Characterization of the rat leukocyte integrin, CD11/CD18, by the use of LFA-1 subunit-specific monoclonal antibodies.
We have attempted to characterize the rat leukocyte integrin, CD11/CD18, by the use of newly generated monoclonal antibodies (mAb) WT.1 (anti-CD11a) and WT.3 (anti-CD18) in conjunction with an mAb, OX42, reactive with a rat integrin-like molecule, with respect to the biochemistry, cellular distribution and function. The conclusion that the mAb WT.1 and WT.3 specifically recognize the rat CD11a and CD18, respectively, was based on: (a) their ability to inhibit homotypic aggregation of splenic concanavalin A (Con A) blasts; (b) sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of the antigens recognized; (c) their ability to inhibit binding of Con A blasts to the purified ligand, namely the ICAM-1 antigen and (d) their blocking abilities in mixed leukocyte reaction. In the rat, CD18 has an apparent molecular mass of 95-100 kDa and can associate with at least three distinct alpha subunits of 160-170 kDa (CD11a), 140-150 kDa and 120-130 kDa. The latter two are precipitated by OX42 from M phi but not from unstimulated lymphocytes. They presumably represent the rat CD11b and CD11c, respectively. Rat thymocytes, PBL, thoracic duct lymphocytes, monocytes and neutrophils expressed differential levels of CD11a and CD18. Peritoneal M phi showed virtually no CD11a expression, although CD18 was expressed at levels similar to those seen on blood monocytes, showing an interesting pattern of LFA-1 expression regulation in this cell lineage. Both WT.1 (anti-CD11a) and WT.3 (anti-CD18) apparently recognize a "low-affinity" as well as a "high-affinity" form of LFA-1 and do not discriminate between the two.
Tamatani
T
T
Department of Immunology, Tokyo Metropolitan Institute of Medical Science, Japan.
Kotani
M
M
Miyasaka
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D018821:CD18 Antigens; D015815:Cell Adhesion Molecules / Q000378:metabolism; D002449:Cell Aggregation; D003179:Complement C3b / Q000276:immunology; D005434:Flow Cytometry; D016023:Integrins / Q000276:immunology*; D018799:Intercellular Adhesion Molecule-1; D016169:Lymphocyte Function-Associated Antigen-1 / Q000737:chemistry / Q000276:immunology*; D011233:Precipitin Tests; D051381:Rats; D016028:Receptors, Leukocyte-Adhesion / Q000737:chemistry / Q000276:immunology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D018821:CD18 Antigens; D015815:Cell Adhesion Molecules; D016023:Integrins; D016169:Lymphocyte Function-Associated Antigen-1; D016028:Receptors, Leukocyte-Adhesion; D018799:Intercellular Adhesion Molecule-1; D003179:Complement C3b; D013755:Tetradecanoylphorbol Acetate