Immunologic Studies in Insulin Resistance. Iii. Measurement of an Insulin Antagonist in the Serum of an Insulin-Resistant Patient by the Blood Sugar Curve Method in Mice

Abstract

The detection of an insulin antagonist in the serum of insulin resistant subjects has been attempted on a number of occasions. In certain instances, the excessive quantities of insulin used in the neutralization experiments in animals may explain the failure to obtain evidence of an insulin antagonist. In the few successful experiments which have been done, little information of a quantitative nature has been obtained. Glen and Eaton (1) and Marble, Fernald and Smith (2) were able to demonstrate decreased responsiveness to insulin in rabbits injected with serum from insulin resistant patients. This change in the experimental animal was still demonstrable a week after injection of the serum. In the same year, Banting et al. (3) described experiments in which the incidence of hypoglycemic convulsions was reduced in mice injected with mixtures of serum and varying amounts of insulin. The serum was obtained from a patient with schizophrenia who developed insulin resistance during insulin shock therapy. This case is especially interesting because diabetes was not present, suggesting that the resistance was directed against prepared insulin only. When the same serum was retested a week later in mice maintained on a high carbohydrate diet, practically no insulin neutralization could be demonstrated. Lerman (4), in studying serum of 5 patients with insulin resistance, obtained evidence for the presence of an insulin antagonist in 2. One of these serums was provided by the writer, and was obtained from a diabetic patient, A. M., the subject of this and previous reports (5,6). A high degree of allergy to insulin, coupled with marked insulin resistance, combined to make the giving of insulin to this patient both disagreeable and ineffective. A further important feature was the presence of a relatively mild degree of diabetes, making possible prolonged periods during which insulin therapy could be omitted without immediate danger to the patient's life. A number of hospital admissions provided opportunity to show that: a. At the end of a period of 5 months or more without insulin therapy, small doses of insulin would give rise to urticaria, constriction in the chest, and, on one occasion, collapse. b. After desensitization with graded doses of insulin, a period of 3 to 5 days followed during which the injection of relatively small doses of insulin would be followed by a fall in blood sugar. c. Within about 11 to 15 days of starting desensitization and treatment with insulin, a solid resistance to insulin developed. These changes were correlated with the absence, and then the appearance, of insulin-neutralizing activity in the patient's serum. The allergic reactions appeared to be mediated by a mechanism quite independent of that giving rise to resistance (5).The method previously used for the demonstration of insulin-neutralizing activity of the patient's serum (6) consisted in observing the incidence of hypoglycemic symptoms in mice injected with a mixture of serum and insulin. This method had a number of disadvantages whic...

Evidence for the Insulin-Directed Specificity of Rabbit Anti-Insulin Serum 1

“…The neutralization of insulin by immune sera has been reported previously (6)(7)(8)(9). However, in the present study the antiserum delayed the hypoglycemic action of insulin only when mixed with insulin for less than fifteen minutes.…”

The Role of Nonprecipitating Insulin Antibodies in Diabetes1

“…It was almost inevitable that once the existence of antibodies to insulin had been suggested, the effect of these on the insulin itself would be questioned. Studies performed on the serum of insulin resistant patients provided further impetus to the concept that antibodies to insulin inhibit the action of insulin, thus necessitating the use of of higher doses than would otherwise have been the case (1,(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). In some instances the methods then available could demonstrate antibodies to insulin by direct skin testing, by passive transfer, by complement fixation or by the use of collodion particles (2,3,18,19).…”

“…7. Studies of the effect of selected plasma and added insulin on the blood sugar of animals (8,9,11,12,16,27).…”

Physical Binding of Insulin by Gamma Globulins of Insulin-Resistant Subjects12

“…However, the immunologic nature of this resistance, which may require the administration of over 1,000 units of insulin per day, has been demonstrated only by indirect means (2). Serum of insulin-resistant patients has been shown to protect mice from the hypoglycemic effect of insulin (3,4), and to prevent the in vitro action of insulin on the rat diaphragm (5). The insulin-neutralizing property of such serum has been shown by electrophoretic (6,7) and salt (8) fractionation to reside in the gamma globulin fraction.…”

“…Yield was 20 per cent of the I"8 used. Less than 4 per cent of the radioactivity of the insulin could be removed by prolonged dialysis. The I"'-labeled insulin was used within a week of its preparation to minimize the possibility of radiation damage to the insulin.…”