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Reliable generation and use of MHC class II:γ2aFc multimers for the identification of antigen-specific CD4+ T cells
- Source :
-
Journal of Immunological Methods . Dec2002, Vol. 271 Issue 1/2, p137. 15p. - Publication Year :
- 2002
-
Abstract
- MHC tetramers have proven to be powerful reagents for the analysis of MHC class I-restricted T cells. However, generating similarly reliable reagents for MHC class II-restricted T cells has been elusive. Here we evaluated the utility of MHC class II:γ2aFc multimers, which contain the MHC class II extracellular domains, with or without recombinantly attached peptides, dimerized via a fos–jun leucine zipper and attached to the hinge of murine IgG2a. We have successfully generated 24 multimers in either myeloma or Drosophila melanogaster S2 cells, with an average yield of 7 mg/L. ‘Empty’ MHC class II:γ2aFc multimers were effectively used in peptide binding assays. Similar versions that contained recombinantly attached peptides stimulated T cells in an antigen-specific, MHC-restricted manner, and identified antigen-specific naı¨ve and effector T cells by flow cytometry. Furthermore, we have successfully used these reagents to stain T cells generated following viral infection. Thus, MHC class II:γ2aFc multimers are robust and reliable reagents for the analysis of MHC class II-restricted T cells. [Copyright &y& Elsevier]
- Subjects :
- *LYSOZYMES
*MAJOR histocompatibility complex
*T cells
Subjects
Details
- Language :
- English
- ISSN :
- 00221759
- Volume :
- 271
- Issue :
- 1/2
- Database :
- Academic Search Index
- Journal :
- Journal of Immunological Methods
- Publication Type :
- Academic Journal
- Accession number :
- 8565996
- Full Text :
- https://doi.org/10.1016/S0022-1759(02)00343-5