Characterization of a murine monoclonal antibody specific for swine beta1 integrin.
Murine monoclonal antibodies were raised against porcine platelets in order to provide tools for investigating interactions of human blood cells and natural antibodies with porcine tissues. Hybridomas were screened by cellular ELISA on porcine platelets and endothelial cells. Positive clones were tested by flow cytometry for reactivity with isolated endothelial cells. One clone, NaM160-1A3, produced an antibody that stained porcine but not human endothelial cells and lymphocytes. The antibody bound to a 116 kDa glycoprotein on Western blot of both platelets and endothelial cells. The antigen was purified from a platelet lysate by affinity chromatography, first on a ConA column and then on a column presenting the immobilized NaM160-1A3 antibody. Two glycoproteins were obtained: one (116 kDa) was recognized by the antibody and one (150 kDa) was not. The 116 kDa protein had an internal decapeptide identical with human beta 1 integrin, and the 150 kDa protein had an internal amino acid sequence belonging to porcine alpha 2 integrin. Therefore, the NaM160-1A3 antibody was directed against porcine beta 1 integrin and allowed the purification of the complex alpha 2 beta 1, also termed Very Late Antigen 2 (VLA-2). It did not recognize human beta 1 integrin.
Richard
C
C
Institut de Transplantation et de Recherche en Transplantation, Institut National de la Santé et de la Recherche Médicale, Unité 437, Nantes, France.
Thibaudeau
K
K
Charreau
B
B
Loirat
M J
MJ
Naulet
J
J
Blanchard
D
D
Soulillou
J P
JP
Bouhours
J F
JF
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001792:Blood Platelets / Q000276:immunology; D004730:Endothelium, Vascular / Q000276:immunology; D006084:Graft Rejection / Q000209:etiology / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D016023:Integrins / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D010980:Platelet Membrane Glycoproteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D038981:Receptors, Collagen; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013552:Swine; D014183:Transplantation, Heterologous / Q000009:adverse effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D019012:Integrin beta1; D016023:Integrins; D010980:Platelet Membrane Glycoproteins; D038981:Receptors, Collagen