ISSN:
1365-3083
Quelle:
Blackwell Publishing Journal Backfiles 1879-2005
Thema:
Medizin
Notizen:
Molecules encoded by the class I major histocompatibility genes bind short (nonameric) peptides produced by inlracellular proleolysis of antigens. These complexes formed intraeellularly are then expressed on membranes of target eells and recognized by the anligen receptor of eytolylic T cells. No binding of externally added peptides could so far be monitored directly on the antigen presenting cells, although cytotoxicity experiments and indirect binding assays provided evidence for its existence. Here we report experiments where specific binding to class I molecules, of externally added peptides, has been monitored on living cells, N-terminal biottn-labelled Kd-restricted peptides (residues 147-155, residues 147-158, and an analogue lacking the arginine at position 156. derived from the sequence of the influenza A virus nucleoprotein) were incubated with murine H-2Kd mastocytoma cells (line P815)dt 4°C. The binding on surface of hve, intact cells was then demonstrated fluoro metrically via ihe inieraction ofa streptavidin-phycoerythrin conjugate with the biotin-labelled peptides. Thus, this binding does not involve processing, and its specificity in terms of peptide structure was established by competition with the respective unmodified peptides. The specificity ofbinding to class I molecules was demonstrated by blocking experiments u.sing monoclonal antibodies specific for H-2Kd. Finally, a correlation was observed between the results of peptide binding measurements and those ofcytotoxieity assays.
Materialart:
Digitale Medien
URL:
http://dx.doi.org/10.1111/j.1365-3083.1992.tb03107.x
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