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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
Author(s) -
Chen Chengcong,
Jiang Xiaotao,
Liu Xuan,
Guo Ling,
Wang Weibin,
Gu Shuqin,
Wen Chunhua,
Yi Xuan,
Tang Libo,
Li Yongyin
Publication year - 2021
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.5ma0620-023rr
Subject(s) - elispot , hbcag , hbsag , hepatitis b virus , biology , virology , flow cytometry , ex vivo , immunology , microbiology and biotechnology , t cell , in vivo , virus , immune system
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.