Characterization of the porcine homologue to human platelet glycoprotein IIb-IIIa (CD41/CD61) by a monoclonal antibody.
Human gpIIb-IIIa or CD41/CD61 is a Ca(2+)-complex dependent heterodimer, abundant on platelets, that plays a key role in hemostasis. This report describes a murine monoclonal antibody, JM2E5, able to recognize and immunoprecipitate the gpIIb/IIIa surface glycoprotein from porcine platelets. Immunoprecipitation analysis showed an antigen molecular weight of 115 and 85 kDa under nonreducing conditions, and of 110, 100 and 25 kDa under reducing conditions. Immunohistochemistry analyses of frozen sections from several porcine lymphoid organs gave specific staining on platelets. EDTA treated platelets were studied by flow cytometry indicating that the epitope recognized was Ca(2+)-complex independent. Western-blotting experiments with porcine platelet extracts gave an antigen molecular weight of 85 kDa under nonreducing conditions, thus localizing the epitope recognized by JM2E5 on the complex light chain gpIIIa or CD61. JM2E5 was also cross-reacting with human, bovine and horse platelets, as shown by flow cytometry. This mAb would allow further studies on this important adhesion molecule on horses, ruminants and pigs, and it should be especially useful as a general anti-porcine platelet reagent.
Pérez de la Lastra
J M
JM
Unidad Mixta CSIC-UCO, Departamento de Genética, Facultad de Veterinaria, Universidad de Córdoba, Spain. gm2pepej@lucano.uco.es
Moreno
A
A
Pérez
J
J
Llanes
D
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D001792:Blood Platelets / Q000187:drug effects / Q000276:immunology*; D002417:Cattle; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D039661:Integrin beta3; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000276:immunology*; D010980:Platelet Membrane Glycoproteins / Q000276:immunology*; D013552:Swine; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D039661:Integrin beta3; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D010980:Platelet Membrane Glycoproteins; D004492:Edetic Acid