HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Reddy, J. Articles by Kuchroo, V. K. Search for Related Content PubMed PubMed Citation Articles by Reddy, J. Articles by Kuchroo, V. K.

Detection of Autoreactive Myelin Proteolipid Protein 139–151-Specific T Cells by Using MHC II (IA^s) Tetramers¹

Jayagopala Reddy^*, Estelle Bettelli^*, Lindsay Nicholson^*, Hanspeter Waldner^*, Mei-Huei Jang, Kai W. Wucherpfennig and Vijay K. Kuchroo^2,*

^* Center for Neurologic Diseases, Brigham and Women’s Hospital, and Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115

Detection of autoreactive T cells using MHC II tetramers is difficult because of the low affinity of their TCR. We have generated a class II tetramer using the IA^s class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP_139–151) and used it to analyze myelin PLP_139–151-reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IA^s tetramers stimulated and stained the PLP_139–151-specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP_139–151, but not a control T cell line specific for PLP_178–191. We used this reagent to optimize conditions to detect low affinity autoreactive T cells. We found that high pH (8.0) and neuraminidase treatment enhances the staining capacity of PLP_139–151 tetramer without compromising specificity. Furthermore, we found that induction of calcium fluxing by tetramers in T cells may be used as a sensitive measure to detect autoreactive T cells with a low affinity. Taken together, the data show that the tetrameric reagent binds and stimulates PLP_139–151-reactive T cells with specificity. This tetrameric reagent will be useful in studying the evolution of PLP_139–151-specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.

This article has been cited by other articles:

				S. Pastor, A. Minguela, W. Mi, and E. S. Ward Autoantigen Immunization at Different Sites Reveals a Role for Anti-Inflammatory Effects of IFN-{gamma} in Regulating Susceptibility to Experimental Autoimmune Encephalomyelitis J. Immunol., May 1, 2009; 182(9): 5268 - 5275. [Abstract] [Full Text] [PDF]

				T. Korn, M. Mitsdoerffer, A. L. Croxford, A. Awasthi, V. A. Dardalhon, G. Galileos, P. Vollmar, G. L. Stritesky, M. H. Kaplan, A. Waisman, et al. IL-6 controls Th17 immunity in vivo by inhibiting the conversion of conventional T cells into Foxp3+ regulatory T cells PNAS, November 25, 2008; 105(47): 18460 - 18465. [Abstract] [Full Text] [PDF]

				D. T. Chung, T. Korn, J. Richard, M. Ruzek, A. P. Kohm, S. Miller, S. Nahill, and M. Oukka Anti-thymocyte globulin (ATG) prevents autoimmune encephalomyelitis by expanding myelin antigen-specific Foxp3+ regulatory T cells Int. Immunol., August 13, 2007; (2007) dxm078v1. [Abstract] [Full Text] [PDF]

				A. Minguela, S. Pastor, W. Mi, J. A. Richardson, and E. S. Ward Feedback Regulation of Murine Autoimmunity via Dominant Anti-Inflammatory Effects of Interferon {gamma} J. Immunol., January 1, 2007; 178(1): 134 - 144. [Abstract] [Full Text] [PDF]

				X. Zhang, J. Reddy, H. Ochi, D. Frenkel, V. K. Kuchroo, and H. L. Weiner Recovery from experimental allergic encephalomyelitis is TGF-{beta} dependent and associated with increases in CD4+LAP+ and CD4+CD25+ T cells Int. Immunol., April 1, 2006; 18(4): 495 - 503. [Abstract] [Full Text] [PDF]

				R. Mallone, S. A. Kochik, H. Reijonen, B. Carson, S. F. Ziegler, W. W. Kwok, and G. T. Nepom Functional avidity directs T-cell fate in autoreactive CD4+ T cells Blood, October 15, 2005; 106(8): 2798 - 2805. [Abstract] [Full Text] [PDF]

				Z. Karabekian, S. D. Lytton, P. B. Silver, Y. V. Sergeev, J. P. Schneck, and R. R. Caspi Antigen/MHC Class II/Ig Dimers for Study of Uveitogenic T Cells: IRBP p161-180 Presented by both IA and IE Molecules Invest. Ophthalmol. Vis. Sci., October 1, 2005; 46(10): 3769 - 3776. [Abstract] [Full Text] [PDF]

				V. Dardalhon, A. S. Schubart, J. Reddy, J. H. Meyers, L. Monney, C. A. Sabatos, R. Ahuja, K. Nguyen, G. J. Freeman, E. A. Greenfield, et al. CD226 Is Specifically Expressed on the Surface of Th1 Cells and Regulates Their Expansion and Effector Functions J. Immunol., August 1, 2005; 175(3): 1558 - 1565. [Abstract] [Full Text] [PDF]

				H. Shao, Y. Peng, T. Liao, M. Wang, M. Song, H. J. Kaplan, and D. Sun A Shared Epitope of the Interphotoreceptor Retinoid-Binding Protein Recognized by the CD4+ and CD8+ Autoreactive T Cells J. Immunol., August 1, 2005; 175(3): 1851 - 1857. [Abstract] [Full Text] [PDF]

				J. N. H. Stern, Z. Illes, J. Reddy, D. B. Keskin, M. Fridkis-Hareli, V. K. Kuchroo, and J. L. Strominger Peptide 15-mers of defined sequence that substitute for random amino acid copolymers in amelioration of experimental autoimmune encephalomyelitis PNAS, February 1, 2005; 102(5): 1620 - 1625. [Abstract] [Full Text] [PDF]

				J. Reddy, Z. Illes, X. Zhang, J. Encinas, J. Pyrdol, L. Nicholson, R. A. Sobel, K. W. Wucherpfennig, and V. K. Kuchroo Myelin proteolipid protein-specific CD4+CD25+ regulatory cells mediate genetic resistance to experimental autoimmune encephalomyelitis PNAS, October 26, 2004; 101(43): 15434 - 15439. [Abstract] [Full Text] [PDF]

				H. Sakuma, K. Kohyama, Y. Jee, and Y. Matsumoto Tracking of V{beta}8.2-Positive Encephalitogenic T Cells by Complementarity-Determining Region 3 Spectratyping and Subsequent Southern Blot Hybridization in Lewis Rats after Neuroantigen Sensitization J. Immunol., October 1, 2004; 173(7): 4516 - 4522. [Abstract] [Full Text] [PDF]

				H. Shao, S. L. Sun, H. J. Kaplan, and D. Sun Characterization of Rat CD8+ Uveitogenic T Cells Specific for Interphotoreceptor Retinal-Binding Protein 1177-1191 J. Immunol., August 15, 2004; 173(4): 2849 - 2854. [Abstract] [Full Text] [PDF]

				J. N. H. Stern, Z. Illes, J. Reddy, D. B. Keskin, E. Sheu, M. Fridkis-Hareli, H. Nishimura, C. F. Brosnan, L. Santambrogio, V. K. Kuchroo, et al. Amelioration of proteolipid protein 139-151-induced encephalomyelitis in SJL mice by modified amino acid copolymers and their mechanisms PNAS, August 10, 2004; 101(32): 11743 - 11748. [Abstract] [Full Text] [PDF]

				A. C. Anderson, J. Reddy, R. Nazareno, R. A. Sobel, L. B. Nicholson, and V. K. Kuchroo IL-10 Plays an Important Role in the Homeostatic Regulation of the Autoreactive Repertoire in Naive Mice J. Immunol., July 15, 2004; 173(2): 828 - 834. [Abstract] [Full Text] [PDF]

				D.-A. Laplaud, C. Ruiz, S. Wiertlewski, S. Brouard, L. Berthelot, M. Guillet, B. Melchior, N. Degauque, G. Edan, P. Brachet, et al. Blood T-cell receptor {beta} chain transcriptome in multiple sclerosis. Characterization of the T cells with altered CDR3 length distribution Brain, May 1, 2004; 127(5): 981 - 995. [Abstract] [Full Text] [PDF]

				F. Bischof, M. Hofmann, T. N. M. Schumacher, F. A. Vyth-Dreese, R. Weissert, H. Schild, A. M. Kruisbeek, and A. Melms Analysis of Autoreactive CD4 T Cells in Experimental Autoimmune Encephalomyelitis after Primary and Secondary Challenge Using MHC Class II Tetramers J. Immunol., March 1, 2004; 172(5): 2878 - 2884. [Abstract] [Full Text] [PDF]

				M Bradl and R Hohlfeld Molecular pathogenesis of neuroinflammation J. Neurol. Neurosurg. Psychiatry, October 1, 2003; 74(10): 1364 - 1370. [Abstract] [Full Text] [PDF]