Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Mar;85(5):1605–1609. doi: 10.1073/pnas.85.5.1605

Identification of a 24-kDa cytokine that is required for development of cytolytic T lymphocytes.

C Cernetti 1, R M Steinman 1, A Granelli-Piperno 1
PMCID: PMC279822  PMID: 3278321

Abstract

It is known that the production of cytolytic T lymphocytes requires growth factors such as interleukins 2 and 4 (IL-2 and IL-4). Elsewhere we have described bioassays that detect a cytokine that operates in concert with growth factor to generate cytolytic T lymphocytes. The factor that is termed cytolytic T-lymphocyte differentiation factor (CDF), together with IL-2 and lectin, mediates the formation of CD8+ killer cells in 2 days from thymocyte or peripheral lymphoid precursors. CDF is not mimicked by natural or recombinant sources of interferons, colony-stimulating factors, and IL-1 to IL-4. Here we use these bioassays to isolate and further characterize a single 24-kDa CDF protein from the conditioned medium of stimulated human blood mononuclear cells. CDF is first enriched by three successive chromatographic procedures that utilize anion exchange, hydroxyapatite, and phenyl-Superose. A single 24-kDa band with CDF activity is then isolated on 12% NaDodSO4/PAGE and clearly distinguished from the 17-kDa band of IL-2. The apparent molecular mass is similar under reducing and nonreducing conditions. After elution from NaDodSO4/PAGE the cytokine is maximally active at 0.25 nM in the CDF assay and has no growth factor activity for T lymphoblasts. To generate cytolytic CD8+, CD4- cells from spleen and lymph node T lymphocytes, IL-2 and small numbers of accessory dendritic cells must be applied together with CDF.

Full text

PDF
1605

Images in this article

1607Image
on p.1607
1608Image
on p.1608
1608Image
on p.1608

Selected References

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

  1. Arya S. K., Wong-Staal F., Gallo R. C. Dexamethasone-mediated inhibition of human T cell growth factor and gamma-interferon messenger RNA. J Immunol. 1984 Jul;133(1):273–276. [PubMed] [Google Scholar]
  2. Austyn J. M., Steinman R. M., Weinstein D. E., Granelli-Piperno A., Palladino M. A. Dendritic cells initiate a two-stage mechanism for T lymphocyte proliferation. J Exp Med. 1983 Apr 1;157(4):1101–1115. doi: 10.1084/jem.157.4.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Azuma C., Tanabe T., Konishi M., Kinashi T., Noma T., Matsuda F., Yaoita Y., Takatsu K., Hammarström L., Smith C. I. Cloning of cDNA for human T-cell replacing factor (interleukin-5) and comparison with the murine homologue. Nucleic Acids Res. 1986 Nov 25;14(22):9149–9158. doi: 10.1093/nar/14.22.9149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baker P. E., Gillis S., Ferm M. M., Smith K. A. The effect of T cell growth factor on the generation of cytolytic T cells. J Immunol. 1978 Dec;121(6):2168–2173. [PubMed] [Google Scholar]
  5. Birkeland M. L., Simpson L., Isakson P. C., Pure E. T-independent and T-dependent steps in the murine B cell response to antiimmunoglobulin. J Exp Med. 1987 Aug 1;166(2):506–519. doi: 10.1084/jem.166.2.506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cernetti C., Steinman R. M., Granelli-Piperno A. Bioassays for the detection of a cytokine that enhances the development of cytolytic T lymphocytes. Cell Immunol. 1987 Oct 1;109(1):148–158. doi: 10.1016/0008-8749(87)90300-5. [DOI] [PubMed] [Google Scholar]
  7. Coffman R. L., Ohara J., Bond M. W., Carty J., Zlotnik A., Paul W. E. B cell stimulatory factor-1 enhances the IgE response of lipopolysaccharide-activated B cells. J Immunol. 1986 Jun 15;136(12):4538–4541. [PubMed] [Google Scholar]
  8. Granelli-Piperno A., Andrus L., Reich E. Antibodies to interleukin 2. Effects on immune responses in vitro and in vivo. J Exp Med. 1984 Sep 1;160(3):738–750. doi: 10.1084/jem.160.3.738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Granelli-Piperno A., Inaba K., Steinman R. M. Stimulation of lymphokine release from T lymphoblasts. Requirement for mRNA synthesis and inhibition by cyclosporin A. J Exp Med. 1984 Dec 1;160(6):1792–1802. doi: 10.1084/jem.160.6.1792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Granelli-Piperno A., Vassalli J. D., Reich E. Purification of murine T cell growth factor. A lymphocyte mitogen with helper activity. J Exp Med. 1981 Aug 1;154(2):422–431. doi: 10.1084/jem.154.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gromo G., Geller R. L., Inverardi L., Bach F. H. Signal requirements in the step-wise functional maturation of cytotoxic T lymphocytes. Nature. 1987 Jun 4;327(6121):424–426. doi: 10.1038/327424a0. [DOI] [PubMed] [Google Scholar]
  12. Inaba K., Steinman R. M. Resting and sensitized T lymphocytes exhibit distinct stimulatory (antigen-presenting cell) requirements for growth and lymphokine release. J Exp Med. 1984 Dec 1;160(6):1717–1735. doi: 10.1084/jem.160.6.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Isakson P. C. Antiimmunoglobulin-treated B cells respond to a B cell differentiation factor for IgG1. J Exp Med. 1986 Jul 1;164(1):303–308. doi: 10.1084/jem.164.1.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Merril C. R., Switzer R. C., Van Keuren M. L. Trace polypeptides in cellular extracts and human body fluids detected by two-dimensional electrophoresis and a highly sensitive silver stain. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4335–4339. doi: 10.1073/pnas.76.9.4335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mier J. W., Gallo R. C. Purification and some characteristics of human T-cell growth factor from phytohemagglutinin-stimulated lymphocyte-conditioned media. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6134–6138. doi: 10.1073/pnas.77.10.6134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nathan C. F., Prendergast T. J., Wiebe M. E., Stanley E. R., Platzer E., Remold H. G., Welte K., Rubin B. Y., Murray H. W. Activation of human macrophages. Comparison of other cytokines with interferon-gamma. J Exp Med. 1984 Aug 1;160(2):600–605. doi: 10.1084/jem.160.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Riendeau D., Harnish D. G., Bleackley R. C., Paetkau V. Purification of mouse interleukin 2 to apparent homogeneity. J Biol Chem. 1983 Oct 25;258(20):12114–12117. [PubMed] [Google Scholar]
  19. Robb R. J., Munck A., Smith K. A. T cell growth factor receptors. Quantitation, specificity, and biological relevance. J Exp Med. 1981 Nov 1;154(5):1455–1474. doi: 10.1084/jem.154.5.1455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shaw J., Caplan B., Paetkau V., Pilarski L. M., Delovitch T. L., McKenzie I. F. Cellular origins of co-stimulator (IL2) and its activity in cytotoxic T lymphocyte responses. J Immunol. 1980 May;124(5):2231–2239. [PubMed] [Google Scholar]
  21. Snapper C. M., Paul W. E. Interferon-gamma and B cell stimulatory factor-1 reciprocally regulate Ig isotype production. Science. 1987 May 22;236(4804):944–947. doi: 10.1126/science.3107127. [DOI] [PubMed] [Google Scholar]
  22. Steinman R. M., Kaplan G., Witmer M. D., Cohn Z. A. Identification of a novel cell type in peripheral lymphoid organs of mice. V. Purification of spleen dendritic cells, new surface markers, and maintenance in vitro. J Exp Med. 1979 Jan 1;149(1):1–16. doi: 10.1084/jem.149.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Takai Y., Herrmann S. H., Greenstein J. L., Spitalny G. L., Burakoff S. J. Requirement for three distinct lymphokines for the induction of cytotoxic T lymphocytes from thymocytes. J Immunol. 1986 Dec 1;137(11):3494–3500. [PubMed] [Google Scholar]
  24. Takatsu K., Kikuchi Y., Takahashi T., Honjo T., Matsumoto M., Harada N., Yamaguchi N., Tominaga A. Interleukin 5, a T-cell-derived B-cell differentiation factor also induces cytotoxic T lymphocytes. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4234–4238. doi: 10.1073/pnas.84.12.4234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Talmage D. W., Woolnough J. A., Hemmingsen H., Lopez L., Lafferty K. J. Activation of cytotoxic T cells by nonstimulating tumor cells and spleen cell factor(s). Proc Natl Acad Sci U S A. 1977 Oct;74(10):4610–4614. doi: 10.1073/pnas.74.10.4610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wagner H., Röllinghoff M., Schawaller R., Hardt C., Pfizenmaier K. T-cell-derived helper factor allows Lyt 123 thymocytes to differentiate into cytotoxic T lymphocytes. Nature. 1979 Aug 2;280(5721):405–406. doi: 10.1038/280405a0. [DOI] [PubMed] [Google Scholar]
  27. Welte K., Wang C. Y., Mertelsmann R., Venuta S., Feldman S. P., Moore M. A. Purification of human interleukin 2 to apparent homogeneity and its molecular heterogeneity. J Exp Med. 1982 Aug 1;156(2):454–464. doi: 10.1084/jem.156.2.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Widmer M. B., Grabstein K. H. Regulation of cytolytic T-lymphocyte generation by B-cell stimulatory factor. Nature. 1987 Apr 23;326(6115):795–798. doi: 10.1038/326795a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES