Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1981 Sep;68(3):792–802. doi: 10.1172/JCI110316

Effect of soluble products from lectin-stimulated lymphocytes on the growth, adhesiveness, and glycosaminoglycan synthesis of cultured synovial fibroblastic cells.

T P Anastassiades, A Wood
PMCID: PMC370862  PMID: 7276172

Abstract

Human blood mononuclear cells exposed to concanavalin A or phytohemagglutinin secrete a soluble factor that arrests the growth of human synovial fibroblastic cells in culture. Once the growth-inhibitory effect is initiated it cannot be reversed by washing the fibroblastic cells, by refeeding with nonconditioned fresh serum-containing medium, by trypsinization, EDTA treatment, or a combination of these procedures. Media from nonstimulated mononuclear cells, fibroblastic cells, or the lectins themselves do not contain similar inhibitory activity that can be detected by the present culture systems. This lectin-dependent, growth-inhibitory activity does not have a cytotoxic effect on the fibroblasts but increases their adhesiveness to plastic or glass surfaces, and the cells tend to assume a less fibroblastic morphology. The growth-inhibitory activity is stable in the cold and is nondialyzable or ultrafilterable, but the activity is rapidly lost at temperature between 60 degrees and 70 degrees C and at pH 2.0. The growth-arrested cells secrete more glycosaminoglycan per cell in the medium and synthesize more cell surface glycosaminoglycan than the controls. However, the increased glycosaminoglycan synthesis cannot be explained as being entirely secondary to a cell density effect as it is also observed when adjustments are made for the differences in growth rates.

Full text

PDF
792

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anastassiades T. P., Ley J., Wood A. Glycosaminoglycan synthesis and glucose uptake by rheumatoid and nonrheumatoid fibroblastic cells in culture. The effect of nutritional factors. Arthritis Rheum. 1979 Aug;22(8):871–876. doi: 10.1002/art.1780220810. [DOI] [PubMed] [Google Scholar]
  2. Anastassiades T. P., Ley J., Wood A., Irwin D. The growth kinetics of synovial fibroblastic cells from inflammatory and noninflammatory arthropathies. Arthritis Rheum. 1978 May;21(4):461–466. doi: 10.1002/art.1780210410. [DOI] [PubMed] [Google Scholar]
  3. Badger A. M., Cooperband S. R., Green J. A. Direct observations on the effect of "proliferation inhibitory factor" on the clonal growth of target cells. J Immunol. 1971 Nov;107(5):1259–1267. [PubMed] [Google Scholar]
  4. Badger A. M., Cooperband S. R., Merluzzi V. J. The production of proliferation inhibitory factor (pif) by mouse spleen cells in vitro. J Immunol. 1974 Apr;112(4):1435–1443. [PubMed] [Google Scholar]
  5. Castor C. W., Ritchie J. C., Scott M. E., Whitney S. L. Connective tissue activation. XI. Stimulation of glycosaminoglycan and DNA formation by a platelet factor. Arthritis Rheum. 1977 Apr;20(3):859–868. doi: 10.1002/art.1780200316. [DOI] [PubMed] [Google Scholar]
  6. Castor C. W. Synovial cell activation induced by a polypeptide mediator. Ann N Y Acad Sci. 1975 Jun 13;256:304–317. doi: 10.1111/j.1749-6632.1975.tb36057.x. [DOI] [PubMed] [Google Scholar]
  7. Castor C. W., Yaron M. Leukocyte-connective tissue cell interaction. II. The specificity, duration, and mechanism of interaction effects. Arthritis Rheum. 1969 Aug;12(4):374–386. doi: 10.1002/art.1780120405. [DOI] [PubMed] [Google Scholar]
  8. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Froehlich J. E., Anastassiades T. P. Possible limitation of growth in human fibroblast cultures by diffusion. J Cell Physiol. 1975 Dec;86 (Suppl 2)(3 Pt 2):567–580. doi: 10.1002/jcp.1040860502. [DOI] [PubMed] [Google Scholar]
  10. Froehlich J. E., Anastassiades T. P. Role of pH in fibroblast proliferation. J Cell Physiol. 1974 Oct;84(2):253–260. doi: 10.1002/jcp.1040840211. [DOI] [PubMed] [Google Scholar]
  11. Gospodarowicz D. Purification of a fibroblast growth factor from bovine pituitary. J Biol Chem. 1975 Apr 10;250(7):2515–2520. [PubMed] [Google Scholar]
  12. Green J. A., Cooperband S. R., Rutstein J. A., Kibrick S. Inhibition of target cell proliferation by supernatants from cultures of human peripheral lymphocytes. J Immunol. 1970 Jul;105(1):48–54. [PubMed] [Google Scholar]
  13. Hiserodt J. C., Fair D. S., Granger G. A. Identification of multiple cytolytic components associated with the beta-LT class of lymphotoxins released by mitogen-activity human lymphocytes in vitro. J Immunol. 1976 Nov;117(5 Pt 1):1503–1506. [PubMed] [Google Scholar]
  14. Hronowski L., Anastassiades T. P. Characterization of glycosaminoglycan-Alcian blue complexes by elution from cellulose acetate utilizing different MgCl2 concentrations. Anal Biochem. 1980 Sep 15;107(2):393–405. doi: 10.1016/0003-2697(80)90401-7. [DOI] [PubMed] [Google Scholar]
  15. Hronowski L., Anastassiades T. P. Quantitation and interaction of glycosaminoglycans with Alcian blue in dimethyl sulfoxide solutions. Anal Biochem. 1979 Feb;93(1):60–72. [PubMed] [Google Scholar]
  16. Hronowski L., Anastassiades T. P. Rates of glycosaminoglycan synthesis and rates of incorporation of radioactive precursors into newly synthesized glycosaminoglycan by confluent rat muscle fibroblasts. J Biol Chem. 1980 Oct 10;255(19):9210–9217. [PubMed] [Google Scholar]
  17. Hronowski L., Anastassiades T. P. The effect of cell density on net rates of glycosaminoglycan synthesis and secretion by cultured rat fibroblasts. J Biol Chem. 1980 Nov 10;255(21):10091–10099. [PubMed] [Google Scholar]
  18. Jeffes E. W., Granger G. A. Relationship of cloning inhibition factor, "lymphotoxin" factor, and proliferation inhibition factor release in vitro by mitogen-activated human lymphoctes. J Immunol. 1975 Jan;114(1 Pt 1):64–69. [PubMed] [Google Scholar]
  19. Kolb W. P., Granger G. A. Lymphocyte in vitro cytotoxicity: characterization of human lymphotoxin. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1250–1255. doi: 10.1073/pnas.61.4.1250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morell B., Froesch E. R. Fibroblasts as an experimental tool in metabolic and hormone studies. II. Effects of insulin and nonsuppressible insulin-like activity (NSILA-S) on fibroblasts in culture. Eur J Clin Invest. 1973 Mar;3(2):119–123. doi: 10.1111/j.1365-2362.1973.tb00338.x. [DOI] [PubMed] [Google Scholar]
  21. Namba Y., Waksman B. H. Regulatory substances produced by lymphocytes. III. Evidence that lymphotoxin and proliferation inhibitory factor are identical and different from the inhibitor of DNA synthesis. J Immunol. 1976 Apr;116(4):1140–1144. [PubMed] [Google Scholar]
  22. Peter J. B., Dawkins R. L. Target cell lysis mediated by soluble cytotoxin released from stimulated lymphocytes. Nat New Biol. 1971 Jul 21;232(29):79–80. doi: 10.1038/newbio232079a0. [DOI] [PubMed] [Google Scholar]
  23. Tomida M., Koyama H., Ono T. A serum factor capable of stimulating hyaluronic acid synthesis in cultured rat fibroblasts. J Cell Physiol. 1977 May;91(2):323–328. doi: 10.1002/jcp.1040910217. [DOI] [PubMed] [Google Scholar]
  24. Walker S. M., Lee S. C., Lucas Z. J. Cytotoxic activity of lymphocytes. VI. Heterogeneity of cytotoxins in supernatants of mitogen-activated lymphocytes. J Immunol. 1976 Mar;116(3):807–815. [PubMed] [Google Scholar]
  25. Williams T. W., Granger G. A. Lymphocyte in vitro cytotoxicity: correlation of derepression with release of lymphotoxin from human lymphocytes. J Immunol. 1969 Aug;103(2):170–178. [PubMed] [Google Scholar]
  26. Yano K., Lucas Z. J. Cytotoxic activity of lymphocytes. VII. cellular origin of alpha-lymphotoxin. J Immunol. 1978 Feb;120(2):385–394. [PubMed] [Google Scholar]
  27. Ziff M. Pathophysiology of rheumatoid arthritis. Fed Proc. 1973 Feb;32(2):131–133. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES