Showing papers by "Andrea J. Tenner published in 1981"

Purification and radiolabeling of human C1q.

Abstract: A new procedure for isolating human C1q from serum or plasma is described. The method, that is highly selective, rapid and involves minimal handling, yields fully active, immunoglobulin-free unaggregated C1q. Several different methods of radiolabeling C1q are compared. These include two methods selective for tyrosine residues, two that label lysine residues, and a method that labels sialic acid residues. The effect of each of the labeling procedures on C1q hemolytic activity was assessed. Also appraised for each method was the ability of the labeled molecules to bind to antibody sensitized cells and to interact with C1r and C1s to form C1. The distribution of each of the radiolabels among the three polypeptide chains of C1q and between the collagenous and globular regions of C1q was determined. Methods were identified that selectively labeled the globular portion of either the A or C polypeptide chain of the C1q molecule without loss of functional activity. Another of the methods labeled all three polypeptide chains relatively uniformly without significant loss of activity.

Identification of types of cells in human peripheral blood that bind C1q.

Abstract: Earlier studies showed that approximately 26% of the cells present in human mononuclear cell preparations had the ability to bind purified monomeric C1q. The present studies were initiated to identify the cell types comprising the C1q binding population. Double marker fluorescence, rosetting, and morphologic studies on cell preparations depleted of or enriched in various cell types were simultaneously employed to identify those subpopulations that bound C1q. C1q binding was detected by fluorescent techniques (with FI-F(ab')2 anti-C1q). Monocytes in mononuclear cell preparations were detected by the ability to phagocytose carbonyl iron. B cells were identified by reactivity with rhodamine-conjugated F(ab')2 anti-human F(ab')2 and by rosetting with erythrocytes bearing C3b. These studies showed that monocytes and B lymphocytes comprised the majority of C1q-binding cells in mononuclear cell preparations, whereas T lymphocytes lacked this property. In addition, a minor population of nonphagocytic cells in such preparations that lacked B and T cell markers also bound C1q. Finally, a high but variable proportion of polymorphonuclear leukocytes bound C1q. Binding of C1q to PMN was concentration-dependent, saturable and specific and exhibited an equilibrium constant of 0.76 X 10(7) M-1. Thus, PMN also possess a specific receptor for C1q.