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. 1990 Nov 1;172(5):1391–1401. doi: 10.1084/jem.172.5.1391

A transforming growth factor beta 2 (TGF-beta 2)-like immunosuppressive factor in amniotic fluid and localization of TGF-beta 2 mRNA in the pregnant uterus

PMCID: PMC2188670  PMID: 1700055

Abstract

This report describes a murine amniotic fluid (MAF) immunosuppressive factor that has properties similar to transforming growth factor beta (TGF-beta). The MAF factor exhibits TGF-beta-like activity in stimulating soft agar colony formation by AKR-2B cells and inhibiting thymidine uptake by Mv1Lu cells. We demonstrate that both the immunosuppressive and TGF-beta-like activities of the MAF factor are completely neutralized by anti-TGF-beta 2-specific antibodies and not by anti-TGF-beta 1-specific antisera. The immunosuppressive factor in MAF is novel in that it appears to be identical or very closely related to TGF-beta 2 and is active in its native state. This active and anti- TGF-beta 2-neutralizable factor chromatographs at approximately 70 kD on Sephadex at neutral pH and appears to be able to complex with alpha- fetoprotein in native amniotic fluid. Chromatography of native MAF under acidic conditions demonstrates a lower molecular mass protein that chromatographs on BioGel in the same position as the mature 25-kD TGF-beta. This protein has the biological properties of TGF-beta and is immunosuppressive. Both of these activities are neutralizable with anti- TGF-beta 2 but not with anti-TGF-beta 1 or other antisera. By Northern analysis, we find high levels of TGF-beta 2 mRNA (with little or no TGF- beta 1) in the pregnant uterus that peak around day 15 of gestation and then fall rapidly by day 19 as birth approaches. The TGF-beta 2-like factor could possibly play a role in maternal immunity, in the retention of the fetal allograft, as well as in regulating fetal and neonatal immunological competence.

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Selected References

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  1. Arceci R. J., Shanahan F., Stanley E. R., Pollard J. W. Temporal expression and location of colony-stimulating factor 1 (CSF-1) and its receptor in the female reproductive tract are consistent with CSF-1-regulated placental development. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8818–8822. doi: 10.1073/pnas.86.22.8818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Assoian R. K., Komoriya A., Meyers C. A., Miller D. M., Sporn M. B. Transforming growth factor-beta in human platelets. Identification of a major storage site, purification, and characterization. J Biol Chem. 1983 Jun 10;258(11):7155–7160. [PubMed] [Google Scholar]
  3. Assoian R. K. Purification of type-beta transforming growth factor from human platelets. Methods Enzymol. 1987;146:153–163. doi: 10.1016/s0076-6879(87)46017-5. [DOI] [PubMed] [Google Scholar]
  4. Bodmer S., Strommer K., Frei K., Siepl C., de Tribolet N., Heid I., Fontana A. Immunosuppression and transforming growth factor-beta in glioblastoma. Preferential production of transforming growth factor-beta 2. J Immunol. 1989 Nov 15;143(10):3222–3229. [PubMed] [Google Scholar]
  5. Carel J. C., Schreiber R. D., Falqui L., Lacy P. E. Transforming growth factor beta decreases the immunogenicity of rat islet xenografts (rat to mouse) and prevents rejection in association with treatment of the recipient with a monoclonal antibody to interferon gamma. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1591–1595. doi: 10.1073/pnas.87.4.1591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cheifetz S., Weatherbee J. A., Tsang M. L., Anderson J. K., Mole J. E., Lucas R., Massagué J. The transforming growth factor-beta system, a complex pattern of cross-reactive ligands and receptors. Cell. 1987 Feb 13;48(3):409–415. doi: 10.1016/0092-8674(87)90192-9. [DOI] [PubMed] [Google Scholar]
  7. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  8. Clark D. A., Falbo M., Rowley R. B., Banwatt D., Stedronska-Clark J. Active suppression of host-vs graft reaction in pregnant mice. IX. Soluble suppressor activity obtained from allopregnant mouse decidua that blocks the cytolytic effector response to IL-2 is related to transforming growth factor-beta. J Immunol. 1988 Dec 1;141(11):3833–3840. [PubMed] [Google Scholar]
  9. Clark D. A., Flanders K. C., Banwatt D., Millar-Book W., Manuel J., Stedronska-Clark J., Rowley B. Murine pregnancy decidua produces a unique immunosuppressive molecule related to transforming growth factor beta-2. J Immunol. 1990 Apr 15;144(8):3008–3014. [PubMed] [Google Scholar]
  10. Crainie M., Semeluk A., Lee K. C., Wegmann T. Regulation of constitutive and lymphokine-induced Ia expression by murine alpha-fetoprotein. Cell Immunol. 1989 Jan;118(1):41–52. doi: 10.1016/0008-8749(89)90356-0. [DOI] [PubMed] [Google Scholar]
  11. Cross D., Cambier J. C. Transforming growth factor beta 1 has differential effects on B cell proliferation and activation antigen expression. J Immunol. 1990 Jan 15;144(2):432–439. [PubMed] [Google Scholar]
  12. Czarniecki C. W., Chiu H. H., Wong G. H., McCabe S. M., Palladino M. A. Transforming growth factor-beta 1 modulates the expression of class II histocompatibility antigens on human cells. J Immunol. 1988 Jun 15;140(12):4217–4223. [PubMed] [Google Scholar]
  13. Danielpour D., Dart L. L., Flanders K. C., Roberts A. B., Sporn M. B. Immunodetection and quantitation of the two forms of transforming growth factor-beta (TGF-beta 1 and TGF-beta 2) secreted by cells in culture. J Cell Physiol. 1989 Jan;138(1):79–86. doi: 10.1002/jcp.1041380112. [DOI] [PubMed] [Google Scholar]
  14. Derynck R., Lindquist P. B., Lee A., Wen D., Tamm J., Graycar J. L., Rhee L., Mason A. J., Miller D. A., Coffey R. J. A new type of transforming growth factor-beta, TGF-beta 3. EMBO J. 1988 Dec 1;7(12):3737–3743. doi: 10.1002/j.1460-2075.1988.tb03257.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dziadek M. A., Andrews G. K. Tissue specificity of alpha-fetoprotein messenger RNA expression during mouse embryogenesis. EMBO J. 1983;2(4):549–554. doi: 10.1002/j.1460-2075.1983.tb01461.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Espevik T., Figari I. S., Ranges G. E., Palladino M. A., Jr Transforming growth factor-beta 1 (TGF-beta 1) and recombinant human tumor necrosis factor-alpha reciprocally regulate the generation of lymphokine-activated killer cell activity. Comparison between natural porcine platelet-derived TGF-beta 1 and TGF-beta 2, and recombinant human TGF-beta 1. J Immunol. 1988 Apr 1;140(7):2312–2316. [PubMed] [Google Scholar]
  17. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  18. Fontana A., Frei K., Bodmer S., Hofer E., Schreier M. H., Palladino M. A., Jr, Zinkernagel R. M. Transforming growth factor-beta inhibits the generation of cytotoxic T cells in virus-infected mice. J Immunol. 1989 Nov 15;143(10):3230–3234. [PubMed] [Google Scholar]
  19. Graycar J. L., Miller D. A., Arrick B. A., Lyons R. M., Moses H. L., Derynck R. Human transforming growth factor-beta 3: recombinant expression, purification, and biological activities in comparison with transforming growth factors-beta 1 and -beta 2. Mol Endocrinol. 1989 Dec;3(12):1977–1986. doi: 10.1210/mend-3-12-1977. [DOI] [PubMed] [Google Scholar]
  20. Heine U., Munoz E. F., Flanders K. C., Ellingsworth L. R., Lam H. Y., Thompson N. L., Roberts A. B., Sporn M. B. Role of transforming growth factor-beta in the development of the mouse embryo. J Cell Biol. 1987 Dec;105(6 Pt 2):2861–2876. doi: 10.1083/jcb.105.6.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hunziker R. D., Wegmann T. G. Placental immunoregulation. Crit Rev Immunol. 1986;6(3):245–285. [PubMed] [Google Scholar]
  22. Ignotz R. A., Heino J., Massagué J. Regulation of cell adhesion receptors by transforming growth factor-beta. Regulation of vitronectin receptor and LFA-1. J Biol Chem. 1989 Jan 5;264(1):389–392. [PubMed] [Google Scholar]
  23. Ikeda T., Lioubin M. N., Marquardt H. Human transforming growth factor type beta 2: production by a prostatic adenocarcinoma cell line, purification, and initial characterization. Biochemistry. 1987 May 5;26(9):2406–2410. doi: 10.1021/bi00383a002. [DOI] [PubMed] [Google Scholar]
  24. Jakowlew S. B., Dillard P. J., Kondaiah P., Sporn M. B., Roberts A. B. Complementary deoxyribonucleic acid cloning of a novel transforming growth factor-beta messenger ribonucleic acid from chick embryo chondrocytes. Mol Endocrinol. 1988 Aug;2(8):747–755. doi: 10.1210/mend-2-8-747. [DOI] [PubMed] [Google Scholar]
  25. Jennings J. C., Mohan S., Linkhart T. A., Widstrom R., Baylink D. J. Comparison of the biological actions of TGF beta-1 and TGF beta-2: differential activity in endothelial cells. J Cell Physiol. 1988 Oct;137(1):167–172. doi: 10.1002/jcp.1041370120. [DOI] [PubMed] [Google Scholar]
  26. Kasid A., Bell G. I., Director E. P. Effects of transforming growth factor-beta on human lymphokine-activated killer cell precursors. Autocrine inhibition of cellular proliferation and differentiation to immune killer cells. J Immunol. 1988 Jul 15;141(2):690–698. [PubMed] [Google Scholar]
  27. Kehrl J. H., Roberts A. B., Wakefield L. M., Jakowlew S., Sporn M. B., Fauci A. S. Transforming growth factor beta is an important immunomodulatory protein for human B lymphocytes. J Immunol. 1986 Dec 15;137(12):3855–3860. [PubMed] [Google Scholar]
  28. Kehrl J. H., Taylor A. S., Delsing G. A., Roberts A. B., Sporn M. B., Fauci A. S. Further studies of the role of transforming growth factor-beta in human B cell function. J Immunol. 1989 Sep 15;143(6):1868–1874. [PubMed] [Google Scholar]
  29. Kehrl J. H., Wakefield L. M., Roberts A. B., Jakowlew S., Alvarez-Mon M., Derynck R., Sporn M. B., Fauci A. S. Production of transforming growth factor beta by human T lymphocytes and its potential role in the regulation of T cell growth. J Exp Med. 1986 May 1;163(5):1037–1050. doi: 10.1084/jem.163.5.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kishi K., Ellingsworth L. R., Ogawa M. The suppressive effects of type beta transforming growth factor (TGF beta) on primitive murine hemopoietic progenitors are abrogated by interleukin-6 and granulocyte colony-stimulating factor. Leukemia. 1989 Oct;3(10):687–691. [PubMed] [Google Scholar]
  31. Labib R. S., Tomasi T. B., Jr Immunosuppressive factors in mouse amniotic fluid and neonate serum. Immunol Commun. 1978;7(2):223–237. doi: 10.3109/08820137809033885. [DOI] [PubMed] [Google Scholar]
  32. Lebman D. A., Lee F. D., Coffman R. L. Mechanism for transforming growth factor beta and IL-2 enhancement of IgA expression in lipopolysaccharide-stimulated B cell cultures. J Immunol. 1990 Feb 1;144(3):952–959. [PubMed] [Google Scholar]
  33. Meyuhas O., Perry R. P. Construction and identification of cDNA clones for mouse ribosomal proteins: application for the study of r-protein gene expression. Gene. 1980 Jul;10(2):113–129. doi: 10.1016/0378-1119(80)90129-8. [DOI] [PubMed] [Google Scholar]
  34. Miller D. A., Lee A., Pelton R. W., Chen E. Y., Moses H. L., Derynck R. Murine transforming growth factor-beta 2 cDNA sequence and expression in adult tissues and embryos. Mol Endocrinol. 1989 Jul;3(7):1108–1114. doi: 10.1210/mend-3-7-1108. [DOI] [PubMed] [Google Scholar]
  35. Ogra S. S., Murgita R. A., Tomasi T. B., Jr Immunosuppressive activity of mouse amniotic fluid. Immunol Commun. 1974;3(5):497–508. doi: 10.3109/08820137409061129. [DOI] [PubMed] [Google Scholar]
  36. Ohta M., Greenberger J. S., Anklesaria P., Bassols A., Massagué J. Two forms of transforming growth factor-beta distinguished by multipotential haematopoietic progenitor cells. Nature. 1987 Oct 8;329(6139):539–541. doi: 10.1038/329539a0. [DOI] [PubMed] [Google Scholar]
  37. Pelton R. W., Nomura S., Moses H. L., Hogan B. L. Expression of transforming growth factor beta 2 RNA during murine embryogenesis. Development. 1989 Aug;106(4):759–767. doi: 10.1242/dev.106.4.759. [DOI] [PubMed] [Google Scholar]
  38. Proper J. A., Bjornson C. L., Moses H. L. Mouse embryos contain polypeptide growth factor(s) capable of inducing a reversible neoplastic phenotype in nontransformed cells in culture. J Cell Physiol. 1982 Feb;110(2):169–174. doi: 10.1002/jcp.1041100210. [DOI] [PubMed] [Google Scholar]
  39. Ranges G. E., Figari I. S., Espevik T., Palladino M. A., Jr Inhibition of cytotoxic T cell development by transforming growth factor beta and reversal by recombinant tumor necrosis factor alpha. J Exp Med. 1987 Oct 1;166(4):991–998. doi: 10.1084/jem.166.4.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Reilly B. D., Goldberg E. H., Tomasi T. B. Prolongation of male skin graft survival in female mice treated with amniotic fluid. Transplantation. 1988 May;45(5):996–998. doi: 10.1097/00007890-198805000-00036. [DOI] [PubMed] [Google Scholar]
  41. Roberts A. B., Flanders K. C., Kondaiah P., Thompson N. L., Van Obberghen-Schilling E., Wakefield L., Rossi P., de Crombrugghe B., Heine U., Sporn M. B. Transforming growth factor beta: biochemistry and roles in embryogenesis, tissue repair and remodeling, and carcinogenesis. Recent Prog Horm Res. 1988;44:157–197. doi: 10.1016/b978-0-12-571144-9.50010-7. [DOI] [PubMed] [Google Scholar]
  42. Rook A. H., Kehrl J. H., Wakefield L. M., Roberts A. B., Sporn M. B., Burlington D. B., Lane H. C., Fauci A. S. Effects of transforming growth factor beta on the functions of natural killer cells: depressed cytolytic activity and blunting of interferon responsiveness. J Immunol. 1986 May 15;136(10):3916–3920. [PubMed] [Google Scholar]
  43. Rosa F., Roberts A. B., Danielpour D., Dart L. L., Sporn M. B., Dawid I. B. Mesoderm induction in amphibians: the role of TGF-beta 2-like factors. Science. 1988 Feb 12;239(4841 Pt 1):783–785. doi: 10.1126/science.3422517. [DOI] [PubMed] [Google Scholar]
  44. Ruegemer J. J., Ho S. N., Augustine J. A., Schlager J. W., Bell M. P., McKean D. J., Abraham R. T. Regulatory effects of transforming growth factor-beta on IL-2- and IL-4-dependent T cell-cycle progression. J Immunol. 1990 Mar 1;144(5):1767–1776. [PubMed] [Google Scholar]
  45. Sharples K., Plowman G. D., Rose T. M., Twardzik D. R., Purchio A. F. Cloning and sequence analysis of simian transforming growth factor-beta cDNA. DNA. 1987 Jun;6(3):239–244. doi: 10.1089/dna.1987.6.239. [DOI] [PubMed] [Google Scholar]
  46. Siepl C., Bodmer S., Frei K., MacDonald H. R., De Martin R., Hofer E., Fontana A. The glioblastoma-derived T cell suppressor factor/transforming growth factor-beta 2 inhibits T cell growth without affecting the interaction of interleukin 2 with its receptor. Eur J Immunol. 1988 Apr;18(4):593–600. doi: 10.1002/eji.1830180416. [DOI] [PubMed] [Google Scholar]
  47. Stoeck M., Miescher S., MacDonald H. R., Von Fliedner V. Transforming growth factors beta slow down cell-cycle progression in a murine interleukin-2 dependent T-cell line. J Cell Physiol. 1989 Oct;141(1):65–73. doi: 10.1002/jcp.1041410111. [DOI] [PubMed] [Google Scholar]
  48. Strober S. Natural suppressor (NS) cells, neonatal tolerance, and total lymphoid irradiation: exploring obscure relationships. Annu Rev Immunol. 1984;2:219–237. doi: 10.1146/annurev.iy.02.040184.001251. [DOI] [PubMed] [Google Scholar]
  49. Suzuki K., Tomasi T. B., Jr Mechanism of immune suppression by murine neonatal fluids. J Immunol. 1980 Oct;125(4):1806–1810. [PubMed] [Google Scholar]
  50. Thompson N. L., Flanders K. C., Smith J. M., Ellingsworth L. R., Roberts A. B., Sporn M. B. Expression of transforming growth factor-beta 1 in specific cells and tissues of adult and neonatal mice. J Cell Biol. 1989 Feb;108(2):661–669. doi: 10.1083/jcb.108.2.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Tomasi T. B., Jr Suppressive factors in amniotic fluid and newborn serum: is alpha-fetoprotein involved? Cell Immunol. 1978 May;37(2):459–466. doi: 10.1016/0008-8749(78)90213-7. [DOI] [PubMed] [Google Scholar]
  52. Torre-Amione G., Beauchamp R. D., Koeppen H., Park B. H., Schreiber H., Moses H. L., Rowley D. A. A highly immunogenic tumor transfected with a murine transforming growth factor type beta 1 cDNA escapes immune surveillance. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1486–1490. doi: 10.1073/pnas.87.4.1486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Tsao S. G., Brunk C. F., Pearlman R. E. Hybridization of nucleic acids directly in agarose gels. Anal Biochem. 1983 Jun;131(2):365–372. doi: 10.1016/0003-2697(83)90185-9. [DOI] [PubMed] [Google Scholar]
  54. Tucker R. F., Shipley G. D., Moses H. L., Holley R. W. Growth inhibitor from BSC-1 cells closely related to platelet type beta transforming growth factor. Science. 1984 Nov 9;226(4675):705–707. doi: 10.1126/science.6093254. [DOI] [PubMed] [Google Scholar]
  55. Wahl S. M., Hunt D. A., Wong H. L., Dougherty S., McCartney-Francis N., Wahl L. M., Ellingsworth L., Schmidt J. A., Hall G., Roberts A. B. Transforming growth factor-beta is a potent immunosuppressive agent that inhibits IL-1-dependent lymphocyte proliferation. J Immunol. 1988 May 1;140(9):3026–3032. [PubMed] [Google Scholar]
  56. Wahl S. M., McCartney-Francis N., Mergenhagen S. E. Inflammatory and immunomodulatory roles of TGF-beta. Immunol Today. 1989 Aug;10(8):258–261. doi: 10.1016/0167-5699(89)90136-9. [DOI] [PubMed] [Google Scholar]
  57. Zuber P., Kuppner M. C., De Tribolet N. Transforming growth factor-beta 2 down-regulates HLA-DR antigen expression on human malignant glioma cells. Eur J Immunol. 1988 Oct;18(10):1623–1626. doi: 10.1002/eji.1830181023. [DOI] [PubMed] [Google Scholar]
  58. van Oers N. S., Cohen B. L., Murgita R. A. Isolation and characterization of a distinct immunoregulatory isoform of alpha-fetoprotein produced by the normal fetus. J Exp Med. 1989 Sep 1;170(3):811–825. doi: 10.1084/jem.170.3.811. [DOI] [PMC free article] [PubMed] [Google Scholar]

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