Detection of Aspergillus fumigatus thioredoxin reductase GliT antigen-specific T cells by ELISPOT assay in healthy individuals and result evaluation
- VernacularTitle:健康人外周血烟曲霉硫氧还蛋白还原酶抗原特异性T 淋巴细胞检测与结果评价
- Author:
Lining SHI
;
Yuan HU
;
Hong LIAO
;
Xue HAN
;
Dandan HAN
;
Xiaojun LI
;
Fangqiu LI
- Publication Type:Journal Article
- Keywords:
Aspergillus fumigates;
Thioredoxin reductase GliT;
T-lymphocytes;
Cytokines;
ELISPOT
- From:
Journal of Medical Postgraduates
2014;(12):1286-1289
- CountryChina
- Language:Chinese
-
Abstract:
Objective The purpose of this study was to evaluate the protective potential of the Aspergillus fumigatus thiore -doxin reductase GliT ( TR) antigen by establishing and optimizing ELISPOT assay for TR antigen-specific T cells ( TR/AST) secreting IFN-γand IL-4 in peripheral blood mononuclear cells ( PBMCs ) and explore the role of TR/AST in invasive aspergillosis ( IA ) . Methods We optimized the reaction conditions of ELISPOT by preliminary checkerboard titration and determined the frequencies of positive spot-forming cells ( SFCs) specifically secreting IFN-γand IL-4 in the PBMCs of 20 healthy individuals with TR as specific stimulant and with PHA and PMA as positive controls ,. Results Checkerboard titration demonstrated the best result of ELISPOT with the TR antigen at the final concentration of 10μg/well and PBMCs at 3 ×105/well.The median frequency of IFN-γSFCs was sig-nificantly higher (15 [3.5, 59.5]) than that of IL-4 SFCs (0 [0, 0]) (P<0.001).TR induced IFN-γresponses in all the 20 healthy donors, including 9 cases of strong IFN-γresponse (SFCs>20/3 ×105 PBMCs), accounting for 45%, but failed to induce IL-4 response in 19 of the healthy individuals . Conclusion The Aspergillus fumigatus TR antigen could induce an immunodominant Th1 response , and therefore might be a potential protective antigen .
