Construction and screening of human immunodeficiency virus-1 negative regulation factor peptide-specific CD4+T lymphocyte clone / 中华传染病杂志

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.