Induced regulatory T cells are phenotypically unstable and do not protect mice from rapidly progressive glomerulonephritis.
Regulatory T (Treg) cells are a suppressive CD4+T-cell subset. We generated induced Treg (iTreg) cells and explored their therapeutic potential in a murine model of rapidly progressive glomerulonephritis. Polyclonal naive CD4+T cells were cultured in vitro with interleukin-2 (IL-2), transforming growth factor-β1, all-trans-retinoic acid and monoclonal antibodies against interferon-γ and IL-4, generating Foxp3+iTreg cells. To enhance their suppressive phenotype, iTreg cultures were modified with the addition of a monoclonal antibody against IL-12p40 or by using RORγt-/-CD4+T cells. Induced Treg cells were transferred into models of delayed-type hypersensitivity and experimental glomerulonephritis. The iTreg cells exhibited comparable surface receptor expression and in vitro suppressive ability to natural Treg cells, but did not regulate antigen-specific delayed-type hypersensitivity or systemic inflammatory immune responses, losing Foxp3 expression in vivo. In glomerulonephritis, transferred iTreg cells did not prevent renal injury or modulate systemic T helper type 1 immune responses. Induced Treg cells cultured with anti-IL-12p40 had an enhanced suppressive phenotype in vitro and regulated dermal delayed-type hypersensitivity in vivo, but were not protective against renal injury, losing Foxp3 expression, especially in the transferred cells recruited to the kidney. Use of RORγt-/-CD4+T cells or iTreg cells generated from sensitized CD4+Foxp3-cells did not regulate renal or systemic inflammatory responses in vivo. In conclusion, iTreg cells suppress T-cell proliferation in vitro, but do not regulate experimental glomerulonephritis, being unstable in this inflammatory milieu in vivo.
Centre for Inflammatory Diseases, Department of Medicine, Monash University, Clayton, Victoria, Australia.|Department of Nephrology, Monash Health, Clayton, Victoria, Australia.
Joanna R
JR
Ghali
Centre for Inflammatory Diseases, Department of Medicine, Monash University, Clayton, Victoria, Australia.
Maliha A
MA
Alikhan
Centre for Inflammatory Diseases, Department of Medicine, Monash University, Clayton, Victoria, Australia.|Department of Nephrology, Monash Health, Clayton, Victoria, Australia.
Stephen R
SR
Holdsworth
Centre for Inflammatory Diseases, Department of Medicine, Monash University, Clayton, Victoria, Australia.|Department of Nephrology, Monash Health, Clayton, Victoria, Australia.|Department of Paediatric Nephrology, Monash Health, Clayton, Victoria, Australia.
A Richard
AR
Kitching
D000818:Animals; D015704:CD4 Antigens / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D060833:Cellular Microenvironment; D004195:Disease Models, Animal; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism; D005921:Glomerulonephritis / Q000276:immunology*; D006801:Humans; D006968:Hypersensitivity, Delayed / Q000276:immunology*; D007165:Immunosuppression Therapy; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3 / Q000235:genetics; D012867:Skin / Q000276:immunology* / Q000473:pathology; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*
D015704:CD4 Antigens; D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D057132:Nuclear Receptor Subfamily 1, Group F, Member 3
Foxp3; all-trans-retinoic acid; induced regulatory T cell; rapidly progressive glomerulonephritis; retoinic acid receptor-related orphan receptor γt