Regulatory T cells expanded from HIV-1-infected individuals maintain phenotype, TCR repertoire and suppressive capacity.
While modulation of regulatory T cell (Treg) function and adoptive Treg transfer are being explored as therapeutic modalities in the context of autoimmune diseases, transplantation and cancer, their role in HIV-1 pathogenesis remains less well defined. Controversy persists regarding their beneficial or detrimental effects in HIV-1 disease, which warrants further detailed exploration. Our objectives were to investigate if functional CD4(+) Tregs can be isolated and expanded from HIV-1-infected individuals for experimental or potential future therapeutic use and to determine phenotype and suppressive capacity of expanded Tregs from HIV-1 positive blood and tissue. Tregs and conventional T cell controls were isolated from blood and gut-associated lymphoid tissue of individuals with HIV-1 infection and healthy donors using flow-based cell-sorting. The phenotype of expanded Tregs was assessed by flow-cytometry and quantitative PCR. T-cell receptor ß-chain (TCR-β) repertoire diversity was investigated by deep sequencing. Flow-based T-cell proliferation and chromium release cytotoxicity assays were used to determine Treg suppressive function. Tregs from HIV-1 positive individuals, including infants, were successfully expanded from PBMC and GALT. Expanded Tregs expressed high levels of FOXP3, CTLA4, CD39 and HELIOS and exhibited a highly demethylated TSDR (Treg-specific demethylated region), characteristic of Treg lineage. The TCRß repertoire was maintained following Treg expansion and expanded Tregs remained highly suppressive in vitro. Our data demonstrate that Tregs can be expanded from blood and tissue compartments of HIV-1+ donors with preservation of Treg phenotype, function and TCR repertoire. These results are highly relevant for the investigation of potential future therapeutic use, as currently investigated for other disease states and hold great promise for detailed studies on the role of Tregs in HIV-1 infection.
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America.
Mathieu
M
Angin
Department of Clinical Immunology and Rheumatology, Academic Medical Center, Amsterdam, The Netherlands.
Paul L
PL
Klarenbeek
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America.
Melanie
M
King
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America.
Siddhartha M
SM
Sharma
HIV Pathogenesis Programme, Doris Duke Medical Research Institute and KwaZulu-Natal Research Institute for TB and HIV, University of KwaZulu-Natal, Durban, South Africa.
Eshia S
ES
Moodley
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America.
Ashley
A
Rezai
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America.
Alicja
A
Piechocka-Trocha
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America.
Ildiko
I
Toth
Massachusetts General Hospital, Gastrointestinal Unit, Boston, Massachusetts, United States of America.
Andrew T
AT
Chan
HIV Pathogenesis Programme, Doris Duke Medical Research Institute and KwaZulu-Natal Research Institute for TB and HIV, University of KwaZulu-Natal, Durban, South Africa ; Department of Paediatrics, University of Oxford, Oxford, United Kingdom.
Philip J
PJ
Goulder
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America ; HIV Pathogenesis Programme, Doris Duke Medical Research Institute and KwaZulu-Natal Research Institute for TB and HIV, University of KwaZulu-Natal, Durban, South Africa.
Thumbi
T
Ndung'u
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America ; Massachusetts General Hospital, Division of Infectious Diseases, Boston, Massachusetts, United States of America.
Douglas S
DS
Kwon
Ragon Institute of MGH, MIT and Harvard, Boston, Massachusetts, United States of America ; Massachusetts General Hospital, Division of Infectious Diseases, Boston, Massachusetts, United States of America ; Department of Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Marylyn M
MM
Addo
D000328:Adult; D049109:Cell Proliferation; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D019175:DNA Methylation / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D041981:Gastrointestinal Tract / Q000276:immunology / Q000821:virology; D015870:Gene Expression / Q000276:immunology; D015658:HIV Infections / Q000097:blood / Q000276:immunology* / Q000821:virology; D015497:HIV-1 / Q000276:immunology* / Q000502:physiology; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D054884:Host-Pathogen Interactions / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D007223:Infant; D008221:Lymphoid Tissue / Q000276:immunology / Q000821:virology; D008297:Male; D008875:Middle Aged; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism
C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D011948:Receptors, Antigen, T-Cell