HLA class-II-mediated homotypic aggregation: involvement of a protein tyrosine kinase and protein kinase C.
Homotypic aggregation of B-lymphocytes, B-cell lines and class-II-positive T cells via HLA class II molecules was examined. Signaling via DR antigens induced rapid aggregation in a dose- and time-dependent manner, maximum and stable aggregation was induced within 20 minutes. On the contrary, rapid signaling via DP or DQ required prestimulation with either PMA or anti-sIg. Aggregation was temperature and energy dependent. [Ca2+] and [Mg2+] concentrations and an intact cytoskeleton were required while neither mRNA or protein synthesis were required. Furthermore, FACS analysis revealed that aggregation was not directly correlated with cell surface expression of HLA class II molecules. Our results demonstrate that aggregation was mediated through a protein tyrosine kinase (PTK)-dependent pathway that preceded activation of protein kinase C (PKC) and failure to generate either the PTK signal or the PKC signal prevented aggregation. The contribution of a tyrosine kinase was further demonstrated by the total inhibition of aggregation following treatment with an anti-CD45 mAb.
Ramirez
R
R
Laboratory of Molecular Immunogenetics, Biomedical Institute of Cordeliers, Paris, France.
Carracedo
J
J
Mooney
N
N
Charron
D
D
D000911:Antibodies, Monoclonal / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D002118:Calcium / Q000502:physiology; D002449:Cell Aggregation; D002460:Cell Line; D003599:Cytoskeleton / Q000502:physiology; D004789:Enzyme Activation; D005434:Flow Cytometry; D019833:Genistein; D006681:HLA-D Antigens / Q000502:physiology*; D006801:Humans; D007529:Isoflavones / Q000494:pharmacology; D017493:Leukocyte Common Antigens / Q000502:physiology*; D008213:Lymphocyte Activation; D008211:Lymphocyte Cooperation / Q000187:drug effects; D008274:Magnesium / Q000502:physiology; D011493:Protein Kinase C / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000494:pharmacology / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D006681:HLA-D Antigens; D007529:Isoflavones; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C; D017493:Leukocyte Common Antigens; D008274:Magnesium; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium