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Identification of the association between HBcAg‐specific T cell and viral control in chronic HBV infection using a cultured ELISPOT assay
Journal of Leukocyte Biology ( IF 5.5 ) Pub Date : 2020-07-03 , DOI: 10.1002/jlb.5ma0620-023rr
Chengcong Chen 1, 2 , Xiaotao Jiang 3, 4 , Xuan Liu 5 , Ling Guo 1 , Weibin Wang 1 , Shuqin Gu 1 , Chunhua Wen 1 , Xuan Yi 1 , Libo Tang 1 , Yongyin Li 1
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Hepatitis B virus (HBV)‐specific T cells play a critical role in determining the outcome of HBV infection. However, T cell response induced by predominant Ag in chronic infection is hardly detectable owing to the lack of a suitable assay. We herein established an optimized method to enumerate HBV‐specific T cells and assessed the association between HBV surface Ag (HBsAg) and HBV DNA. Sixty chronic HBV infection patients were enrolled. HBV‐specific T cells were expanded by using overlapping peptide pools covering the entire sequence of HBV genotypes B and C. IFN‐γ‐producing HBV‐specific T cells were detected by a cultured enzyme‐linked immunospot (ELISPOT) assay, ex vivo ELISPOT assay, or flow cytometry staining. The association between HBV‐specific T cells and serum levels of HBsAg and HBV DNA were analyzed. Cultured ELISPOT assay had a higher sensitivity than ex vivo ELISPOT in the detection of HBV‐specific T cells. Moreover, consistent results were acquired by flow cytometry analysis and cultured ELISPOT assay, but the latter required only a limited number of cells for detection. Interestingly, HBV core peptide pool induced a robust HBV‐specific T cell response in patients with lower levels of HBV DNA and HBsAg. Specifically, the frequency of HBV core Ag‐specific IFN‐γ+ spot‐forming cells was inversely correlated with serum levels of HBV DNA and HBsAg. An optimized cultured ELISPOT assay reveals the association between HBV core Ag‐induced T cell response and HBV control; this method may favor the investigation of HBV‐specific T cell in chronic HBV infection.

中文翻译:

通过培养的ELISPOT分析鉴定HBcAg特异性T细胞与慢性HBV感染中的病毒控制之间的关联

乙肝病毒(HBV)特异性T细胞在确定HBV感染的结果中起关键作用。然而,由于缺乏合适的检测方法,在慢性感染中主要由Ag诱导的T细胞反应很难检测到。我们在此建立了一种枚举HBV特异性T细胞的优化方法,并评估了HBV表面Ag(HBsAg)与HBV DNA之间的关联。招募了60名慢性HBV感染患者。通过使用覆盖整个HBV基因型B和C序列的重叠肽池来扩增HBV特异性T细胞。通过培养的酶联免疫斑点法(ELISPOT),离体ELISPOT检测到产生IFNγ的HBV特异性T细胞。分析或流式细胞仪染色。分析了HBV特异性T细胞与血清HBsAg和HBV DNA的相关性。培养的ELISPOT分析在检测HBV特异性T细胞方面比离体ELISPOT具有更高的灵敏度。此外,通过流式细胞仪分析和培养的ELISPOT分析获得了一致的结果,但后者仅需要有限数量的细胞进行检测。有趣的是,HBV核心肽库在HBV DNA和HBsAg水平较低的患者中引起强烈的HBV特异性T细胞反应。具体来说,HBV核心Ag特异性IFN-γ的频率+点形成细胞与血清HBV DNA和HBsAg呈负相关。优化的培养ELISPOT分析揭示了HBV核心Ag诱导的T细胞反应与HBV对照之间的联系。这种方法可能有助于研究慢性HBV感染中HBV特异性T细胞。
更新日期:2020-07-03
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