Homotypic cell aggregation via conformational change of CD44 molecule induced by anti-CD44 monoclonal antibodies.
The homotypic cell aggregation of leukocytes is an unique adhesive event which is caused by cellular activation. Anti-CD44 monoclonal antibody (mAb) induces homotypic cell aggregation of hematopoietic cell lines expressing CD44, but the mechanism of homotypic cell aggregation is poorly understood. We used four mAbs against CD44: TL-1 which was newly developed and seemed to react with a non-hyaluronate binding site, OS/37 and BU52 which recognized a hyaluronate binding site, and Hermes-3 which recognized a non-hyaluronate binding site. TL-1 treatment induced strong homotypic cell aggregation in several types of cell lines including a B cell line from a patient with leukocyte adhesion deficiency syndromes (LAD) and normal peripheral blood lymphocytes (PBL). OS/37 and BU52 also induced weak homotypic cell aggregation. None of these anti-CD44 mAbs-induced homotypic cell aggregations was blocked by antibodies against LFA-1, ICAM-1, VLA-4, or L-selectin. Interestingly, the TL-1-induced homotypic cell aggregation was blocked by Hermes-3 or OS/37, but not by BU52. BU52-induced homotypic cell aggregation was blocked by Hermes-3 or OS/37, but not by TL-1. OS/37-induced homotypic cell aggregation was blocked by Hermes-3, TL-1 or BU52. The blocking experiments with anti-metabolic agents revealed that the induced homotypic cell aggregation was energy-dependent and associated with intracytoplasmic actin filaments. This homotypic cell aggregation did not require de novo protein synthesis, because it was not affected by pretreatment with either cycloheximide or actinomycin D. FACS analysis revealed that TL-1 binding did not affect the intensity of expression of the CD44 molecule on the cell surface.
Cao
L
L
Department of Pathology, Okayama University Medical School, Japan.
Yoshino
T
T
Nishiuchi
R
R
Yamadori
I
I
Akagi
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology*; D001667:Binding, Competitive / Q000276:immunology; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D002449:Cell Aggregation / Q000187:drug effects / Q000276:immunology*; D002460:Cell Line; D005434:Flow Cytometry; D018960:Hyaluronan Receptors / Q000737:chemistry / Q000276:immunology*; D006820:Hyaluronic Acid / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation / Q000187:drug effects
D000911:Antibodies, Monoclonal; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid