Binding sites for peptidoglycan on mouse lymphocytes.
Binding of peptidoglycan (PG), a B-cell mitogen and polyclonal activator, to mouse lymphocytes was studied using rosetting with PG-sensitized erythrocytes and a direct binding assay with 125I-labeled PG. Thirty-four percent of splenic lymphocytes formed PG rosettes, 62% of which were inhibited by preincubation of lymphocytes with free PG. Less than 1 or 3% of spleen cells formed rosettes with uncoated or albumin-coated red cells. The formation of rosettes was not inhibited by 0.1% azide and was not dependent on the presence of complement or immunoglobulins. The 125I-PG bound both specifically and nonspecifically to the lymphocytes. The binding was completed within 15-20 min, was proportional to the cell concentration, and was not inhibited by 0.1% azide or treatment of lymphocytes with formalin. The cells had one set of specific binding sites of low affinity (KD = 1.2-4.6 X 10(-7) M +/- 9% SE, based on competitive) experiments. The binding, however, was complex, probably involving interaction of multiple binding sites on PG with the cell surface. The EC50 (920 micrograms/ml) was similar to the optimal lymphocyte-activating concentration of PG (400-1000 micrograms/ml). The binding correlated with the ability of different PG preparations to stimulate lymphocytes, since only high Mr PG (not low Mr PG preparations, muramyl dipeptide (MDP), or PG pentapeptide) had the ability to specifically bind to lymphocytes, to compete with PG binding, and to stimulate lymphocytes. Also, low Mr PG preparations, MDP, or PG pentapeptide did not inhibit the mitogenic stimulation of lymphocytes by high Mr PG. These results indicate the presence of specific binding sites for PG on the surface of murine lymphocytes and suggest that the binding of PG to these binding sites is involved in lymphocyte activation by PG.
Dziarski
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Department of Microbiology and Immunology, Indiana University School of Medicine, Gary 46408.
D000818:Animals; D001665:Binding Sites; D005260:Female; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D010457:Peptidoglycan / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000032:analysis*; D012397:Rosette Formation; D013154:Spleen / Q000166:cytology
D010457:Peptidoglycan; D011956:Receptors, Cell Surface; C051223:peptidoglycan receptor