Viral competition assay to assess the role of HIV-1 proteins in immune evasion

Gabriel Duette; Samantha Cronin; Anthony D Kelleher; Sarah Palmer

doi:10.1016/j.xpro.2022.102025

Viral competition assay to assess the role of HIV-1 proteins in immune evasion

STAR Protoc. 2023 Mar 17;4(1):102025. doi: 10.1016/j.xpro.2022.102025. Epub 2023 Jan 14.

Authors

Gabriel Duette¹, Samantha Cronin², Anthony D Kelleher³, Sarah Palmer⁴

Affiliations

¹ Centre for Virus Research, The Westmead Institute for Medical Research, Westmead, NSW 2145, Australia; Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2050, Australia. Electronic address: gabriel.duette@sydney.edu.au.
² Centre for Virus Research, The Westmead Institute for Medical Research, Westmead, NSW 2145, Australia; Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2050, Australia.
³ The Kirby Institute, University of New South Wales, Sydney, NSW 2052, Australia.
⁴ Centre for Virus Research, The Westmead Institute for Medical Research, Westmead, NSW 2145, Australia; Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2050, Australia. Electronic address: sarah.palmer@sydney.edu.au.

Abstract

CD8+ T lymphocytes can recognize and eliminate cells infected by viruses. However, the human immunodeficiency virus (HIV-1) has developed mechanisms to evade CD8+ T-cell-mediated clearance. Here, we describe a protocol to assess the role of the HIV-1 protein Nef in immune evasion. The viral competition assay reveals the preferential killing of HIV-1-infected cells unable to express Nef. This methodology can be extended to study HIV-1 proteins involved in immune evasion and viral variants encoding cytotoxic T lymphocyte escape mutations. For complete details on the use and execution of this protocol, please refer to Duette et al. (2022).¹.

Keywords: Cell Biology; Flow Cytometry/Mass Cytometry; Immunology; Microbiology.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

CD8-Positive T-Lymphocytes
HIV-1* / genetics
HIV-1* / metabolism
Humans
Immune Evasion*
T-Lymphocytes, Cytotoxic
nef Gene Products, Human Immunodeficiency Virus / genetics
nef Gene Products, Human Immunodeficiency Virus / metabolism

Substances

nef Gene Products, Human Immunodeficiency Virus

Abstract

Publication types

MeSH terms

Substances

Grants and funding