloma is an amorphous group of monocytes, macrophages, and neutrophils; nevertheless, the macrophages differentiate into numerous specialized cell types, including foamy macrophages, multinucleated giant cells, and epithelioid macrophages. The development of an acquired immune response through arrival of lymphocytes, the granuloma acquires a more planned, stratified structure. The macrophage rich core is surrounded by a mantle of lymphocytes that may be enclosed inside a fibrous cuff which marks the periphery of the structure. Appearance of the Mtb-specific lymphocytes in about 2–3 weeks post infection marks an end of the phase of fast bacterial replication, and the onset of a ‘‘containment’’ state that is characterized by comparatively stable bacterial numbers in mice. At this moment, the granuloma is comprehensively vascularized, and cells are vigorously recruited to the site of infection. There is also an increase in the number of foamy macrophages, which may be accountable for the increase in the cellular debris in granuloma core (Russell et al., 2009). At late stages the caseous portion of the granuloma has become hypoxic (Via et al., 2008), a condition that can encourage a state of non-replicative persistence in the culture of Mtb. Infected tissues histology from immune competent patients with active TB reveals granulomas in all states of development from containment to active disease, which imply that the providence of each granuloma is determined locally, not systemically. Active form of granulomas expose extensive pathology, and ultimately, rupturing of granuloma spills thousands of viable, infectious bacilli into the airways (Kaplan et al., 2003), which results in the development of a productive cough that facilitates

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

3

Table 1 Protective immune response against tuberculosis. Protective innate and adaptive responses

Possible response on tuberculosis

Mucosal immunitya Serum and secretory Ig CD4+ ab T cellsb

All of the below functions at site of initial infection and source of transmission

CD8+ ab T cellsb

CTL recognition and destruction of Mtb-infected macrophages; inhibition of intracellular mycobacterial replication Cytokine-mediated macrophage activation; CTL recognition and destruction of infected macrophages; and helper function for CTL and antibody Cytokine-mediated macrophage activation; CTL recognition and destruction of infected macrophages; and perhaps helper function for CTL and antibody

Prevent or reduce primary infection and secondary transmission Prevent disease dissemination and infection; prevent or reduce transmission Prevent disease dissemination and reactivation; and maintenance of latency Inhibition of disease reactivation and clearance of latent infection Prevent disease dissemination and reactivation; clearance of latent infection Prevent disease dissemination and reactivation; clearance of latent infection

CD1-restricted ab T cellsc cd T cellsd

Enhanced innate immune killing of extracellular Mtb; enhanced APC stimulation of T-cell responses Cytokine-mediated macrophage activation and helper function for CTL and antibody

CTL = cytolytic T lymphocyte. APC = antigen-presenting cell. a All of the below T-cell and B-cell responses. b MHC-restricted and peptide-specific. c MHC-unrestricted and lipid-specific. d MHC-unrestricted and phosphoantigen-specific.

161 162 163 164 165 166 167 168 169

170 171 172 173 174 175 176

aerosol dissemination of infectious bacilli. One observation in human TB patients indicated that neutrophil influx in late-stage disease may also contribute to the tissue damage and the diffusion of infectious bacteria into the airways (Eum et al., 2010). Modern imaging observations on human TB stress that the equilibrium between containment and disease progression is complex and extremely dynamic, and appears to be a local phenomenon involving degree of difference in progression of granulomas within Q5 a single individual (Barry et al., 2009) (see Table 1).

3. Protective immune responses against M. Tuberculosis infection Mtb infection initially can be blocked at the surface of mucosa by secretory IgG or IgA (Hoft, 2008), or protective responses by immunoglobulins are capable of preventing binding between the surface components of mycobacterium and host receptors implicated in the initial uptake of mycobacterium lungs (Hoft, 2008). In addition, T

cells within the mucosal lumen and superficial layers of the lung mucosa could recognize and inhibit mycobacteria within the initial alveolar macrophages infected by aerosol exposure with the pathogen. Interferon-c (IFN- c) production by CD4+ Th-1 cells could initially activate the alveolar macrophages infected with aerosolized Mtb to produce microbicidal activities intracellularly such as nitric-oxide superoxide and superoxide production. CD8+ T cells could lyse infected alveolar macrophages and could secrete granulysin or induce apoptosis of infected cells. cd T cells tend to concentrate in the surface of mucosa surfaces of the lung and quickly respond to non-peptide phosphor antigen components of Mtb, could also participate to early protection against infection at initial stage after aerosol exposure to pathogen. CD1-restricted ab T cells could be recognized by unique lipids of mycobacterium and further inhibit early mycobacterial growth intracellularly. All these immune responses, which are important for protection against Mtb infection at initial stage, could also be protective against transmission of Mtb (Hoft, 2008). Protective immune responses, against Mtb, is summarized in Table 2 (Hoft, 2008).

Fig. 2. Low-dose aerosol infection of Mtb and development of immunity through dendritic cell, which bridge between lung and lymph node.

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 4

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

Table 2 Classification, origin and function of cytokines, chemokines against Mtb. S. No.

Cytokine

Pro-inflammatory cytokines 1 TNF-a (prototype)

Origin

Functions/Major Roles

 Monocytes  Macrophages  Dendritic cells

 Control of acute and critical Mtb infection  Mediator of macrophage activation  Granuloma formation, Synergy with IFN-c induces NOS2 expression  Containment of latent infection in granuloma  Inflammatory effects like fever and wasting  Affects cell migration to and localization within tissues in Mtb infection  Influences expression of adhesion molecules as well as chemokines and chemokine receptors

 T cells

2

IL-1

 Monocytes  Macrophages  Dendritic cells

 Acute phase response such as fever and cachexia  T lymphocyte expression of IL-2 receptors and IL-2 release  Implicated in immunosuppressive mechanisms

3

IL-1b

 Monocytes  Macrophages  Dendritic cells

 Granuloma formation  Increased IL-1b/IL-1Ra ratio protects against tuberculosis

4

IL-6

 Monocytes  Macrophages  Dendritic cells

 Pro and anti-inflammatory properties  Multiple role in immune response, inflammation, hematopoiesis and differentiation of T cells  Suppression of T cell responses  Inhibits the production of TNF-a and IL-1b

5

IL-12

 Macrophages  Dendritic cells

 Production of IFN-c, and controlling Mtb infection  Connects the innate and adaptive host response to Mtb  Development of a Th1 response

6

IL-18

 Monocytes  Macrophages  Dendritic cells

 Novel proinflammatory cytokine  Stimulates the production of other proinflammatory cytokines, chemokines, and transcription factors  Induces IFN-c synergistically with IL-12  Protective against mycobacterial infections

7

IL-15

 Monocytes  Macrophages

 T-cell and NK-cell proliferation and activation  IL-15 resembles IL-2 in its biologic activities

8

IFN-c

 CD4+ T cells  CD8+ T cells  NK cells

 Augment antigen presentation  Recruitment of CD4+ T-lymphocytes and/or cytotoxic T lymphocytes  Macrophage activation  Macrophage mediated killing of mycobacteria either alone or in synergy IL-4, IL-6 and GM-CSF with IFN-c  Used as surrogate marker of infection

Anti-inflammatory cytokines 9 IL-2

 CD4 Th1 cells

 Expansion of the lymphocytes & protective CD4 Th1 cells  Can influence the course of mycobacterial infections, either alone or in combination with other cytokines

10

IL-4

 TH2 cells

 Suppression of IFN-c production  Reduce granuloma formation  Potent inducer of IL-12, suppression of IFN-c production, and macrophage activation

11

IL-10

 Macrophages  T cells

 Macrophage-deactivating properties  Down regulation of IL-12 production and IFN-c production by T cells  IL-10 directly inhibits CD4+ T cell responses  Inhibiting APC function of cells infected with mycobacteria  Counter the macrophage activating properties of IFN-c

12

TGF-b

 Monocytes

 Deactivation of macrophage production of ROI and RNI  Inhibition of T cell proliferation  Interference with NK and CTL function and down regulation of IFN-c, TNF-a and IL-1 release  Suppression of T cell responses in tuberculosis patients  Counteract protective immunity in tuberculosis & induce IL-10 production

 Alveolar Macrophages  Pulmonary epithelial cells  Monocytes  Fibroblasts  Keratinocytes/ Lymphocytes

 Recruits neutrophils, T lymphocytes, possibly monocytes  And basophils in response to stimuli

Chemokines 14 IL-8

 It is the neutrophil activating factor

15

MCP-1

 Monocytes  Macrophages

 Increase/promote the granuloma formation  Increase Th1-type cytokine production  Higher concentration of MCP-1 is found in alveolar lavage fluid, serum, and pleural fluid from tuberculosis patients

16

RANTES

 Alveolar macrophages  Monocytes

 Associated with development of M. bovis induced pulmonary granulomas  May be involved in cell trafficking in tuberculosis

196

3.1. B lymphocytes

197

B cells are well known as professional antigen presenting cells (APCs) and play different roles for the development of immunity

198

such as production of antibody, regulation and differentiation of T cells, and development of CMI. These cells can stimulate the survival, proliferation with differentiation of T cells. The activated B cells produce different type of cytokines and chemokines. Some

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

199 200 201 202

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

5

Fig. 3. The productive teamwork between T-cell populations and macrophages (M/) for control of Mtb. Mtb can survive inside the phagosomal section of macrophages and dendritic cells (DCs). B: B cell; CD4 Th: CD4 T helper cells; PRR: pattern recognition receptors; IL-2: Interleukin 2; IFN-c: Interferon-c; TNF-a: tumor necrosis factor-a; PNGs: polymorphonuclear granulocytes; Treg: regulatory T cells; TGF: transforming growth factor; Teff: effector T cells; CTL: cytolytic T lymphocytes; Tm: memory T cells; Ags: Antigens.

203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220

effector B cells type 1 (Be1) are helpful for the secretion of cytokines related usually with Th type 1 responses like IFN-c and IL-12, while effector B cell type 2 (Be2) secrete the IL-4 cytokine, characteristic of Th2 cells. Moreover, activated B cells can secrete huge amounts of IL-6 and IL-10 that play an influential role in the development of T cell-dependent immunity. IL-6 is an essential co-stimulator of T cell responses, and IL-10 suppresses immune reactions by inhibiting APCs such as macrophages and DCs. B cells stimulate inflammatory diseases and suppress autoimmune diseases through production of cytokines such as IL-10. B cell-derived pro-inflammatory IL-10 also suppresses IL-6 and IL-12 production by DCs, and they in turn can inhibit differentiation of Th1 and Th17 respectively. B cells do have regulatory role and which is not limited to autoimmune disorders. For instance chronic infection with Schistosoma mansoni induces production of IL-10 by B cells, which can suppress anaphylaxis. Likewise some viruses have developed mechanisms to stimulate production of IL-10 by B cells, and thereby subvert the immune system (Hoehlig et al., 2008).

221

3.2. T lymphocytes

222

There are several studies in humans and animal models demonstrating the requirement of acquired immunity against Mtb which needs contributions by multiple subsets of T cell, which include a dominant role for CD4+ T cells and significant roles for CD8+, CD1 restricted T cells and cd T cells (Boom, 1996). The reasons for involvement of these multiple T cell subsets are not well understood yet. The diversity in T cells that are different in antigen processing mechanisms and molecules used for antigen presentation greatly expand repertoire of the recognition of mycobacterial antigens (Fig. 3). The competence of Mtb to stay alive and persist in one of the major APCs, macrophage, participates further to T cell

223 224 225 226 227 228 229 230 231 232

activation. The leisurely growth and chronic nature of the infection of Mtb resulting in prolonged contact to a large diversity of antigens. Though most of the research was focused on CD4+ T cell response against tuberculosis, there has been increased interest in roles of CD8+ T cells in evoking substantial immune response against this pathogen. The results as documented in in vivo mouse models has shown increase in the number of activated CD4+ and CD8+ T cells in the lung-draining lymph nodes within one week of infection by virulent Mtb (Park et al., 2005). Two and four weeks of post-infection with Mtb transmigration of CD4+ and CD8+ T cells to the lungs, and stable effector/memory phenotype (CD44hiCD45loCD62L) with 50% of these cells were discovered CD69C (Newport et al., 1995; Park et al., 2005). This experiment indicates that over the period of pathogenesis the activated T cells move towards the site of infection and interact with APCs. The granuloma of tuberculosis contains both CD4 and CD8 T subsets (Feng and Britton, 2000; Ladel et al., 1995) possibly help containing infection in the granuloma from being eliminated by immune effectors of host.

233

3.3. Role of immunogens

252

The role of antibody and its understanding mechanism in protection against TB infectious disease is still under investigation. Numerous studies show that many cells other than helper T cells and antibodies are involved in immune regulation. One of the early indications of the pivotal role of antibodies in regulating the induction of T cell immunity against intracellular microbial pathogen was obtained from analysis of genital chlamydial infection in antibody-deficient mice. Conventionally, cell-mediated immune responses have been thought to be the only important ones implicated in protective

253

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251

254 255 256 257 258 259 260 261 262

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 6

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

Table 3 TB vaccine candidates in the clinical phase. Status

Product

Product description

Sponsors

Type of vaccine

Target populations

Indication

Phase III

Mw [M. indicus pranii (MIP)]

Whole cell saprophytic non-TB mycobacterium

DBT (Ministry of Science & Technology, Government of India), M/s. Cadila Pharmaceuticals Ltd.

Whole cell, Inactivated or Disrupted

–

IT

Phase III completed

M. vaccae

Inactivated whole cell non-TB mycobacterium; phase III in BCGprimed HIV+ population completed; reformulation pending

NIH, Immodulon

Whole cell, Inactivated or Disrupted

BCG-vaccinated HIV+ adults

B, PI, IT

Phase IIb

MVA85A/ AERAS-485

Modified vaccinia Ankara vector expressing Mtb antigen 85A

Oxford-Emergent Tuberculosis Consortium (OETC), Aeras, EDCTP, Wellcome Trust

Viral Vectored

B, PI, IT

AERAS402/Crucell Ad35

Replication-deficient adenovirus 35 vector expressing Mtb antigens 85A, 85B, TB10.4

Crucell, Aeras, EDCTP, NIH

Viral Vectored

BCG-vaccinated infants and adolescents; HIV infected adults BCG-vaccinated infants, children and adults

M72 + AS01

Recombinant protein composed of a fusion of Mtb antigens Rv1196 and Rv0125 & adjuvant AS01 Adjuvanted recombinant protein composed of Mtb antigens 85B and ESAT-6 rBCG Prague strain expressing listeriolysin and carries a urease deletion mutation Fragmented Mtb cells

GSK, Aeras

Recombinant Protein

Adolescents/adults, infants

B, PI

Statens Serum Institute (SSI), TBVI, EDCTP, Intercell

Recombinant Protein

Adolescents; adults

P, B, PI,

Max Planck, Vakzine Projekt Management GmbH, TBVI

Recombinant Live

–

P, B

Archivel Farma, S.L.

Whole cell, Inactivated or Disrupted

HIV+ adults, LTBI diagnosed

B, PI, IT

Aeras

Recombinant Live

Infants

P

UCLA, NIH, NIAID, Aeras

Recombinant Live

Newborns, adolescents, and adults –

B, PI

Phase II

HybridI + IC31 VPM 1002

RUTI

Phase 1 completed

AERAS-422

rBCG30

Phase I

Recombinant BCG expressing mutated PfoA and over expressing antigens 85A, 85B, and Rv3407 rBCG Tice strain expressing 30 kDa Mtb antigen 85B

B

B, PI, IT

M. smegmatis

Whole cell extract

–

Whole cell, Inactivated or Disrupted

AdAg85A

Replication-deficient adenovirus 5 vector expressing Mtb antigen 85A Adjuvanted recombinant protein composed of Mtb antigens 85B and ESAT-6 Adjuvanted recombinant protein composed of Mtb antigens 85B, ESAT-6 and Rv2660 Adjuvanted recombinant protein composed of a fusion of Mtb antigens 85B and TB10.4 Subunit fusion protein composed of 4 Mtb antigens

McMaster University

Viral Vectored Recombinant Protein

Infants; adolescents; HIV+ Adolescents, adults

P, B, PI

SSI, Aeras, Intercell

Recombinant Protein

Adolescents, adults

P, B, PI

SSI, sanofi-pasteur, Aeras, Intercell

Recombinant Protein

Infants

B

Infectious Disease Research Institute (IDRI), Aeras

Recombinant Protein

Adolescents, adults

P, B, IT

HybridI + CAF01 Hybrid 56 + IC31 HyVac 4/ AERAS404, + IC31 ID93/GLASE

SSI, TBVI

P, B, IT

DBT: Department of Biotechnology; P: Prime; B: Boost; PI: Candidate is indicated post-infection; IT: Candidate is indicated for immunotherapy.

263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278

tuberculosis immunity. Mtb is thought to replicate predominantly inside macrophage phagosomes, and intracellular pathogens in general are recognized and targeted by cellular and non-humoral immune mechanisms. The antibody responses are induced by BCG vaccination and Mtb infection (Brown et al., 2003) but these immunogen based responses are considered irrelevant for control of mycobacterial replication. Antibodies, however, could provide mechanisms of protection against infection under some circumstances. The extracellular mycobacteria, implicated in propagation of Mtb infection and present in areas of active mycobacterial replication within cavitary pulmonary lesions, could be targets for protective antibody responses. The antibodies bound to live Mtb could alter the type of phagocytic cell internalizing the organism or alter the specific uptake pathway used for phagocytosis. First, opsonization of mycobacteria might improve their phagocytosis by neutrophils that are more effective in killing intracellular

Mtb. Second, macrophage uptake of mycobacteria through immunoglobulin (Ig) receptors could increase intracellular killing by the usual host for mycobacterial persistence. Third, Ig-induced redirection of uptake or cellular activation, or both, in APCs could increase the induction of mycobacteria-specific T-cell responses. We have noted that human mycobacteria-specific antibodies induced by BCG vaccination are capable of mediating all three of above-mentioned protective mechanisms (de Valliere et al., 2005; Hoff et al., 2007). The MBS43 (IgG2b isotype), monoclonal antibody has shown protective response to MPB83 at both low and high dose test with M. bovis (Chambers et al., 2004). There are, however, a few studies state that passive transfer of antibodies in animal models does not render protection against Mtb (Dunlap and Briles, 1993). Moreover, there is a correlation exists between specific antibody profiles and TB state in humans (Davidow et al., 2005).

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

303

Recognition of Mtb by phagocytic cells leads to cell activation and production of cytokines and chemokines, which themselves induce further activation and production in a much complex process of regulation and cross-regulation. Consistent to what has been accounted above, cytokines and chemokines bear a critical role in provoking inflammatory response and affect the outcome of Mtb infections. Detailed information on pro-and anti-inflammatory cytokines as well as chemokines is provided in Table 2 (Raja, 2004; van Crevel et al., 2002).

304

3.4. Inflammasome

295 296 297 298 299 300 301 302

305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358

The studies of non-adaptive immune responses to infectious pathogens have uncovered the nucleotide binding oligomerization domain-like receptors or NLRs. NLRs are expressed predominantly in the cytoplasm, whereas Toll-like receptors (TLRs) are present on the cell membrane. Besides recognizing pathogen-associated molecular patterns released by intracellular organisms into the cytoplasm of infected cells, NLRs also detect host molecules released into the cytoplasm in response to molecular stress or cell damage so-called ‘‘danger signals’’ or danger-associated molecular patterns, such as ATP (Eisenbarth and Flavell, 2009). The activation of NLRs initiates a cascade of intracellular events that result in activation of NF-jB, which in turn leads to the expression of pro-inflammatory cytokine precursors, pro-IL-1b and pro-IL-18, and assembly of the inflammasomes, a tripartite protein complex that includes a sensor protein (NLR), an adaptor protein (apoptosis associated speck-like protein containing a caspase recruitment domain), and an effector molecule (procaspase-1). The assembly of inflammasome is required for conversion of procaspase-1 into active caspase-1, which processes pro-IL-1band pro-IL-18 into mature, active and secreted forms of IL-1b and IL-18. Therefore, activation of inflammasome by NLR controls secretion of IL-1b and IL-18, and is important for regulating macrophage activation and secretion of pro-inflammatory cytokines (Mariathasan and Monack, 2007). So far, four different inflammasomes have been identified, and Nlrp3 is the most widely used. The aberrant activation of inflammasome, due to genetic mutations in Nlrp3, causes enhanced activation of caspase-1 and elevated IL-1b levels in humans resulting in an auto-inflammatory disorder known as cryopyrin-associated periodic syndrome (Verma et al., 2008). The activation of inflammasome, followed by production of IL-1b and IL-18, is also critical for innate immunity against many human pathogens (Mariathasan and Monack, 2007). The Mtb infection drives macrophages to produce IL-1b and IL-18, (Giacomini et al., 2001; Tsao et al., 1999) which may contribute to chronic inflammation. Therefore it is imperative to understand the mechanism of production of IL-1b and IL-18 by Mtb infection. The infecQ6 tion of macrophages with wild type Mtb and Mycobacterium marinum induced potent IL-1b secretion, compared to that seen with their respective ESX-1 gene knock-outs (Giacomini et al., 2001; Koo et al., 2008; Kurenuma et al., 2009). This is suggestive of the requirement the secretion of ESX-1b substrate proteins for the production of IL-1b by macrophages. Indeed, recombinant ESAT-6 directly activates Nlrp3 inflammasome and elicits secretion of IL-1b by macrophages (Mishra et al., 2010). The ESAT-6 activates Nlrp3 inflammasome probably through permeabilization of phagosomes and activation of Syk tyrosine kinase in Mtb infected macrophages, and activation of Nlrp3 may be associated with necrotic death of Mtb infected macrophages. These findings further single out the importance of ESAT-6 which contributes to the initiation and regulation of inflammatory responses in tuberculosis infection by activating inflammasome in macrophages. All research so far has surfaced an inevitable role of ESAT-6 in lung pathology and chronic inflammation by the activation of inflammasome in alveolar macrophages during acute tuberculosis infection.

7

4. Various novel delivery systems in vaccine development

359

Over the past 20 years the technology of vaccine development has seen radical changes and numerous vaccines entering clinical phase (Table 3). However, most of the developed vaccines are themselves not sufficiently immunogenic due to depleted innate immune stimulus, and therefore use of adjuvant to these types of vaccines has become an urgent need. In addition subunit vaccines themselves do not have an inherent ability to be delivered to appropriate sites for optimal immune stimulation. Thus in order to ensure the optimum presentation to both innate and adaptive type of immune system, alternate forms of vaccine delivery systems such as use of liposomes, microparticles (MPs), nanoparticles (NPs) have been deployed. Also, use of new adjuvants as well as prime-boost concept (Li et al., 2006) have been explored in order to have an enhanced, long lasting and consistent effect on the immune response. The carriers such as MPs, emulsions, immune stimulating complexes [ISCOMS], liposomes, virosomes and virus-like particles (VLP) offer several attributes for vaccine delivery. Although DNA vaccines have been shown to induce a potent CTL responses (Srivastava and Liu, 2003), but they are not capable enough in eliciting antibody responses (Wang et al., 1998), and low immunogenicity has been observed in human clinical trials as well (Bivas-Benita et al., 2009). This low immunogenicity of DNA vaccines may be addressed by the use of various novel delivery systems coupled with suitable adjuvants for DNA vaccines (Bivas-Benita et al., 2004, 2009; de la Torre et al., 2009; Meerak et al., 2013; Rosada et al., 2008, 2012; Wang et al., 2010; Yu et al., 2012) (Table 4). Thus strategies are needed to enhance the Q7 immunogenicity of DNA vaccines (Yu et al., 2012). A few strategies which might be adopted are as follows; improving transfection of host cells and antigen expression; augmenting antigen presentation; enhancing co-stimulation and increasing T lymphocyte expansion.

360

4.1. Liposomes

393

The versatile lipid vesicle (liposome) delivery vehicle was introduced by A, Bangham (Wagner and Vorauer-Uhl, 2011; Wang et al., 2010) about five decades ago. These vesicles are microscopic spherical structures with an aqueous core surrounded by one or more outer shell(s) consisting of lipids systematize in a bilayer configuration. The value of liposomes as carriers for drugs has been recognized years ago and has shown an enormous potential (Sessa and Weissmann, 1968) in various pharmaceutical applications. The advantages of liposomes are their ability to encapsulate various materials combined with their structural flexibility and versatility. The predominance in drug delivery and targeting have enabled these carriers to be used as therapeutic tool in tumor targeting, gene silencing, anti-sense therapy, immunomodulation and most importantly in genetic vaccination (Tyagi et al., 2012).The rationale behind the usage of liposomes as delivery systems in intracellular infections such as mycobacteria is their tendency of being taken up by the macrophages, following systemic administration (Couvreur and Vauthier, 2006). There are considerable evidence suggestive of immunomodulatory effects of liposome when administered into body as a vaccine adjuvant (Agger et al., 2008; Derrick et al., 2012). The cationic liposomes, especially DNA delivery vesicles in vaccines, are well defined prescripts to enhance the immune recognition efficiency against inert or poorly immunogenic subunits of proteins. Some of the comprehensively investigated liposomeforming lipids associate are; 1,2-dioleoyl-3-trimethylammonium propane (DOTAP), dimethyldioctadecylammonium (DDA), and 3b[N-(N0 ,N0 -dimethylaminoethane)carbomyl] cholesterol (DCChol).

394

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392

395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 8

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

Table 4 Different carrier systems (Drug Delivery System) used for TB vaccine development. Author/Ref

Material

Microparticles/Microspheres Zhu et al., Chitosan (2007) de Paula et al. (2007)

PLGA

Ajdary et al. (2007)

Sodium Alginate

Nature of vaccine/ antigen

Animal/dose/rout(s)

Immune response (achievement)

Protein vaccine Fusion protein, AMM

Female C57BL/6 mice Three dose (1, 3, 5 week)/ s.c.

Chitosan microspheres when used as a carrier for fusion protein AMM obtained strong humoral and cell-mediated immune responses

DNA vaccine

Female Hartley guinea pigs & BALB/c mice Single-dose/i.m.

The single-shot prime-boost formulation presented excellent efficacy and diminished lung pathology in both mice and guinea pigs

Live attenuated vaccine 108 CFU of BCG

BALB/c mice Single dose/oral (intragastric administration)

Oral administration of BCG in alginate microspheres results in strong systemic protective immune responses; however, mucosal immunity and protection against pulmonary infection after oral vaccination with alginate-encapsulated BCG remains to be determined

pDNAhsp65 and the adjuvant TDM

Lu et al. (2007)

PLGA

Protein vaccine Recombinant protein rAg85B

In-vitro study performed Specific CD4 T-hybridoma cells DB-1. DB-1 cells, THP1 cells

PLGA microspheres in respirable sizes were successful in delivering rAg85B in an immunologically appropriate manner to macrophages. The result was a base for further investigation into the prospective use of PLGA particles for delivery of vaccines to prevent Mtb infection

Coelho et al. (2006)

PLGA

DNA vaccine DNA-hsp65 or DNAhsp65 and TDM

Bagg Albino/c female mice Two dose at 3 weeks interval/i.m.

Encapsulated MHSP/TDM was more immunogenic than naked hsp65 DNA, and has great potential to improve vaccine effectiveness against leishmaniasis and tuberculosis

Ha et al. (2006)

PLGA

Protein vaccine IL-12EM + AS01B

C57BL/6 mice Two dose at 0 and 8 weeks/ dorsal, s.c.

Codelivered with IL-12EM are designed for a sustained release of IL-12 and induced strong Th1 immune responses specific to Ag85A and ESAT-6. The adjuvant combination of IL-12EM plus AS01B was a more efficient way to induce a sustained Th1 immunity and protection against Mtb

Cai et al. (2005)

PLGA

DNA vaccine DNA (Ag85B, MPT-64 and MPT-83) + DDA

C57BL/6 female mice Single/i.m.

Mice receiving a single dose of PLGA encapsulated DNA were protected against Mtb challenge at levels comparable to groups of mice immunized with three doses of non-encapsulated DNA vaccine or with Mycobacterium bovis BCG

Evans et al. (2004)

PLGA

Protein vaccine Recombinant Mtb8.4 protein

Female C57Bl/6 mice Single/s.c. i.d.

The CD8 T-cells response elicited by Mtb8.4 protein-microspheres was far superior to the adjuvant formulations examined and was comparable to that elicited by Mtb8.4-DNA following only a single subcutaneous or intradermal microsphere immunization

Lima et al. (2003a,b)

Gold

DNA vaccine DNA–hsp65

Female BALB/c mice Three dose (3 occasions at 2-week intervals)/i.m.

Immunization by gene gun induced immune response with plasmid doses 100-fold lower than those required for intramuscular immunization. However, in contrast to intramuscular immunization, which was protective in these studies, gene gun immunization did not protect BALB/c mice against challenge infection

Lima et al. (2003a,b)

PLGA

DNA vaccine pCDNA3-hsp65 + TDM

BALB/c mice Single dose/i.m.

Higher levels of IgG2a subtype antibody and IFN-c in the supernatant of spleen cell cultures. DNA-hsp65/TDM-encapsulated microspheres were also capable to stimulate higher IFN-c production in bulk lung cells from challenged mice and give protection as effective as that attained after three doses of naked DNA administration

Lima et al. (2001)

PLGA, charcoal

Protein vaccine TDM

BALB/c mice Single/Intratracheal or i.p.

High levels of IL-6, TNF-a, IL-12, IL-10, IFN-c, & IL-4 production were detected in lung cells, and nitric oxide (NO) production was high in culture supernatants of bronchoalveolar lavage cells. TDM contributes to persistence of infection through production of cytokines, which are essential for the recruitment of inflammatory cells and maintenance of a granulomatous reaction

Wilkinson et al. (2000)

Polystyrene

Protein vaccine CFF antigen 85 A, B, and C (Ag85A, B & C)

In vitro study Mtb strain H37Rv Intranasal

BAA induced activation of both CD4+ and CD8+ T cell subsets. However, CD4+ responses in general were higher and their antigenic repertoire was wider than the CD8+ responses. By contrast, CD8+ responses were strongest to the lower molecular weight BAA. When CFF were chemically attached to carboxyl modified microspheres (BCA), stimulation of IFN-c was similar to BAA

Dhiman and Khuller (1998)

DL-PLG

Protein vaccine 71-kDa cell wall associated protein emulsified in FIA

Mice of NMRI strain Three dose (2 s.c. on days 0 & 7, followed by an i.m. on day 14.

The 71-kDa-PLG immunized group showed a notably higher clearance of bacterial load from the lungs and livers in contrast to the 71-kDa-FIA immunized group. The results advocate the long-term protective potential of a PLG-microparticle based antigen delivery system for tuberculosis

Cationic lipid (octa-/hexadecane)

Oligonucleotide and protein vaccine 1. P407 + CpG 2. Ag85A protein

BALB/c mice

P407 and CpG all increased immune responses to Ag85A.

Three times at 3 weeks intervals/UA or DL

CpG led to a T-helper type-I phenotype and the combination with the carrier P407 generated a more polyfunctional T cells phenotype

Chitosan

DNA vaccines Ag85B

Female BALB/c mice Three dose 0, 14 and 28 days/s.c

Autophagy-inducing plasmid into a DNA vaccine improved host immune responses to a DNA vaccine against the MTB antigen delivered by chitosan particles in mice

Nanoparticles Todoroff et al. (2013) Meerak et al. (2013)

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 9

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx Table 4 (continued) Author/Ref

Material

Nature of vaccine/ antigen

Animal/dose/rout(s)

Immune response (achievement)

Yu et al. (2012)

Fe3O4-Glupolyethyleneimin

DNA vaccine Ag85A-ESAT-6-IL-21

Male C57BL/6 mice Three dose with 3-week intervals/i.m.

The improved DNA vaccine Ag85A-ESAT-6-IL-21-coated Fe3O4 NPs transported plasmid DNA vaccine into cells effectively and induced powerful immune responses and protection against Mtb challenge in contrast to using the DNA vaccine Ag85AESAT-6-IL-21 alone in a mouse model

Ballester et al. (2011)

Pluronicstabilized nanoparticles (NPs),

Protein vaccine Ag85A protein

C57BL/6 mice Three dose at 0, 7, 21 days/ i.d. or pulmonary administration

Pulmonary NP administration beneficially influences the quality of the immune response, leading to the generation of polyfunctional Th1 and Th17 responses, correlating with protection from Mtb challenge

Bivas-Benita et al. (2009)

PLGA–PEI

DNA vaccine Encoding the Mtb latency antigen Rv1733c

Balb/c female mice Three times at 3 weeks intervals/i.m. or endotracheal aerosol

The strongest immunogenicity was obtained by pulmonary priming with NPs-adsorbed Rv1733c DNA followed by boosting with Rv1733c protein. These results prove that PLGA–PEI NPs are proficient DNA vaccine delivery system to enhance T cell responses through pulmonary delivery in a DNA prime/protein boost vaccine regimen

Bivas-Benita et al. (2004)

Chitosan

DNA vaccine encoding HLA-A0201restricted T-cell epitopes

HLA-A0201/Kb (HLA-A2/ Kb) female transgenic mice

DNA plasmid expressing different Mtb epitopes that are recognized by patients with the HLA-A0201 polymorphic class I molecule is superior to the common intramuscular application. Chitosan nanoparticles are a suitable delivery system for DNA vaccines, protecting DNA from degradation by nucleases, inducing DCs maturation and resulting in increased IFN-c secretion from T cells after pulmonary mucosal immunization

PC Cardiolipin

Peptide Bcn1, Bcn2, Bcn3, Bcn4, Bcn 5

C57BL/6JCit (B6) mice Single/i.v.

Bcn (Class IIa) produced by bacteria constitute a set of molecules that are of great concern as potential anti-TB drugs, mainly in combination with liposomal vectors conferring intracellular delivery

CAF01

Peptide

Female BABL/c, C57BL/6 mice Three dose at 2-week intervals/s.c.

CAF01 is a potentially appropriate adjuvant for a broad range of diseases including targets requiring both CMI and humoral immune responses for defense

Female CB6F1 (C57BL/ 66BALB/c) mice Twice at two-week intervals/s.c.

Heterologous prime boost based on H4, formed an additive outcome on the priming of CD4 and CD8 cells and in terms of the defending capacity of the vaccine, and therefore represent an attractive new vaccine strategy against Mtb

Liposomes Sosunov et al. (2007) Agger et al. (2008)

Ovalbumin with DDA, TDB Elvang et al. (2009)

CAF01

Protein vaccine Two fusion protein antigens Ag85B and TB10.4

Three dose in 3 weeks intervals/i.m. endotracheal aerosol

HenriksenLacey et al. (2011)

DDA, DC-Chol and DOTAP

Protein vaccine Ag85B-ESAT-6

C57BL/6 mice Three dose with 2 week interval/i.m.

A long term retention and slow release of liposome and vaccine antigen from the injection site hence appears to favor a stronger Th1 immune response

Derrick et al. (2012)

DDA/TDB

Attenuated live vaccine DmmaA4BCG mutant was derived from BCG

C57BL/6 female mice and SCID mice One immunization or Three immunizations 2 weeks apart./s.c.

BCG Pasteur/DmmaA4 mutant BCG strain with DDA/TDB adjuvant yielded safer formulations that induced drastically more anti-tuberculosis protective immunity than BCG controls

Rosada et al. (2008)

PC, DOPE, DOTAP

DNA-hsp65

Female 6-week-old BALB/c mic One or two doses/i.m. or nasal

Sixteen-fold reduction in the pDNA amount administered in only one dose with the additional advantage of using a non-invasive route of administration (intranasal route)

Aagaard et al. (2009)

CAF01

Protein vaccine

Female CB6F1 (BALB/ C  C57BL/6) mice Three times, 2 week apart/ s.c.

The T cell epitopes could differ significantly depending on the host HLA haplotype, it would be ideal to expose all epitopes, subdominant and dominant, without the need for identifying and modifying the dominant epitope

Rosada et al. (2012)

L-a-PC, DOTAP, DOPE

DNA vaccine DNAhsp65

In vitro test and Mice Four dose/Intranasal route

Pseudo-ternary complex is a promising gene vaccine for Mtb treatment. It contributes to the development of multifunctional nanostructures in the search for strategies for in vivo DNA delivery

Wang et al. (2010)

Lipofectamine TM 2000

DNA Vaccine pcDNA3.1+/Ag85A DNA

C57BL/6 mice Three times each 14 days interval/Oral

de la Torre et al. (2009)

EPC/DOTAP/ DOPE

DNA Vaccine Plasmid pVAX-hsp65 (6 kb)

BALB/c Mice Four times at 2-week intervals/i.m.

Cellular compartment in the epithelium of small intestine acts a major role on the regulation of immune response to eliminate Mtb. This is important understanding and possible implications for the design of new strategies based on oral DNA vaccine on regulation of immune response in protection against Mtb EPC inclusion in the DOTAP/DOPE-DNA structure has changed the physicochemical properties and as a consequence, decreased the in vitro cytotoxicity and delayed antibody production’s length of time

CTA1-DD Quil A saponin

Protein vaccine Fusion protein Ag85BESAT-6

Female C57BL/6 mice Three dose with a 2-week interval/i.n.

ISCOMS Andersen et al. (2007)

ESAT-6 contains numerous potential T cell epitopes

Intra nasal administration of the antigen Ag85B-ESAT-6 mixed with CTA1DD-ISCOMs as adjuvant invokes a Th1 immune response and enhances the preferential recruitment of antigen-specific IFN-c-secreting T cells to the lungs following a live M. tuberculosis challenge (continued on next page)

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 10

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

Table 4 (continued) Author/Ref

Material

Nature of vaccine/ antigen

Animal/dose/rout(s)

Immune response (achievement)

da Fonseca et al. (2000)

PA-NHS

Protein vaccine

Immunogenic by inducing antibodies, Th1 and CTL responses

NaDOC Quil A saponin

38-kDa Mtb protein

C57BL/6 (H-2b) female mice Three dose/s.c.

AMM: Ag85B–PT64190–198–Mtb8.4; BAA: bead-adsorbed antigens; BCA: bead-coupled antigens; Bcn: Bacteriocins; CAF01: Cordfactor; CFF: culture filtrate fractions; CTA1DD: cholera toxin-derived fusion protein; DC-Chol: 3b-[N-(N0 ,N0 -dimethylaminoethane)carbomyl] Cholesterol; DDA: N,N0 -dimethyl-N,N0 -dioctadecylammonium bromide; DL: deep lung; DNA Ag85A: DNA vaccine expressing antigen 85A; DOPE: 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine; DOTAP: 1,2-Dioleoyl-3-trimethylammonium propane; EPC: egg phosphatidylcholine ESAT: early secretory antigenic target; ESAT-6: 6-kDa early secretory antigen target; FIA: Freund’s incomplete adjuvant; hsp65: heatshock protein 65; IL-12 EM: IL-12 encapsulated microspheres; NPs: nanoparticles; NaDOC: sodium deoxycholate; P407: Poloxamer 407; PC: phosphatidylcholin; pDNAhsp65: plasmid DNA encoding the hsp65; PA-NHS: N-(palmitoyloxy) succinimide; PEI: polyethyleneimine; PPS: polypropylene sulfide; TDB: a,a’-trehalose 6,6’-dibeheneate; TDM: trehalose dimycolate; UA: upper airways; IgG2a: immunoglobulin G2a.

421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461

462 463 464 465 466 467 468 469

DDA has been broadly studied in combination with different immunomodulators like trehalose 6,60 -dibehenate (TDB) (Table 4 and discussed latter). The CAF01, designed as combination of DDA and TDB, has been proven successful at inducing protective immune responses against different infectious pathogens like influenza, chlamydia and erythrocytic-stage malaria, and tuberculosis (TB) (Henriksen-Lacey et al., 2011). However, given the complexity of the events leading to a protective immune response, the latest generation of adjuvants is unlikely to be based on a single component. Relatively, more complex adjuvant formulations based on combinations of numerous mono therapeutic agents being able of targeting multiple different receptors, and hence more likely to induce complex and sufficient immune responses, are seen as the way forward (Moingeon et al., 2001). An example of this development is represented by the development of the archaeosomes based on the polar lipid fraction from archaea. The archaeosome adjuvant system has been Q8 characterized and its proficiency in inducing a humoral as well as cell-mediated immune response (Krishnan et al., 2000) is well documented. The pulmonary retention of therapeutic agents has also been achieved through the use of liposomes. The liposomes have also been shown to enhance the potency of DNA vaccines as evidenced by the mediated plasmid uptake by APC (Perrie et al., 2001). In the context of gene therapy, lipid/DNA complexes have been successfully aerosolized and delivered to the lungs of monkeys (McDonald et al., 1998). The membrane fusion and endocytosis are discovered the main mechanism for transporting liposomal content into the cells (Tyagi et al., 2012). Thus all these developments are suggestive of addressing many of the problems encountered in the administration and low immune response of conventional vaccines may be circumvented by genetic immunization approach in conjunction and usage with novel delivery vehicles (Tyagi et al., 2012). There are number of reports regarding the design and development of liposomal aerosols as vaccine carriers for intracellular pathogens like Mtb (Vyas et al., 2004, 2005). Liposomal aerosols offer various advantages including sustained release, reduced cytotoxicity, prevention of local irritation, improved stability in large aqueous core, and possibility to manipulate release in the area of targeting specific site by altering the bilayer constituents and improving formulation techniques. 4.1.1. DDA DDA liposomes have been used in development of vaccine against tuberculosis (Table 4). This quaternary ammonium compound has been previously reported as an effective adjuvant for eliciting cell-mediated and humoral responses. DDA was discovered as an adjuvant (Gall, 1966), and has been tested in combination with a number of different viral and bacterial antigens in different animal species (Brandt et al., 2000; Holten-Andersen

et al., 2004). It is a synthetic amphiphilic lipid compound consisting of a hydrophilic positively charged head–group of dimethylammonium attached to two 18-carbon alkyl chains (hydrophobic tail). In aqueous environment DDA form self-assembled closed bilayers vesicles i.e. liposome. The adjuvant efficiency and stability of the liposome (DDA) was improved by anchoring synthetic glycolipid TDB (trehalose 6,60 -dibehenate), a synthetic analogue to immune stimulatory constituent of the mycobacterial cell wall, referred to as the cord factor or trehalose dimycolate (Carmona-Ribeiro and Chaimovich, 1986). In one study M. bovis BCG lipid extract were explored for their immunostimulatory properties. These lipid extracts were formulated in a cationic liposome composed of DDA (Rosenkrands et al., 2005). It was found that administering these lipids in DDA liposome (mycosome) induced a powerful Th1 response characterized by marked elevated antigen-specific immunoglobulin G2a (IgG2a) isotype antibodies, and substantial production of IFN-c. Furthermore, these mycosomes also induced immune responses to protein antigens from several sources such as Mtb, Chlamydia muridarum and tetanus toxoid. A more sustained immunological memory was observed when the mycosomes were combined with the Ag85-ESAT-6 fusion protein and a level of protection was found to be superior to that seen with live BCG (Rosenkrands et al., 2005).

470

4.1.2. CAF01 (DDA:TDB) CAF01 is an extremely novel, versatile adjuvant system inducing concurrently a CMI response and an antibody response with elevated levels of IgG2 antibodies. These antibody responses were maintained extended periods (>1 year) post-immunization of subunit TB vaccine (Gram et al., 2009). CD4 cells (compare to CD8 T cells) were more poly-functional and antigen specific, recruited at a higher rate, and proliferated more following Mtb infection. While both CD4 and CD8 epitopes are clearly presented at very onset of infection, CD4 and CD8 T cells take action very differently to live infection in a way which supports the consensus that CD4 T cells bear the most critical role during the early stages of Mtb infection (Elvang et al., 2009). Nevertheless, TDB having the synergic effect in enhancing immune responses, and it also has a great stabilizing effect on the formulation of DDA liposome and maintains the particle size distribution of same formulation for more than 1.5 years at 4 °C. This preparation was scheduled to enter clinical trials together with the TB vaccine candidate Ag85BESAT6 (Agger et al., 2008). Vaccines prepared by the combination of either BCG Pasteur or a DmmaA4 mutant BCG strain with DDA) – D(+) TDB (CAF01) adjuvant yield safer formulations that induced significantly more anti-tuberculosis protective immunity compare to plain BCG. The low-cost, safer, and more immunogenic and compatible TB vaccination strategies are urgently needed to confirm the increased safety and immunogenicity of these adjuvanted BCG formulations (Derrick et al., 2012).

493

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492

494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

548

4.1.3. DOTAP DOTAP is well known cationic lipid used for the development of cationic liposome deployed to developing TB vaccine (Table 4). DOTAP and DOPE are new synthetic peptide containing a NLS (based on the classical Simian Virus SV 40T sequence) and their incorporation into the cationic liposome/DNAhsp65 gene vaccine. This pseudo-ternary complex presented therapeutic effects against TB, even greater than the cationic liposome/DNAhsp65 gene vaccine and similar to naked DNA, effectively reducing the total amount of DNAhsp65 administered and circumvented needle phobia. The pseudo-ternary NLS/DNAhsp65/cationic liposome is a promising gene vaccine for TB treatment. In addition multifunctional NPs with different domains (DNA delivery inside cell and nuclear transport) were able to contributing in vivo applications and gene delivery (Rosada et al., 2012). Henriksen-Lacey et al., compared three well-known cationic lipids, which are used for the formulation of liposome, as potential adjuvants and the delivery of protein subunit. The potential of DC-Cholesterol, DOTAP, and DDA Liposome incorporating immunomodulating TDB to form an antigen depot at the site of injection (SOI), and to induce immunological recall responses against co-administered tuberculosis vaccine antigen (Ag85B-ESAT-6) is well established. Liposome composition plays a significant role in vaccine retention at the site of injection, and its ability to provoking immune system to induce a vaccine specific recall response. While all these cationic liposome facilitated increased antigen presentation by APCs, the monocyte permeation to the SOI and the production of IFN-c upon antigen recall were markedly higher for DDA and DC-Chol based liposome which exhibited a longer retention profile at the SOI (Henriksen-Lacey et al., 2011).

549

4.2. Microparticles

550

Microparticles have various application and advantages in the field of vaccinology and drug delivery. Firstly, they have been suggested to have similar size to that of the pathogens that the immune system has evolved to recognize, combat and, consequently, efficiently internalize by APCs. The uptake of MPs (<5 lm) by APC is likely to be important in the ability of particles to be used as vaccine adjuvants. It has been reported that macrophages that carry MPs to the lymph nodes can mature into dendritic cells (DC) (Randolph et al., 1999). In addition, uptake of biodegradable MPs directly into DC has been demonstrated both in vitro (Lutsiak et al., 2002) and in vivo (Newman et al., 2002). The appropriate size for MPs appears to be in the range of 1–3 lm (Tabata and Ikada, 1988), and cationic MPs are particularly effective for being taken up by macrophages and DC (Thiele et al., 2003). A second useful property of MPs which makes them more optimal for B cells activation is their ability to present multiple copies of antigens on their surfaces (Fehr et al., 1998). These organized arrays of antigen surface are able to efficiently cross-link B cell receptors and constitute a strong activation signal (Fehr et al., 1998). It has been documented that the duration of antigen persistence in the body should be long enough in order to elicit long-lasting and effective protective T cell response (Storni et al., 2003). Therefore, persistence of antigens in the body may be enhanced by associating it with MPs that has been shown to protect the antigens from degradation. Moreover, particulate delivery systems are thought to promote trapping and retention of antigens in local lymph nodes for generating effective immune responses. The immunopotentiators/adjuvants may be included in the particulate delivery systems to elicit enhanced immune response and to process them by the APC. Therefore various particulate carriers have been explored for their potential to be used as delivery vehicle for vaccine constructs. Several biodegradable polymers namely polyanhydrides, polyorthoesters, hyaluronic acid, chitosan, and starch, polymers that self-assemble into particulates (poloxamers),

519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 541 542 543 544 545 546 547

551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582

11

or soluble polymers (polyphosphazenes) (Payne et al., 1998) have been used to prepare MPs for antigen delivery. Another approach to enhance delivery of target antigens into the APC is by using recombinant proteins which are known to self-assemble into the particulate structures. The immunogenicity of MPs is affected by particle size as smaller particles are more immunogenic than the larger ones. It has been observed from studies carried out in mouse models that MPs induce antibodies predominantly of the IgG2a isotype, indicating a Th1 response (Vordermeier et al., 1995). PLG based delivery system has also been evaluated for the development of vaccine against tuberculosis. A greater antigen specific IgG1/IgG2a and IFN-c secretion was observed when immunodominant 38 kDa Mtb antigen entrapped in PLG MPs was delivered as compared to those with FIA. PLG has also been reported as an ideal vaccine delivery system for the two highly protective vaccine candidates (30 kDa secretory protein and 71 kDa cell wall protein of Mtb H37Ra) compared to other delivery systems consisting of FIA, Liposome and DDA. Moreover, single shot PLG MPs based subunit vaccines to these antigens were not only equally protective than consecutive three shot schedule (Sharma et al., 1999), but protection induced was also found to be long lasting as compared to FIA. Microspheres prepared from biodegradable PLGA also have the potential to act as mediators of DNA transfection targeted to phagocytic cells such as macrophages or DCs (Lima et al., 2001, 2003a,b), and to protect them against the degradative action of nucleases (Barry et al., 1999). To overcome the problems associated with microencapsulation, DNA had been adsorbed on to the surface of cationic microparticle based on a biodegradable & biocompatible polymer PLGA (Lima et al., 2001, 2003a,b). It was observed that not only DNA adsorbed efficiently but also induced an enhanced immune response in comparison to that seen with naked DNA. The heightened immune responses were apparent in all species evaluated, including nonhuman primates. In addition cationic MPs could deliver several plasmids simultaneously at high loading levels. The cationic PLG MPs with adsorbed DNA also showed protective efficacy in a rodent colon cancer model, increased antibody and cellular responses to HBsAg, and greater protective efficacy of a DNA-based tuberculosis vaccine (Mollenkopf et al., 2004).It is now known that DNA adsorbed onto the cationic PLG MPs are efficiently delivered to DCs(Denis-Mize et al., 2000). Also, these PLG MPs have the potential to recruit the DC to the injection site, and also protect the adsorbed DNA from degradation in vivo.

583

4.3. Nanoparticles

627

The uses of polymeric NPs for vaccine and adjuvant delivery have been investigated for a few reasons. The first reason is that NPs have a higher surface area for adsorption, allowing higher antigen-polymer ratio (Wendorf et al., 2006). Second, possible enhanced immunogenicity (the effectiveness of submicron particles [200–500 nm] compared with 1–2 lm particle is not definitive and still an area of current research in progress (Table 4). Third, NPs have an advantage over MPs i.e. ease of preparation and benign conditions during preparation (Wendorf et al., 2006).

628

4.3.1. PLGA & PLA Delivery of nucleic acids into cells using cationic polymers has recently attracted remarkable interest in the field of non-viral gene therapy, due to their structural diversity, easy production, non-immunogenicity, and safety. Most effective and widely used synthetic polymers are poly(lactide) (PLA) and poly(lactide-coglycolide) (PLGA) (Bivas-Benita et al., 2009). Both have been approved by FDA for several therapeutic applications, and exhibit biodegradability, biocompatibility and safety in humans (Garg

637

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626

629 630 631 632 633 634 635 636

638 639 640 641 642 643 644 645

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 12 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 681 682 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699 700 701 702 703 704 705 706 707 708 709

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

et al., 2010). The potential of PLGA-based delivery system relies on their capacity to release entrapped antigen over an extended period of time and their ability to deliver antigen directly to the phagocytic APCs (Bivas-Benita et al., 2009). In brief, PLGA polymers have generated immense interest as delivery systems due to their excellent biocompatibility and biodegradability. These systems being versatile in terms of nature of the agent encapsulated, time period of release and diverse route of their delivery, represent an exciting technology for delivery of different biomolecules and therapeutic agents (Garg et al., 2010). Biodegradable NPs of PLA and PLGA, have been studied extensively as materials for sutures in surgery, as vaccine adjuvants as well as for vaccine delivery systems in particular for mucosal immunization via oral route (Putney and Burke, 1998). It has been previously demonstrated that MPs with entrapped antigens had comparable immunogenicity to the Freund’s adjuvant. There has been little investigation of alternative delivery systems and administration routes for vaccines to elicit effective immunity against Mtb particulates to improve vaccine antigen delivery (Binjawadagi et al., 2014). Micro and nano-system of PLGA copolymers have been major focus as carriers for antigens (Table 4). PLGA or PLGA-associated antigen particles enter the phagosome/phagolysosome, and there is potential for some antigen to traffic to the cytoplasm. Moreover, PLGA microspheres have the ability to elicit CTL responses (Binjawadagi et al., 2014; de Paula et al., 2007; Lima et al., 2003a,b) and the potential for mucosal immunization (Garg et al., 2010). Macrophages pulsed with antigens encapsulated in small particles can present antigen 100 to 1000-fold more efficiently than macrophages pulsed with soluble antigen (Lu et al., 2007). 4.3.2. Chitosan Over the last decade, chitosan is another biodegradable and biocompatible natural occurring polymer. Chitosan and its nanoand micro-carrier system were also investigated for delivery of hydrophilic bioactive molecules such as peptide, protein, and drugs. Chitosan also has been used for vaccine development. It is a cationic polysaccharide made of repeating units of N-acetylD-glucosamine and D-glucosamine derived by the partial de-acetylation of chitin. The term chitosan includes a series of chitosan polymers with different molecular weights (50–2000 kDa) with 40–98% degree of acetylation. Chitosan and its microspheres and NPs have many benefits for vaccine delivery. First, chitosan can open the intercellular tight junctions and favor the paracellular transport of bioactive molecules. Second, nanoparticle and MPs of chitosan are perfect delivery system for sustained and controlled release of bioactive molecules. Third, chitosan particles are most professionally taken up by phagocytotic/APC cells. Thus, chitosan and its derivatives could induce strong systemic and mucosal immune responses against antigens (Zhu et al., 2007). One study reveals a novel approach based on chitosan NPs to improve the immune response to a DNA based vaccine against tuberculosis through incorporation of an autophagy-inducing plasmid into the vaccine. Autophagy-inducing plasmid DNA loaded chitosan NPs were prepared and determined immunity for TB in mice (Bivas-Benita et al., 2004). It was therefore concluded from the study that incorporating an autophagy-inducing element into a DNA loaded chitosan vaccine may help improving strong immune response (Meerak et al., 2013). A very recent study investigated that mucosal delivery of gene vaccine in chitosan formulation enhanced particular SIgA level and mucosal IFN-c (+) T cell response which were optimistically correlated with immunological protection against tuberculosis (Ai et al., 2013). 4.3.3. Alginate Alginate is a naturally occurring biodegradable polysaccharide which can be easily cross-linked into a solid matrix by addition

of bivalent cations. Alginates are safe, non-immunogenic and inexpensive natural polymers with high mucoadhesive properties. It consists of varying proportions of 1, 4-linked b-D-mannuronic acid (M), a-L-glucuronic acid (G) and alternating (MG) blocks. The cross linking in water-in-oil emulsion results in the formation of microspheres. Alginate microspheres are biodegradable and safe to use in animals with minimum toxicity. They have been used to encapsulate proteins, live viruses and bacteria, and plasmid DNA, and have been recognized as safe and approved as a food and pharmaceutical ingredient by the U.S. Food and Drug Administration (FDA) in the early 1970s. It is known as a non-toxic and nonirritant material, and has many ideal characteristics which make it appropriate polymer to use in vaccine developments, such as non-immunogenicity, stability with long shelf life, biodegradability and low production cost. Sodium alginate has been found as adjuvanticity effect on subcutaneously injected BCG in BALB/c mice (Ajdary et al., 2007). BCG loaded alginate microspheres have been prepared and used for oral delivery for strong systemic protective immune responses. However, mucosal immunity and protection against pulmonary tuberculosis infection after oral vaccination with alginate-encapsulated BCG remains to be determined. We understand this is the first report to evaluate immunogenicity and protective efficacy of alginate encapsulated BCG used as oral BCG vaccine. Apart from above mentioned polymers (PLGA, PLA, chitosan, and alginate), various polymers have been used for formulation of micro and nano-system such as gold, charcoal, and cationic lipid FeO4-Glupolyethylen (Table 4).

710

4.4. ISCOMS

738

ISCOMS are negatively charged cage-like structures with the size of 30 ± 40 nm. They can be formed spontaneously after mixing antigens with cholesterol, phospholipids and the saponin. Quil A is the saponin adjuvant extracted from the bark of tree Quillaja saponaria Molina, and is composed of a mixture of structurally similar triterpenoids. The complexes formulated without protein antigen are called iscomatrix (matrix), and have the same morphology as ISCOMS. Efficient incorporation of protein antigens into ISCOMS requires accessible hydrophobic regions. The antigen lacking these regions can be still incorporated into ISCOMS by covalent attachment of lipid tails. The ISCOM was created with the aim of presenting antigens and adjuvant in the same particle, and thus to increase immunogenicity of incorporated antigen and reduce the dose of both antigen and adjuvant. The ISCOMS induce long lasting antibody responses and cellular immune responses in vivo. When ISCOMS was immunized with 38-kDa mycobacterial protein (ISCOMS; 38-kDa ISCOMS), it was found immunogenic by inducing antibodies, Th1 and CTL responses (da Fonseca et al., 2000). It has also been proven as an effective mucosal vaccine against Mtb, when Ag85B-ESAT-6 mixed with CTA1-DD/ISCOMs (as adjuvant) and nasally immunized (Andersen et al., 2007).

739

5. Development of liposome based novel mucosal vaccine

760

There are several logistic and immunological merits of mucosal vaccination. One understandable but very significant asset of mucosal vaccines is that it can be administered without the use of needles and syringes. Therefore, this approach makes immunization perform more acceptable, safer and improved suited for mass administration. Nevertheless, so far only some vaccines approved for human being use are administered mucosally (Garg et al., 2010).For oral vaccine including the live attenuated polio vaccine, rotavirus vaccines, various cholera vaccines (killed whole-cell/ cholera toxin (CT) B subunit or live-attenuated), oral typhoid

761

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

711 712 713 714 715 716 717 718 719 720 721 722 723 724 725 726 727 728 729 730 731 732 733 734 735 736 737

740 741 742 743 744 745 746 747 748 749 750 751 752 753 754 755 756 757 758 759

762 763 764 765 766 767 768 769 770

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

13

Fig. 4. Schematic representation of designed mechanism of immune induction: (1) Targeting to NALT using claudin-4 ligand followed by nasal immunization with (2) cationic liposomal DNA.

771 772 773 774 775 776 777 778 779 780 781 782

783

vaccine (live attenuated) and for nasal rout influenza are the greatest example for nasal vaccination. The best suitable example of the useful success of mucosal vaccination is the polio oral vaccine. Millions of people were immunized with polio vaccine in a single day because of the oral route is very convenient to children (Bloom and Widdus, 1998). Such mob vaccination efforts are just possible in terms of both price and practicability when oral or nasal vaccines are used for vaccination to people. Also markedly, a main benefit of using the mucosal immunization route is the stimulation of local secretory IgA (SIgA) antibodies for immune exclusion of pathogens at the site of entrance before attack can take place route (Andersen et al., 2007; Ritz et al., 2004; Rosada et al., 2008). 5.1. Specialized mucosal sites for induction of immunity

804

The body’s mucosal surfaces are extensive and, due to direct or indirect contact to the environment, they behave as an immunological barrier. Antigen stimulation through mucosal surfaces leads to initiation of local synthesis of dimeric IgA which can be exported by the polymeric Ig receptor (pIgR) as SIgA antibodies. By dispersion of activated B cells from mucosa-associated lymphoid tissue (MALT) (Garg et al., 2010), SIgA may be locally generated at the surface of all mucosa (Garg et al., 2010). Nevertheless, there is no completely common but rather an included mucosal immune system, which is compartmentalized. Following mucosal antigen stimulation, in addition to the production of SIgA, a compound and powerful chain of immune actions is initiated by mucosal vaccination. This leads to the selection and potential deployment of other types of particular antibodies, Th cells and cytotoxic T lymphocytes (CTLs) with the probability to attack host cells infected with Mtb. Therefore vaccines can be given mucosally together with an appropriate adjuvant may acquire about different types of local and systemic immunity. Mucosal vaccination in the worldwide, and intranasal in meticulous, has been revealed to give up as good, or even better, systemic immune responses compare to the corresponding parenteral administration (Kallenius et al., 2007).

805

5.2. Nasal DNA vaccination: opportunities and impediments

806

The presently available vaccine, M. bovis BCG, is defending only against a severe form of childhood TB, but does not decrease the worldwide TB burden in adults (Meerak et al., 2013). Thus, a novel

784 785 786 787 788 789 790 791 792 793 794 795 796 797 798 799 800 801 802 803

807 808

vaccine against Mtb is urgently needed, and numerous types of new TB vaccines are in the early stages of development and, also a few TB vaccines are in different phase of clinical trials (Meerak et al., 2013; Ritz et al., 2004). Presently, DNA vaccines are one of the many types of new TB vaccine candidates under investigation. Some studies have discovered the potency of plasmid DNAs (pDNAs) carrying different antigen-encoding genes of Mtb in initiation of a defensive immune response in mice as well as in non-human primates (Meerak et al., 2013). Genetic immunization has evolved to be an attractive alternative to live attenuated pathogens in terms of safety, ease of mass production and low cost (Donnelly et al., 1997). DNA vaccination is capable of inducing a broad spectrum of immunity (humoral and cellular) against the pathogen. Plasmid DNA (pDNA) vaccines have an established record of efficacy in pre-clinical studies, and can be safely used in human even in immune-compromised individuals. A DNA immunization through nasal route can effectively induce the specific immunity (humoral and cellular) at respiratory tract (Brun et al., 2008; Kallenius et al., 2007). It has been observed that specific immunity at respiratory tract can effectively block the transmission and offer the significant protection against disease reactivation and clearance of actively dividing bacilli. However, despite these encouraging characteristics, naked antigen encoding plasmids are usually unable to stimulate immune responses following intranasal administration. Firstly, their ineffective delivery to immune NALT due to mucus, mucociliary clearance, low permeation. Secondly, the transport of pDNA to the nucleus is generally restricted by extensive lysosomal and phagosomal degradation of the internalized pDNA ‘‘in vivo’’ (Wang et al., 2010, 2011). Therefore, it is believed that the success of nasal vaccine depends on effective delivery of the antigen encoding pDNA to the target cells i.e. APCs. Thus, the antigens encoding pDNA should be delivered effectively to NALT, and subsequently to resident APCs for the development of a potent nasal vaccine (Goonetilleke et al., 2003; Kallenius et al., 2007).

809

5.3. Mechanism of vaccine delivery via nasal route

844

The antigen encoding pDNA can be delivered effectively through nasal route for efficient induction of cellular and humoral immune response when two approaches are coupled together. These two approaches involve efficient delivery of pDNA firstly, to NALT (nasal

845

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

810 811 812 813 814 815 816 817 818 819 820 821 822 823 824 825 826 827 828 829 830 831 832 833 834 835 836 837 838 839 840 841 842 843

846 847 848

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 14 849 850 851 852 853 854 855 856 857 858 859 860 861

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

associated lymphoid tissue) and secondly, to the nucleus of APCs. It is considered that single layer of epithelial cell sheet follicle-associated epithelium (FAE) covers NALT and FAE plays a pivotal role in mucosal immunological response (Garg et al., 2010; Neutra et al., 1996). The antigen delivery using a ligand for the FAE would be a potent strategy for the effective mucosal delivery of vaccine. Claudin-4 is a tight junction transmembrane protein that plays a role in establishing trans-epithelial electrical resistance in the FAE. These findings strongly indicate that claudin-4-targeting may be useful for nasal vaccines. The claudin-4 using Clostridium perfringens enterotoxin (CPE) may be targeted for effective delivery of DNA encoding antigen to the immune inductive sites of nasal mucosal (Ebihara et al., 2006).

862

5.4. Delivery to nucleus

863

879

The second pre-requisite following the delivery of pDNA to NALT is the delivery of pDNA to nucleus through delivery vehicle, which is capable of overcoming barriers to gene transfection. In comparison with viral systems, non-viral systems have advantages such as safety, possibility of multiple deliveries, delivery of larger pieces of DNA, and effective costs. In addition, these non-viral carriers avoid DNA degradation and facilitate targeted delivery to APCs (Brun et al., 2008). It has been well-known that pDNA encapsulated into positively charged (cationic) liposome (discussed above) leads to greatly improved delivery of pDNA to the nucleus, and has an intrinsic immune adjuvant ability that results into the induction of strong humoral and cell mediated immunity (Saeki et al., 2009). Based upon the knowledge about liposome, it may be the best alternative to engineer ligand anchored cationic liposome containing TB antigen encoding pDNA such as claudin-4 for targeted and sustained release of vaccine formulation in order to achieving heightened and long lasting immune responses (Fig. 4).

880

6. Conclusion

881

Up until now lots of research has been undertaken towards the development of a new tuberculosis vaccine which can either replace the current available vaccine, BCG or further boost its effect. The first breakthrough came by the M. Horwitz group of researchers in 2003 who tried to develop a new tuberculosis vaccine based on the recombinant approach by constructing an rBCG which over expresses the immunodominant antigens of vaccine (Horwitz and Harth, 2003). The study was a great success; however, it failed to meet its predefined end points. Therefore, novel delivery systems such as Liposome, MPs and NPs have been explored in this area. These systems have been further engineered to elicit a desired and specific immune response. However, role of novel carrier(s) in targeted and controlled delivery of antigens, proteins and pDNA needs more attention to explore the possibility of effective vaccine against Mtb. The use of adjuvants has played a critical role in the development of such delivery systems which are thought to enhance immune response by a significant value. Apart from adjuvants, the specific ligand anchoring has been explored in order to impart targetability to specific areas in the body. The administration of antigen-encoding plasmid DNA via engineered Liposome could circumvent the need of muscle involvement and facilitate its uptake by APC. This offers the effective delivery of their contents including DNA through non-invasive routes (nasal) thus eliminate the disadvantages and phobia associated with needle injections. There are many alternative approaches to the nasal delivery (non-invasive route). The versatility and delivery potential of engineered liposome has been revealed and its role in intracellular cytosolic delivery of DNA has been advocated. The potential approach to the cytosolic delivery of vaccine is the encapsulation/

864 865 866 867 868 869 870 871 872 873 874 875 876 877 878

882 883 884 885 886 887 888 889 890 891 892 893 894 895 896 897 898 899 900 901 902 903 904 905 906 907 908 909

entrapment of antigen/DNA into the ligand anchored lipid-based formulation of pH sensitive liposome (Fig. 4). The lipid based delivery systems can be manipulated to enhance the efficacy of nasally administered vaccine in number of ways: they can protect antigen or DNA from degradation, concentrate them in one area of tissue for better presentation, targeted delivery by coating their surface with ligands specific for the dendritic cell/macrophages receptors. In conclusion, an efficient ligand anchored delivery vehicle combined with an effective adjuvant administered through a most favorable route of immunization, will finally allow for the development of an unbeaten needle free (nasal) TB vaccine in humans.

910

Acknowledgements

921

The research work is supported by various grants from Council of Scientific and Industrial Research-Human Resource Development Group (CSIR-HRDG), Department of Biotechnology (DBT), New Delhi, India. The authors have no other relevant affiliations or financial involvement with any other organization or entity with a financial interest in or financial conflict with the subject matter or material discussed in the manuscript apart from those disclosed. No writing assistance was taken in this manuscript.

922

References

930

Agger, E.M., Rosenkrands, I., Hansen, J., Brahimi, K., Vandahl, B.S., Aagaard, C., Werninghaus, K., Kirschning, C., Lang, R., Christensen, D., et al., 2008. Cationic liposomes formulated with synthetic mycobacterial cordfactor (CAF01): a versatile adjuvant for vaccines with different immunological requirements. PLoS One 3 (9), e3116. Ai, W., Yue, Y., Xiong, S., Xu, W., 2013. Enhanced protection against pulmonary mycobacterial challenge by chitosan-formulated polyepitope gene vaccine was associated with elevated pulmonary SIgA and IFN-gamma(+) T cell response. Microbiol. Immunol.. http://dx.doi.org/10.1111/j.1348-0421.12027. Ajdary, S., Dobakhti, F., Taghikhani, M., Riazi-Rad, F., Rafiei, S., Rafiee-Tehrani, M., 2007. Oral administration of BCG encapsulated in alginate microspheres induces strong Th1 response in BALB/c mice. Vaccine 25 (23), 4595–4601. Andersen, C.S., Dietrich, J., Agger, E.M., Lycke, N.Y., Lovgren, K., Andersen, P., 2007. The combined CTA1-DD/ISCOMs vector is an effective intranasal adjuvant for boosting prior Mycobacterium bovis BCG immunity to Mycobacterium tuberculosis. Infect. Immun. 75 (1), 408–416. Barry, M.E., Pinto-Gonzalez, D., Orson, F.M., McKenzie, G.J., Petry, G.R., Barry, M.A., 1999. Role of endogenous endonucleases and tissue site in transfection and CpG-mediated immune activation after naked DNA injection. Hum. Gene Ther. 10 (15), 2461–2480. Barry 3rd, C.E., Boshoff, H.I., Dartois, V., Dick, T., Ehrt, S., Flynn, J., Schnappinger, D., Wilkinson, R.J., Young, D., 2009. The spectrum of latent tuberculosis: rethinking the biology and intervention strategies. Nat. Rev. Microbiol. 7 (12), 845–855. Binjawadagi, B., Dwivedi, V., Manickam, C., Ouyang, K., Wu, Y., Lee, L.J., Torrelles, J.B., Renukaradhya, G.J., 2014. Adjuvanted poly(lactic-co-glycolic) acid nanoparticleentrapped inactivated porcine reproductive and respiratory syndrome virus vaccine elicits cross-protective immune response in pigs. Int. J. Nanomed. 9, 679–694. Bivas-Benita, M., van Meijgaarden, K.E., Franken, K.L., Junginger, H.E., Borchard, G., Ottenhoff, T.H., Geluk, A., 2004. Pulmonary delivery of chitosan-DNA nanoparticles enhances the immunogenicity of a DNA vaccine encoding HLAA⁄0201-restricted T-cell epitopes of Mycobacterium tuberculosis. Vaccine 22 (13–14), 1609–1615. Bivas-Benita, M., Lin, M.Y., Bal, S.M., van Meijgaarden, K.E., Franken, K.L., Friggen, A.H., Junginger, H.E., Borchard, G., Klein, M.R., Ottenhoff, T.H., 2009. Pulmonary delivery of DNA encoding Mycobacterium tuberculosis latency antigen Rv1733c associated to PLGA-PEI nanoparticles enhances T cell responses in a DNA prime/ protein boost vaccination regimen in mice. Vaccine 27 (30), 4010–4017. Bloom, B.R., Widdus, R., 1998. Vaccine visions and their global impact. Nat. Med. 4 (5 Suppl.), 480–484. Boom, W.H., 1996. The role of T-cell subsets in Mycobacterium tuberculosis infection. Infect. Agents Dis. 5 (2), 73–81. Brandt, L., Elhay, M., Rosenkrands, I., Lindblad, E.B., Andersen, P., 2000. ESAT-6 subunit vaccination against Mycobacterium tuberculosis. Infect. Immun. 68 (2), 791–795. Brown, R.M., Cruz, O., Brennan, M., Gennaro, M.L., Schlesinger, L., Skeiky, Y.A., Hoft, D.F., 2003. Lipoarabinomannan-reactive human secretory immunoglobulin A responses induced by mucosal bacille Calmette-Guerin vaccination. J. Infect. Dis. 187 (3), 513–517. Brun, P., Zumbo, A., Castagliuolo, I., Delogu, G., Manfrin, F., Sali, M., Fadda, G., GrillotCourvalin, C., Palu, G., Manganelli, R., 2008. Intranasal delivery of DNA encoding antigens of Mycobacterium tuberculosis by non-pathogenic invasive Escherichia

931 932 933 934 935 936 937 938 939 940 941 942 943 944 945 946 947 948 949 950 951 952 953 954 955 956 957 958 959 960 961 962 963 964 965 966 967 968 969 970 971 972 973 974 975 976 977 978 979 980 981 982

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

911 912 913 914 915 916 917 918 919 920

923 924 925 926 927 928 929

Q9

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx 983 984 985 986 987 988 989 990 991 992 993 994 995 996 997 998 999 1000 1001 1002 1003 1004 1005 1006 1007 1008 1009 1010 1011 1012 1013 1014 1015 1016 1017 1018 1019 1020 1021 1022 1023 1024 1025 1026 1027 1028 1029 1030 1031 1032 1033 1034 1035 1036 1037 1038 1039 1040 1041 1042 1043 1044 1045 1046 1047 1048 1049 1050 1051 1052 1053 1054 1055 1056 1057 1058 1059 1060 1061 1062 1063 1064 1065 1066 1067 1068

coli. Vaccine 26(16), 1934–1941. doi:http://dx.doi.org/10.1016/ j.vaccine.2008.02.023. Epub 2008 Feb 25. Carmona-Ribeiro, A.M., Chaimovich, H., 1986. Salt-induced aggregation and fusion of dioctadecyldimethylammonium chloride and sodium dihexadecylphosphate vesicles. Biophys. J. 50 (4), 621–628. Chambers, M.A., Gavier-Widen, D., Hewinson, R.G., 2004. Antibody bound to the surface antigen MPB83 of Mycobacterium bovis enhances survival against high dose and low dose challenge. FEMS Immunol. Med. Microbiol. 41 (2), 93–100. Couvreur, P., Vauthier, C., 2006. Nanotechnology: intelligent design to treat complex disease. Pharm. Res. 23 (7), 1417–1450. da Fonseca, D.P., Frerichs, J., Singh, M., Snippe, H., Verheul, A.F., 2000. Induction of antibody and T-cell responses by immunization with ISCOMS containing the 38-kilodalton protein of Mycobacterium tuberculosis. Vaccine 19 (1), 122–131. Davidow, A., Kanaujia, G.V., Shi, L., Kaviar, J., Guo, X., Sung, N., Kaplan, G., Menzies, D., Gennaro, M.L., 2005. Antibody profiles characteristic of Mycobacterium tuberculosis infection state. Infect. Immun. 73 (10), 6846–6851. de la Torre, L.G., Rosada, R.S., Trombone, A.P., Frantz, F.G., Coelho-Castelo, A.A., Silva, C.L., Santana, M.H., 2009. The synergy between structural stability and DNAbinding controls the antibody production in EPC/DOTAP/DOPE liposomes and DOTAP/DOPE lipoplexes. Colloids Surf. B: Biointerfaces 73 (2), 175–184. de Paula, L., Silva, C.L., Carlos, D., Matias-Peres, C., Sorgi, C.A., Soares, E.G., Souza, P.R., Blades, C.R., Galleti, F.C., Bonato, V.L., et al., 2007. Comparison of different delivery systems of DNA vaccination for the induction of protection against tuberculosis in mice and guinea pigs. Genet. Vaccines Ther. 5, 2. de Valliere, S., Abate, G., Blazevic, A., Heuertz, R.M., Hoft, D.F., 2005. Enhancement of innate and cell-mediated immunity by antimycobacterial antibodies. Infect. Immun. 73 (10), 6711–6720. Denis-Mize, K.S., Dupuis, M., MacKichan, M.L., Singh, M., Doe, B., O’Hagan, D., Ulmer, J.B., Donnelly, J.J., McDonald, D.M., Ott, G., 2000. Plasmid DNA adsorbed onto cationic microparticles mediates target gene expression and antigen presentation by dendritic cells. Gene Ther. 7 (24), 2105–2112. Derrick, S.C., Dao, D., Yang, A., Kolibab, K., Jacobs, W.R., Morris, S.L., 2012. Formulation of a mmaA4 gene deletion mutant of Mycobacterium bovis BCG in cationic liposomes significantly enhances protection against tuberculosis. PLoS One 7 (3), e32959. Donnelly, J.J., Ulmer, J.B., Shiver, J.W., Liu, M.A., 1997. DNA vaccines. Annu. Rev. Immunol. 15, 617–648. Dunlap, N.E., Briles, D.E., 1993. Immunology of tuberculosis. Med. Clin. North Am. 77 (6), 1235–1251. Ebihara, C., Kondoh, M., Hasuike, N., Harada, M., Mizuguchi, H., Horiguchi, Y., Fujii, M., Watanabe, Y., 2006. Preparation of a claudin-targeting molecule using a Cterminal fragment of Clostridium perfringens enterotoxin. J. Pharmacol. Exp. Ther. 316 (1), 255–260. Eisenbarth, S.C., Flavell, R.A., 2009. Innate instruction of adaptive immunity revisited: the inflammasome. EMBO Mol. Med. 1 (2), 92–98. Elvang, T., Christensen, J.P., Billeskov, R., Thi Kim Thanh Hoang, T., Holst, P., Thomsen, A.R., Andersen, P., Dietrich, J., 2009. CD4 and CD8 T cell responses to the M. tuberculosis Ag85B-TB10.4 promoted by adjuvanted subunit, adenovector or heterologous prime boost vaccination. PLoS One 4(4), e5139. Eum, S.Y., Kong, J.H., Hong, M.S., Lee, Y.J., Kim, J.H., Hwang, S.H., Cho, S.N., Via, L.E., Barry 3rd, C.E., 2010. Neutrophils are the predominant infected phagocytic cells in the airways of patients with active pulmonary TB. Chest 137 (1), 122–128. Fehr, T., Skrastina, D., Pumpens, P., Zinkernagel, R.M., 1998. T cell-independent type I antibody response against B cell epitopes expressed repetitively on recombinant virus particles. Proc. Natl. Acad. Sci. USA 95 (16), 9477–9481. Feng, C.G., Britton, W.J., 2000. CD4+ and CD8+ T cells mediate adoptive immunity to aerosol infection of Mycobacterium bovis bacillus Calmette-Guerin. J. Infect. Dis. 181 (5), 1846–1849. Flynn, J.L., Chan, J., 2005. What’s good for the host is good for the bug. Trends Microbiol. 13 (3), 98–102. Gall, D., 1966. The adjuvant activity of aliphatic nitrogenous bases. Immunology 11 (4), 369–386. Garg, N.K., Mangal, S., Khambete, H., Tyagi, R.K., 2010. Mucosal delivery of vaccines: role of mucoadhesive/biodegradable polymers. Recent Patents Drug Deliv. Form. 4 (2), 114–128. Giacomini, E., Iona, E., Ferroni, L., Miettinen, M., Fattorini, L., Orefici, G., Julkunen, I., Coccia, E.M., 2001. Infection of human macrophages and dendritic cells with Mycobacterium tuberculosis induces a differential cytokine gene expression that modulates T cell response. J. Immunol. 166 (12), 7033–7041. Goonetilleke, N.P., McShane, H., Hannan, C.M., Anderson, R.J., Brookes, R.H., Hill, A.V., 2003. Enhanced immunogenicity and protective efficacy against Mycobacterium tuberculosis of bacille Calmette-Guerin vaccine using mucosal administration and boosting with a recombinant modified vaccinia virus Ankara. J. Immunol. 171 (3), 1602–1609. Gram, G.J., Karlsson, I., Agger, E.M., Andersen, P., Fomsgaard, A., 2009. A novel liposome-based adjuvant CAF01 for induction of CD8(+) cytotoxic Tlymphocytes (CTL) to HIV-1 minimal CTL peptides in HLA-A⁄0201 transgenic mice. PLoS One 4 (9), e6950. Henriksen-Lacey, M., Christensen, D., Bramwell, V.W., Lindenstrom, T., Agger, E.M., Andersen, P., Perrie, Y., 2011. Comparison of the depot effect and immunogenicity of liposomes based on dimethyldioctadecylammonium (DDA), 3beta-[N-(N0 ,N0 -dimethylaminoethane)carbomyl] cholesterol (DCChol), and 1,2-dioleoyl-3-trimethylammonium propane (DOTAP): prolonged liposome retention mediates stronger Th1 responses. Mol. Pharm. 8 (1), 153– 161.

15

Hoehlig, K., Lampropoulou, V., Roch, T., Neves, P., Calderon-Gomez, E., Anderton, S.M., Steinhoff, U., Fillatreau, S., 2008. Immune regulation by B cells and antibodies a view towards the clinic. Adv. Immunol. 98, 1–38. Hoff, S.T., Abebe, M., Ravn, P., Range, N., Malenganisho, W., Rodriques, D.S., Kallas, E.G., Soborg, C., Mark Doherty, T., Andersen, P., et al., 2007. Evaluation of Mycobacterium tuberculosis-specific antibody responses in populations with different levels of exposure from Tanzania, Ethiopia, Brazil, and Denmark. Clin. Infect. Dis. 45 (5), 575–582. Hoft, D.F., 2008. Tuberculosis vaccine development: goals, immunological design, and evaluation. Lancet 372 (9633), 164–175. Holten-Andersen, L., Doherty, T.M., Korsholm, K.S., Andersen, P., 2004. Combination of the cationic surfactant dimethyl dioctadecyl ammonium bromide and synthetic mycobacterial cord factor as an efficient adjuvant for tuberculosis subunit vaccines. Infect. Immun. 72 (3), 1608–1617. Horwitz, M.A., Harth, G., 2003. A new vaccine against tuberculosis affords greater survival after challenge than the current vaccine in the guinea pig model of pulmonary tuberculosis. Infect. Immun. 71 (4), 1672–1679. Kallenius, G., Pawlowski, A., Brandtzaeg, P., Svenson, S., 2007. Should a new tuberculosis vaccine be administered intranasally? Tuberculosis (Edinb) 87 (4), 257–266. Kaplan, G., Post, F.A., Moreira, A.L., Wainwright, H., Kreiswirth, B.N., Tanverdi, M., Mathema, B., Ramaswamy, S.V., Walther, G., Steyn, L.M., et al., 2003. Mycobacterium tuberculosis growth at the cavity surface: a microenvironment with failed immunity. Infect. Immun. 71 (12), 7099–7108. Koo, I.C., Wang, C., Raghavan, S., Morisaki, J.H., Cox, J.S., Brown, E.J., 2008. ESX-1dependent cytolysis in lysosome secretion and inflammasome activation during mycobacterial infection. Cell Microbiol 10(9), 1866–1878. doi:http://dx.doi.org/ 10.1111/j.1462-5822.2008.01177.x. Epub 2008 Jun 28. Krishnan, L., Sad, S., Patel, G.B., Sprott, G.D., 2000. Archaeosomes induce long-term CD8+ cytotoxic T cell response to entrapped soluble protein by the exogenous cytosolic pathway, in the absence of CD4+ T cell help. J. Immunol. 165 (9), 5177–5185. Kurenuma, T., Kawamura, I., Hara, H., Uchiyama, R., Daim, S., Dewamitta, S.R., Sakai, S., Tsuchiya, K., Nomura, T., Mitsuyama, M., 2009. The RD1 locus in the Mycobacterium tuberculosis genome contributes to activation of caspase-1 via induction of potassium ion efflux in infected macrophages. Infect. Immun. 77 (9), 3992–4001. Ladel, C.H., Daugelat, S., Kaufmann, S.H., 1995. Immune response to Mycobacterium bovis bacille Calmette Guerin infection in major histocompatibility complex class I- and II-deficient knock-out mice: contribution of CD4 and CD8 T cells to acquired resistance. Eur. J. Immunol. 25 (2), 377–384. Li, Z., Zhang, H., Fan, X., Zhang, Y., Huang, J., Liu, Q., Tjelle, T.E., Mathiesen, I., Kjeken, R., Xiong, S., 2006. DNA electroporation prime and protein boost strategy enhances humoral immunity of tuberculosis DNA vaccines in mice and nonhuman primates. Vaccine 24 (21), 4565–4568. Lima, V.M., Bonato, V.L., Lima, K.M., Dos Santos, S.A., Dos Santos, R.R., Goncalves, E.D., Faccioli, L.H., Brandao, I.T., Rodrigues-Junior, J.M., Silva, C.L., 2001. Role of trehalose dimycolate in recruitment of cells and modulation of production of cytokines and NO in tuberculosis. Infect. Immun. 69 (9), 5305–5312. Lima, K.M., dos Santos, S.A., Santos, R.R., Brandao, I.T., Rodrigues Jr., J.M., Silva, C.L., 2003a. Efficacy of DNA-hsp65 vaccination for tuberculosis varies with method of DNA introduction in vivo. Vaccine 22 (1), 49–56. Lima, K.M., Santos, S.A., Lima, V.M., Coelho-Castelo, A.A., Rodrigues Jr., J.M., Silva, C.L., 2003b. Single dose of a vaccine based on DNA encoding mycobacterial hsp65 protein plus TDM-loaded PLGA microspheres protects mice against a virulent strain of Mycobacterium tuberculosis. Gene Ther. 10 (8), 678–685. Lu, D., Garcia-Contreras, L., Xu, D., Kurtz, S.L., Liu, J., Braunstein, M., McMurray, D.N., Hickey, A.J., 2007. Poly(lactide-co-glycolide) microspheres in respirable sizes enhance an in vitro T cell response to recombinant Mycobacterium tuberculosis antigen 85B. Pharm. Res. 24 (10), 1834–1843. Lutsiak, M.E., Robinson, D.R., Coester, C., Kwon, G.S., Samuel, J., 2002. Analysis of poly(D,L-lactic-co-glycolic acid) nanosphere uptake by human dendritic cells and macrophages in vitro. Pharm. Res. 19 (10), 1480–1487. Mariathasan, S., Monack, D.M., 2007. Inflammasome adaptors and sensors: intracellular regulators of infection and inflammation. Nat. Rev. Immunol. 7 (1), 31–40. McDonald, R.J., Liggitt, H.D., Roche, L., Nguyen, H.T., Pearlman, R., Raabe, O.G., Bussey, L.B., Gorman, C.M., 1998. Aerosol delivery of lipid:DNA complexes to lungs of rhesus monkeys. Pharm. Res. 15 (5), 671–679. Meerak, J., Wanichwecharungruang, S.P., Palaga, T., 2013. Enhancement of immune response to a DNA vaccine against Mycobacterium tuberculosis Ag85B by incorporation of an autophagy inducing system. Vaccine 31 (5), 784–790. Mishra, B.B., Moura-Alves, P., Sonawane, A., Hacohen, N., Griffiths, G., Moita, L.F., Anes, E., 2010. Mycobacterium tuberculosis protein ESAT-6 is a potent activator of the NLRP3/ASC inflammasome. Cell. Microbiol. 12 (8), 1046–1063. Moingeon, P., Haensler, J., Lindberg, A., 2001. Towards the rational design of Th1 adjuvants. Vaccine 19 (31), 4363–4372. Mollenkopf, H.J., Dietrich, G., Fensterle, J., Grode, L., Diehl, K.D., Knapp, B., Singh, M., O’Hagan, D.T., Ulmer, J.B., Kaufmann, S.H., 2004. Enhanced protective efficacy of a tuberculosis DNA vaccine by adsorption onto cationic PLG microparticles. Vaccine 22 (21–22), 2690–2695. Neutra, M.R., Frey, A., Kraehenbuhl, J.P., 1996. Epithelial M cells: gateways for mucosal infection and immunization. Cell 86 (3), 345–348. Newman, K.D., Elamanchili, P., Kwon, G.S., Samuel, J., 2002. Uptake of poly(D,Llactic-co-glycolic acid) microspheres by antigen-presenting cells in vivo. J. Biomed. Mater. Res. 60 (3), 480–486.

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

1069 1070 1071 1072 1073 1074 1075 1076 1077 1078 1079 1080 1081 1082 1083 1084 1085 1086 1087 1088 1089 1090 1091 1092 1093 1094 1095 1096 1097 1098 1099 1100 1101 1102 1103 1104 1105 1106 1107 1108 1109 1110 1111 1112 1113 1114 1115 1116 1117 1118 1119 1120 1121 1122 1123 1124 1125 1126 1127 1128 1129 1130 1131 1132 1133 1134 1135 1136 1137 1138 1139 1140 1141 1142 1143 1144 1145 1146 1147 1148 1149 1150 1151 1152 1153 1154

PHASCI 3035

No. of Pages 16, Model 5G

10 June 2014 16 1155 1156 1157 1158 1159 1160 1161 1162 1163 1164 1165 1166 1167 1168 1169 1170 1171 1172 1173 1174 1175 1176 1177 1178 1179 1180 1181 1182 1183 1184 1185 1186 1187 1188 1189 1190 1191 1192 1193 1194 1195 1196 1197 1198 1199 1200 1201 1202 1203 1204 1205 1206 1207 1208 1209 1210 1211

N.K. Garg et al. / European Journal of Pharmaceutical Sciences xxx (2014) xxx–xxx

Newport, M., Levin, M., Blackwell, J., Shaw, M.A., Williamson, R., Huxley, C., 1995. Evidence for exclusion of a mutation in NRAMP as the cause of familial disseminated atypical mycobacterial infection in a Maltese kindred. J. Med. Genet. 32 (11), 904–906. Park, H., Li, Z., Yang, X.O., Chang, S.H., Nurieva, R., Wang, Y.H., Wang, Y., Hood, L., Zhu, Z., Tian, Q., et al., 2005. A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat. Immunol. 6 (11), 1133–1141. Payne, L.G., Jenkins, S.A., Woods, A.L., Grund, E.M., Geribo, W.E., Loebelenz, J.R., Andrianov, A.K., Roberts, B.E., 1998. Poly[di(carboxylatophenoxy)phosphazene] (PCPP) is a potent immunoadjuvant for an influenza vaccine. Vaccine 16 (1), 92–98. Perrie, Y., Frederik, P.M., Gregoriadis, G., 2001. Liposome-mediated DNA vaccination: the effect of vesicle composition. Vaccine 19 (23–24), 3301–3310. Putney, S.D., Burke, P.A., 1998. Improving protein therapeutics with sustainedrelease formulations. Nat. Biotechnol. 16 (2), 153–157. Raja, A., 2004. Immunology of tuberculosis. Indian J. Med. Res. 120 (4), 213–232. Randolph, G.J., Inaba, K., Robbiani, D.F., Steinman, R.M., Muller, W.A., 1999. Differentiation of phagocytic monocytes into lymph node dendritic cells in vivo. Immunity 11 (6), 753–761. Ritz, S.A., Cundall, M.J., Gajewska, B.U., Swirski, F.K., Wiley, R.E., Alvarez, D., Coyle, A.J., Stampfli, M.R., Jordana, M., 2004. The lung cytokine microenvironment influences molecular events in the lymph nodes during Th1 and Th2 respiratory mucosal sensitization to antigen in vivo. Clin. Exp. Immunol. 138 (2), 213–220. Rosada, R.S., de la Torre, L.G., Frantz, F.G., Trombone, A.P., Zarate-Blades, C.R., Fonseca, D.M., Souza, P.R., Brandao, I.T., Masson, A.P., Soares, E.G., et al., 2008. Protection against tuberculosis by a single intranasal administration of DNAhsp65 vaccine complexed with cationic liposomes. BMC Immunol. 9(38), 1–13, 9–38. Rosada, R.S., Silva, C.L., Santana, M.H., Nakaie, C.R., de la Torre, L.G., 2012. Effectiveness, against tuberculosis, of pseudo-ternary complexes: peptideDNA-cationic liposome. J. Colloid Interface Sci. 373 (1), 102–109. Rosenkrands, I., Agger, E.M., Olsen, A.W., Korsholm, K.S., Andersen, C.S., Jensen, K.T., Andersen, P., 2005. Cationic liposomes containing mycobacterial lipids: a new powerful Th1 adjuvant system. Infect. Immun. 73 (9), 5817–5826. Russell, D.G., 2007a. Who puts the tubercle in tuberculosis? Nat. Rev. Microbiol. 5 (1), 39–47, Epub 2006 Dec 11. Russell, D.G., 2007b. Who puts the tubercle in tuberculosis? Nat. Rev. Microbiol. 5 (1), 39–47. Russell, D.G., Cardona, P.J., Kim, M.J., Allain, S., Altare, F., 2009. Foamy macrophages and the progression of the human tuberculosis granuloma. Nat. Immunol. 10 (9), 943–948. Russell, D.G., Barry 3rd, C.E., Flynn, J.L., 2010. Tuberculosis: what we don’t know can, and does, hurt us. Science 328 (5980), 852–856. Saeki, R., Kondoh, M., Kakutani, H., Tsunoda, S., Mochizuki, Y., Hamakubo, T., Tsutsumi, Y., Horiguchi, Y., Yagi, K., 2009. A novel tumor-targeted therapy using a claudin-4-targeting molecule. Mol. Pharmacol. 76 (4), 918–926. Schijns, V.E., 2003. Mechanisms of vaccine adjuvant activity: initiation and regulation of immune responses by vaccine adjuvants. Vaccine 21 (9–10), 829–831. Sessa, G., Weissmann, G., 1968. Phospholipid spherules (liposomes) as a model for biological membranes. J. Lipid Res. 9 (3), 310–318. Sharma, A.K., Verma, I., Tewari, R., Khuller, G.K., 1999. Adjuvant modulation of T-cell reactivity to 30-kDa secretory protein of Mycobacterium tuberculosis H37Rv and its protective efficacy against experimental tuberculosis. J. Med. Microbiol. 48 (8), 757–763. Srivastava, I.K., Liu, M.A., 2003. Gene vaccines. Ann. Intern. Med. 138 (7), 550–559. Storni, T., Ruedl, C., Renner, W.A., Bachmann, M.F., 2003. Innate immunity together with duration of antigen persistence regulate effector T cell induction. J. Immunol. 171 (2), 795–801.

Tabata, Y., Ikada, Y., 1988. Macrophage phagocytosis of biodegradable microspheres composed of L-lactic acid/glycolic acid homo- and copolymers. J. Biomed. Mater. Res. 22 (10), 837–858. Thiele, L., Merkle, H.P., Walter, E., 2003. Phagocytosis and phagosomal fate of surface-modified microparticles in dendritic cells and macrophages. Pharm. Res. 20 (2), 221–228. Tsao, T.C., Hong, J., Huang, C., Yang, P., Liao, S.K., Chang, K.S., 1999. Increased TNFalpha, IL-1 beta and IL-6 levels in the bronchoalveolar lavage fluid with the upregulation of their mRNA in macrophages lavaged from patients with active pulmonary tuberculosis. Tuber. Lung Dis. 79 (5), 279–285. Tyagi, R.K., Garg, N.K., Sahu, T., 2012. Vaccination strategies against malaria: novel carrier(s) more than a tour de force. J. Controlled Release 162 (1), 242–254. van Crevel, R., Ottenhoff, T.H., van der Meer, J.W., 2002. Innate immunity to Mycobacterium tuberculosis. Clin. Microbiol. Rev. 15 (2), 294–309. Verma, D., Lerm, M., Blomgran Julinder, R., Eriksson, P., Soderkvist, P., Sarndahl, E., 2008. Gene polymorphisms in the NALP3 inflammasome are associated with interleukin-1 production and severe inflammation: relation to common inflammatory diseases? Arthritis Rheum. 58 (3), 888–894. Via, L.E., Lin, P.L., Ray, S.M., Carrillo, J., Allen, S.S., Eum, S.Y., Taylor, K., Klein, E., Manjunatha, U., Gonzales, J., et al., 2008. Tuberculous granulomas are hypoxic in guinea pigs, rabbits, and nonhuman primates. Infect. Immun. 76 (6), 2333– 2340. Vordermeier, H.M., Coombes, A.G., Jenkins, P., McGee, J.P., O’Hagan, D.T., Davis, S.S., Singh, M., 1995. Synthetic delivery system for tuberculosis vaccines: immunological evaluation of the M. tuberculosis 38 kDa protein entrapped in biodegradable PLG microparticles. Vaccine 13 (16), 1576–1582. Vyas, S.P., Kannan, M.E., Jain, S., Mishra, V., Singh, P., 2004. Design of liposomal aerosols for improved delivery of rifampicin to alveolar macrophages. Int. J. Pharm. 269 (1), 37–49. Vyas, S.P., Quraishi, S., Gupta, S., Jaganathan, K.S., 2005. Aerosolized liposome-based delivery of amphotericin B to alveolar macrophages. Int. J. Pharm. 296 (1–2), 12–25. Wagner, A., Vorauer-Uhl, K., 2011. Liposome technology for industrial purposes. J. Drug Deliv. 2011, 591325. Wang, R., Doolan, D.L., Le, T.P., Hedstrom, R.C., Coonan, K.M., Charoenvit, Y., Jones, T.R., Hobart, P., Margalith, M., Ng, J., et al., 1998. Induction of antigen-specific cytotoxic T lymphocytes in humans by a malaria DNA vaccine. Science 282 (5388), 476–480. Wang, D., Xu, J., Feng, Y., Liu, Y., McHenga, S.S., Shan, F., Sasaki, J., Lu, C., 2010. Liposomal oral DNA vaccine (mycobacterium DNA) elicits immune response. Vaccine 28 (18), 3134–3142. Wang, Y., Huang, Y., Xue, C., He, Y., He, Z.G., 2011. ClpR protein-like regulator specifically recognizes RecA protein-independent promoter motif and broadly regulates expression of DNA damage-inducible genes in mycobacteria. J. Biol. Chem. 286 (36), 31159–31167. Wendorf, J., Singh, M., Chesko, J., Kazzaz, J., Soewanan, E., Ugozzoli, M., O’Hagan, D., 2006. A practical approach to the use of nanoparticles for vaccine delivery. J. Pharm. Sci. 95 (12), 2738–2750. Yu, F., Wang, J., Dou, J., Yang, H., He, X., Xu, W., Zhang, Y., Hu, K., Gu, N., 2012. Nanoparticle-based adjuvant for enhanced protective efficacy of DNA vaccine Ag85A-ESAT-6-IL-21 against Mycobacterium tuberculosis infection. Nanomedicine 8 (8), 1337–1344. Zhu, B., Qie, Y., Wang, J., Zhang, Y., Wang, Q., Xu, Y., Wang, H., 2007. Chitosan microspheres enhance the immunogenicity of an Ag85B-based fusion protein containing multiple T-cell epitopes of Mycobacterium tuberculosis. Eur. J. Pharm. Biopharm. 66 (3), 318–326.

Please cite this article in press as: Garg, N.K., et al. Development of novel carrier(s) mediated tuberculosis vaccine: More than a tour de force. Eur. J. Pharm. Sci. (2014), http://dx.doi.org/10.1016/j.ejps.2014.05.028

1212 1213 1214 1215 1216 1217 1218 1219 1220 1221 1222 1223 1224 1225 1226 1227 1228 1229 1230 1231 1232 1233 1234 1235 1236 1237 1238 1239 1240 1241 1242 1243 1244 1245 1246 1247 1248 1249 1250 Q101251 1252 1253 1254 1255 1256 1257 1258 1259 1260 1261 1262 1263 1264 1265 1266 1267 1268