Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology · 2008
[Development of a CFSE-based flow cytometry for evaluating EIAV-stimulated proliferation of T lymphocytes].
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Lin Yue-zhi, Deng Xi-lin, Shen Nan, Lü Xiao-ling, Zhao Li-ping, Kong Xian-gang, Shao Yi-ming, Zhou Jian-hua
Аннотация
AIM: To develop a flow cytometry using (5-carboxyfluorescein diacetate succinmidyl ester, CFSE) to detect the proliferation of specific T lymphocytes from equine infectious anemia virus (EIAV).
METHODS: Peripheral blood mononuclear cells (PBMC) were isolated, stained with CFSE and incubated with EIAV for 5 days. After interacted with either CD4(+) or CD8(+) antibody, the cells were detected for proliferated population, which contained serially 2-fold reduced CFSE in CD4(+) and CD8(+) T lymphocytes.
RESULTS: The concentration of CFSE, and the type, concentration and reaction time of EIAV-specific antigens were tested and optimized to improve the sensitivity and specificity of this flow cytometry-based assay. PBMCs isolated from the horses infected with either virulent or vaccine strains of EIAV and health controls were analyzed to evaluate this technique. The proliferation rates of CD4(+) and CD8(+) T lymphocytes were observed in the cells from differently treated horses.
CONCLUSION: The CFSE-based flow cytometry is a reliable tool to analyze antigen-specific proliferation of lymphocytes, which plays an important role in evaluating the immune responses induced by EIAV and other viruses.
METHODS: Peripheral blood mononuclear cells (PBMC) were isolated, stained with CFSE and incubated with EIAV for 5 days. After interacted with either CD4(+) or CD8(+) antibody, the cells were detected for proliferated population, which contained serially 2-fold reduced CFSE in CD4(+) and CD8(+) T lymphocytes.
RESULTS: The concentration of CFSE, and the type, concentration and reaction time of EIAV-specific antigens were tested and optimized to improve the sensitivity and specificity of this flow cytometry-based assay. PBMCs isolated from the horses infected with either virulent or vaccine strains of EIAV and health controls were analyzed to evaluate this technique. The proliferation rates of CD4(+) and CD8(+) T lymphocytes were observed in the cells from differently treated horses.
CONCLUSION: The CFSE-based flow cytometry is a reliable tool to analyze antigen-specific proliferation of lymphocytes, which plays an important role in evaluating the immune responses induced by EIAV and other viruses.