Construction and screening of human immunodeficiency virus-1 negative regulation factor peptide-specific CD4+T lymphocyte clone

VernacularTitle:人类免疫缺陷病毒-1型负向调节因子多肽特异性CD4+T淋巴细胞克隆的建立和筛选
Author: Peixin SONG ; Yanlei LI ; Nan ZHENG
Publication Type:Journal Article
Keywords: HIV-1; Negative regulation factor peptide; CD4+ T cell clone
From: Chinese Journal of Infectious Diseases 2015;33(4):215-219
CountryChina
Language:Chinese
Abstract: Objective To construct and screen the human immunodeficiency virus-1 (HIV-1) negative regulation factor (Nef) peptide-specific CD4+ T lymphocyte clone.Methods Peripheral blood mononuclear cells (PBMC) from five asymptomatic HIV-1 infected patients were collected and Bulkcultured with Nef end peptides.The CD4 molecule and intracellular interferon (IFN)-gamma of cultured cells were detected by two-color flow cytometry.The Nef end peptide-specific T cell clone was then constructed by limited dilution and confirmed through enzyme linked immunospot assay (ELISPOT).The best grown cells were selected and cultured as the final clone.Results The Nef end peptide-specific-T lymphocyte clone was successfully constructed from PBMC of one HIV-infected patient and confirmed by ELISPOT.The detection of human leukocyte antigen (HLA)-DRB1 type showed that the epitope of this peptide was probably HLA-DRB1 * 0406.Conclusion The Nef end specific-T cell clone is successfully constructed,and a new epitope in the C-terminus of Nef protein and its HLA restriction are identified.