Chicken coccidiosis is a major parasitic disease with substantial economic burden to the poultry industry.Enhanced green fluorescent protein(EGFP) tagged recombinant Bacille Calmette-Guerin(rBCG),as a fusion protein w...Chicken coccidiosis is a major parasitic disease with substantial economic burden to the poultry industry.Enhanced green fluorescent protein(EGFP) tagged recombinant Bacille Calmette-Guerin(rBCG),as a fusion protein with coccidian rhomboid antigen was constructed to track rBCG in vivo in chickens in this study.Immunization of chickens with one dose of rBCG pMV361-Rho/EGFP induced humoral immune response.The colonization of rBCG in liver,spleen,lung,kidney and caecum was observed by laser confocal microscopy.Real-time quantitative RT-PCR showed a rise expression level of rhomboid protein on the 7th day and a peak on the 14th day and disappearance on the 28th day after immunization.These results have significant implications for the development of rBCG vaccines against avian coccidiosis.展开更多
Objective: To determine the effects of toll-like receptor 2(TLR-2) agonist, Pam3 CSK4, on cellular and humoral immune response against recombinant Mycobacterium bovis bacille Calmette-Guérin(rBCG) expressing the ...Objective: To determine the effects of toll-like receptor 2(TLR-2) agonist, Pam3 CSK4, on cellular and humoral immune response against recombinant Mycobacterium bovis bacille Calmette-Guérin(rBCG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum.Methods: Six groups of mice(n=6 per group) received intraperitoneal phosphate buffered saline T80(PBS-T80), BCG or rBCG in the presence or absence of Pam3 CSK4. Enzymelinked immunosorbent assay was carried out to measure serum total IgG, IgG1, IgG2 a, and Ig G2 b production. Spleens were also harvested and splenocytes were co-cultured with rBCG antigen for in vitro determination of IL-4 and IFN-γ via enzyme-linked immunosorbent assay.Results: The production of total IgG and the isotype IgG1, IgG2 a and IgG2 b was significantly higher in rBCG-immunised mice than in the BCG and PBS-T80-immunised mice, and Pam3 CSK4 further enhanced their productions. A similar pattern was also observed in IFN-γ production. Moreover, there was no significant difference in IL-4 production in all groups either in the presence or absence of Pam3 CSK4.Conclusions: We present evidence of the adjuvant effects of TLR-2 agonist in enhancing the production of total IgG, IgG1, IgG2 a, IgG2 b, as well as IFN-γ in response to rBCG. However, the presence or absence of Pam3 CSK4 had no effect on IL-4 production.展开更多
基金Supported by Grants from the Foundation of the Jilin Provincial Science and Technology Department of China (Grant No.20050211-2)the National Natural Science Foundation of China (Grant No.30170696,30500370 and 30671580)
文摘Chicken coccidiosis is a major parasitic disease with substantial economic burden to the poultry industry.Enhanced green fluorescent protein(EGFP) tagged recombinant Bacille Calmette-Guerin(rBCG),as a fusion protein with coccidian rhomboid antigen was constructed to track rBCG in vivo in chickens in this study.Immunization of chickens with one dose of rBCG pMV361-Rho/EGFP induced humoral immune response.The colonization of rBCG in liver,spleen,lung,kidney and caecum was observed by laser confocal microscopy.Real-time quantitative RT-PCR showed a rise expression level of rhomboid protein on the 7th day and a peak on the 14th day and disappearance on the 28th day after immunization.These results have significant implications for the development of rBCG vaccines against avian coccidiosis.
基金supported by the Universiti Sains Malaysia(USM)Research University(RU)Grants(No.1001/PPSK/8011100)
文摘Objective: To determine the effects of toll-like receptor 2(TLR-2) agonist, Pam3 CSK4, on cellular and humoral immune response against recombinant Mycobacterium bovis bacille Calmette-Guérin(rBCG) expressing the C-terminus of merozoite surface protein-1 of Plasmodium falciparum.Methods: Six groups of mice(n=6 per group) received intraperitoneal phosphate buffered saline T80(PBS-T80), BCG or rBCG in the presence or absence of Pam3 CSK4. Enzymelinked immunosorbent assay was carried out to measure serum total IgG, IgG1, IgG2 a, and Ig G2 b production. Spleens were also harvested and splenocytes were co-cultured with rBCG antigen for in vitro determination of IL-4 and IFN-γ via enzyme-linked immunosorbent assay.Results: The production of total IgG and the isotype IgG1, IgG2 a and IgG2 b was significantly higher in rBCG-immunised mice than in the BCG and PBS-T80-immunised mice, and Pam3 CSK4 further enhanced their productions. A similar pattern was also observed in IFN-γ production. Moreover, there was no significant difference in IL-4 production in all groups either in the presence or absence of Pam3 CSK4.Conclusions: We present evidence of the adjuvant effects of TLR-2 agonist in enhancing the production of total IgG, IgG1, IgG2 a, IgG2 b, as well as IFN-γ in response to rBCG. However, the presence or absence of Pam3 CSK4 had no effect on IL-4 production.
