PMID- 36532063 OWN - NLM STAT- MEDLINE DCOM- 20221220 LR - 20230117 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 13 DP - 2022 TI - Assessment of a diverse panel of transmitted/founder HIV-1 infectious molecular clones in a luciferase based CD8 T-cell mediated viral inhibition assay. PG - 1029029 LID - 10.3389/fimmu.2022.1029029 [doi] LID - 1029029 AB - INTRODUCTION: 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. METHODS: 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 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 & DISCUSSION: 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. CI - Copyright (c) 2022 Fernandez, Hayes, Makinde, Hare, King, Xu, Rehawi, Mezzell, Kato, Mugaba, Serwanga, Chemweno, Nduati, Price, Osier, Ochsenbauer, Yue, Hunter, Gilmour and The IAVI protocol C investigators. FAU - Fernandez, Natalia AU - Fernandez N AD - IAVI Human Immunology Laboratory, Imperial College, London, United Kingdom. FAU - Hayes, Peter AU - Hayes P AD - IAVI Human Immunology Laboratory, Imperial College, London, United Kingdom. FAU - Makinde, Julia AU - Makinde J AD - IAVI Human Immunology Laboratory, Imperial College, London, United Kingdom. FAU - Hare, Jonathan AU - Hare J AD - IAVI Human Immunology Laboratory, Imperial College, London, United Kingdom. AD - IAVI, New York, NY, United States. FAU - King, Deborah AU - King D AD - IAVI Human Immunology Laboratory, Imperial College, London, United Kingdom. FAU - Xu, Rui AU - Xu R AD - Emory Vaccine Center at Yerkes National Primate Research Center, Emory University, Atlanta, GA, United States. FAU - Rehawi, Ola AU - Rehawi O AD - University of Alabama at Birmingham, Birmingham, AL, United States. FAU - Mezzell, Allison T AU - Mezzell AT AD - University of Alabama at Birmingham, Birmingham, AL, United States. FAU - Kato, Laban AU - Kato L AD - Uganda Virus Research Institute, Entebbe, Uganda. AD - Medical Research Council, Uganda Virus Research Institute and London School of Hygiene and Tropical Medicine Uganda Research Unit, Entebbe, Uganda. FAU - Mugaba, Susan AU - Mugaba S AD - Uganda Virus Research Institute, Entebbe, Uganda. AD - Medical Research Council, Uganda Virus Research Institute and London School of Hygiene and Tropical Medicine Uganda Research Unit, Entebbe, Uganda. FAU - Serwanga, Jennifer AU - Serwanga J AD - Uganda Virus Research Institute, Entebbe, Uganda. AD - Medical Research Council, Uganda Virus Research Institute and London School of Hygiene and Tropical Medicine Uganda Research Unit, Entebbe, Uganda. FAU - Chemweno, James AU - Chemweno J AD - Kenya Medical Research Institute (KEMRI) Wellcome Trust Research Programme, Kilifi, Kenya. FAU - Nduati, Eunice AU - Nduati E AD - Kenya Medical Research Institute (KEMRI) Wellcome Trust Research Programme, Kilifi, Kenya. FAU - Price, Matt A AU - Price MA AD - IAVI, New York, NY, United States. AD - Department of Epidemiology and Biostatistics, University of California at San Francisco, San Francisco, CA, United States. FAU - Osier, Faith AU - Osier F AD - IAVI Human Immunology Laboratory, Imperial College, London, United Kingdom. FAU - Ochsenbauer, Christina AU - Ochsenbauer C AD - University of Alabama at Birmingham, Birmingham, AL, United States. FAU - Yue, Ling AU - Yue L AD - Emory Vaccine Center at Yerkes National Primate Research Center, Emory University, Atlanta, GA, United States. FAU - Hunter, Eric AU - Hunter E AD - Emory Vaccine Center at Yerkes National Primate Research Center, Emory University, Atlanta, GA, United States. FAU - Gilmour, Jill AU - Gilmour J AD - Department of Infectious Diseases, Imperial College, London, United Kingdom. CN - IAVI protocol C investigators LA - eng PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20221201 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - EC 1.13.12.- (Luciferases) SB - IM MH - Humans MH - *HIV-1 MH - *HIV Infections MH - CD8-Positive T-Lymphocytes MH - Luciferases MH - Clone Cells PMC - PMC9751811 OTO - NOTNLM OT - CD8 T-cells OT - HIV OT - T-cell response OT - infection OT - infectious molecular clones OT - transmitted founder OT - viral inhibition COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2022/12/20 06:00 MHDA- 2022/12/21 06:00 PMCR- 2022/01/01 CRDT- 2022/12/19 04:05 PHST- 2022/08/26 00:00 [received] PHST- 2022/11/14 00:00 [accepted] PHST- 2022/12/19 04:05 [entrez] PHST- 2022/12/20 06:00 [pubmed] PHST- 2022/12/21 06:00 [medline] PHST- 2022/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2022.1029029 [doi] PST - epublish SO - Front Immunol. 2022 Dec 1;13:1029029. doi: 10.3389/fimmu.2022.1029029. eCollection 2022.