Regulation of MHC Class II and Interleukin-1 Gene Expression in Differentiating Macrophages
The expression and regulation of the MHC class II and interleukin-1 genes was studied during the differentiation of bone marrow stem cells into macrophages. Bone marrow cells were cultured in the presence of CSF-1 and allowed to differentiate for 3, 5, 7, and 9 d. Cells were stimulated with either medium, endotoxin (1 gg/ml) or interferon-r (IFN-r, 100 U/ml) 24 hr prior to the end of the culture period. Upon completion of culture, total cytoplasmic RNA was isolated from the adherent bone marrow-derived macrophages (BMDM) and subjected to Northern blot analysis. The A and E mRNA was constitutively expressed by medium treated BMDM after 3, g, and/d, but by 9 d the mRNA had declined to values lower than those observed on d 3. IFN-r treatment augmented A and E mRNA at each time point, whereas endotoxin treatment decreased values after 5 and 7 d. Additional studies demonstrated that IFN-r was not being produced by either the non-adherent or adherent cells nor was the production of PGE responsible for the decline in MHC class II gene expression. IL-1 mRNA was only detected in those cultures stimulated with endotoxin; medium alone or IFN-r treatment did not induce IL-1 mRNA transcription. Antigen-primed lymph node T cells and T cell hybridomas demonstrated that the ability of BMDM to function as antigen presenting cells was limited by the differentiation-specific regulation of both class II and IL-1 gene products. Exogenous Eα and Aβ genes were inserted into bone marrow stem cells using either a modified calcium phosphate transfection or an electroporation procedure and their expression was monitored using Northern blot and immunofluorescence analysis. However, there was no alteration in the expression of membrane IL-1 following trans-fection. We currently are examining the expression of regulatory factors which are responsible for the differentiation-specific regulation of MHC class II and IL-1 gene expression.
KeywordsBone Marrow Cell Northern Blot Analysis Thymidine Kinase Bone Marrow Stem Cell Nonadherent Cell
Unable to display preview. Download preview PDF.
- Bender, J. G., Van Epps, D. E., and Stewart, C. C.: A model for the regulation of myelopoiesis by specific factors. J. Leuk. Biol. 39: 101 - 111, 1986.Google Scholar
- Collart, M. A., Belin, D., Vassalli, J.-D., de Kossodo, S., and Vassalli, P.: 7-Interferon enhances macrophage transcription of the tumor necro sis factor/cachectin, interleukin 1, and urokinase genes, which are controlled by short-lived repressors. J. Exp. Med. 164: 2113 - 2118, 1986.Google Scholar
- Eustis-Turf, E. P., Wang, X.-M., and Schook, L. B.: Electroporation of bone marrow cells: parameters and applications. Bio-Radiation, in press, 1987a.Google Scholar
- Eustis-Turf, E. P., Pullen, J. K., Myers, M. J., Wang, X.-M., and Schook, L. B.: Regulation of class II MHC gene expression during macrophage differentiation. In, L. B. Schook and J. G. Tew (ed.), Antigen Presenting Cells: Diversity, Differentiation, and Regulation. Alan R. Liss Publications, Inc., (in press), 1987b.Google Scholar
- Grabstein, K., Mochizaki, D., Kronheim, S., Price, V., Cosman, D., Urdal, D., Gillis, S., and Conlow, P.: Regulation of antibody production in vitro by granulocyte-macrophage colony stimulating factor. J. Mol. Cell. Immunol. 2:199-207, 1986.Google Scholar
- Kawasaki, E. S., Ladner, M. B., Wang, A. M., Van Arsdell, J., Warren, M. K., Coyne, M. Y., Schweickart, V. L., Lee, M.-T., Wilson, K. J., Boosman, A., Stanley, E. R., and David, D. F.: Molecular cloning of a complementary DNA encoding human macrophage-specific colony-stimulating factor (CSF-1). Science 230: 291 - 296, 1985.Google Scholar
- Stern, A. A Erb, P., and Gisler, R. H.: Ia-bearing bone marrow-cultured macrophages induce antigen-specific helper T cells for antibody synthesis. J. Immunol. 123:612-615, 1979.Google Scholar