Intracellular localization of the human receptor for the globular domains of C1q.
This study was performed to determine the localization of the recently described receptor for the globular domain of C1q, gC1qR. In contrast to previous reports, we were not able to detect significant surface expression of gC1qR on Raji cells, monocytes, neutrophils, human or rat mesangial cells, the endothelial cell line EA.hy 926, or HUVEC using FACS analysis. Only by using digoxigenin-conjugated Abs could some surface staining of gC1qR be observed on rat mesangial cells and neutrophils. However, after permeabilizing these cells with saponin, a strong positive intracellular staining for gC1qR was observed by FACS, fluorescence microscopy on coverslips, and confocal laser scanning microscopic analysis. By reflection contrast microscopy and electron microscopy on ultrathin sections of permeabilized Raji cells, it was shown that gC1qR is present in double membranous cytoplasmic vesicles located in the proximity of the plasma membrane. To determine whether certain conditions could induce surface expression of gC1qR, Raji cells were either stimulated with T cell growth factor, LPS, or driven to apoptosis by incubation with fenretinide or by serum depletion. None of the conditions resulted in significant surface expression of gC1qR. Our hypothesis that gC1qR is not a surface molecule but a soluble molecule that is secreted by cells is supported by the observation that gC1qR is found in significant concentrations in supernatants of several cultured cells and in normal human and rat sera. Our results suggest that the recently described gC1qR is not a cell surface receptor, but a soluble binding protein with affinity for the globular heads of C1q. Excreted gC1qR might act as a potential fluid phase regulator of complement activation.
van den Berg
R H
RH
Department of Nephrology, Leiden University Hospital, The Netherlands.
Prins
F
F
Faber-Krol
M C
MC
Lynch
N J
NJ
Schwaeble
W
W
van Es
L A
LA
Daha
M R
MR
D000818:Animals; D002460:Cell Line; D015922:Complement C1q / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D018613:Microscopy, Confocal; D051381:Rats; D011951:Receptors, Complement / Q000032:analysis* / Q000378:metabolism / Q000648:ultrastructure; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis* / Q000378:metabolism / Q000648:ultrastructure
D011951:Receptors, Complement; D018160:Receptors, Cytoplasmic and Nuclear; D015922:Complement C1q