SEARCH

SEARCH BY CITATION

Abstract

  1. Top of page
  2. Abstract
  3. REFERENCES

Marijuana and many of its constituent cannabinoids influence the central nervous system (CNS), probably through the cannabinoid receptor, which has recently been cloned in rat and human. While numerous reports have also described effects of cannabinoids on the immune system, the observation of both mRNA and cannabinoid receptor has hitherto been exclusively confined to the brain, a reported detection in the testis being the sole example of its presence at the periphery. Here we report the expression of the cannabinoid receptor on human immune tissues using a highly sensitive polymerase-chain-reaction-based method for mRNA quantification. We show that, although present in a much lower abundance than in brain, cannabinoid receptor transcripts are found in human spleen, tonsils and peripheral blood leukocytes. The distribution pattern displays important variations of the mRNA level for the cannabioid receptor among the main human blood cell subpopulations. The rank order of mRNA levels in these cells is B cells > natural killer cells ≥ polymorphonuclear neutrophils ≥ T8 cells > monocytes > T4 cells. Cannabinoid-receptor mRNA, which is also found in monocytic, as well as T and B leukemia cell lines but not in Jurkat cells, presents a great diversity of expression on these cells as well, B-cell lines expressing a much higher level than T-cell lines. The cannabinoid receptor PCR products from leukocytes and brain are identical both in size and sequence suggesting a strong similarity between central and peripheral cannabinoid receptors. The expression of this receptor was demonstrated on membranes of the myelomonocytic U937 cells using the synthetic cannabinoid [3H]CP-55940 as ligand. The Kd determined from Scatchard analysis was 0.1 nM and the Bmax for membranes was 525 fmol/mg protein. The demonstration of cannabinoid-receptor expression at both mRNA and protein levels on human leukocytes provides a molecular basis for cannabinoid action on these cells.

Abbreviations
G protein

guanine-nucleotide-binding regulatory protein

CNS

central nervous system

RT

reverse transcription

9-THC

-9-tetrahydrocannabinol

PCR

polymerase chain reaction

PBMC

peripheral blood mononuclear cells

PMN

polymorphonuclear neutrophils

NK cells

natural killer cells

REFERENCES

  1. Top of page
  2. Abstract
  3. REFERENCES
  • Baczynsky, W. O. T. & Zimmerman, A. M. (1983) Effects of δ9-tetrahydrocannabinol, cannabinol and cannabidiol on the immune system in mice, Pharmacology (Basel) 26, 1219.
  • Bell, M. R., D'Ambra, T. E., Kumar, V., Eissenstat, M. A., Herrmann, J. L., Wetzel, J. R., Rosi, D., Philion, R. E., Daum, S. J., Hlasta, D. J., Kullnig, R. K., Ackerman, J. H., Haubrich, D. R., Luttinger, D. A., Baizman, E. R., Miller, M. S. & Ward, S. J. (1991) Antinociceptive (Aminoalkyl)indoles, J. Med. Chem. 34, 10991110.
  • Bidaut-Russell, M., Devane, W. A. & Howlett A. C. (1990) Cannabinoid receptors and modulation of cyclic AMP accumulation in the rat brain, J. Neurochem. 55, 2126.
  • Blanchard, D. K., Newton, C., Klein, T. W., Stewart, W. E. II & Friedman, H. (1986) In vitro and in vivo suppressive effects of delta-9-tetrahydrocannabinol on interferon production by murine spleen cells, Int. J. Immunopharmacol. 8, 819824.
  • Bouaboula, M., Legoux, P., Pessegue, B., Delpech, B., Dumont, X., Piechaczyk, M., Casellas, P. & Shire, D. (1992) Standardization of mRNA titration using a polymerase chain reaction method involving co-amplification with a multispecific internal control, J. Biol. Chem. 267, 2183021838.
  • Canat, X., Carayon, P., Bouaboula, M., Cahard, D., Shire, D., Roque, C., Le Fur, G. & Casellas, P. (1993) Distribution profile and properties of peripheral-type benzodiazepine receptors on human hemopoietic cells, Life Sci. 52, 107118.
  • Carayon, P., Bord, A. & Raymond, M. (1991) Simultaneous identification of eight leucocyte subsets of human peripheral blood using three-colour immunofluorescence flow cytometric analysis, J. Immunol. Methods 138, 257264.
  • Cheng, Y. & Prussof, W. (1973) Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50% inhibition (150) of an enzymatic reaction, Biochem. Pharmacol. 22, 30993108.
  • Chirgwin, J. M., Przybyla, A. E., Mac Donald, R. J. & Rutter, W. J. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease, Biochemistry 18, 52945299.
  • Devane, W. A., Dysarz III, F. A., Johnson, M. R., Melvin, L. S. & Howlett, A. C. (1988) Determination and characterization of a cannabinoid receptor in rat brain, Mol. Pharmacol. 34, 605613.
  • Fischer-Stenger, K., Updegrove, A. W. & Cabral, G. A. (1992) Delta 9-tetrahydrocannabinol decreases cytotoxic T lymphocyte activity to herpes simplex virus type 1-infected cells, Proc. Soc. Exp. Biol. Med. 200, 422430.
  • Géard, C., Mollereau, C., Vassart, G. & Parmentier, M. (1990) Nucleotide sequence of a human cannabinoid receptor cDNA, Nucleic Acids Res. 29, 7142.
  • Gérard, C. M., Mollereau, C., Vassart, G. & Parmentier, M. (1991) Molecular cloning of a human cannabinoid receptor which is also expressed in testis, Biochem. J. 279, 129134.
  • Howlett, A. C., Qualy, J. M. & Khachatrian, L. L. (1985) Involvement of Gi in the inhibition of adenylate cyclase by cannabimimetic drugs, Mol. Pharmacol. 29, 307313.
  • Jansen, E. M., Haycock, D. A., Ward, S. J. & Seybold, V. S. (1992) Distribution of cannabinoid receptors in rat brain determined with aminoalkylindoles, Brain Res. 575, 93102.
  • Kaminsky, N. E., Abood, M. E., Kessler, F. K., Martin, B. R. & Schatz, A. R. (1992) Identification of a functionally relevant cannabinoid receptor on mouse spleen cells that is involved in cannabinoid-mediated immune modulation. Mol. Pharmacol. 42, 736742.
  • Klein, E., Klein, G., Nadkarni, J. S., Nadkarni, J. J., Wigzell, H. & Clifford P. (1968) Surface IgM-kappa specificity on a Burkitt lymphoma cell in vivo and in derived culture lines, Cancer Res. 28, 13001310.
  • Klein, T. W., Kawakami, Y., Newton, C. & Freidman H. (1991) Marijuana components suppress induction and cytolytic function of murine cytotoxic T cells in vitro and in vivo, J. Toxicol. Environ. Health 32, 465477.
  • Krug, M. S. & Berger, S. L. (1987) First-strand cDNA synthesis primed with oligo(dT), Methods Enzymol. 112, 316324.
  • Luo, Y. D., Patel, M. K., Wiederhold, M. D. & Ou, D. W. (1992) Effects of cannabinoids and cocaine on the mitogen-induced transformations of lymphocytes of human and mouse origins, Int. J. Immunopharmacol. 14, 4956.
  • Mackie, K. & Hille, B. (1992) Cannabinoids inhibit N-type calcium channels in neuroblastoma-glioma cells, Proc. Natl Acad. Sci. USA 89, 38253829.
  • Mailleux, P. & Vanderhaeghen, J. J. (1992) Distribution of neuronal cannabinoid receptor in the adult rat brain: a comparative receptor binding radioautography and in situ hybridization histochemistry, Neuroscience 48, 655668.
  • Marx, J. (1990) Marijuana receptor gene cloned, Science 249, 624626.
  • Matsuda, L. A., Lolait, S. J., Brownstein, M. J., Young, A. C. & Bonner, T. I. (1990) Structure of a cannabinoid receptor and functional expression of the cloned cDNA, Nature 346, 561564.
  • Michalevicz, R., Taheri, M. R., Katz, E. F., Blacklock, H. A., Ivory, K. I. & Hoffbrand, V. A. (1987) JM, a Thy-ALL cell line produces both inhibitory and stimulatory lymphokines for bone marrow progenitor cells, Leuk. Res. 11, 649654.
  • Murison, G., Chubb, C. B. H., Maeda, S., Gemmell, M. A. & Huberman, E. (1987) Cannabinoids induce incomplete maturation of cultured human leukemia cell, Proc. Natl Acad. Sci. USA 84, 54145418.
  • Murphy, L. D., Herzog, C. E., Rudick, J. B., Fojo, A. T. & Bates, S. E. (1990) Use of the polymerase chain reaction in the quantitation of mdr-1 gene expression, Biochemistry 29, 1035110356.
  • Novak, T. J. & Rothenberg, E. V. (1990) cAMP inhibits induction of interleukin 2 but not of interleukin 4 in T cells, Proc. Natl Acad. Sci. USA 87, 93539357.
  • Pacheco, M., Childers, S. R., Arnold, R., Casiano, F. & Ward, S. J. (1991) Aminoalkylindoles: actions on specific G-protein-linked receptors, J. Pharmacol. Exp. Ther. 257, 170183.
  • Pross, S. H., Klein, T. W., Newton, C. A. & Friedman, H. (1987) Differential effects of marijuana components on proliferation of spleen, lymph node and thymus cells in vitro, Int. J. Immunopharmacol. 9, 363370.
  • Pross, S. H., Klein, T. W., Newton, C. A., Smith, J., Widden, R. & Friedman, H. (1990) Differential suppression of T-cell subpopulations by THC (Delta-9-tetrahydrocannabinol) Int. J. Immunopharmacol. 12, 539544.
  • Saiki, R. K., Gelfaud, D. H., Stoffel, S., Scharf, S. J., Higuchi, R., Horn, G. Y., Mullis, K. & Erlich, H. A. (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase, Science 239, 487491.
  • Sanger, F., Nicklen, S. & Coulson, A. T. (1977) DNA sequencing with chain-terminating inhibitors, Proc. Natl Acad. Sci. USA 74, 54635467.
  • Spector, T. (1978) Refinement of the Coomassie blue method of protein quantitation, Anal. Biochem. 86, 142146.
  • Sun-Yu, N. G., Gunnin, G. P., Eddy, R., Ponte, P., Leavitt, J., Shows, T. & Kedes, L. (1985) Evolution of the functional human betaactin gene and its multi-pseudogene family: conservation of noncoding regions and chromosomal dispersion of pseudogenes, Mol. Cell. Biol. 5, 27202732.
  • Sundstrom, C. & Nilsson, K. (1976) Establishment and characterization of a human histiocytic lymphoma cell line (U-937), Int. J. Cancer 17, 565577.
  • Tahir, S. K. & Zimmerman, A. (1991) Influence of marijuana on cellular structures and biochemical activites, Pharmacol. Biochem. Behav. 40, 617623.
  • Titishov, N., Mechoulam, R. & Zimmerman, A. M. (1989) Stereospecific effects of (-)- and (+)-7-hydroxy-delta-6-tetrahydrocannabinol-dimethylheptyl on the immune system of mice, Pharmacology 39, 337349.
  • Tsuchiya, S., Kobayashi, Y., Goto, Y., Okumura, H., Nakae, S., Konno, T. & Tada, K. (1982) Induction of maturation in cultured human monocvtic leukemia cells by a phorbol diester, Cancer Res. 42, 15301536.
  • Van Boxel, J. A. & Buell, D. N. (1974) IgD on cell membranes of human lymphoid cell lines with multiple immunoglobulin classes, Nature 251, 443444.
  • Watzl, B., Scuderi, P. & Watson, R. R. (1991) Marijuana components stimulate human peripheral blood mononuclear cell secretion of interferon-gamma and suppress interleukin-1 alpha in vitro, Int. J. Immunopharmacol. 13, 10911097.
  • Weiss, A., Wiskocil, R. L. & Stobo, J. D. (1984) The role of T3 surface molecules in the activation of human T cells: a two-stimulus requirement for IL2 production reflects events occurring at a pre-translational level, J. Immunol. 133, 123128.
  • Whisler, R. L., Beiqing, L., Grants, I. S. & Newhouse, Y. G. (1992) Cyclic AMP modulation of human B cell proliferative responses: role of cAMP-dependent protein kinases in enhancing B cell responses to phorbol diesters and ionomycin, Cell. Immunol. 142, 398415.
  • Yamamoto, Y., Mitsui, H. & Nishikawa, Y. (1984) Purification of a major sialoglycoprotein (SGP140) on P12/Ichikawa cells and its expression on differentiated HL-60 cells, J. Immunol. 132, 340346.