In Vivo Tracking of 'color-coded' Effector, Natural and Induced Regulatory T Cells in the Allograft Response

Overview

Journal Nat Med

Specialties General Medicine
Molecular Biology

Date 2010 May 25

PMID 20495571

Citations 74

Authors

Zhigang Fan

Joel A Spencer

Yan Lu

Costas M Pitsillides

Gurbakhshish Singh

Pilhan Kim

Seok H Yun

Vasilis Toxavidis

Terry B Strom

Charles P Lin

Maria Koulmanda

Affiliations

Soon will be listed here.

Abstract

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.

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References

Koulmanda M, Budo E, Bonner-Weir S, Qipo A, Putheti P, Degauque N . Modification of adverse inflammation is required to cure new-onset type 1 diabetic hosts. Proc Natl Acad Sci U S A. 2007; 104(32):13074-9. PMC: 1941818. DOI: 10.1073/pnas.0705863104. View

Novak J, Georgakoudi I, Wei X, Prossin A, Lin C . In vivo flow cytometer for real-time detection and quantification of circulating cells. Opt Lett. 2004; 29(1):77-9. PMC: 2801600. DOI: 10.1364/ol.29.000077. View

Kim P, Puorishaag M, Cote D, Lin C, Yun S . In vivo confocal and multiphoton microendoscopy. J Biomed Opt. 2008; 13(1):010501. PMC: 2752311. DOI: 10.1117/1.2839043. View

Kamradt T, Mitchison N . Tolerance and autoimmunity. N Engl J Med. 2001; 344(9):655-64. DOI: 10.1056/NEJM200103013440907. View

Gao W, Lu Y, El Essawy B, Oukka M, Kuchroo V, Strom T . Contrasting effects of cyclosporine and rapamycin in de novo generation of alloantigen-specific regulatory T cells. Am J Transplant. 2007; 7(7):1722-32. PMC: 3831357. DOI: 10.1111/j.1600-6143.2007.01842.x. View

Toso C, Zaidi H, Morel P, Armanet M, Wojtusciszyn A, Mai G . Assessment of 18F-FDG-leukocyte imaging to monitor rejection after pancreatic islet transplantation. Transplant Proc. 2006; 38(9):3033-4. DOI: 10.1016/j.transproceed.2006.08.135. View

Wu Y, Ye Q, Foley L, Hitchens T, Sato K, Williams J . In situ labeling of immune cells with iron oxide particles: an approach to detect organ rejection by cellular MRI. Proc Natl Acad Sci U S A. 2006; 103(6):1852-7. PMC: 1413627. DOI: 10.1073/pnas.0507198103. View

Medarova Z, Tsai S, Evgenov N, Santamaria P, Moore A . In vivo imaging of a diabetogenic CD8+ T cell response during type 1 diabetes progression. Magn Reson Med. 2008; 59(4):712-20. DOI: 10.1002/mrm.21494. View

Joffre O, Santolaria T, Calise D, Al Saati T, Hudrisier D, Romagnoli P . Prevention of acute and chronic allograft rejection with CD4+CD25+Foxp3+ regulatory T lymphocytes. Nat Med. 2007; 14(1):88-92. PMC: 2443705. DOI: 10.1038/nm1688. View

10.

Korn T, Bettelli E, Gao W, Awasthi A, Jager A, Strom T . IL-21 initiates an alternative pathway to induce proinflammatory T(H)17 cells. Nature. 2007; 448(7152):484-487. PMC: 3805028. DOI: 10.1038/nature05970. View

11.

Zheng X, Sanchez-Fueyo A, Domenig C, Strom T . The balance of deletion and regulation in allograft tolerance. Immunol Rev. 2003; 196:75-84. DOI: 10.1046/j.1600-065x.2003.00089.x. View

12.

Fontenot J, Rasmussen J, Williams L, Dooley J, Farr A, Rudensky A . Regulatory T cell lineage specification by the forkhead transcription factor foxp3. Immunity. 2005; 22(3):329-41. DOI: 10.1016/j.immuni.2005.01.016. View

13.

Strom T, Roy-Chaudhury P, Manfro R, Zheng X, Nickerson P, Wood K . The Th1/Th2 paradigm and the allograft response. Curr Opin Immunol. 1996; 8(5):688-93. DOI: 10.1016/s0952-7915(96)80087-2. View

14.

Bettelli E, Carrier Y, Gao W, Korn T, Strom T, Oukka M . Reciprocal developmental pathways for the generation of pathogenic effector TH17 and regulatory T cells. Nature. 2006; 441(7090):235-8. DOI: 10.1038/nature04753. View

15.

Strom T . Is transplantation tolerable?. J Clin Invest. 2004; 113(12):1681-3. PMC: 420515. DOI: 10.1172/JCI22153. View

16.

Wells A, Li X, Li Y, Walsh M, Zheng X, Wu Z . Requirement for T-cell apoptosis in the induction of peripheral transplantation tolerance. Nat Med. 1999; 5(11):1303-7. DOI: 10.1038/15260. View

17.

Zhong X, Gao W, Degauque N, Bai C, Lu Y, Kenny J . Reciprocal generation of Th1/Th17 and T(reg) cells by B1 and B2 B cells. Eur J Immunol. 2007; 37(9):2400-4. DOI: 10.1002/eji.200737296. View

18.

Fontenot J, Gavin M, Rudensky A . Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat Immunol. 2003; 4(4):330-6. DOI: 10.1038/ni904. View

19.

Li Y, Li X, Zheng X, Wells A, Turka L, Strom T . Blocking both signal 1 and signal 2 of T-cell activation prevents apoptosis of alloreactive T cells and induction of peripheral allograft tolerance. Nat Med. 1999; 5(11):1298-302. DOI: 10.1038/15256. View

20.

Edinger M, Hoffmann P, Contag C, Negrin R . Evaluation of effector cell fate and function by in vivo bioluminescence imaging. Methods. 2003; 31(2):172-9. DOI: 10.1016/s1046-2023(03)00127-0. View