Detection of autoreactive myelin proteolipid protein 139-151-specific T cells by using MHC II (IAs) tetramers

N R Jayagopala Reddy, Estelle Bettelli, Lindsay Nicholson, Hanspeter Waldner, Mei Huei Jang, Kai W. Wucherpfennig, Vijay K. Kuchroo

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

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 IAs class II molecule combined with an autoantigenic peptide from myelin proteolipid protein (PLP; PLP139-151) and used it to analyze myelin PLP139-151-reactive T cells. Using monomers and multimerized complexes labeled with PE, we confirmed the specificity of the reagent by bioassay and flow cytometry. The IAs tetramers stimulated and stained the PLP139-151-specific 5B6 TCR transgenic T cells and a polyclonal cell line specific for PLP139-151, but not a control T cell line specific for PLP178-191. 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 PLP139-151 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 PLP139-151-reactive T cells with specificity. This tetrameric reagent will be useful in studying the evolution of PLP139-151-specific repertoire in naive mice and its expansion during the autoimmune disease experimental autoimmune encephalomyelitis.

Original languageEnglish (US)
Pages (from-to)870-877
Number of pages8
JournalJournal of Immunology
Volume170
Issue number2
DOIs
StatePublished - Jan 15 2003

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T-Lymphocytes
Myelin Proteolipid Protein
T-Cell Antigen Receptor Specificity
Cell Line
Autoimmune Experimental Encephalomyelitis
Neuraminidase
Myelin Sheath
myelin proteolipid protein (139-151)
Biological Assay
Autoimmune Diseases
Flow Cytometry
Staining and Labeling
Calcium
Peptides

ASJC Scopus subject areas

  • Immunology

Cite this

Reddy, N. R. J., Bettelli, E., Nicholson, L., Waldner, H., Jang, M. H., Wucherpfennig, K. W., & Kuchroo, V. K. (2003). Detection of autoreactive myelin proteolipid protein 139-151-specific T cells by using MHC II (IAs) tetramers. Journal of Immunology, 170(2), 870-877. https://doi.org/10.4049/jimmunol.170.2.870

Detection of autoreactive myelin proteolipid protein 139-151-specific T cells by using MHC II (IAs) tetramers. / Reddy, N R Jayagopala; Bettelli, Estelle; Nicholson, Lindsay; Waldner, Hanspeter; Jang, Mei Huei; Wucherpfennig, Kai W.; Kuchroo, Vijay K.

In: Journal of Immunology, Vol. 170, No. 2, 15.01.2003, p. 870-877.

Research output: Contribution to journalArticle

Reddy, NRJ, Bettelli, E, Nicholson, L, Waldner, H, Jang, MH, Wucherpfennig, KW & Kuchroo, VK 2003, 'Detection of autoreactive myelin proteolipid protein 139-151-specific T cells by using MHC II (IAs) tetramers', Journal of Immunology, vol. 170, no. 2, pp. 870-877. https://doi.org/10.4049/jimmunol.170.2.870
Reddy, N R Jayagopala ; Bettelli, Estelle ; Nicholson, Lindsay ; Waldner, Hanspeter ; Jang, Mei Huei ; Wucherpfennig, Kai W. ; Kuchroo, Vijay K. / Detection of autoreactive myelin proteolipid protein 139-151-specific T cells by using MHC II (IAs) tetramers. In: Journal of Immunology. 2003 ; Vol. 170, No. 2. pp. 870-877.
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