Analysis of cell-free human alpha1 integrin with a monoclonal antibody to the I-domain: detection in ocular fluid and function as an adhesion substrate.
The alpha1 beta1 integrin, an inserted (1) domain containing collagen receptor, is expressed in the cell surface membrane of normal and malignant cells, and may play a role in their migration through tissues or in metastatic spread. Here we report that a functional anti-human alpha1beta1 integrin monoclonal antibody (mAb) (1B3.1) directly and specifically binds plastic bound recombinant human alpha1 I-domain protein containing the collagen binding site. Detection was diminished by acidification of the I-domain protein but was enhanced by increasing concentrations of Mg2+ cation. Furthermore, we detected binding of the mAb to proteins from the ocular fluids of 6 patients, with the highest concentration, corresponding to 22.1 ng/ml of I-domain, found in a sample from the eye of a patient with metastatic lung adenocarcinoma. Interestingly, we found that both SKNSH neuroblastoma cells and virally transformed human T cells adhered specifically to plastic wells coated with either immobilized collagen IV or alpha1 I-domain. MAb I B3.1 inhibited adhesion to collagen IV but not to immobilized I-domain. These results suggest a novel function for cell free alpha1 I-domain as a substrate for cellular adhesion, which may have relevance in tumor spread in vivo.
Bank
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I
Department of Medicine, Chaim Sheba Medical Center, Tel Hashomer, Israel. ibank@post.tau.ac.il
Kapyla
J
J
Grinbaum
A
A
Doolman
R
R
Bank
J
J
Sela
B A
BA
D000230:Adenocarcinoma / Q000175:diagnosis / Q000556:secondary; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001082:Aqueous Humor / Q000737:chemistry*; D002386:Cataract / Q000175:diagnosis; D002412:Cations / Q000737:chemistry; D015815:Cell Adhesion Molecules / Q000276:immunology*; D002474:Cell-Free System; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005134:Eye Neoplasms / Q000556:secondary; D006801:Humans; D039423:Integrin alpha1 / Q000737:chemistry / Q000276:immunology* / Q000502:physiology; D039222:Integrin alpha1beta1 / Q000276:immunology; D008175:Lung Neoplasms / Q000175:diagnosis / Q000473:pathology; D017434:Protein Structure, Tertiary
D000911:Antibodies, Monoclonal; D002412:Cations; D015815:Cell Adhesion Molecules; D039423:Integrin alpha1; D039222:Integrin alpha1beta1