Antibodies against the
insulin receptor (Anti-R), which are found in the serum of type B patients with the syndrome of
insulin resistance and
acanthosis nigricans, inhibit the binding of
insulin to its receptor and mimic the actions of
insulin when studied acutely in vitro. After prolonged exposure of 3T3-L1 cells to Anti-R, the insulinomimetic activity is lost, and the cells show a marked decrease in their maximal response to
insulin (antibody-induced desensitization), thus providing a model for the
insulin resistance seen in vivo. This study explores in detail the mechanism and specificity of desensitization in 3T3-L1 cells.Desensitization, like the insulinomimetic activity of Anti-R, requires bivalence. Monovalent preparations of Anti-R inhibit
insulin binding and shift the
insulin biological dose-response curve to the right, but do not decrease the maximal
insulin response. The affinity of monovalent Anti-R is less than that of the native antibody. Cross-linking of monovalent Anti-R reconstitutes its insulinomimetic activity and partially reconstitutes desensitization. Desensitized cells are resistant to the insulinomimetic actions of
concanavalin A, which interacts with the
insulin receptor, but are not desensitized to
spermine and
vitamin K(5), insulinomimetic agents that are thought to act independently of the
insulin receptor.
Glucose,
pyruvate, or certain
hexoses are required in the incubation media for desensitization to occur. Although Anti-R is taken up into cells and degraded by lysosomes,
chloroquine,
cycloheximide,
colchicine, and
cytochalasin E have little influence on the induction of or recovery from antibody-induced desensitization. These data suggest that desensitization is not merely due to the inhibition of
insulin binding, but is a complex process involving a decreased ability of the receptor to generate a
biological response.