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Functional and immunological responses of Jurkat lymphocytes transfected with the substance P receptor

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Summary

  1. 1.

    We have transfected the rat substance P receptor (SPR) cDNA into the leukemic T-lymphocyte cell line Jurkat (J-wt) in order to study the effects of substance P (SP) on lymphocyte signaling mechanisms and the resultant neuropeptide-induced immunological changes.

  2. 2.

    The SPR cDNA was transfected into J-wt by the method of electroporation. Clones expressing SPRs were selected using a functional assay that measured SP-induced mobilization of intracellular Ca2+([Ca2+]i) in a fluorescence activated cell sorter (FACS) and by their expression of specific125I-SP binding.

  3. 3.

    One clone, J-SPR, was identified and shown by Northern blot and125I-SP saturation binding techniques to express the 2.2-kb SPR message and approximately 50,000 SPRs/cell with aK d of 0.3 nM, respectively. Stimulation of J-SPR by SP resulted in the rapid mobilization of [Ca2+]i. This response was dose dependent in the range 10−11–10−6 M SP and was maximal at 10−7 M SP, with an EC50 of 0.3–0.5 nM SP. We further demonstrated that the SPR is rapidly desensitized following SP stimulation and by activation of the cell's T-cell receptor (TCR). Whole-cell patch-clamp experiments on J-SPR show that SP stimulation induces a Cl− current by a Ca2+ mediated process dependent on Ca2+/calmodulin-dependent protein kinase (CaMK).

  4. 4.

    Stimulation of J-SPR by SP results in changes in the cell surface expression of a number of molecules that play important roles in cell adhesion and activation: the expression of LFA-1 is decreased, and CD2 and IL-2 receptors are increased by 30 min, 6 hr, and 24 hr, respectively, following stimulation, as assessed by antibody staining in a FACS.

  5. 5.

    The expression of functional SPRs in Jurkat lymphocytes will now permit a detailed examination of how the activation of SPRs result in altered immune responses and further elucidate the role this neuropeptide receptor plays in inflammation.

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Authors and Affiliations

  1. Department of Medicine and Microbiology and Immunology, University of California, San Francisco, School of Medicine, 94143, San Francisco, California, USA

    Julie Sudduth-Klinger & Donald G. Payan

  2. The Howard Hughes Medical Institute, University of California, San Francisco, School of Medicine, 94143, San Francisco, California, USA

    Julie Sudduth-Klinger & Donald G. Payan

  3. Departments of Medicine and Pharmacology, Stanford University School of Medicine, 94305, Stanford, California, USA

    Muhammad Schumann & Phyllis Gardner

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  1. Julie Sudduth-Klinger
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  2. Muhammad Schumann
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  3. Phyllis Gardner
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  4. Donald G. Payan
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Sudduth-Klinger, J., Schumann, M., Gardner, P. et al. Functional and immunological responses of Jurkat lymphocytes transfected with the substance P receptor. Cell Mol Neurobiol 12, 379–395 (1992). https://doi.org/10.1007/BF00711540

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