The equine homologue of LFA-1 (CD11a/CD18): cellular distribution and differential determinants.
The equine homologue of the leucocyte integrin LFA-1 (CD11a/CD18) has been characterized using a panel of four monoclonal antibodies (mAbs). The antibodies labelled almost all leukocytes, thymocytes and lymph node cells from normal horses, and immunoprecipitated two noncovalently associated polypeptides with molecular weights of 180 kDa and 100 kDa, respectively. The antigen recognized by one mAb could be precipitated by another in this cluster in a sequential immunoprecipitation assay. The mAbs, however, did not block the activities on lymphocyte function of one another. A mAb to the beta subunit of human LFA-1 cross-reacted with equine LFA-1, but an antibody to its alpha subunit did not, suggesting that the beta subunit of the leukocyte integrin may be more highly-conserved. Functionally, H20A and a human CD18 antibody (MHM23) inhibited phorbol ester-mediated homotypic lymphocyte aggregation, whereas mAb CZ3.2 induced rather than inhibited the homotypic cell aggregation. The formation of lymphocyte aggregates induced by CZ3.2 was not blocked by the inhibitory antibodies H20A or MHM23. CZ3.1 seemed to have little inducible or inhibitory effects on homotypic cell aggregation. The mAb CZ3.1 defined a unique LFA-1 determinant present on granulocytes, but absent on lymphocytes in members of an extended horse family, in contrast to the other antibodies which labelled both granulocytes and lymphocytes from these animals. In all other horses tested, no differences in reactivity of CZ3.1 and the other LFA-1 antibodies were observed when the antibodies were tested on lymphocytes or granulocytes. Our results indicate that common epitopes are shared' between human and equine LFA-1, and that the described panel of monoclonal antibodies identifies distinct determinants present on the equine LFA-1 molecule. The following monoclonal antibodies used in this study were given official workshop designations at the Second International Workshop on Equine Leukocyte Antigens (Lunn et al., 1998) CZ3.1 (Cor) = W45; CZ3.2 (Cor) = W77.
Zhang
C H
CH
James A. Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Davis
W C
WC
Grünig
G
G
Antczak
D F
DF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D018821:CD18 Antigens / Q000276:immunology; D002449:Cell Aggregation / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006736:Horses / Q000276:immunology*; D016169:Lymphocyte Function-Associated Antigen-1 / Q000276:immunology*; D008214:Lymphocytes / Q000276:immunology; D011233:Precipitin Tests
D000911:Antibodies, Monoclonal; D018821:CD18 Antigens; D016169:Lymphocyte Function-Associated Antigen-1