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. 1985 Aug;82(16):5490–5494. doi: 10.1073/pnas.82.16.5490

Purification to homogeneity and characterization of human B-cell differentiation factor (BCDF or BSFp-2).

T Hirano, T Taga, N Nakano, K Yasukawa, S Kashiwamura, K Shimizu, K Nakajima, K H Pyun, T Kishimoto
PMCID: PMC391148  PMID: 2410927

Abstract

Human B-cell differentiation factor (BCDF) was purified to homogeneity by sequential filtration and chromatography of culture supernatants from TCL-Na1 cells on an AcA34 gel column and then on a Mono P column with fast protein liquid chromatography and reversed-phase HPLC. A 5300-fold enrichment in specific activity of BCDF with about 25% recovery was attained. The homogeneity of purified BCDF was evidenced by the following: (i) the specific activity was 1.7 X 10(7) units/mg of protein, (ii) only two bands, Mr 19,000 and 21,000, were identified by NaDodSO4/PAGE under reduced as well as nonreduced conditions, and (iii) BCDF activity was recovered from the gel after NaDodSO4/PAGE in the fractions corresponding to protein bands of Mr 19,000 or 21,000. Purified BCDF induced Ig secretion in Epstein-Barr virus-transformed cell lines; as little as 3 pM gave 50% of the maximum reaction achieved by 30-80 pM BCDF. Purified BCDF induced Ig production in activated B cells without any effect on cell growth. Purified BCDF did not show any activity of interleukin 1 or 2, B-cell stimulatory factor (BSF)p-1, B-cell growth factor II (BCGF-II), or interferon. Since BCDF was isolated and characterized as described, we propose that the BCDF that induces the final differentiation of B cells into high-rate Ig-secreting cells be designated BSFp-2.

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Selected References

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  1. Familletti P. C., Rubinstein S., Pestka S. A convenient and rapid cytopathic effect inhibition assay for interferon. Methods Enzymol. 1981;78(Pt A):387–394. doi: 10.1016/0076-6879(81)78146-1. [DOI] [PubMed] [Google Scholar]
  2. Goldsmith P. K. A highly sensitive enzyme-linked immunosorbent assay for human immunoglobulin E: comparison of microtiter plate and disk methodologies. Anal Biochem. 1981 Oct;117(1):53–60. doi: 10.1016/0003-2697(81)90690-4. [DOI] [PubMed] [Google Scholar]
  3. Hirano T., Fujimoto K., Teranishi T., Nishino N., Onoue K., Maeda S., Shimada K. Phorbol ester increases the level of interleukin 2 mRNA in mitogen-stimulated human lymphocytes. J Immunol. 1984 May;132(5):2165–2167. [PubMed] [Google Scholar]
  4. Hirano T., Teranishi T., Lin B., Onoue K. Human helper T cell factor(s). IV. Demonstration of a human late-acting B cell differentiation factor acting on Staphylococcus aureus Cowan I-stimulated B cells. J Immunol. 1984 Aug;133(2):798–802. [PubMed] [Google Scholar]
  5. Hirano T., Teranishi T., Onoue K. Human helper T cell factor(s). III. Characterization of B cell differentiation factor I (BCDF I). J Immunol. 1984 Jan;132(1):229–234. [PubMed] [Google Scholar]
  6. Howard M., Farrar J., Hilfiker M., Johnson B., Takatsu K., Hamaoka T., Paul W. E. Identification of a T cell-derived b cell growth factor distinct from interleukin 2. J Exp Med. 1982 Mar 1;155(3):914–923. doi: 10.1084/jem.155.3.914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Howard M., Paul W. E. Regulation of B-cell growth and differentiation by soluble factors. Annu Rev Immunol. 1983;1:307–333. doi: 10.1146/annurev.iy.01.040183.001515. [DOI] [PubMed] [Google Scholar]
  8. Isakson P. C., Puré E., Vitetta E. S., Krammer P. H. T cell-derived B cell differentiation factor(s). Effect on the isotype switch of murine B cells. J Exp Med. 1982 Mar 1;155(3):734–748. doi: 10.1084/jem.155.3.734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kikutani H., Taga T., Akira S., Kishi H., Miki Y., Saiki O., Yamamura Y., Kishimoto T. Effect of B cell differentiation factor (BCDF) on biosynthesis and secretion of immunoglobulin molecules in human B cell lines. J Immunol. 1985 Feb;134(2):990–995. [PubMed] [Google Scholar]
  10. 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]
  11. Leibson H. J., Gefter M., Zlotnik A., Marrack P., Kappler J. W. Role of gamma-interferon in antibody-producing responses. 1984 Jun 28-Jul 4Nature. 309(5971):799–801. doi: 10.1038/309799a0. [DOI] [PubMed] [Google Scholar]
  12. Leibson H. J., Marrack P., Kappler J. W. B cell helper factors. I. Requirement for both interleukin 2 and another 40,000 mol wt factor. J Exp Med. 1981 Nov 1;154(5):1681–1693. doi: 10.1084/jem.154.5.1681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Muraguchi A., Kasahara T., Oppenheim J. J., Fauci A. S. B cell growth factor and T cell growth factor produced by mitogen-stimulated normal human peripheral blood T lymphocytes are distinct molecules. J Immunol. 1982 Dec;129(6):2486–2489. [PubMed] [Google Scholar]
  14. Muraguchi A., Kishimoto T., Miki Y., Kuritani T., Kaieda T., Yoshizaki K., Yamamura Y. T cell-replacing factor- (TRF) induced IgG secretion in a human B blastoid cell line and demonstration of acceptors for TRF. J Immunol. 1981 Aug;127(2):412–416. [PubMed] [Google Scholar]
  15. Nakagawa T., Hirano T., Nakagawa N., Yoshizaki K., Kishimoto T. Effect of recombinant IL 2 and gamma-IFN on proliferation and differentiation of human B cells. J Immunol. 1985 Feb;134(2):959–966. [PubMed] [Google Scholar]
  16. Nakanishi K., Howard M., Muraguchi A., Farrar J., Takatsu K., Hamaoka T., Paul W. E. Soluble factors involved in B cell differentiation: identification of two distinct T cell-replacing factors (TRF). J Immunol. 1983 May;130(5):2219–2224. [PubMed] [Google Scholar]
  17. Okada M., Sakaguchi N., Yoshimura N., Hara H., Shimizu K., Yoshida N., Yoshizaki K., Kishimoto S., Yamamura Y., Kishimoto T. B cell growth factors and B cell differentiation factor from human T hybridomas. Two distinct kinds of B cell growth factor and their synergism in B cell proliferation. J Exp Med. 1983 Feb 1;157(2):583–590. doi: 10.1084/jem.157.2.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Paige C. J., Schreier M. H., Sidman C. L. Mediators from cloned T helper cell lines affect immunoglobulin expression by B cells. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4756–4760. doi: 10.1073/pnas.79.15.4756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pike B. L., Vaux D. L., Clark-Lewis I., Schrader J. W., Nossal G. J. Proliferation and differentiation of single hapten-specific B lymphocytes is promoted by T-cell factor(s) distinct from T-cell growth factor. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6350–6354. doi: 10.1073/pnas.79.20.6350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Puré E., Isakson P. C., Takatsu K., Hamaoka T., Swain S. L., Dutton R. W., Dennert G., Uhr J. W., Vitetta E. S. Induction of B cell differentiation by T cell factors. I. Stimulation of IgM secretion by products of a T cell hybridoma and a T cell line. J Immunol. 1981 Nov;127(5):1953–1958. [PubMed] [Google Scholar]
  21. Ralph P., Welte K., Levi E., Nakoinz I., Litcofsky P. B., Mertelsmann R. H., Moore M. A. Human B cell-inducing factor(s) for production of IgM, IgG and IgA: independence from IL 2. J Immunol. 1984 Apr;132(4):1858–1862. [PubMed] [Google Scholar]
  22. Robb R. J., Smith K. A. Heterogeneity of human T-cell growth factor(s) due to variable glycosylation. Mol Immunol. 1981 Dec;18(12):1087–1094. doi: 10.1016/0161-5890(81)90024-9. [DOI] [PubMed] [Google Scholar]
  23. Saiki O., Ralph P. Clonal differences in response to T cell replacing factor (TRF) for IgM secretion and TRF receptors in a human B lymphoblast cell line. Eur J Immunol. 1983 Jan;13(1):31–34. doi: 10.1002/eji.1830130108. [DOI] [PubMed] [Google Scholar]
  24. Schimpl A., Wecker E. Replacement of T-cell function by a T-cell product. Nat New Biol. 1972 May 3;237(70):15–17. doi: 10.1038/newbio237015a0. [DOI] [PubMed] [Google Scholar]
  25. Shimizu K., Hirano T., Ishibashi K., Nakano N., Taga T., Sugamura K., Yamamura Y., Kishimoto T. Immortalization of BGDF (BCGF II)- and BCDF-producing T cells by human T cell leukemia virus (HTLV) and characterization of human BGDF (BCGF II). J Immunol. 1985 Mar;134(3):1728–1733. [PubMed] [Google Scholar]
  26. Swain S. L., Dennert G., Warner J. F., Dutton R. W. Culture supernatants of a stimulated T-cell line have helper activity that acts synergistically with interleukin 2 in the response of B cells to antigen. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2517–2521. doi: 10.1073/pnas.78.4.2517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Swain S. L., Howard M., Kappler J., Marrack P., Watson J., Booth R., Wetzel G. D., Dutton R. W. Evidence for two distinct classes of murine B cell growth factors with activities in different functional assays. J Exp Med. 1983 Sep 1;158(3):822–835. doi: 10.1084/jem.158.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Takatsu K., Harada N., Hara Y., Takahama Y., Yamada G., Dobashi K., Hamaoka T. Purification and physicochemical characterization of murine T cell replacing factor (TRF). J Immunol. 1985 Jan;134(1):382–389. [PubMed] [Google Scholar]
  29. Takatsu K., Tanaka K., Tominaga A., Kumahara Y., Hamaoka T. Antigen-induced T cell-replacing factor (TRF). III. Establishment of T cell hybrid clone continuously producing TRF and functional analysis of released TRF. J Immunol. 1980 Dec;125(6):2646–2653. [PubMed] [Google Scholar]
  30. Teranishi T., Hirano T., Arima N., Onoue K. Human helper T cell factor(s) (ThF). II. Induction of IgG production in B lymphoblastoid cell lines and identification of T cell-replacing factor- (TRF) like factor(s). J Immunol. 1982 Apr;128(4):1903–1908. [PubMed] [Google Scholar]
  31. Teranishi T., Hirano T., Lin B. H., Onoue K. Demonstration of the involvement of interleukin 2 in the differentiation of Staphylococcus aureus Cowan I-stimulated B cells. J Immunol. 1984 Dec;133(6):3062–3067. [PubMed] [Google Scholar]
  32. Yoshizaki K., Nakagawa T., Fukunaga K., Tseng L. T., Yamamura Y., Kishimoto T. Isolation and characterization of B cell differentiation factor (BCDF) secreted from a human B lymphoblastoid cell line. J Immunol. 1984 Jun;132(6):2948–2954. [PubMed] [Google Scholar]
  33. Yoshizaki K., Nakagawa T., Kaieda T., Muraguchi A., Yamamura Y., Kishimoto T. Induction of proliferation and Ig production in human B leukemic cells by anti-immunoglobulins and T cell factors. J Immunol. 1982 Mar;128(3):1296–1301. [PubMed] [Google Scholar]
  34. Zubler R. H., Lowenthal J. W., Erard F., Hashimoto N., Devos R., MacDonald H. R. Activated B cells express receptors for, and proliferate in response to, pure interleukin 2. J Exp Med. 1984 Oct 1;160(4):1170–1183. doi: 10.1084/jem.160.4.1170. [DOI] [PMC free article] [PubMed] [Google Scholar]

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