- Email: [email protected]
B vaccine
24 June 1997 - Poster presentations
glutinin peptide HA1 188-205 were investigated for their cytokine secretion profiles. MatarIaIaand Mathods:T cells were maintained in vitro by stimulation with A/x31/88 virus and syngeneic APC under identical culture conditions. T cell supematants were analysed for cyiokine secretion after antigenic stimulation, IL-3 by the ability to stimulate an IL-3 dependent cell line, IFN-y and IL-5 by ELISA capture assays. T cell receptor a and @chains were sequenced by standard procedures. Resufts~ Two clones B19 and 817 established by limiting dilution from a single polyclonal cell line expressed identical T cell receptor (I and fi chains but exhibited differences in cytokine profiles. Here we find that T cell clones established under identical conditions exhibit phenotypic diversity. Concluaiona:It is considered that a THI or THYphenotype is a consequence of route of infection or immunisation or of exposure to lymphokines in vitro (IL-4 or IL-10 or IFN-y). However we find that T cell clones established under identical condiions exhibit phenotypic diversity. These results suggest that individual cells may exhibit complex and heterogenous combinations of cytokine secretion. Many cells may not be classified into THI, THY subsets based on the original ctiteria.
Znununityto viruses
ReauitazThe group of RSV positive infants had a significantly higher expression of CD23 antigen on CD20+ B lymphocytes when compared to healthy children and to their RSV positive mothers. Three color immunofluorescence analysis revealed that in RSV infected, infants and mothers, elevated CD23 antigen expression was observed on particular CD20+ CD21-subpopulation of B cells. There was no difference in the percentage of CD21+ B lymphocytes in infants with RSV infection in comparison to healthy infants. The amount of CD23 antigen expression in RSV pdsitive mothers ~8s higher then in healthy age- and sex-matched historical controls. Conclualon: Present results, obtained on additional larger group of tested children with RSV infection, confirmed that RSV infection in infants provoked elevated expression of low-affinity IgE receptor (CD23) on B cells. The same changes were obtained in the group of RSV positive mothers. Therefore, it seems that induction of CD23 expression after RSV infection could be attributed to the characteristics of the virus, rather than to the functional immaturity of immune cells. The exact relationship between increased CD23 antigen and RSV infection remains to be determined. We currently investigate the predominant cytokine profile in CD4+ T lymphocyte in RSV infected infants. P.4.04.27
P.4.04.25
Epltope specificity of Thflh2 CD4+ T lymphocyte cla~l~;nduced by vacclnatlon with rHBsAg
MC. Honorati I**, P. Dolzani ‘, E. Matiani ‘.*, A. Piacentini ‘, G. Lisignoli ‘, C. Ferrari 3+‘,A. Facchini I.*. ’ Laboratorio di lmmunologia e Genetica, lstituto di Ricema Codivilla Pufti, 1.0.R., Bologna, Ifa& * Di~timento di Medicina lntema e Gastmentemlogia, Univers& di Bokgna,’ Ita& 3Cattedta di Malattie Infetffve, Univwsit’d di Parma, Itak 4Divkione di Ma/aft& InfWve e lmmunopatologia Vi&e, Azienda Ospedaliera di Panna, Italy
Introduction:Different amino acid sequences of HBsAg are involved in the activation of CD4+ lymphocytes needed to induce an optimal antiviral function. Aim of the present study was to characterize the CDCmediated response to immunodominant HBsAg epitopes by defining minimal sequences recognized by T cells, cytokine profiles and HLA restriction of peptide recognition. Matarlalaand Methods:T lymphocyte lines and clones specific for HBsAg were isolated from the peripheral blood of subjects immunized with recombinant HBsAgand stimulated in vitrowith synthetic peptides spanning the whole HBsAg sequence. Reeults: Four immunodominant epitopes (sequences 21-40, 136-155, 156-175 and 211-226) were identified. Using panels of truncated peptides of different length, sequences 21-28, 165-172 and 215-223 were shown to correspond to the minimal epitopes recognized by T cells. The antigen-specific T lymphocyte proliferation was HLA cl& II-rest&ted and each p&tide could bs presented in association with different HLA class II determinants. Th01Th2 cytokine patterns were induced upon peptide stimulation. Conclualon:These results indicate the presence of at least four immundominant epitopes within HBsAg which represent potential candidates for the design of anti-HBV synthetic vaccines. This work was supported by the Italian Ministry of Health and University of Bologna, Italy.
S. RabatC’, A. Gagro ‘, V. K&uloviC-HreSiC*, R. Lokar-Kdbas *, V. Dr&enoviC 3, G. MinariC-GalinoviC4. ’ Institute of Immunology Zagreb, Croatia, *University Children’s Hospital, Zagreb, Croatia, 3Diviskm of Virolog): Croatian Institute of Public Health, Zagreb, Croatia, 4Department of Microbiollogy;” A. &aF School of kkiic Health, Za&eb, Croatia
Introduction: Acute respiratory tract infection with RSV may induce asthma-like symptoms followed by an RSV-specific IgE response in infants. We have previously observed (Rabatib et. al. J. Infect.Dis. 175: 32, 1997). that some infants with RSV infection had increased percentage of CD23+ B lymphocytes and that these children had RSV-specific IgE and lgG4 antibodies detectable at the acute stage of infection. These effects may be a result of Th2-like response to RSV infection. The aim of present study was to investigate weather the observed changes in CD23 expression in RSV infected infants could be related to the immature immune response or are a consequence of direct RSV impact on B cells. Materlala and Mathoda:Twenty-one children (mean age 4 months) with bronchiolitis and RSV infection, 21 mothers (eight RSV positive) and 9 healthy infants (mean age 4 months) were included in the study. Two and three color immunofluorescence analysis of CD23 and CD21 antigen on CD20+ B cells were analyzed on flow-cytometer. Rapid viral antigen detection and isolation were used to diagnose the RSV infection.
lmmunogeniclty of two injectlons of a combined hepatitis A/B vaccine
X.Q.Chen’, G.J. Boland’, G.C. deGast*, R.P.M. van Kesse13,J.E. van Steenbergen 3, J. van Hattum ’ ’ Deparfment of Gasfroenferology, Ufrecbt, The Netherlands, *Department of Immunohaematology, University Hospital of Utrecht, Utmcht, The Netherlands, 3Municipal Health Service. Utrecht. The Netherlands Introduction:To investigate the immunogenicity of two injections (0 and 6 months) of combined hepatitis A/hepatitis B vaccine and that of simultaneous separate injection of hepatitis A and B vaccines. Mathoda:Healthy volunteers (n = 103), aged 14 to 59 years, negative for anti-HAV and anti-HBs were randomly allocated into four groups: group 1 (n = 34), group 2 (n = 31) and group 3 (n = 29) received combined hepatitis A/B vaccine lot 1, 2 and 3 respectively, and group 4 (n = 9), received simultaneous injections in separate arms of hepatitis A and hepatitis B vaccines, as control. Serum anti-HAV and anti-HBs were determined before and 1, 3, 6, 7 and 12 months after the first injection. Results:At month 7, anti-HAV seroconversion was 100% in all orouos. AntiHAV GMTs were respectively 7,464, 11,020, 11,272, and 7,316 mlIj/mi without significant differences between the groups. The anti-HBaSeroconversion rates of combined hepatitis AB vaccine were respectively 94% for lot 1, 93% for lot 2, 100% for both lot 3 and for separate injections of HA and HB vaccines. Lot 3 even retained 100% seroconversion at month 12. Anti-HBs GMTs at month 7 were 342, 313,778 and I,918 mlU/mL in the four groups, respectively. Conclusion: Two injections of combined hepatitis AiB vaccine were well tolerated in healthy adults and adolescents. Lot 3 of combined vaccine induced an effective and adequate protection against both hepatitis A and hepatitis B which was comparable to simultaneous separate injection of hepatitis A and B vaccines.
P.4.04.26 1P.4.04.26 1 The influence of respiratory syncytial virus (RSV) infectlon on the expression of low affinity IgE receptor (CD29 in infants and their mothers
301
Characterization of herpes simplex virus (HSV) specific T cells isolated from vitreous fluid of patients with HSV-mediated acute retinal necrosis
G.M.G.M. Verjans’.*, E.J. Feron’, J.G.C. Comelissen’, G.S. Baarsma’, A. van der Lelij 3, A.D.M.E. Osterhaus *. ’ The Rotterdam Eye Hospital, Rotterdam, The Netherlands. * Depatiment of Virology, Erasmus Univetsiw Rotterdam, The Nether/an&, 3Depament of Ophthalmology, Academic Hospital utrecht, Utmcht, The Netherlands
Objactlvaa:Acute retinal necrosis (ARN) is an inflammatory eye disease which often leads to severe impairment of vision within days or weeks after onset. In most cases, ARN is caused by an infection with either varicella zoster virus (VZV) or HSV. Not only the direct cytopathic effect of the virus, but also the (cellular) immune response against the virus and/or retinal autoantigens are causing the disease. To address the possible involvement of virus-specific T cells in the immunopathology of ARN we have analyzed the antigen-specificity of intraocular T cells from 3 patients with a defined HSV-mediated ARN. Methods:We designed a strategy to isolate and characterize Tcells obtained from vitreous fluid (VF) of patients with HSV mediated ARN. T cell lines were generated by mitogenic stimulation of vitreal cells and PBMC in the oresence of 11-2and ail&neic feeders. Subsequently, T cell clones (XC) wile obtained by limiting dilution. Autologous Epstein Barr virus transfoned B ceils (BLCL) were used as antigen presenting cells (APC). The virus-specificity of theT cells was analyzed in standard proliferation and [51Crl_releaseassays using either HSV-1, HSV-2 or mock infected BLCL. The cytokine secretion levels of the TCC was determined by ELISA in the culture supematants of antigenic stimulated TCC.
glutinin peptide HA1 188-205 were investigated for their cytokine secretion profiles. MatarIaIaand Mathods:T cells were maintained in vitro by stimulation with A/x31/88 virus and syngeneic APC under identical culture conditions. T cell supematants were analysed for cyiokine secretion after antigenic stimulation, IL-3 by the ability to stimulate an IL-3 dependent cell line, IFN-y and IL-5 by ELISA capture assays. T cell receptor a and @chains were sequenced by standard procedures. Resufts~ Two clones B19 and 817 established by limiting dilution from a single polyclonal cell line expressed identical T cell receptor (I and fi chains but exhibited differences in cytokine profiles. Here we find that T cell clones established under identical conditions exhibit phenotypic diversity. Concluaiona:It is considered that a THI or THYphenotype is a consequence of route of infection or immunisation or of exposure to lymphokines in vitro (IL-4 or IL-10 or IFN-y). However we find that T cell clones established under identical condiions exhibit phenotypic diversity. These results suggest that individual cells may exhibit complex and heterogenous combinations of cytokine secretion. Many cells may not be classified into THI, THY subsets based on the original ctiteria.
Znununityto viruses
ReauitazThe group of RSV positive infants had a significantly higher expression of CD23 antigen on CD20+ B lymphocytes when compared to healthy children and to their RSV positive mothers. Three color immunofluorescence analysis revealed that in RSV infected, infants and mothers, elevated CD23 antigen expression was observed on particular CD20+ CD21-subpopulation of B cells. There was no difference in the percentage of CD21+ B lymphocytes in infants with RSV infection in comparison to healthy infants. The amount of CD23 antigen expression in RSV pdsitive mothers ~8s higher then in healthy age- and sex-matched historical controls. Conclualon: Present results, obtained on additional larger group of tested children with RSV infection, confirmed that RSV infection in infants provoked elevated expression of low-affinity IgE receptor (CD23) on B cells. The same changes were obtained in the group of RSV positive mothers. Therefore, it seems that induction of CD23 expression after RSV infection could be attributed to the characteristics of the virus, rather than to the functional immaturity of immune cells. The exact relationship between increased CD23 antigen and RSV infection remains to be determined. We currently investigate the predominant cytokine profile in CD4+ T lymphocyte in RSV infected infants. P.4.04.27
P.4.04.25
Epltope specificity of Thflh2 CD4+ T lymphocyte cla~l~;nduced by vacclnatlon with rHBsAg
MC. Honorati I**, P. Dolzani ‘, E. Matiani ‘.*, A. Piacentini ‘, G. Lisignoli ‘, C. Ferrari 3+‘,A. Facchini I.*. ’ Laboratorio di lmmunologia e Genetica, lstituto di Ricema Codivilla Pufti, 1.0.R., Bologna, Ifa& * Di~timento di Medicina lntema e Gastmentemlogia, Univers& di Bokgna,’ Ita& 3Cattedta di Malattie Infetffve, Univwsit’d di Parma, Itak 4Divkione di Ma/aft& InfWve e lmmunopatologia Vi&e, Azienda Ospedaliera di Panna, Italy
Introduction:Different amino acid sequences of HBsAg are involved in the activation of CD4+ lymphocytes needed to induce an optimal antiviral function. Aim of the present study was to characterize the CDCmediated response to immunodominant HBsAg epitopes by defining minimal sequences recognized by T cells, cytokine profiles and HLA restriction of peptide recognition. Matarlalaand Methods:T lymphocyte lines and clones specific for HBsAg were isolated from the peripheral blood of subjects immunized with recombinant HBsAgand stimulated in vitrowith synthetic peptides spanning the whole HBsAg sequence. Reeults: Four immunodominant epitopes (sequences 21-40, 136-155, 156-175 and 211-226) were identified. Using panels of truncated peptides of different length, sequences 21-28, 165-172 and 215-223 were shown to correspond to the minimal epitopes recognized by T cells. The antigen-specific T lymphocyte proliferation was HLA cl& II-rest&ted and each p&tide could bs presented in association with different HLA class II determinants. Th01Th2 cytokine patterns were induced upon peptide stimulation. Conclualon:These results indicate the presence of at least four immundominant epitopes within HBsAg which represent potential candidates for the design of anti-HBV synthetic vaccines. This work was supported by the Italian Ministry of Health and University of Bologna, Italy.
S. RabatC’, A. Gagro ‘, V. K&uloviC-HreSiC*, R. Lokar-Kdbas *, V. Dr&enoviC 3, G. MinariC-GalinoviC4. ’ Institute of Immunology Zagreb, Croatia, *University Children’s Hospital, Zagreb, Croatia, 3Diviskm of Virolog): Croatian Institute of Public Health, Zagreb, Croatia, 4Department of Microbiollogy;” A. &aF School of kkiic Health, Za&eb, Croatia
Introduction: Acute respiratory tract infection with RSV may induce asthma-like symptoms followed by an RSV-specific IgE response in infants. We have previously observed (Rabatib et. al. J. Infect.Dis. 175: 32, 1997). that some infants with RSV infection had increased percentage of CD23+ B lymphocytes and that these children had RSV-specific IgE and lgG4 antibodies detectable at the acute stage of infection. These effects may be a result of Th2-like response to RSV infection. The aim of present study was to investigate weather the observed changes in CD23 expression in RSV infected infants could be related to the immature immune response or are a consequence of direct RSV impact on B cells. Materlala and Mathoda:Twenty-one children (mean age 4 months) with bronchiolitis and RSV infection, 21 mothers (eight RSV positive) and 9 healthy infants (mean age 4 months) were included in the study. Two and three color immunofluorescence analysis of CD23 and CD21 antigen on CD20+ B cells were analyzed on flow-cytometer. Rapid viral antigen detection and isolation were used to diagnose the RSV infection.
lmmunogeniclty of two injectlons of a combined hepatitis A/B vaccine
X.Q.Chen’, G.J. Boland’, G.C. deGast*, R.P.M. van Kesse13,J.E. van Steenbergen 3, J. van Hattum ’ ’ Deparfment of Gasfroenferology, Ufrecbt, The Netherlands, *Department of Immunohaematology, University Hospital of Utrecht, Utmcht, The Netherlands, 3Municipal Health Service. Utrecht. The Netherlands Introduction:To investigate the immunogenicity of two injections (0 and 6 months) of combined hepatitis A/hepatitis B vaccine and that of simultaneous separate injection of hepatitis A and B vaccines. Mathoda:Healthy volunteers (n = 103), aged 14 to 59 years, negative for anti-HAV and anti-HBs were randomly allocated into four groups: group 1 (n = 34), group 2 (n = 31) and group 3 (n = 29) received combined hepatitis A/B vaccine lot 1, 2 and 3 respectively, and group 4 (n = 9), received simultaneous injections in separate arms of hepatitis A and hepatitis B vaccines, as control. Serum anti-HAV and anti-HBs were determined before and 1, 3, 6, 7 and 12 months after the first injection. Results:At month 7, anti-HAV seroconversion was 100% in all orouos. AntiHAV GMTs were respectively 7,464, 11,020, 11,272, and 7,316 mlIj/mi without significant differences between the groups. The anti-HBaSeroconversion rates of combined hepatitis AB vaccine were respectively 94% for lot 1, 93% for lot 2, 100% for both lot 3 and for separate injections of HA and HB vaccines. Lot 3 even retained 100% seroconversion at month 12. Anti-HBs GMTs at month 7 were 342, 313,778 and I,918 mlU/mL in the four groups, respectively. Conclusion: Two injections of combined hepatitis AiB vaccine were well tolerated in healthy adults and adolescents. Lot 3 of combined vaccine induced an effective and adequate protection against both hepatitis A and hepatitis B which was comparable to simultaneous separate injection of hepatitis A and B vaccines.
P.4.04.26 1P.4.04.26 1 The influence of respiratory syncytial virus (RSV) infectlon on the expression of low affinity IgE receptor (CD29 in infants and their mothers
301
Characterization of herpes simplex virus (HSV) specific T cells isolated from vitreous fluid of patients with HSV-mediated acute retinal necrosis
G.M.G.M. Verjans’.*, E.J. Feron’, J.G.C. Comelissen’, G.S. Baarsma’, A. van der Lelij 3, A.D.M.E. Osterhaus *. ’ The Rotterdam Eye Hospital, Rotterdam, The Netherlands. * Depatiment of Virology, Erasmus Univetsiw Rotterdam, The Nether/an&, 3Depament of Ophthalmology, Academic Hospital utrecht, Utmcht, The Netherlands
Objactlvaa:Acute retinal necrosis (ARN) is an inflammatory eye disease which often leads to severe impairment of vision within days or weeks after onset. In most cases, ARN is caused by an infection with either varicella zoster virus (VZV) or HSV. Not only the direct cytopathic effect of the virus, but also the (cellular) immune response against the virus and/or retinal autoantigens are causing the disease. To address the possible involvement of virus-specific T cells in the immunopathology of ARN we have analyzed the antigen-specificity of intraocular T cells from 3 patients with a defined HSV-mediated ARN. Methods:We designed a strategy to isolate and characterize Tcells obtained from vitreous fluid (VF) of patients with HSV mediated ARN. T cell lines were generated by mitogenic stimulation of vitreal cells and PBMC in the oresence of 11-2and ail&neic feeders. Subsequently, T cell clones (XC) wile obtained by limiting dilution. Autologous Epstein Barr virus transfoned B ceils (BLCL) were used as antigen presenting cells (APC). The virus-specificity of theT cells was analyzed in standard proliferation and [51Crl_releaseassays using either HSV-1, HSV-2 or mock infected BLCL. The cytokine secretion levels of the TCC was determined by ELISA in the culture supematants of antigenic stimulated TCC.