Detection of cellular immunity to rabies antigens in human vaccinees
Detection of cellular immunity to rabies antigens in human vaccinees
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DOI：
10.1007/s10875-006-9044-0
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发表时间：
2006-11-01
影响因子：
9.1
通讯作者：
Briggs, Deborah J.
中科院分区：
文献类型：
--
作者：
Moore, Susan M.;Wilkerson, Melinda J.;Briggs, Deborah J.
建立了一种非放射性多参数流式细胞术，用于鉴定先前接种狂犬病毒疫苗的人类受试者的抗原特异性淋巴细胞，并随后与标准氚化胸腺嘧啶方法进行比较。细胞跟踪染料羧基荧光素琥珀酰酯与CD4和CD8细胞的表面标记结合使用，以便在抗原召回试验中确定淋巴细胞对杀死的狂犬病毒的反应。狂犬病病毒特异性淋巴细胞反应在10名接种疫苗和5名未接种疫苗的受试者中分别与体液免疫反应进行了比较。所有10名接种者均观察到淋巴细胞对狂犬病毒的反应；有的早在刺激后3天就出现，有的则在刺激后7天才出现。CFSE法的增殖指数与[H-3]胸腺嘧啶法的模拟指数吻合较好（kappa统计量=0.73）。狂犬病病毒中和抗体（RVNA）水平与接种者淋巴细胞对灭活狂犬病病毒的反应呈反比关系。细胞因子的产生与体液和细胞反应水平之间的关系在四个有代表性的受试者中进行了研究。两名高增殖指数和低RVNA滴度的疫苗接种者对狂犬病毒刺激产生Th1型细胞因子，而两名低增殖指数和高RVNA滴度的疫苗接种者不产生这些细胞因子。
A nonradioactive multi-parameter flow cytometry assay was developed to identify antigen-specific lymphocytes in human subjects previously vaccinated against rabies virus and was subsequently compared to the standard tritiated thymidine method. A cell tracking dye, carboxyfluorescein succinimidyl ester, was used in combination with surface label for CD4 and CD8 cells in order to determine the response of lymphocytes to killed rabies virus in an antigen recall assay. The rabies virus-specific lymphocyte response was compared to the humoral immune response in each of ten vaccinated and five non-vaccinated subjects. Lymphocyte responses to rabies virus were observed in all ten vaccinated subjects; some noted as early as 3 days after stimulation while others were not until 7 days after stimulation. There was good agreement between the proliferation index of the CFSE assay and the simulation index of the [H-3]thymidine assay (kappa statistic=0.73). An inverse relationship was detected between the level of rabies virus neutralizing antibody (RVNA) and the lymphocyte response to inactivated rabies virus in the vaccinated subjects. The association between cytokines production and level of humoral and cellular response was investigated in four representative subjects. Two vaccinated subjects with high proliferation indices and low RVNA titers produced Th1 type cytokines to rabies virus stimulation, whereas two vaccinated individuals with low proliferation indices and high RVNA titers responses did not produce these cytokines.