Effect of cryopreservation on CD4+ T cell subsets in foreskin tissue

Authors

Omar Almomani, Department of Microbiology and Immunology, Western University, London, Canada.
James Nnamutete, Rakai Health Sciences Program, Kalisizo, Uganda.
Zhongtian Shao, Department of Microbiology and Immunology, Western University, London, Canada.
Victoria Menya Biribawa, UVRI-IAVI HIV Vaccine Program Limited, Entebbe, Uganda.
HenryRoger Ssemunywa, UVRI-IAVI HIV Vaccine Program Limited, Entebbe, Uganda.
Annemarie Namuniina, UVRI-IAVI HIV Vaccine Program Limited, Entebbe, Uganda.
Brenda Okech, UVRI-IAVI HIV Vaccine Program Limited, Entebbe, Uganda.
Sofya Ulanova, Department of Microbiology and Immunology, Western University, London, Canada.
David Zuanazzi, Department of Microbiology and Immunology, Western University, London, Canada.
Cindy M. Liu, Department of Environmental and Occupational Health, Milken Institute School of Public Health, George Washington University, Washington, DC, United States of America.
Aaron A. Tobian, Department of Pathology, Johns Hopkins University School of Medicine, Johns Hopkins University, Baltimore, Maryland, United States of America.
Ronald M. Galiwango, Rakai Health Sciences Program, Kalisizo, Uganda.
Rupert Kaul, Departments of Medicine and Immunology, University of Toronto, Toronto, Canada.
Jessica L. Prodger, Department of Microbiology and Immunology, Western University, London, Canada.

Document Type

Journal Article

Publication Date

1-1-2024

Journal

PloS one

Volume

19

Issue

3

DOI

10.1371/journal.pone.0297884

Abstract

Voluntary medical male circumcision (VMMC) reduces HIV acquisition by at least 60%, but the determinants of HIV susceptibility in foreskin tissues are incompletely understood. Flow cytometry is a powerful tool that helps us understand tissue immune defenses in mucosal tissue like the inner foreskin, but foreskin flow cytometry has only been validated using fresh tissue samples. This restricts immune analyses to timepoints immediately after surgical acquisition and hinders research in this area. We compared fresh analysis with whole tissue cryopreservation and later thawing and digestion to analyze CD4+ T cell populations relevant to HIV susceptibility (CCR5, CD25, CD127, CCR4, CXCR3, CCR6, CCR10, HLA-DR, and CD38). Eight foreskin samples from HIV-negative males aged >18 years were collected after VMMC. For each sample, half the foreskin was immediately cryopreserved for later digestion and flow cytometry analysis, while the remaining tissues were analyzed fresh. We demonstrate no significant impact of cryopreservation on CD4+ T cell expression of CD25, CCR4, CCR6, HLA-DR, CCR10, or CD127. Although expression levels of CCR5, CD38, and CXCR3 were increased after cryopreservation, the relative ranking of participants was retained. In conclusion, cryopreserved foreskin tissues may be suitable for subsequent digestion and flow cytometry phenotyping of HIV-susceptible T cell populations.

APA Citation

Almomani, Omar; Nnamutete, James; Shao, Zhongtian; Biribawa, Victoria Menya; Ssemunywa, HenryRoger; Namuniina, Annemarie; Okech, Brenda; Ulanova, Sofya; Zuanazzi, David; Liu, Cindy M.; Tobian, Aaron A.; Galiwango, Ronald M.; Kaul, Rupert; and Prodger, Jessica L., "Effect of cryopreservation on CD4+ T cell subsets in foreskin tissue" (2024). GW Authored Works. Paper 4589.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/4589

Department

Environmental and Occupational Health