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

• Published in: Frontiers in immunology Vol. 13; p. 1029029
• Main Authors: Fernandez, Natalia, Hayes, Peter, Makinde, Julia, Hare, Jonathan, King, Deborah, Xu, Rui, Rehawi, Ola, Mezzell, Allison T., Kato, Laban, Mugaba, Susan, Serwanga, Jennifer, Chemweno, James, Nduati, Eunice, Price, Matt A., Osier, Faith, Ochsenbauer, Christina, Yue, Ling, Hunter, Eric, Gilmour, Jill
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 01.12.2022
• Subjects: CD8 T-cells; CD8-Positive T-Lymphocytes; Clone Cells; HIV; HIV Infections; HIV-1; Humans; Immunology; infection; Luciferases; T-cell response; transmitted founder; viral inhibition; infection; viral inhibition; HIV; transmitted founder; infectious molecular clones; CD8 T-cells; T-cell response
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2022.1029029

Immunological 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. The 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 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. CD4 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.