TY - JOUR
T1 - Monoclonal antibodies against porcine LFA-1
T2 - Species cross-reactivity and functional effects of β-subunit-specific antibodies
AU - Hildreth, James
AU - Holt, Valerie
AU - August, J. Thomas
AU - Pescovitz, Mark D.
PY - 1989
Y1 - 1989
N2 - As a first step in developing a porcine model system to study the effect of LFA-1-specific monoclonal antibodies on allograft rejection, we have identified anti-human LFA-1 monoclonal antibodies that cross-react with porcine leukocytes and have used one of these antibodies against the LFA-1 β-subunit to purify porcine LFA-1. Immunization of Balb/c mice with the purified antigen yielded eight monoclonal antibodies directed against the β-subunit of porcine LFA-1. These antibodies immunoprecipitated pig proteins corresponcling to the human α-subunits of LFA-1 (CD11a), Mac-1 (CD11b) and LeuM5 (CD11c), and their common β-subunit (CD18). Under both reducing and non-reducing conditions, the porcine β-subunit showed a different electrophoretic mobility from that of the human β-subunit. In addition, results of peptide mapping studies revealed structural differences between the β-subunits of human and porcine LFA-1. The anti-LFA-1 β-chain antibodies were found to cross-react with antigens expressed on rat, mouse, hamster, rabbit, dog, cow and human leukocytes, indicating that the β-subunit contains highly conserved epitopes. Competitive binding studies showed that these antibodies and three anti-human LFA-1 antibodies defined at least four distinct epitopes on the porcine β-subunit, three of which were also found on human β-subunits. Whereas the cross-reactive anti-human β-subunit antibody H52 profoundly inhibited mitogenic and allogenic stimulation of porcine T lymphocytes as well as cytotoxic T cell and natural killer (NK) cell function, the anti-porcine LFA-1 monoclonal antibodies had very limited effect on porcine lymphocyte function. Our results demonstrate that porcine leukocytes express the LFA-1 molecule and that this antigen is involved in T cell and NK cell functions, as has previously been shown in human and murine systems. We conclude that the pig may serve as a useful model in which to test the effect of anti-LFA-1 antibodies on allograft rejection.
AB - As a first step in developing a porcine model system to study the effect of LFA-1-specific monoclonal antibodies on allograft rejection, we have identified anti-human LFA-1 monoclonal antibodies that cross-react with porcine leukocytes and have used one of these antibodies against the LFA-1 β-subunit to purify porcine LFA-1. Immunization of Balb/c mice with the purified antigen yielded eight monoclonal antibodies directed against the β-subunit of porcine LFA-1. These antibodies immunoprecipitated pig proteins corresponcling to the human α-subunits of LFA-1 (CD11a), Mac-1 (CD11b) and LeuM5 (CD11c), and their common β-subunit (CD18). Under both reducing and non-reducing conditions, the porcine β-subunit showed a different electrophoretic mobility from that of the human β-subunit. In addition, results of peptide mapping studies revealed structural differences between the β-subunits of human and porcine LFA-1. The anti-LFA-1 β-chain antibodies were found to cross-react with antigens expressed on rat, mouse, hamster, rabbit, dog, cow and human leukocytes, indicating that the β-subunit contains highly conserved epitopes. Competitive binding studies showed that these antibodies and three anti-human LFA-1 antibodies defined at least four distinct epitopes on the porcine β-subunit, three of which were also found on human β-subunits. Whereas the cross-reactive anti-human β-subunit antibody H52 profoundly inhibited mitogenic and allogenic stimulation of porcine T lymphocytes as well as cytotoxic T cell and natural killer (NK) cell function, the anti-porcine LFA-1 monoclonal antibodies had very limited effect on porcine lymphocyte function. Our results demonstrate that porcine leukocytes express the LFA-1 molecule and that this antigen is involved in T cell and NK cell functions, as has previously been shown in human and murine systems. We conclude that the pig may serve as a useful model in which to test the effect of anti-LFA-1 antibodies on allograft rejection.
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U2 - 10.1016/0161-5890(89)90145-4
DO - 10.1016/0161-5890(89)90145-4
M3 - Article
C2 - 2481234
AN - SCOPUS:0024453893
VL - 26
SP - 883
EP - 895
JO - Molecular Immunology
JF - Molecular Immunology
SN - 0161-5890
IS - 9
ER -