title: Mouse CD40-transfected cell lines cannot exhibit the binding and RANTES-stimulating activity of exogenous heat shock protein 70.
abstract: Here we demonstrate the inducible mouse Hsp72 binds markedly to lymphoid neoplastic macrophage-like P388D1 cells. To examine whether mouse CD40 can play a role in signaling exogenously administered HSP70 in a fashion similar to that of human CD40, we established mouse CD40-transfectants of both human 293 cells and murine-pro-B cell line Ba/F3. A small portion of mouse CD40 expressed on 293-derived transfectants was the mature form with a signal-transducible C-terminal domain, whereas a majority of expressed antigen showed the molecular size smaller than we expect. Flow cytometry showed that mouse Hsp72, but neither its deletion variants nor the related Escherichia coli DnaK, bound to the 293-derived transfectants regardless of CD40 expression. CD40 molecules expressed on the transfectants showed the binding of soluble form of CD40L but this binding was not inhibited by excess amount of HSP70. CD40L, but not any HSP70 recombinant proteins, stimulated the production of chemokine RANTES in the transfectants. Furthermore, no RANTES production was induced by HSP70-RCMLA complex in the transfectants, although it binds to 293-derived cells in a CD40-independent manner. No interaction between mouse CD40 and HSP70 recombinant proteins was detected by using the Ba/F3-derived transfectants that express the mature form of mouse CD40. The present results imply that mouse CD40 expressed on the transfectants differs from its human homolog in the binding of exogenously administered HSP70.
authors.affiliation: Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Goka-sho, Uji, Kyoto 611-0011, Japan.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Tao
authors.forename: Masayo
authors.initials: M
authors.lastname: Nomura
authors.forename: Naofumi
authors.initials: N
authors.lastname: Kitabatake
authors.forename: Fumito
authors.initials: F
authors.lastname: Tani
mesh_terms: D000818:Animals; D019013:CD40 Antigens / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D002460:Cell Line; D045744:Cell Line, Tumor; D018946:Chemokine CCL5 / Q000235:genetics / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism / Q000502:physiology*; D050884:HSP72 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007941:Leukemia P388 / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D014162:Transfection
chemical_list: D019013:CD40 Antigens; D018946:Chemokine CCL5; D018840:HSP70 Heat-Shock Proteins; D050884:HSP72 Heat-Shock Proteins