Immune responses specific for ESAT-6 expressed by recombinant Salmonella typhimurium
10.3760/cma.j.issn.0254-5101.2009.06.012
- VernacularTitle:重组沙门菌表达结核分枝杆菌ESAT-6及其特异性免疫应答分析
- Author:
Hui ZHANG
;
Liu LIU
;
Shizhong GENG
;
Maozhi HU
;
Ke WEN
;
Zhiming PAN
;
Xinan JIAO
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis;
ESAT-6;
Intranasal immunization;
Cellular immune re-sponse
- From:
Chinese Journal of Microbiology and Immunology
2009;29(6):528-532
- CountryChina
- Language:Chinese
-
Abstract:
Objective To determine the immune responses induced by recombinant Salmonella ty-phimurium expressing the secreting antigen ESAT-6 of Mycobacterium tuberculosis. Methods ESAT-6 cod-ing gene was cloned and identified by PCR and sequencing. Prokaryotic expression plasmid pYA33-esat car-rying the ESAT-6 coding sequence was constructed firstly and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria was named as X4550(33-esat). C57BL/6 mice were immunized intranasally (I. N) with 108 CFU recombinant bacteria at day 0 and 18. Cells from spleen, lung, mesenteric lymph node (MLN) and Peyer's patch (PP) were collected from mice after second immu-nization, and the specific IFN-γ-secreting cells and IL-4-secreting cells were detected by ELISPOT assay u-sing ESAT-6 peptide as stimulus. Furthermore, CTL effects were in vivo evaluated by CFSE assay. Results The results showed that cellular immune responses specific for ESAT-6 could be detected by ELISPOT assay. In lung and PP cells, immune responses against ESAT-6 were biased toward Th1 type, the frequency of IFN-γ-secreting cells was much higher than that of IL-4-secreting cells. In splenocytes and MLN cells, the anti-gen specific immune responses acted as Thl and Th2 balance, the frequency of IFN-γ-secreting cells was close to that of IL-4-secreting cells. CFSE assay indicated that recombinant bacteria could induce the high level of CTL effects specific for ESAT-6 peptide. Conclusion These results suggested that recombinant Sal-monella typhimurium X4550(33-esat) not only can induce cellular immune responses, but also can elicit specific CTL responses after I. N immunization. It also provided the useful information for the control of infec-tious disease of tuberculosis.
