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C.4 Characterization of the carp, Cyprinus carpio L., interleukin-1-like factor
The Scientific and Social Program o f the Vf n ISDCI Congress
C.3 INTERACTIONS OF A TUNICATE MAMMALIAN CELLS
IMMUNOMODULATORY
PROTEIN
967
WITH
David A. Raftos, Robert L. Raison Immunobiology Unit, University of Technology Sydney, P.O. Box 123, Broadway, Sydney, NSW 2007, Australia The ability of a tunicate immunomodulatory protein , tunlLl, to interact with mammalian cells was investigated. In tunicates, tunlL1 regulates inflammatory defense reactions. It can also stimulate the proliferation of mammalian thymocytes and enhance the vascular permeability of rabbit skin. In the current study, tunlL1 has been shown to enhance L929 fibroblast proliferation, induce interleukin-2 (IL2) secretion from human mononuclear cells and enhance IL2 receptor expression by EL-4 murine lymphoma cells. These data show that tunlL1 has a number of biological activities that are analogous to those of the mammalian inflammatory cytokine, interleukin-I (ILl). However, tunlL1 does not appear to stimulate these ILl-like effects by interacting with ILl cell surface receptors. TunlLl cannot block the binding of anti-ILl receptor antibodies to EL-4 cells, nor can anti-ILl receptor antibodies inhibit the capacity of tunlL1 to stimulate fibroblast proliferation. This indicates that tunlL 1 does not induce its IL l-like effects via structural homology to mammalian ILl.
C.4 CHARACTERIZATION LIKE FACTOR
OF THE CARP, CYPRINUS CARPIO L., INTERLEUKIN-1-
F.A.A. Weyts 1, B.M.L. Verburg-van Kemenade t, G. Flik2 ~Dept. of Experimental Animal Morphology & Cell Biology, Agricultural University, P.O. Box 338, 6700 AH Wageningen, The Netherlands, and 2Dept. of Animal Physiology, Faculty of Science, University of Nijmegen, P.O. Box 9010, 6525 ED Nijmegen, The Netherlands Cytokine regulation of the immune response of teleost fish has been a point of research in our lab for several years. Macrophage- and neutrophilic granulocyte-enriched fractions of carp headkidney were isolated and showed respiratory burst activity in vitro. Culture supernatants of these cell fractions are co-stimulatory with PHA in a lymphocyte proliferation assay. Moreover, these supernatants were found to enhance proliferation of the II-1 dependent, murine DI0(N4)M cell line. Bioactivity of the phagocyte culture supernatants to stimulate proliferation in both fish lymphocytes and murine D10(N4)M cells could be blocked using polyclonal sera against recombinant human interleukin-l~ and /3. Western blots of concentrated macrophage- and granulocyte-culture supernatants incubated with the same anti 11-1 polyclonal sera, revealed immunoreactive molecular species of 20-23 kDa and of 14-15 kDa. Currently, research is focussed on the primary structure of the carp II-l-like factor via molecular approaches. RNA of macrophage enriched fractions is isolated and PCR reactions are performed with primers based on conserved mammalian I1-1 sequences. A macrophage/granulocyte eDNA library has been constructed and will be screened for interleukin sequences.
C.3 INTERACTIONS OF A TUNICATE MAMMALIAN CELLS
IMMUNOMODULATORY
PROTEIN
967
WITH
David A. Raftos, Robert L. Raison Immunobiology Unit, University of Technology Sydney, P.O. Box 123, Broadway, Sydney, NSW 2007, Australia The ability of a tunicate immunomodulatory protein , tunlLl, to interact with mammalian cells was investigated. In tunicates, tunlL1 regulates inflammatory defense reactions. It can also stimulate the proliferation of mammalian thymocytes and enhance the vascular permeability of rabbit skin. In the current study, tunlL1 has been shown to enhance L929 fibroblast proliferation, induce interleukin-2 (IL2) secretion from human mononuclear cells and enhance IL2 receptor expression by EL-4 murine lymphoma cells. These data show that tunlL1 has a number of biological activities that are analogous to those of the mammalian inflammatory cytokine, interleukin-I (ILl). However, tunlL1 does not appear to stimulate these ILl-like effects by interacting with ILl cell surface receptors. TunlLl cannot block the binding of anti-ILl receptor antibodies to EL-4 cells, nor can anti-ILl receptor antibodies inhibit the capacity of tunlL1 to stimulate fibroblast proliferation. This indicates that tunlL 1 does not induce its IL l-like effects via structural homology to mammalian ILl.
C.4 CHARACTERIZATION LIKE FACTOR
OF THE CARP, CYPRINUS CARPIO L., INTERLEUKIN-1-
F.A.A. Weyts 1, B.M.L. Verburg-van Kemenade t, G. Flik2 ~Dept. of Experimental Animal Morphology & Cell Biology, Agricultural University, P.O. Box 338, 6700 AH Wageningen, The Netherlands, and 2Dept. of Animal Physiology, Faculty of Science, University of Nijmegen, P.O. Box 9010, 6525 ED Nijmegen, The Netherlands Cytokine regulation of the immune response of teleost fish has been a point of research in our lab for several years. Macrophage- and neutrophilic granulocyte-enriched fractions of carp headkidney were isolated and showed respiratory burst activity in vitro. Culture supernatants of these cell fractions are co-stimulatory with PHA in a lymphocyte proliferation assay. Moreover, these supernatants were found to enhance proliferation of the II-1 dependent, murine DI0(N4)M cell line. Bioactivity of the phagocyte culture supernatants to stimulate proliferation in both fish lymphocytes and murine D10(N4)M cells could be blocked using polyclonal sera against recombinant human interleukin-l~ and /3. Western blots of concentrated macrophage- and granulocyte-culture supernatants incubated with the same anti 11-1 polyclonal sera, revealed immunoreactive molecular species of 20-23 kDa and of 14-15 kDa. Currently, research is focussed on the primary structure of the carp II-l-like factor via molecular approaches. RNA of macrophage enriched fractions is isolated and PCR reactions are performed with primers based on conserved mammalian I1-1 sequences. A macrophage/granulocyte eDNA library has been constructed and will be screened for interleukin sequences.