Abstract
gamma-Interferon (IFN-gamma) is a well characterized lymphokine known to regulate many mononuclear phagocyte functions, including expression of class I and class II major histocompatibility complex genes. The second component of complement (C2) and factor B are major histocompatibility complex class III gene products synthesized in mononuclear phagocytes. Recombinant IFN-gamma increased the synthesis of C2 and factor B in primary cultures of human mononuclear phagocytes and in murine fibroblasts transfected with cosmid DNA bearing the human C2 and factor B genes. In both cell types the increases in C2 and factor B protein synthesis were detected at concentrations of IFN-gamma less than 1 unit/ml and the regulation of each was pretranslational. The IFN-gamma-induced increases in C2 and factor B mRNA did not require new protein synthesis. In primary cultures of human monocytes, the kinetics of induction of C2 and factor B synthesis differed, but in the transfected L-cells the kinetics were similar, suggesting differences in transduction of the IFN-gamma signal, transcriptional, and/or post-transcriptional events in the two cell types. The small size of the factor B 5' flanking region, which is bounded by the 3' terminus of the IFN-gamma-regulated C2 gene, provides a well defined region to probe the structural basis for IFN-gamma regulation of gene expression.
Highlights
?-Interferon Increases Expression of ClassI11 Complement Genes C2 and FactorB in Human Monocytes andin Murine Fibroblasts Transfected with Human C2 and FactorB Genes*
IFN-y increased the synthesis of C2 and factor B in primary cultures of human mononuclear phagocytes and in murine fibroblasts transfewctietdh cosmid DNA
In bothcell types the increases Cin2 andfactor B protein synthesis were detected at concentrations of IFN-y less than 1 unitlmlandtheregulationofeach was pretranslational
Summary
After 1-10 days in culture, the monolayers were washed three times to remove spent medium and serum and were cultured for periods ranging from 4 to 24 h in Medium 199 containing bovine serum albumin (1 mg/ml) alone (control) omr edium containing interferon.Mouse L-cells transfected with human cosmid DNA containing the genes for C2 and factor B (class I11 transfectants) [16] and control L-cells transfected with cosmid DNA containing a murine class I gene (class I transfectants) [16] were maintained as monolayers in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum, 1.36mg/ml hypoxanthine, 17.4 mg/ml aminopterin, and 388 ,ug/ml thymidine These cells were subcultured into 35-mm dishes and grown to confluence before incubation in control or IFN-y-containingmedium. Nick-translated 32P-labeledBamHI digest of X phage was included on each gel as a molecular mass marker
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