The hemopoietic progenitor 32DCl3(G) cells synthesize C3 molecules and acquire C3b acceptor sites upon differentiation or neoplastic transformation.
32DCl3(G) cells are a diploid, nontumorigenic, IL-3-dependent hemopoietic progenitor cell line, which undergoes terminal differentiation into neutrophilic granulocytes when cultured in presence of G-CSF. The infection with BALB-Moloney murine sarcoma virus, containing a v-HA-ras oncogene, renders this cell line IL-3 independent and continuously growing in the presence of G-CSF, nontumorigenic and with an apparent block at the level of promyelocyte/myelocyte (32D-Ha-ras). After infection with Abelson murine leukemia virus containing a v-abl oncogene, the cell line originates (32D-abl) that is also IL-3 independent but is tumorigenic and unable to differentiate in the presence of G-CSF. This cellular model allowed us to study the relationship between distinct steps of cell differentiation, neoplastic transformation, and C3 synthesis, activation, and characteristics of binding. We demonstrated that C3 synthesis, release, and cleavage are properties already present in the progenitor 32DCl3(G) cells. The more differentiated 32D-Ha-ras cells acquired C3 acceptor sites, apparently completely saturated by the constitutively released molecules. The transformation with Abelson murine leukemia virus, although it did not affect all these properties, let the cells bind considerable amounts of C3-related fragments activated in normal murine serum through the alternative pathway. This last event was a result of the acquisition of PMSF-sensitive serine proteases associated with the plasma membrane.
Department of Experimental Medicine, University, La Sapienza, Rome, Italy.
L
L
Di Renzo
A
A
Zicari
A
A
Longo
M
M
Realacci
G
G
Naso
G
G
Pontieri
M
M
Lipari
D000818:Animals; D001665:Binding Sites; D002454:Cell Differentiation; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000276:immunology*; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D003176:Complement C3 / Q000096:biosynthesis*; D003179:Complement C3b / Q000378:metabolism*; D003170:Complement Pathway, Alternative; D011905:Genes, ras; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008809:Mice, Inbred C3H
D003176:Complement C3; D003179:Complement C3b