Abstract
The Nb2 rat lymphoma cell line has the unique property that its growth is dependent on lactogenic pituitary hormones. Cell surface staining with monoclonal antibodies showed expression of class I MHC alloantigens of the RT1u haplotype, but no expression of class II MHC antigens. Staining for differentiation markers was strongly positive with antibodies OX52, W3/13 and OX44. Partial and weaker staining was obtained with CD2, P4/16 and the transferrin receptor. Nb2 cells were negative with CD5, OX40 and CD4, whilst CD8 stained only a minor fraction (1%) and certain variant clones of the cell line. This general pattern of staining is consistent with the phenotype of a small subpopulation of immature T cells. Nb2 cells proliferated in response to recombinant human IL-2, although they did not stain with antibodies against the IL-2 receptor. Enhancement of the stimulation by IL-2 in the presence of a submitogenic concentration of hGH indicated a synergism between these two hormones, and responses were suppressed by a similar dose of cyclosporin A (ID50=2 microg/ml). Although IL-2 could not be identified in culture supernatants, the presence of mRNA for IL-2, IL-2R and IL-4 was demonstrated by dot blot analysis. Finally, evidence that the Nb2 lymphoma is of T-cell lineage was given by Northern blot detection of mRNA for the alpha and beta chains of the T-cell receptor.
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- Bernton E. W., Meltzer M. S., Holaday J. W. Suppression of macrophage activation and T-lymphocyte function in hypoprolactinemic mice. Science. 1988 Jan 22;239(4838):401–404. doi: 10.1126/science.3122324. [DOI] [PubMed] [Google Scholar]
- Bresser J., Hubbell H. R., Gillespie D. Biological activity of mRNA immobilized on nitrocellulose in NaI. Proc Natl Acad Sci U S A. 1983 Nov;80(21):6523–6527. doi: 10.1073/pnas.80.21.6523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brideau R. J., Carter P. B., McMaster W. R., Mason D. W., Williams A. F. Two subsets of rat T lymphocytes defined with monoclonal antibodies. Eur J Immunol. 1980 Aug;10(8):609–615. doi: 10.1002/eji.1830100807. [DOI] [PubMed] [Google Scholar]
- Ceredig R. Proliferation in vitro and interleukin production by 14 day fetal and adult Lyt-2-/L3T4- mouse thymocytes. J Immunol. 1986 Oct 1;137(7):2260–2267. [PubMed] [Google Scholar]
- Crispe I. N., Bevan M. J. Expression and functional significance of the J11d marker on mouse thymocytes. J Immunol. 1987 Apr 1;138(7):2013–2018. [PubMed] [Google Scholar]
- Croze F., Walker A., Friesen H. G. Stimulation of growth of Nb2 lymphoma cells by interleukin-2 in serum-free and serum-containing media. Mol Cell Endocrinol. 1988 Feb;55(2-3):253–259. doi: 10.1016/0303-7207(88)90140-2. [DOI] [PubMed] [Google Scholar]
- Dallman M. J., Thomas M. L., Green J. R. MRC OX-19: a monoclonal antibody that labels rat T lymphocytes and augments in vitro proliferative responses. Eur J Immunol. 1984 Mar;14(3):260–267. doi: 10.1002/eji.1830140311. [DOI] [PubMed] [Google Scholar]
- Depper J. M., Leonard W. J., Drogula C., Krönke M., Waldmann T. A., Greene W. C. Interleukin 2 (IL-2) augments transcription of the IL-2 receptor gene. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4230–4234. doi: 10.1073/pnas.82.12.4230. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gause W. C., Mountz J. D., Steinberg A. D. Characterization and differentiation of CD4-, CD8- thymocytes sorted with the Ly-24 marker. J Immunol. 1988 Jan 1;140(1):1–7. [PubMed] [Google Scholar]
- Gout P. W., Beer C. T., Noble R. L. Prolactin-stimulated growth of cell cultures established from malignant Nb rat lymphomas. Cancer Res. 1980 Jul;40(7):2433–2436. [PubMed] [Google Scholar]
- Hiestand P. C., Mekler P., Nordmann R., Grieder A., Permmongkol C. Prolactin as a modulator of lymphocyte responsiveness provides a possible mechanism of action for cyclosporine. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2599–2603. doi: 10.1073/pnas.83.8.2599. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hunt S. V., Fowler M. H. A repopulation assay for B and T lymphocyte stem cells employing radiation chimaeras. Cell Tissue Kinet. 1981 Jul;14(4):445–464. doi: 10.1111/j.1365-2184.1981.tb00551.x. [DOI] [PubMed] [Google Scholar]
- Ivanyi J. Study of antigenic structure and inhibition of activity of human growth hormone and chorionic somatomammotropin by monoclonal antibodies. Mol Immunol. 1982 Dec;19(12):1611–1618. doi: 10.1016/0161-5890(82)90272-3. [DOI] [PubMed] [Google Scholar]
- Jefferies W. A., Brandon M. R., Williams A. F., Hunt S. V. Analysis of lymphopoietic stem cells with a monoclonal antibody to the rat transferrin receptor. Immunology. 1985 Feb;54(2):333–341. [PMC free article] [PubMed] [Google Scholar]
- Kemp J. D., Thorson J. A., McAlmont T. H., Horowitz M., Cowdery J. S., Ballas Z. K. Role of the transferrin receptor in lymphocyte growth: a rat IgG monoclonal antibody against the murine transferrin receptor produces highly selective inhibition of T and B cell activation protocols. J Immunol. 1987 Apr 15;138(8):2422–2426. [PubMed] [Google Scholar]
- Killeen N., Barclay A. N., Willis A. C., Williams A. F. The sequence of rat leukosialin (W3/13 antigen) reveals a molecule with O-linked glycosylation of one third of its extracellular amino acids. EMBO J. 1987 Dec 20;6(13):4029–4034. doi: 10.1002/j.1460-2075.1987.tb02747.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koch F., Thiele H. G., Low M. G. Release of the rat T cell alloantigen RT-6.2 from cell membranes by phosphatidylinositol-specific phospholipase C. J Exp Med. 1986 Oct 1;164(4):1338–1343. doi: 10.1084/jem.164.4.1338. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lamb J. R., Zanders E. D., Sewell W., Crumpton M. J., Feldmann M., Owen M. J. Antigen-specific T cell unresponsiveness in cloned helper T cells mediated via the CD2 or CD3/Ti receptor pathways. Eur J Immunol. 1987 Nov;17(11):1641–1644. doi: 10.1002/eji.1830171118. [DOI] [PubMed] [Google Scholar]
- Larson D. F. Cyclosporin. Mechanism of action: antagonism of the prolactin receptor. Prog Allergy. 1986;38:222–238. [PubMed] [Google Scholar]
- Lesniak M. A., Gorden P., Roth J., Gavin J. R., 3rd Binding of 125I-human growth hormone to specific receptors in human cultured lymphocytes. Characterization of the interaction and a sensitive radioreceptor assay. J Biol Chem. 1974 Mar 25;249(6):1661–1667. [PubMed] [Google Scholar]
- Love R. R., Rose D. P. Elevated bioactive prolactin in women at risk for familial breast cancer. Eur J Cancer Clin Oncol. 1985 Dec;21(12):1553–1554. doi: 10.1016/0277-5379(85)90251-2. [DOI] [PubMed] [Google Scholar]
- Mascardo R. N., Barton R. W., Sherline P. Somatostatin has an antiproliferative effect on concanavalin A-activated rat thymocytes. Clin Immunol Immunopathol. 1984 Oct;33(1):131–138. doi: 10.1016/0090-1229(84)90299-x. [DOI] [PubMed] [Google Scholar]
- Montgomery D. W., Zukoski C. F., Shah G. N., Buckley A. R., Pacholczyk T., Russell D. H. Concanavalin A-stimulated murine splenocytes produce a factor with prolactin-like bioactivity and immunoreactivity. Biochem Biophys Res Commun. 1987 Jun 15;145(2):692–698. doi: 10.1016/0006-291x(87)91020-5. [DOI] [PubMed] [Google Scholar]
- Nagy E., Berczi I., Friesen H. G. Regulation of immunity in rats by lactogenic and growth hormones. Acta Endocrinol (Copenh) 1983 Mar;102(3):351–357. doi: 10.1530/acta.0.1020351. [DOI] [PubMed] [Google Scholar]
- Noble R. L., Beer C. T., Gout P. W. Evidence in vivo and in vitro of a role for the pituitary in the growth of malignant lymphomas in Nb rats. Cancer Res. 1980 Jul;40(7):2437–2440. [PubMed] [Google Scholar]
- Osawa H., Diamantstein T. The characteristics of a monoclonal antibody that binds specifically to rat T lymphoblasts and inhibits IL 2 receptor functions. J Immunol. 1983 Jan;130(1):51–55. [PubMed] [Google Scholar]
- Paterson D. J., Green J. R., Jefferies W. A., Puklavec M., Williams A. F. The MRC OX-44 antigen marks a functionally relevant subset among rat thymocytes. J Exp Med. 1987 Jan 1;165(1):1–13. doi: 10.1084/jem.165.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Paterson D. J., Jefferies W. A., Green J. R., Brandon M. R., Corthesy P., Puklavec M., Williams A. F. Antigens of activated rat T lymphocytes including a molecule of 50,000 Mr detected only on CD4 positive T blasts. Mol Immunol. 1987 Dec;24(12):1281–1290. doi: 10.1016/0161-5890(87)90122-2. [DOI] [PubMed] [Google Scholar]
- Robinson A. P., Puklavec M., Mason D. W. MRC OX-52: a rat T-cell antigen. Immunology. 1986 Apr;57(4):527–531. [PMC free article] [PubMed] [Google Scholar]
- Russell D. H., Kibler R., Matrisian L., Larson D. F., Poulos B., Magun B. E. Prolactin receptors on human T and B lymphocytes: antagonism of prolactin binding by cyclosporine. J Immunol. 1985 May;134(5):3027–3031. [PubMed] [Google Scholar]
- Shevach E. M. The effects of cyclosporin A on the immune system. Annu Rev Immunol. 1985;3:397–423. doi: 10.1146/annurev.iy.03.040185.002145. [DOI] [PubMed] [Google Scholar]
- Snow E. C., Feldbush T. L., Oaks J. A. The effect of growth hormone and insulin upon MLC responses and the generation of cytotoxic lymphocytes. J Immunol. 1981 Jan;126(1):161–164. [PubMed] [Google Scholar]
- Tanaka T., Shiu R. P., Gout P. W., Beer C. T., Noble R. L., Friesen H. G. A new sensitive and specific bioassay for lactogenic hormones: measurement of prolactin and growth hormone in human serum. J Clin Endocrinol Metab. 1980 Nov;51(5):1058–1063. doi: 10.1210/jcem-51-5-1058. [DOI] [PubMed] [Google Scholar]
- Taniguchi T., Matsui H., Fujita T., Takaoka C., Kashima N., Yoshimoto R., Hamuro J. Structure and expression of a cloned cDNA for human interleukin-2. Nature. 1983 Mar 24;302(5906):305–310. doi: 10.1038/302305a0. [DOI] [PubMed] [Google Scholar]
- Williams A. F., Barclay A. N., Clark S. J., Paterson D. J., Willis A. C. Similarities in sequences and cellular expression between rat CD2 and CD4 antigens. J Exp Med. 1987 Feb 1;165(2):368–380. doi: 10.1084/jem.165.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williams A. F., Galfrè G., Milstein C. Analysis of cell surfaces by xenogeneic myeloma-hybrid antibodies: differentiation antigens of rat lymphocytes. Cell. 1977 Nov;12(3):663–673. doi: 10.1016/0092-8674(77)90266-5. [DOI] [PubMed] [Google Scholar]
- Yokota T., Otsuka T., Mosmann T., Banchereau J., DeFrance T., Blanchard D., De Vries J. E., Lee F., Arai K. Isolation and characterization of a human interleukin cDNA clone, homologous to mouse B-cell stimulatory factor 1, that expresses B-cell- and T-cell-stimulating activities. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5894–5898. doi: 10.1073/pnas.83.16.5894. [DOI] [PMC free article] [PubMed] [Google Scholar]


