Identification of the Association Between HBcAg-specific T Cell and Viral Control in Chronic HBV Infection Using a Cultured ELISPOT Assay
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
Affiliations
Affiliations
1
State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Viral Hepatitis Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China.
2
Department of Radiation Oncology, Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou, China.
3
Department of Immunology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
4
Guangdong Provincial Key Laboratory of Proteomic, Guangzhou, China.
5
Department of Pediatrics, Nanfang Hospital, Southern Medical University, Guangzhou, China.
PMID: 32620046 DOI: 10.1002/JLB.5MA0620-023RR
Abstract
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.