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Baculovirus induces protective antiviral innate immune responses in neonatal chicken
S256
Abstracts / Journal of Biotechnology 136S (2008) S252–S260
IV5-P-012
IV5-P-013
Chitosan-alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY): In vivo evaluation
Baculovirus induces protective antiviral innate immune responses in neonatal chicken
Xiaoyu Li 1,∗ , Liji Jin 1 , Junjun Liu 1 , Huaqiang Li 2 , Yuhong Zhen 1,3 , Yanan Lu 1 , Linhui Wang 1 , Yongping Xu 1
Mingshan Niu ∗ , Wenli Li, Yan Han, Lijia An
1
Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China 2 College of Life Sciences, Liaoning Normal University, Dalian 116029, China 3 Department of Pharmacy, Dalian Medical University, Dalian 116044, China E-mail address: [email protected] (X. Li).
The protective effect of microencapsulated IgY obtained from hens immunized with enterotoxigenic Escherichia coli (ETEC) K88 was evaluated in 40-day-old pigs in which ETEC diarrhea was induced. All pigs were challenged with 5 ml of ETEC K88 at a dose of 1011 cfu/ml per pig at time 0 h. The bacterial suspension was delivered orally to each pig with a syringe in the pig’s oral cavity (Yokoyama et al., 1992). The pigs in treatment groups were treated three times (−1, 4 and 9 h after the challenge) on the first day and twice a day for following two consecutive days each time. The pigs in Treatment I were each given 0.4 g non-encapsulated IgY, suspended in 30 ml of water, delivered as described bacterial suspension; Those pigs in Treatment II were given 2 g of microencapsulated IgY (equivalent to 0.4 g of IgY), flushed down the oral cavity with 30 ml of water; those pigs in Treatment III were given 0.25 g of aureomycin, mixed with proper feed by 30 mL of water whereas the control pigs received 30 mL of saline. The clinical response of each pig was monitored throughout the experiment in terms of occurrence of diarrhea, fecal consistency score, weight loss and cure rate (Marquardt et al., 1998). The results demonstrated that the ETEC K88 induced diarrhea was cured 24 h after treated with microencapsulated IgY, and the body weight gain was the fastest (P < 0.05) during the period of the experiment while those treated with non-encapsulated IgY were cured within 72 h after infection. Whereas control pigs treated with normal saline had severe diarrhea and dehydration after infection. These results indicated that the stability of IgY under gastric conditions was greatly improved by encapsulation in chitosan-alginate microcapsules, and pigs that received microencapsulated IgY were better protected against ETEC infection.
School of Environmental & Biological Science & Technology, Dalian University of Technology, Dalian, China E-mail address: [email protected] (M. Niu). Baculovirus has been used widely for the expression of recombinant proteins and as a biopesticide (Szewczyk et al., 2006). It has been recently demonstrated that Autographa californica nuclear polyhedrosis virus (AcNPV) can stimulate cytokine production in mammalian cells in vitro and protect mice from a lethal challenge of encephalomyocarditis virus (ECMV) and influenza viruses in vivo (Gronowski et al., 1999; Abe et al., 2003). However, there is no information regarding stimulatory activity of BV in avian immune cells. In this study, Antheraea pernyi nuclear polyhedrosis virus, a member of the baculovirus family, is evaluated for its in vitro stimulation of chicken peripheral blood mononuclear cells (PBMC) and macrophage cell line HD 11, and in vivo protection against infectious bronchitis virus (IBV) in neonatal chickens. Our studies showed that ApNPV significantly enhanced inflammatory cytokines mRNA expression in chicken PBMC and HD 11 cells via a clathrin-dependent endocytic and endosomal maturation pathway, and up regulated nitric oxide production in HD 11 cells. Further, it was found that budded virus (BV) can induce antiviral effects in HD 11 cells, while occlusion-derived virus (ODV) cannot. These results indicate that immunostimulatory BV stimulated innate immune activities and enhanced the resistance to infectious virus in neonatal chickens. References Abe, T., Takahashi, H., Hamazaki, H., Miyano-Kurosaki, N., Matsuura, Y., Takaku, H., 2003. Baculovirus induces an innate immune response and confers protection from lethal influenza virus infection in mice. J. Immunol. 171, 1133–1139. Gronowski, A.M., Hilbert, D.M., Sheehan, K.C., Garotta, G., Schreiber, R.D., 1999. Baculovirus stimulates antiviral effects in mammalian cells. J. Virol. 73, 9944–9951. Szewczyk, B., Hoyos-Carvajal, L., Paluszek, M., Skrzecz, W., Souza, M.L., 2006. Baculoviruses-re-emerging biopesticides. Biotech. Adv. 24, 143–160.
doi:10.1016/j.jbiotec.2008.07.547 IV5-P-014
References Marquardt, R.R., Jin, L.Z., Kim, J.W., Fang, L., Frohlich, A.A., Baidoo, S.K., 1998. Passive protective effect of egg-yolk antibodies against enterotoxigenic Escherichia coli K88+ infection in neonatal and early-weaned pigs. FEMS Immunol. Med. Microbiol. 23, 283–288. Yokoyama, H., Peralta, R.C., Diaz, R., Sendo, S., Ikemori, Y., Kodama, Y., 1992. Passive protective effect of chicken egg yolk immunoglobulins against experimental enterotoxigenic Escherichia coli infection in neonatal pigs. Infect. Immun. 60, 998–1007.
doi:10.1016/j.jbiotec.2008.07.546
Insect baculoviruse selectively up-regulates TLRS expression and induces the innate immune response via p38-dependent, ERK-independent pathways in avian macrophage cells (HD11) Han Yan ∗ , Mingshan Niu, Wenli Li, Lijia An Department of Bioscience and Biotechnology, School of Environment and Life, Dalian University of Technology, Dalian, China Our previously work had shown that Antheraea pernyi nuclear polyhedrosis virus (ApNPV), a member of the baculovirus (BV) family is capable of stimulating innate immune response in avian macrophage cells (HD11) (Wang et al., 2007). However, the signal transduction mechanism following BV infection remains unclear. Toll-like receptor (TLR) family plays a key role in pathogen recognition and in the induction of the innate immune response. In mammals, macrophage cells recognized BV by a TLR9 signaling pathway (Abe et al., 2005). While, in avian, the orthologue of TLR9 has not been identified or chicken may not have a TLR9 (Jenkins et al., 2007). In this study, we investigated all the identified TLRs gene expression in HD11 and detected the downstream
Abstracts / Journal of Biotechnology 136S (2008) S252–S260
IV5-P-012
IV5-P-013
Chitosan-alginate microcapsules for oral delivery of egg yolk immunoglobulin (IgY): In vivo evaluation
Baculovirus induces protective antiviral innate immune responses in neonatal chicken
Xiaoyu Li 1,∗ , Liji Jin 1 , Junjun Liu 1 , Huaqiang Li 2 , Yuhong Zhen 1,3 , Yanan Lu 1 , Linhui Wang 1 , Yongping Xu 1
Mingshan Niu ∗ , Wenli Li, Yan Han, Lijia An
1
Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China 2 College of Life Sciences, Liaoning Normal University, Dalian 116029, China 3 Department of Pharmacy, Dalian Medical University, Dalian 116044, China E-mail address: [email protected] (X. Li).
The protective effect of microencapsulated IgY obtained from hens immunized with enterotoxigenic Escherichia coli (ETEC) K88 was evaluated in 40-day-old pigs in which ETEC diarrhea was induced. All pigs were challenged with 5 ml of ETEC K88 at a dose of 1011 cfu/ml per pig at time 0 h. The bacterial suspension was delivered orally to each pig with a syringe in the pig’s oral cavity (Yokoyama et al., 1992). The pigs in treatment groups were treated three times (−1, 4 and 9 h after the challenge) on the first day and twice a day for following two consecutive days each time. The pigs in Treatment I were each given 0.4 g non-encapsulated IgY, suspended in 30 ml of water, delivered as described bacterial suspension; Those pigs in Treatment II were given 2 g of microencapsulated IgY (equivalent to 0.4 g of IgY), flushed down the oral cavity with 30 ml of water; those pigs in Treatment III were given 0.25 g of aureomycin, mixed with proper feed by 30 mL of water whereas the control pigs received 30 mL of saline. The clinical response of each pig was monitored throughout the experiment in terms of occurrence of diarrhea, fecal consistency score, weight loss and cure rate (Marquardt et al., 1998). The results demonstrated that the ETEC K88 induced diarrhea was cured 24 h after treated with microencapsulated IgY, and the body weight gain was the fastest (P < 0.05) during the period of the experiment while those treated with non-encapsulated IgY were cured within 72 h after infection. Whereas control pigs treated with normal saline had severe diarrhea and dehydration after infection. These results indicated that the stability of IgY under gastric conditions was greatly improved by encapsulation in chitosan-alginate microcapsules, and pigs that received microencapsulated IgY were better protected against ETEC infection.
School of Environmental & Biological Science & Technology, Dalian University of Technology, Dalian, China E-mail address: [email protected] (M. Niu). Baculovirus has been used widely for the expression of recombinant proteins and as a biopesticide (Szewczyk et al., 2006). It has been recently demonstrated that Autographa californica nuclear polyhedrosis virus (AcNPV) can stimulate cytokine production in mammalian cells in vitro and protect mice from a lethal challenge of encephalomyocarditis virus (ECMV) and influenza viruses in vivo (Gronowski et al., 1999; Abe et al., 2003). However, there is no information regarding stimulatory activity of BV in avian immune cells. In this study, Antheraea pernyi nuclear polyhedrosis virus, a member of the baculovirus family, is evaluated for its in vitro stimulation of chicken peripheral blood mononuclear cells (PBMC) and macrophage cell line HD 11, and in vivo protection against infectious bronchitis virus (IBV) in neonatal chickens. Our studies showed that ApNPV significantly enhanced inflammatory cytokines mRNA expression in chicken PBMC and HD 11 cells via a clathrin-dependent endocytic and endosomal maturation pathway, and up regulated nitric oxide production in HD 11 cells. Further, it was found that budded virus (BV) can induce antiviral effects in HD 11 cells, while occlusion-derived virus (ODV) cannot. These results indicate that immunostimulatory BV stimulated innate immune activities and enhanced the resistance to infectious virus in neonatal chickens. References Abe, T., Takahashi, H., Hamazaki, H., Miyano-Kurosaki, N., Matsuura, Y., Takaku, H., 2003. Baculovirus induces an innate immune response and confers protection from lethal influenza virus infection in mice. J. Immunol. 171, 1133–1139. Gronowski, A.M., Hilbert, D.M., Sheehan, K.C., Garotta, G., Schreiber, R.D., 1999. Baculovirus stimulates antiviral effects in mammalian cells. J. Virol. 73, 9944–9951. Szewczyk, B., Hoyos-Carvajal, L., Paluszek, M., Skrzecz, W., Souza, M.L., 2006. Baculoviruses-re-emerging biopesticides. Biotech. Adv. 24, 143–160.
doi:10.1016/j.jbiotec.2008.07.547 IV5-P-014
References Marquardt, R.R., Jin, L.Z., Kim, J.W., Fang, L., Frohlich, A.A., Baidoo, S.K., 1998. Passive protective effect of egg-yolk antibodies against enterotoxigenic Escherichia coli K88+ infection in neonatal and early-weaned pigs. FEMS Immunol. Med. Microbiol. 23, 283–288. Yokoyama, H., Peralta, R.C., Diaz, R., Sendo, S., Ikemori, Y., Kodama, Y., 1992. Passive protective effect of chicken egg yolk immunoglobulins against experimental enterotoxigenic Escherichia coli infection in neonatal pigs. Infect. Immun. 60, 998–1007.
doi:10.1016/j.jbiotec.2008.07.546
Insect baculoviruse selectively up-regulates TLRS expression and induces the innate immune response via p38-dependent, ERK-independent pathways in avian macrophage cells (HD11) Han Yan ∗ , Mingshan Niu, Wenli Li, Lijia An Department of Bioscience and Biotechnology, School of Environment and Life, Dalian University of Technology, Dalian, China Our previously work had shown that Antheraea pernyi nuclear polyhedrosis virus (ApNPV), a member of the baculovirus (BV) family is capable of stimulating innate immune response in avian macrophage cells (HD11) (Wang et al., 2007). However, the signal transduction mechanism following BV infection remains unclear. Toll-like receptor (TLR) family plays a key role in pathogen recognition and in the induction of the innate immune response. In mammals, macrophage cells recognized BV by a TLR9 signaling pathway (Abe et al., 2005). While, in avian, the orthologue of TLR9 has not been identified or chicken may not have a TLR9 (Jenkins et al., 2007). In this study, we investigated all the identified TLRs gene expression in HD11 and detected the downstream