Expression of mRNA of co-stimulatory molecules B7-1 and B7- 2 in cultured rat pulmonary dendritic cells is enhanced by exposure to specific cytokines

R. E. Goodman, G. H. Chen, P. J. Christensen, L. Pastoriza, J. J. Fak, R. A. McDonald, G. B. Toews

Research output: Contribution to journalArticlepeer-review

Abstract

Dendritic cells (DC) are potent primary antigen presenting cells (APC). Previously we demonstrated that GM-CSF enhances proliferation in an allogeneic MLR of rat pulmonary DC and resting T cells. Since B7 molecules expressed on APC contribute to their capacity to stimulate T cells, we are studying regulation of their expression in pulmonary DC. We cloned and sequenced a partial cDNA of the previously unreported rat B7-2 gene and used semi-quantitative RT-PCR (reverse transcriptase-polymerase chain reaction) to measure changes in mRNA expression of B7-1 and B7-2. Primers for PCR amplification of the rat gene were designed from mouse and human sequences. These were used to amplify cDN A products synthesized from RN A of cultured rat lung DC. The longest clone (449bp) sequenced (GenBank ace. RNU31330) corresponds to 50% of the coding region of mouse B7-2. It shares 89% identity with the mouse nucleic acid sequence and 83% with the coded amino acid sequence. Pulmonary DC (>90% pure; low density, non-phagocytic, MHC class II bright) were cultured in the presence or absence of recombinant mGM-CSF, rGM-CSF, mM- CSF, mTNFa, mIL-1/3 or mIFN7. RNA was isolated and amplified by RT-PCR. Analytical PCR primers and probes were designed from rat B7-1, B7-2, /Jactin and GAPDH sequences. Products were analyzed from hybridized Southern blots. Expression of both B7-1 and B7-2 was enhanced by GM- CSF, TNFα or IL-1β, but not by M-CSF or IFNγ.

Original languageEnglish (US)
Pages (from-to)A1484
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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