Inducible binding of human lymphocytes to hyaluronate via CD44 does not require cytoskeleton association but does require new protein synthesis.
We have examined molecular mechanisms of the PMA-inducible HA binding ability of human lymphocytes. Newly established OS/6 and OS/37, specific for human CD44, specifically inhibited PMA-induced HA binding of several human cell lines, suggesting that both mAb detect HA binding epitope(s) on CD44. Sequential staining revealed that these mAb cross-blocked each other's binding to Molt-4, T lymphoblast lines, and that neither of them interfered with staining of Molt-4 by other anti-CD44 mAb which induced significant homotypic cell aggregation. Biochemical and PCR analyses did not provide any evidence that PMA stimulation induced dramatic changes in molecular weight or molecular isoforms of CD44. Interestingly, HA binding was not affected and rather slightly increased by cytochalasin B which disrupts F-actin microfilament integrity. This suggests that the ability of CD44 to bind to HA does not correlate with the association of CD44 with the cytoskeleton. On the other hand, protein synthesis inhibitors, cycloheximide and anisomycin clearly inhibited the induction of HA binding of PMA-activated Molt-4 without affecting the expression of CD44 at the same time after stimulation. The same treatment had no effect on PMA-induced FN binding of the cells, which was mediated by VLA integrins. These results suggest that the adhesion functions of CD44 and integrins are differently regulated despite the fact that both are induced by PMA stimulation, and that new protein synthesis is essential for the PMA-induced HA binding by CD44.
Murakami
S
S
Department of Periodontology and Endodontology, Osaka University Faculty of Dentistry, Japan.
Shimabukuro
Y
Y
Miki
Y
Y
Saho
T
T
Hino
E
E
Kasai
D
D
Nozaki
T
T
Kusumoto
Y
Y
Okada
H
H
D000841:Anisomycin / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D000939:Epitopes; D015870:Gene Expression; D006801:Humans; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000378:metabolism*; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000166:cytology*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D016201:Receptors, Lymphocyte Homing / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015854:Up-Regulation
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D000939:Epitopes; D018960:Hyaluronan Receptors; D012333:RNA, Messenger; D016201:Receptors, Lymphocyte Homing; D003571:Cytochalasin B; D000841:Anisomycin; D006820:Hyaluronic Acid; D003513:Cycloheximide; D013755:Tetradecanoylphorbol Acetate