In vivo tracking of 'color-coded' effector, natural and induced regulatory T cells in the allograft response.
Here we present methods to longitudinally track islet allograft-infiltrating T cells in live mice by endoscopic confocal microscopy and to analyze circulating T cells by in vivo flow cytometry. We developed a new reporter mouse whose T cell subsets express distinct, 'color-coded' proteins enabling in vivo detection and identification of effector T cells (T(eff) cells) and discrimination between natural and induced regulatory T cells (nT(reg) and iT(reg) cells). Using these tools, we observed marked differences in the T cell response in recipients receiving tolerance-inducing therapy (CD154-specific monoclonal antibody plus rapamycin) compared to untreated controls. These results establish real-time cell tracking as a powerful means to probe the dynamic cellular interplay mediating immunologic rejection or transplant tolerance.
Fan
Zhigang
Z
Transplant Institute, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
Spencer
Joel A
JA
Lu
Yan
Y
Pitsillides
Costas M
CM
Singh
Gurbakhshish
G
Kim
Pilhan
P
Yun
Seok H
SH
Toxavidis
Vasilis
V
Strom
Terry B
TB
Lin
Charles P
CP
Koulmanda
Maria
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D003116:Color; D005434:Flow Cytometry / Q000379:methods; D006084:Graft Rejection / Q000276:immunology* / Q000503:physiopathology; D006085:Graft Survival / Q000276:immunology* / Q000502:physiology; D007694:Killer Cells, Natural / Q000276:immunology / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal / Q000379:methods; D016176:T-Lymphocyte Subsets / Q000502:physiology; D050378:T-Lymphocytes, Regulatory / Q000276:immunology / Q000502:physiology*; D014184:Transplantation, Homologous
D000911:Antibodies, Monoclonal