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In vitro processing of insulin for recognition by murine T cells results in the generation of A chains with free CysSH.

J Hampl, G Gradehandt, H Kalbacher and E Rüde
J Immunol May 1, 1992, 148 (9) 2664-2671;
J Hampl
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G Gradehandt
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H Kalbacher
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E Rüde
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Abstract

Studies on the processing of insulin as an Ag for the presentation to MHC class II-restricted T cells revealed that the amino acid residues 1-14 of the insulin A chain are recognized by insulin-specific T cells. An A1-14 peptide containing three cys-residues that were protected by S-sulfonate groups still needed processing by APC for efficient presentation similar to native insulin. We suspected that reductive deblocking or opening of disulfide bonds that generates CysSH-residues may be an essential processing step for these Ag. Due to the instability of SH-groups it was not possible to test A chain peptides with free SH-groups in the usual way for processing-independent presentation by fixed APC. However, under acidic conditions (pH 5) during APC pulsing with the Ag we could demonstrate that the freshly reduced A1-14 fragment as well as reduced insulin are able to bind to Ia Ag and to stimulate appropriate T cells without further processing. Various substitutions of cys-residues by Ser within this peptide revealed that only CysA7 is critical for Ia binding and/or T cell recognition. In intact insulin, this residue links the A chain containing the T cell epitope to the B chain. Therefore, we propose that insulin processing is not dependent on proteolysis or on the generation of a conformational determinant but on the separation of A and B chains resulting in A chains whose cys-residues are converted into CysSH.

  • Copyright © 1992 by American Association of Immunologists

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The Journal of Immunology
Vol. 148, Issue 9
1 May 1992
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In vitro processing of insulin for recognition by murine T cells results in the generation of A chains with free CysSH.
J Hampl, G Gradehandt, H Kalbacher, E Rüde
The Journal of Immunology May 1, 1992, 148 (9) 2664-2671;

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In vitro processing of insulin for recognition by murine T cells results in the generation of A chains with free CysSH.
J Hampl, G Gradehandt, H Kalbacher, E Rüde
The Journal of Immunology May 1, 1992, 148 (9) 2664-2671;
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Print ISSN 0022-1767        Online ISSN 1550-6606