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. 1992 Oct;12(5):379–395. doi: 10.1007/BF00711540

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

Julie Sudduth-Klinger 1,2, Muhammad Schumann 3, Phyllis Gardner 3, Donald G Payan 1,2,✉
PMCID: PMC11567415  PMID: 1281754

Abstract

  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. 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. 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. 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. 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.

Key words: substance P, substance P receptor, Ca2+/calmodulin-dependent protein kinase, chloride channel, neuroimmulogy

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