Two human tumor necrosis factor receptor genes were cloned into eukaryotic expression vectors pcDNA3, pDR2 and pXJ-41, respectively. Those vectors were transfected into BHK-21 cells and the expression of hTNFRs was identified and demonstrated by binding of (125)I hTNF-alpha and Scatchard analysis. The expression of two hTNFRs in BHK cells at the RNA transcriptional and the protein translation levels were also demonstrated by RT-PCR and indirect ELISA. However, hTNF-alpha did not appear cytotoxic to these cells. The competitive binding activity of mutant hTNF-alpha-R2K and wild type hTNF-alpha with these two receptors, respectively, were assayed. These results laid basis for further studies on structure-function relationships of these receptors as well as the hTNF-alpha.
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