Abstract
A fibroblast growth regulator, which inhibits the growth and division of proliferating fibroblasts, has been isolated from medium conditioned by exposure to density-inhibited mouse 3T3 cells. This partially purified preparation of the growth-inhibitory activity, termed FGR-s, contained two major polypeptides (Mrs, 10,000 and 13,000). Using FGR-s as the immunogen, we have carried out in vitro immunization of rat splenocytes and have generated hybridoma lines, each secreting an antibody directed against components of the FGR-s preparation. One such monoclonal antibody, designated 2A4, specifically bound the Mr 13,000 polypeptide of FGR-s. Antibody 2A4 also neutralized the growth-inhibitory effect of FGR-s in a concentration-dependent fashion. These results strongly suggest that the Mr 13,000 polypeptide carries growth-inhibitory activity.
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- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Böhmer F. D., Lehmann W., Schmidt H. E., Langen P., Grosse R. Purification of a growth inhibitor for Ehrlich ascites mammary carcinoma cells from bovine mammary gland. Exp Cell Res. 1984 Feb;150(2):466–476. doi: 10.1016/0014-4827(84)90591-3. [DOI] [PubMed] [Google Scholar]
- Datta P., Natraj C. V. Fibroblast growth regulatory factor inhibits DNA synthesis in BALB/c 3T3 cells by arresting in G1. Exp Cell Res. 1980 Feb;125(2):431–439. doi: 10.1016/0014-4827(80)90137-8. [DOI] [PubMed] [Google Scholar]
- Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
- Holley R. W., Böhlen P., Fava R., Baldwin J. H., Kleeman G., Armour R. Purification of kidney epithelial cell growth inhibitors. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5989–5992. doi: 10.1073/pnas.77.10.5989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ikeda Y., Steiner M. Isolation of platelet microtubule protein by an immunosorptive method. J Biol Chem. 1976 Oct 10;251(19):6135–6141. [PubMed] [Google Scholar]
- Kinders R. J., Johnson T. C. Isolation of cell-surface glycopeptides from bovine cerebral cortex that inhibit cell growth and protein synthesis in normal but not in transformed cells. Biochem J. 1982 Sep 15;206(3):527–534. doi: 10.1042/bj2060527. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kinders R. J., Milenkovic A. G., Nordin P., Johnson T. C. Characterization of cell-surface glycopeptides from mouse cerebral cortex that inhibit cell growth and protein synthesis. Biochem J. 1980 Sep 15;190(3):605–614. doi: 10.1042/bj1900605. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lieberman M. A., Keller-McGandy C. E., Woolsey T. A., Glaser L. Binding of isolated 3T3 surface membranes to growing 3T3 cells and their effect on cell growth. J Cell Biochem. 1982;20(1):81–93. doi: 10.1002/jcb.240200109. [DOI] [PubMed] [Google Scholar]
- Luben R. A., Mohler M. A. In vitro immunization as an adjunct to the production of hybridomas producing antibodies against the lymphokine osteoclast activating factor. Mol Immunol. 1980 May;17(5):635–639. doi: 10.1016/0161-5890(80)90161-3. [DOI] [PubMed] [Google Scholar]
- McMahon J. B., Farrelly J. G., Iype P. T. Purification and properties of a rat liver protein that specifically inhibits the proliferation of nonmalignant epithelial cells from rat liver. Proc Natl Acad Sci U S A. 1982 Jan;79(2):456–460. doi: 10.1073/pnas.79.2.456. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McMahon J. B., Iype P. T. Specific inhibition of proliferation of nonmalignant rat hepatic cells by a factor from rat liver. Cancer Res. 1980 Apr;40(4):1249–1254. [PubMed] [Google Scholar]
- Natraj C. V., Datta P. Control of DNA synthesis in growing BALB/c 3T3 mouse cells by a fibroblast growth regulatory factor. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6115–6119. doi: 10.1073/pnas.75.12.6115. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nilsen-Hamilton M., Holley R. W. Rapid selective effects by a growth inhibitor and epidermal growth factor on the incorporation of [35S]methionine into proteins secreted by African green monkey (BSC-1) cells. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5636–5640. doi: 10.1073/pnas.80.18.5636. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pardue R. L., Brady R. C., Perry G. W., Dedman J. R. Production of monoclonal antibodies against calmodulin by in vitro immunization of spleen cells. J Cell Biol. 1983 Apr;96(4):1149–1154. doi: 10.1083/jcb.96.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peterson S. W., Lerch V. Inhibition of DNA synthesis in SV3T3 cultures by isolated 3T3 plasma membranes. J Cell Biol. 1983 Jul;97(1):276–279. doi: 10.1083/jcb.97.1.276. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peterson S. W., Lerch V., Moynahan M. E., Carson M. P., Vale R. Partial characterization of a growth-inhibiting protein in 3T3 cell plasma membranes. Exp Cell Res. 1982 Dec;142(2):447–451. doi: 10.1016/0014-4827(82)90386-x. [DOI] [PubMed] [Google Scholar]
- Steck P. A., Blenis J., Voss P. G., Wang J. L. Growth control in cultured 3T3 fibroblasts II. Molecular properties of a fraction enriched in growth inhibitory activity. J Cell Biol. 1982 Feb;92(2):523–530. doi: 10.1083/jcb.92.2.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steck P. A., Voss P. G., Wang J. L. Growth control in cultured 3T3 fibroblasts. Assays of cell proliferation and demonstration of a growth inhibitory activity. J Cell Biol. 1979 Dec;83(3):562–575. doi: 10.1083/jcb.83.3.562. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stoker M. G., Rubin H. Density dependent inhibition of cell growth in culture. Nature. 1967 Jul 8;215(5097):171–172. doi: 10.1038/215171a0. [DOI] [PubMed] [Google Scholar]
- TODARO G. J., GREEN H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol. 1963 May;17:299–313. doi: 10.1083/jcb.17.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Voss P. G., Steck P. A., Calamia J. C., Wang J. L. Growth control in cultured 3T3 fibroblasts. III. Binding interactions of a growth inhibitory activity with target cells. Exp Cell Res. 1982 Apr;138(2):397–407. doi: 10.1016/0014-4827(82)90188-4. [DOI] [PubMed] [Google Scholar]
- Wells V., Mallucci L. Properties of a cell growth inhibitor produced by mouse embryo fibroblasts. J Cell Physiol. 1983 Nov;117(2):148–154. doi: 10.1002/jcp.1041170203. [DOI] [PubMed] [Google Scholar]
- Whittenberger B., Glaser L. Inhibition of DNA synthesis in cultures of 3T3 cells by isolated surface membranes. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2251–2255. doi: 10.1073/pnas.74.6.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]