Bone marrow-derived macrophage expression of endogenous and transfected class II MHC genes during differentiation in vitro.

C57BL/6 (H-2b) mice fail to express I-E molecules on the surface of their cells and thus are unable to respond to I-E-restricted antigens such as GL phi and cytochrome c. Previous experiments in our laboratory have involved developing a system for studying differentiation of bone marrow cells into mature macrophage to gain a better understanding of class II MHC gene expression and function. In this study, we have used this system to transfect the E alpha d gene (cosmid 17.2) into C57BL/6 bone marrow cells and subsequently observed I-E expression on bone marrow-derived macrophages (BMDM) after differentiation in vitro. By using a modified calcium phosphate protocol, we found that the optimal period for transfection of the bone marrow cells was after 2 days of culture in vitro. By using the anti-I-E monoclonal antibody (Ia.7) derived from hybridoma 14-4-4, we detected the I-E molecule on the surface of transfected macrophages by a radiobinding assay and immunoprecipitation. BMDM expressed the I-E product maximally at 5 days of differentiation, and expression then declined. Furthermore, we have found that the expression of the I-E molecule on transfected macrophage was dependent upon exposure to interferon-gamma. Expression of I-E molecules was also detected by the generation of an allogeneic response. Transfected BMDM were compared with (CB6)F1 BMDM for their ability to stimulate C57BL/6 T cells and they were found to be equally effective. By using these initial findings, we hope to further optimize the conditions for insertion and expression of class II MHC genes in bone marrow cells.

• Publication year

  1986

• Venue

  Journal of Immunology

• Publication date

  1986-08-15

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.4049/jimmunol.137.4.1359PMID 3090147
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Correcting an immune-response deficiency by creating E alpha gene transgenic mice.

  1985cited by this paper
• Functional expression of a microinjected Ed alpha gene in C57BL/6 transgenic mice.

  1985cited by this paper
• Dissection of macrophage tumoricidal and protozoacidal activities using T-cell hybridomas and recombinant lymphokines

  1985cited by this paper
• A trans-acting class II regulatory gene unlinked to the MHC controls expression of HLA class II genes

  1985cited by this paper
• Genetics and expression of mouse Ia antigens.

  1985cited by this paper
• Transfer of genes into hematopoietic cells using recombinant DNA viruses.

  1985cited by this paper
• The generation of macrophage-like cell lines by transfection with SV40 origin defective DNA.

  1984cited by this paper
• Effect of IFN-gamma on the immune response in vivo and on gene expression in vitro.

  1984cited by this paper
• Differential requirements for antigen processing by macrophages for lysozyme-specific T cell hybridomas.

  1984cited by this paper
• Functional and inducible expression of a transfected murine class II major histocompatibility complex gene.

  1984cited by this paper
• Effect of glucocorticosteroids on epidermal cell-induced immune responses.

  1984cited by this paper
• In-vitro-derived bone marrow macrophages. Expression of Ia antigens during macrophage differentiation.

  1984cited by this paper
• Alpha-fetoprotein inhibits macrophage expression of Ia antigens.

  1984cited by this paper
• Genetic transformation of murine bone marrow cells to methotrexate resistance.

  1983cited by this paper
• Induction of Ia and H-2 antigens on a macrophage cell line by immune interferon.

  1983cited by this paper
• Bone marrow-derived macrophage as accessory cells in antigen-induced T cell proliferation. H-2I region requirements for L-glutamic60-L-alanine30-L-tyrosine10 response.

  1983cited by this paper
• CSF‐1—A mononuclear phagocyte lineage‐specific hemopoietic growth factor

  1983cited by this paper
• Differential regulation of HLA‐DR mRNAs and cell surface antigens by interferon.

  1983cited by this paper
• Several mechanisms can account for defective E alpha gene expression in different mouse haplotypes.

  1983cited by this paper
• Characterization and expression of H-21 region gene products on bone marrow-derived macrophages.

  1982cited by this paper
• Regulation of murine macrophage Ia antigen expression by an immune interferon-like lymphokine: inhibitory effect of endotoxin.

  1982cited by this paper
• A molecular map of the immune response region from the major histocompatibility complex of the mouse

  1982cited by this paper
• In vivo assembly of regularly spaced nucleosomes on mouse βmaj-globin DNA cloned in an SV40 recombinant

  1982cited by this paper
• Functional heterogeneity of culture-grown bone marrow-derived macrophages. II. Lymphokine stimulation of antigen-presenting function.

  1982cited by this paper
• Prostaglandins modulate macrophage Ia expression

  1982cited by this paper
• Analysis of Ir gene function using monoclonal antibodies: independent regulation of GAT and GLPhe T cell responses by I-A and I-E subregion products on a single accessory cell population.

  1981cited by this paper
• Variable synthesis and expression of E alpha and Ae (E beta) Ia polypeptide chains in mice of different H-2 haplotypes.

  1981cited by this paper
• Monoclonal antibody against an Ir gene product?

  1980cited by this paper
• Demonstration of a soluble mediator that induces exudates rich in Ia- positive macrophages

  1980cited by this paper
• Regulation of macrophage populations. I. Preferential induction of Ia-rich peritoneal exudates by immunologic stimuli.

  1980cited by this paper
• Regulation of murine macrophage Ia-antigen expression by products of activated spleen cells

  1980cited by this paper
• Gene transfer in intact animals

  1980cited by this paper
• Secretion of plasminogen activator by bone marrow-derived mononuclear phagocytes and its enhancement by colony-stimulating factor

  1979cited by this paper
• Gene complementation in the T-lymphocyte proliferative response to poly (Glu55Lys36Phe9)n. A demonstration that both immune response gene products must be expressed in the same antigen-presenting cell

  1979cited by this paper
• Lymphocyte specificity to protein antigens. I. Characterization of the antigen-induced in vitro T cell-dependent proliferative response with lymph node cells from primed mice.

  1977cited by this paper
• A rapid method for the isolation of functional thymus‐derived murine lymphocytes

  1973cited by this paper
• Histocompatibility-linked immune response genes.

  1972cited by this paper
• Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

  1970cited by this paper

• The opioid antagonist naltrexone blocks acute endotoxic shock by inhibiting tumor necrosis factor-α production☆

  2004cites this paper
• The opioid antagonist naltrexone blocks acute endotoxic shock by inhibiting tumor necrosis factor-alpha production.

  2004cites this paper
• CD28 ligation prevents bacterial toxin-induced septic shock in mice by inducing IL-10 expression.

  1997cites this paper
• Endotoxin and interferon-gamma differentially regulate the transcriptional levels of proto-oncogenes and cytokine genes during the differentiation of colony-stimulating factor type-1-derived macrophages.

  1995cites this paper
• 6 – Gene Transfer Technology: Alternative Techniques and Applications

  1994cites this paper
• Mechanisms generating functionally heterogeneous macrophages: chaos revisited

  1993cites this paper
• Transfer of MHC genes into hematopoietic stem cells by electroporation: A model for monitoring gene expression

  1990cites this paper
• Regulation of IL-1 and TNF-alpha expression during the differentiation of bone marrow derived macrophage.

  1989cites this paper
• Regulation of colony-stimulating factor 1-dependent macrophage precursor proliferation by type beta transforming growth factor.

  1988cites this paper
• Bone marrow derived macrophages have polyamine and ectoenzyme phenotypes distinct from resident macrophages.

  1988cites this paper
• Modification of macrophage differentiation: dimethylnitrosamine induced alteration in the responses towards the regulatory signals controlling myelopoiesis.

  1987cites this paper
• Regulation of MHC Class II and Interleukin-1 Gene Expression in Differentiating Macrophages

  1987cites this paper
• Alteration of macrophage anti-tumor activity and transferrin receptor expression by exposure to dimethylnitrosamine in vivo.

  1987cites this paper