Role of C1q in phagocytosis of Salmonella minnesota by pulmonary endothelial cells.
The Re mutant of Salmonella minnesota adheres in much greater numbers than the wild type to endothelial cells derived from the bovine pulmonary artery. Since the Re mutant is distinguished from wild-type S. minnesota by its ability to bind C1q and since endothelial cells possess receptors for C1q, we examined the role of C1q in the phagocytosis of the S. minnesota Re mutant. First, preincubating endothelial cells with C1q-enriched medium resulted in increased adherence of the Re mutant (17.9 x 10(4) versus 6.6 x 10(4]. Second, preincubating the Re mutant with C1q-enriched medium resulted in increased numbers of adherent bacteria (62.1 x 10(4) versus 6.6 x 10(4]. Preincubation of both endothelial cells and bacteria with C1q-enriched medium resulted in increased adherence above control levels but less adherence than when either cells or bacteria were preincubated separately in C1q-enriched medium. If serum depleted of C1q was used for preincubation of endothelial cells or bacteria, adherence was reduced below control levels. Thus, C1q plays an important role in the initial steps (recognition, binding, and ingestion) of phagocytosis. Next, the role of C1q was investigated in the respiratory burst response. Levels of superoxide anion released from endothelial cells 15 min after phagocytosis of the Re mutant (100 bacteria per endothelial cell) were assayed by measurement of the superoxide dismutase-inhibitable reduction of ferricytochrome c. Superoxide anion release was increased during phagocytosis of the Re mutant (35 nmol of O2- per 3 x 10(6) endothelial cells) and was also elevated above control values by incubation with soluble C1q (10 nmol of O2- per 3 x 10(6) endothelial cells). These results indicate a role for C1q in both the ingestion and the response of endothelial cells to the S. minnesota Re mutant.
Ryan
U S
US
Department of Medicine, University of Miami School of Medicine, Florida 33101.
Schultz
D R
DR
Goodwin
J D
JD
Vann
J M
JM
Selvaraj
M P
MP
Hart
M A
MA
D000818:Animals; D001422:Bacterial Adhesion; D002417:Cattle; D002478:Cells, Cultured; D003166:Complement Activating Enzymes / Q000502:physiology*; D003172:Complement C1 / Q000502:physiology*; D015922:Complement C1q; D004730:Endothelium, Vascular / Q000382:microbiology*; D005838:Genotype; D066298:In Vitro Techniques; D008855:Microscopy, Electron, Scanning; D010587:Phagocytosis; D011651:Pulmonary Artery / Q000382:microbiology*; D012475:Salmonella / Q000235:genetics / Q000276:immunology*; D013481:Superoxides / Q000378:metabolism
D003172:Complement C1; D013481:Superoxides; D015922:Complement C1q; D003166:Complement Activating Enzymes