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Use of Single Chain MHC Technology to Investigate Co-agonism in Human CD8+ T Cell Activation
Published on: February 28, 2019
Expression of HLA-DR mRNA in T cells following activation is early and can precede DNA synthesis
K Zier1, B Gansbacher, N Ikegaki
1Department of Medicine and Microbiology, Mt. Sinai School of Medicine, New York, NY 10029.
Abstract:
The stimulation of T cells is accompanied by the appearance of a number of activation antigens not found on resting T cells. We have studied the expression of one group of such antigens, the HLA class II antigens (DR, DQ and DP), following stimulation with PHA and PMA. Immunofluorescence studies using monoclonal antibodies indicated that cell surface class II determinants were detectable as little as 30 minutes following stimulation, and that their levels remained constant for about 24 hours, after which they began to increase. Studies on the molecule level demonstrated an increase in the steady state levels of mRNA for DR beta by 15 minutes. Immunoblot analysis of resting T cell lysates using a monoclonal antibody reactive with DR detected a polypeptide of approximately 28 KDa, which agrees with the known molecular mass of the light chain of class II molecules on SDS-PAGE, suggesting that the rapid surface expression might have been due to the presence of pre-formed class II polypeptides. These results demonstrate 1) that T cells defined as resting by conventional criteria contain class II polypeptides and 2) that increases in steady state levels of class II mRNA is a very early event which precedes the initiation of DNA synthesis.
Insights
Resting T cells contain HLA class II polypeptides. Stimulation rapidly increases HLA class II mRNA levels, preceding DNA synthesis, indicating early T cell activation.
Area of Science:
- Immunology
- Molecular Biology
Background:
- T cell activation involves the expression of specific activation antigens.
- HLA class II antigens (DR, DQ, DP) are key markers of T cell activation.
Purpose of the Study:
- To investigate the expression kinetics of HLA class II antigens on T cells post-stimulation.
- To determine the early molecular events, including mRNA changes, during T cell activation.
Main Methods:
- Immunofluorescence using monoclonal antibodies to detect cell surface HLA class II.
- Analysis of mRNA steady-state levels for DR beta.
- Immunoblot analysis of T cell lysates to identify HLA class II polypeptides.
Main Results:
- Cell surface HLA class II determinants appeared within 30 minutes of stimulation.
- Steady-state levels of DR beta mRNA increased by 15 minutes post-stimulation.
- Resting T cells possess detectable HLA class II polypeptides, suggesting pre-formed molecules.
Conclusions:
- T cells, conventionally considered resting, contain HLA class II polypeptides.
- Increased HLA class II mRNA levels are an early event in T cell activation, preceding DNA synthesis.
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