Acute regulation of human lymphocyte insulin receptors. Analysis by the glucose clamp.

The T lymphocyte insulin receptor model has been used to explore the regulation of insulin receptor appearance in that lymphocytes do not bear the insulin receptor in the circulation and thus are not amenable to regulation by virtue of ligand binding. Such cells synthesize insulin receptors when stimulated by antigen in vivo or in vitro. In these studies, the glucose clamp technique was employed to isolate perturbations in plasma glucose and plasma insulin as potential mediators of the regulation of the mitogen-induced T lymphocyte insulin receptor. Nondiabetic, normal weight individuals volunteered for 10 hyperglycemic clamp studies and nine euglycemic clamp studies with five individuals studied by both protocols. Hyperglycemia and hyperinsulinemia were created by the hyperglycemic clamp (basal plasma glucose was increased from 89 +/- 2 mg/dl to 230 +/- 2 mg/dl and an insulin of 99 +/- 8 microU/ml was reached). Blood was removed for isolation of T lymphocytes at 0, 1, 3, and 4 h of the clamped condition. After 1 h of hyperglycemia accompanied by an elevated plasma insulin, T cell insulin binding fell from 9.9 +/- 0.9 pg/10(6) lymphocytes to 8.5 +/- 0.9 pg/10(6), and reached a nadir of 19 +/- 4% at the conclusion of the clamp. Scatchard analysis of binding data from two of the subjects who underwent the hyperglycemic clamp demonstrated a reduction of the number of binding sites per cell without a change in the affinity of ligand for receptor. To separate the effects of glucose and insulin and the manner in which insulin is provided, the 4-h euglycemic clamp was performed in which fasting plasma glucose was maintained (95 +/- 2 mg/dl) while constant hyperinsulinemia was created (80 +/- 3 microU/ml). Insulin binding to activated, cultured T lymphocytes demonstrated a similar fall in insulin binding. Scatchard analysis of three additional studies again revealed a reduction in receptor number to approximately 40% of base line. These studies reveal that T cell insulin receptor regulation is achieved by hyperinsulinemia independent of the glucose level achieved. The reduction in insulin binding and receptor number could not be accounted for by variations in the strength of lectin stimulation, the time course of lectin response, or by the stress of the clamp itself. The effect of the clamp was specific for the lymphocyte insulin receptor in that the clamp had little effect on the interleukin II receptor activation marker. Acute changes in plasma insulin by the glucose clamp technique are perceived by the T lymphocyte and displayed in tissue culture by an alteration in lectin-induced insulin receptors. One can conclude that rapid changes in ambient in vivo insulin concentrations can regulate the synthesis of T lymphocyte insulin receptors generated in vitro.

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