Fibronectin and serum amyloid P component stimulate C3b- and C3bi-mediated phagocytosis in cultured human monocytes.

Fibronectin (FN) and serum amyloid P component (SAP) markedly enhance phagocytosis mediated by the C3b and C3bi receptors of cultured human monocytes but not of granulocytes. (The C3b and C3bi receptors of granulocytes can be activated by treatment of these phagocytes with PMA.) Activation of monocyte C3 receptors by FN is developmentally regulated: Freshly explanted monocytes respond to FN with a small increase in C3 receptor-mediated phagocytosis while monocytes matured in culture exhibit a much greater response. The mechanism of action of FN on C3 receptors of cultured monocytes is unique in two respects. First, while substrate-bound FN or SAP activate monocyte C3 receptors, soluble FN does not. Second, stimulation of the basal surface of monocyte plasma membranes by substrate-bound FN activates C3b and C3bi receptors on the apical surface of the plasma membrane, i.e., at sites remote from the segments of membrane in contact with the FN or SAP.

Communication between receptors for different ligands on a single cell: ligation of fibronectin receptors induces a reversible alteration in the function of complement receptors on cultured human monocytes.

Receptors for the third component of complement (C3) on cultured human monocytes (MO) bind ligand-coated particles but do not initiate phagocytosis. The function of these receptors, however, is altered dramatically after MO attach to surfaces coated with fibronectin (FN) or after MO are exposed to phorbol esters. FN and phorbol esters "activate" C3 receptors such that they promote vigorous phagocytosis. Here we show that activation of C3 receptors requires the continuous presence of FN or phorbol esters and is rapidly reversible when these stimuli are removed. Activation does not change the number or distribution of C3 receptors on the surface of MO. We conclude that the function of C3 receptors is regulated by reversible reactions that are initiated by ligation of a different class of receptors on the surface of the same cell.

Dehydration inhibits suckling behavior in weanling rats.

The relation between dehydration and suckling behavior was determined in weanling rats 14, 20, and 23 days of age. After 15 days of age, intracellular and extracellular dehydration sharply reduced both the number of rats that suckled and the amount of milk consumed. Rehydration returned both behaviors to control levels. Thus, during the weaning period, the internal determinants of suckling are not homologous with those of drinking but are more homologous with those governing feeding.

Identification of the C3bi receptor of human monocytes and macrophages by using monoclonal antibodies.

We have obtained four monoclonal antibodies, IB4, OKM1, OKM9, and OKM10, all directed against the C3bi receptor of human monocytes and macrophages (M phi). Two criteria were used to determine the specificity of these antibodies. First, culture surfaces coated with the antireceptor antibodies caused specific down modulation of C3bi receptor activity on M phi adherent to these substrates. Second, receptor protein purified by using IB4 or OKM1 retained the ability to bind selectively to C3bi-coated erythrocytes. Each of the antibodies recognizes a distinct epitope on the C3bi receptor; they do not compete with one another for binding to monocytes. Further, when immobilized on a solid support, each of the antibodies binds a molecule from M phi lysates that can simultaneously bind one of the other monoclonal anti-C3bi receptor antibodies. OKM10 binds and masks the ligand-binding site of the C3bi receptor, while IB4, OKM1, and OKM9 bind to sites remote from the C3bi binding site. All four antibodies immunoprecipitated polypeptides of Mr 185,000 and 105,000 from 125I-surface-labeled M phi. IB4 also precipitates polypeptides of Mr 185,000, 153,000, and 105,000. We conclude that the C3bi receptor of human M phi is a complex composed of two polypeptides, Mr 185,000 and 105,000. We have identified monoclonal antibodies reacting with four distinct antigenic determinants of this complex. The determinant recognized by antibody OKM10 is at or near the ligand-binding site of the receptor. The determinant recognized by antibody IB4 is shared by at least two other leukocyte surface proteins.