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一种用于分析有自然麻疹儿童病史的健康成年人中麻疹病毒特异性记忆T淋巴细胞频率的新型灵敏方法。

A novel sensitive approach for frequency analysis of measles virus-specific memory T-lymphocytes in healthy adults with a childhood history of natural measles.

作者信息

Nanan R, Rauch A, Kämpgen E, Niewiesk S, Kreth H W

机构信息

Children's Hospital, University of Würzburg and Department of Dermatology, Josef-Schneider-Str. 2, D-97080 Würzburg, Germany.

出版信息

J Gen Virol. 2000 May;81(Pt 5):1313-9. doi: 10.1099/0022-1317-81-5-1313.

DOI:10.1099/0022-1317-81-5-1313
PMID:10769074
Abstract

Measles virus (MV), a single-stranded negative-sense RNA virus, is an important pathogen causing almost 1 million deaths annually. Acute MV infection induces immunity against disease throughout life. The immunological factors which are responsible for protection against measles are still poorly understood. However, T-cell-mediated immune responses seem to play a central role. The emergence of new single-cell methods for quantification of antigen-specific T-cells directly ex vivo has prompted us to measure frequencies of MV-specific memory T-cells. As an indicator for T-cell activation IFN-gamma production was measured. PBMC were analysed by intracellular staining and ELISPOT assay after stimulation with MV-infected autologous B-lymphoblastoid cell lines or dendritic cells. T-cell responses were exclusively seen with PBMC from MV-seropositive healthy adults with a history of natural measles in childhood. The median frequency of MV-specific T-cells was 0.35% for CD3(+)CD4(+) and 0.24% for the CD3(+)CD8(+) T-cell subset. These frequencies are comparable with T-cell numbers reported by other investigators for persistent virus infections such as Epstein-Barr virus, cytomegalovirus or human immunodeficiency virus. Hence, this study illustrates that MV-specific CD4(+) and CD8(+) T-cells are readily detectable long after the acute infection, and thus are probably contributing to long-term immunity. Furthermore, this new approach allows efficient analysis of T-cell responses from small samples of blood and could therefore be a useful tool to further elucidate the role of cell-mediated immunity in measles as well as in other viral infections.

摘要

麻疹病毒(MV)是一种单链负义RNA病毒,是一种重要的病原体,每年导致近100万人死亡。急性MV感染可诱导终身抵抗该疾病的免疫力。负责预防麻疹的免疫因素仍知之甚少。然而,T细胞介导的免疫反应似乎起着核心作用。用于直接在体外定量抗原特异性T细胞的新单细胞方法的出现促使我们测量MV特异性记忆T细胞的频率。作为T细胞活化的指标,测量了干扰素-γ的产生。在用MV感染的自体B淋巴母细胞系或树突状细胞刺激后,通过细胞内染色和ELISPOT测定法分析外周血单核细胞(PBMC)。仅在有儿童期自然麻疹病史的MV血清阳性健康成年人的PBMC中观察到T细胞反应。CD3(+)CD4(+)的MV特异性T细胞的中位频率为0.35%,CD3(+)CD8(+)T细胞亚群的中位频率为0.24%。这些频率与其他研究人员报道的持续性病毒感染(如爱泼斯坦-巴尔病毒、巨细胞病毒或人类免疫缺陷病毒)的T细胞数量相当。因此,本研究表明,急性感染后很长时间仍可轻易检测到MV特异性CD4(+)和CD8(+)T细胞,因此它们可能有助于长期免疫。此外,这种新方法允许对少量血液样本中的T细胞反应进行有效分析,因此可能是进一步阐明细胞介导的免疫在麻疹以及其他病毒感染中的作用的有用工具。

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