Analysis of insulin receptors on human lymphoblastoid cell lines by flow cytometry.
Antibodies to the insulin receptor have provided important experimental probes of receptor structure and function. In the present study, we have characterized the insulin receptor on human lymphoblastoid cell lines using polyclonal and monoclonal anti-receptor antibodies and fluorescence flow cytometry. The cell lines were derived by Epstein-Barr virus transformation of peripheral mononuclear leucocytes from normal subjects or patients with disorders that affect the insulin receptor. Fluorescence analysis revealed a high level of specific fluorescence on lymphoid cell lines from normal individuals (mean peak fluorescence 30-50 units above the control) and was similar to the labelling of the spontaneously transformed lymphoblastoid cell line IM-9. Transformed cells from patients with syndromes of insulin resistance, such as the Rabson Mendenhall syndrome, leprechaunism and the type A syndrome of insulin resistance and acanthosis nigricans, exhibited little or no specific fluorescence. In all cases, there was an unimodal distribution of receptors on cells. In addition, there was a good correlation between specific binding of 125I-insulin and percentage peak fluorescence. The data indicate that fluorescence flow cytometry can be used to study the distribution of insulin receptor on different cell lines and to study cells derived from patients with disease states.
D000911:Antibodies, Monoclonal; D002460:Cell Line; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D011972:Receptor, Insulin / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D011972:Receptor, Insulin