Measurement of Interleukin 7

Anthony E. Namen1, Philip J. Morrissey1

1 Immunex Corporation, Seattle, Washington
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 6.7
DOI:  10.1002/0471142735.im0607s17
Online Posting Date:  May, 2001
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Abstract

This unit describes an assay system to detect biologically active interleukin 7 (IL‐7). The target or indicator cells are a clonally derived long‐term pre‐B cell line designated IxN/2bx (or, more simply, 2bx). These cells demonstrate an absolute specificity for IL‐7 for continued growth and viability and are used to establish a thymidine‐uptake proliferation assay similar to those established for other interleukins. A describes maintenance of the IxN/2bx cell line.

     
 
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Table of Contents

  • Support Protocol 1: Maintenance of IxN/2bx Cell Line
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1:

  Materials
  • IxN/2bx cells (see protocol 2; Immunex)
  • Complete IMDM‐5 medium ( appendix 2A)
  • 0.1 mCi/ml [3H]thymidine ( appendix 3D)
  • 96‐well flat‐bottom microtiter plate
  • Additional reagents and equipment for determination of [3H]thymidine uptake ( appendix 3D)
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Figures

Videos

Literature Cited

Literature Cited
   Chazen, G.D., Pereira, G.M.B., Le Gros, G., Gillis, S., and Shevach, E.M. 1989. Interleukin 7 is a T cell growth factor. Proc. Natl. Acad. Sci. U.S.A. 86:5923‐5927.
   Davis, B., Huffman, M., Knoblock, K., and Baker, P. 1983. Logit analysis of interleukin‐2 activity using a BASIC program. Comput. Appl. Laboratory 4:265‐268.
   Friend, S.L., Hosier, S., Nelson, A., Foxworthe, D., Williams, D.E., and Farr, A. 1994. A thymic stromal cell line supports in vitro development of surface IgM+ B cells and produces a novel growth factor affecting B and T lineage cells. Exp. Hematol. 22:321‐328.
   Grabstein, K.H., Namen, A.E., Shanebeck, K., Voice, R.F., Reed, S.G., and Widmer, M.B. 1990. Regulation of T cell proliferation by IL‐7. J. Immunol. 144:3015‐3020.
   Grabstein, K.H., Waldschmidt, T.J., Finkelman, F.D., Hess, B.W., Alpert, A.R., Boiani, N.E., Namen, A.E., and Morrissey, P.J. 1993. Inhibition of murine B and T lymphopoiesis in vivo by an anti–interleukin 7 monoclonal antibody. J. Exp. Med. 178:257‐264.
   Ishihara, K., Medina, K., Hayashi, S., Pietrangeli, C., Namen, A.E., Miyake, K., and Kincade, P.W. 1991. Stromal‐cell and cytokine‐dependent lymphocyte clones which span the pre‐B‐ to B‐cell transition. Devel. Immunol. 1:149‐161.
   Morrissey, P.J., Goodwin, R.G., Nordan, R.P., Anderson, D., Grabstein, K.H., Cosman, D., Sims, J., Lupton, S., Acres, B., Reed, S.G., Mochizuki, D., Eisenman, J., Conlon, P.J., and Namen, A.E. 1989. Recombinant interleukin‐7, pre–B cell growth factor, has costimulatory activity on purified mature T cells. J. Exp. Med. 169:707‐716.
   Namen, A.E., Lupton, S., Hjerrild, K., Wignall, J., Mochizuki, D.Y., Schmierer, A., Mosley, B., March, C.J., Urdal, D.L., Gillis, S., Cosman, D., and Goodwin, R.G. 1988a. Stimulation of B cell progenitors by interleukin 7. Nature 333:571‐573.
   Namen, A.E., Schmierer, A.E., March, C.J., Overell, R.W., Park, L.S., Urdal, D.L., and Mochizuki, D.Y. 1988b. B cell precursor growth promoting activity. Purification and characterization of a growth factor active on lymphocyte precursors. J. Exp. Med. 167:988‐1002.
   Nishikawa, S.I., Ogawa, M., Nishikawa, S., Kunisada, T., and Kodama, H. 1988. B lymphopoiesis on stromal cell clone: Stromal cell clones acting on different stages of B cell differentiation. Eur. J. Immunol. 18:1767‐1771.
   Okazaki, H., Ito, M., Sudo, T., Hattori, M., Kano, S., Katsura, Y., and Minato, N. 1989. IL‐7 promotes thymocytes bearing α/β or γ/δ T cell receptors in vitro: Synergism with IL‐2. J. Immunol. 143:2917‐2922.
   Park, L.S., Friend, D.J., Schmierer, A.S., Dower, S.K., and Namen, A.E. 1990. Murine interleukin 7 receptor. Characterization on an IL‐7‐dependent cell line. J. Exp. Med. 171:1073‐1089.
   Sudo, T., Nishikawa, S., Ohno, N., Akiyama, N., Tamakoshi, M., Yoshida, H., and Nishikawa, S.I. 1993. Expression and function of the interleukin 7 receptor in murine lymphocytes. Proc. Natl. Acad. Sci. U.S.A. 90:9125‐9129.
   Welch, P.A., Namen, A.E., Goodwin, R.G., Armitage, R., and Cooper, M.D. 1989. Human IL‐7: A novel T cell growth factor. J. Immunol. 143:3562‐3567.
   Whitlock, C.A. and Witte, O.N. 1982. Long term culture of B lymphocytes and their precursors from murine bone marrow. Proc. Natl. Acad. Sci. U.S.A. 79:3608‐3612.
   Whitlock, C.A., Robertson, D.A., and Witte, O.N. 1984. Murine B cell lymphopoiesis in long term culture. J. Immunol. Methods 67:333‐337.
   Willcocks, J.L., Hales, A., Page, T.H., and Foxwell, B.M.J. 1993. The murine T cell line CT6 provides a novel bioassay for interleukin‐7. Eur. J. Immunol. 23:716‐720.
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