Assessment of a diverse panel of transmitted/founder HIV-1 infectious molecular clones in a luciferase based CD8 T-cell mediated viral inhibition assay

• DOI: 10.3389/fimmu.2022.1029029
• Publication Date: 2022-12-01T17:57:50Z
• Authors: Natalia Fernandez; Peter Hayes; Julia Makinde; Jonathan Hare; Deborah King; Rui Xu; Ola Rehawi; Allison T. Mezzell; Laban Kato; Susan Mugaba; Jennifer Serwanga; James Chemweno; Eunice Nduati; Matt A. Price; Faith Osier; Christina Ochsenb...; Ling Yue; Eric Hunter; Jill Gilmour
• Keywords: Immunology; 610; HIV; HIV Infections; viral inhibition; RC581-607; CD8-Positive T-Lymphocytes; infection; CD8 T-cells; Clone Cells; T-cell response; 616; HIV-1; Humans; Immunologic diseases. Allergy; Luciferases; transmitted founder

IntroductionImmunological protection against human immunodeficiency virus-1 (HIV-1) infection is likely to require both humoral and cell-mediated immune responses, the latter involving cytotoxic CD8 T-cells. Characterisation of CD8 T-cell mediated direct anti-viral activity would provide understanding of potential correlates of immune protection and identification of critical epitopes associated with HIV-1 control.MethodsThe present report describes a functional viral inhibition assay (VIA) to assess CD8 T-cell-mediated inhibition of replication of a large and diverse panel of 45 HIV-1 infectious molecular clones (IMC) engineered with a Renilla reniformis luciferase reporter gene (LucR), referred to as IMC-LucR. HIV-1 IMC replication in CD4 T-cells and CD8 T-cell mediated inhibition was characterised in both ART naive subjects living with HIV-1 covering a broad human leukocyte antigen (HLA) distribution and compared with uninfected subjects.Results & discussionCD4 and CD8 T-cell lines were established from subjects vaccinated with a candidate HIV-1 vaccine and provided standard positive controls for both assay quality control and facilitating training and technology transfer. The assay was successfully established across 3 clinical research centres in Kenya, Uganda and the United Kingdom and shown to be reproducible. This IMC-LucR VIA enables characterisation of functional CD8 T-cell responses providing a tool for rational T-cell immunogen design of HIV-1 vaccine candidates and evaluation of vaccine-induced T-cell responses in HIV-1 clinical trials.