Journal article
Multiparameter immunohistochemistry analysis of HIV DNA, RNA and immune checkpoints in lymph node tissue
ZA Richardson, C Deleage, CSA Tutuka, M Walkiewicz, PM Del Río-Estrada, RD Pascoe, VA Evans, G Reyesteran, M Gonzales, S Roberts-Thomson, M González-Navarro, F Torres-Ruiz, JD Estes, SR Lewin, PU Cameron
Journal of Immunological Methods | Published : 2022
Abstract
The main barrier to a cure for HIV is the persistence of long-lived and proliferating latently infected CD4+ T-cells despite antiretroviral therapy (ART). Latency is well characterized in multiple CD4+ T-cell subsets, however, the contribution of regulatory T-cells (Tregs) expressing FoxP3 as well as immune checkpoints (ICs) PD-1 and CTLA-4 as targets for productive and latent HIV infection in people living with HIV on suppressive ART is less well defined. We used multiplex detection of HIV DNA and RNA with immunohistochemistry (mIHC) on formalin-fixed paraffin embedded (FFPE) cells to simultaneously detect HIV RNA and DNA and cellular markers. HIV DNA and RNA were detected by in situ hybrid..
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Awarded by National Institutes of Health
Funding Acknowledgements
We acknowledge the participants who donated their LNs for this study, Dr. Mauricio Gonz ' alez-Navarro and Dr. Fernanda Torres-Ruiz for performing the LN biopsies (Centro de Investigaci ' on en Enfermdades Infecciosas, Mexico city, Mexico). We thank Caroline Tumpach and Michael Roche for providing us with cells (RMIT, Melbourne Australia and Peter Doherty Institute, Melbourne Australia), Dr. Andreas Behren (Olivia Newton John Cancer Institute) for using his lab and imaging facility, Metta Jana, Rejhan Idriza and Cameron Skinner for the analysis and imaging facility (Petermac, Melbourne, Australia), Dr. Byron Martina for proof-reading (Erasmus MC, Rotterdam, The Netherlands), T. Luka, C. Li for flow cytometric cell sorting (University of Melbourne Flow Cytometry Facility, Melbourne, Australia). This work was funded by grants to Sharon Lewin from National Institutes of Health Delaney AIDS Research Enterprise to Find a Cure Collaboratory (Grant UM1AI126611-01). JD Este was funded from the National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases and National Institute of Allergy and Infectious Diseases grants R01 DK119945 (JDE) and R01 AI143411-01A1 (JDE). SRL is an NHMRC Practitioner Fellow