This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by TATSUNO, I. Articles by ARIMURA, A. Search for Related Content PubMed PubMed Citation Articles by TATSUNO, I. Articles by ARIMURA, A.

Inhibition of Mitogen-Stimulated Proliferation of Murine Splenocytes by a Novel Neuropeptide, Pituitary Adenylate Cyclase Activating Polypeptide: A Comparative Study with Vasoactive Intestinal Peptide^*

U.S.-Japan Biomedical Research Laboratories, Tulane University Hebert Center Belle Chasse, Louisiana, 70037
Departments of Medicine, Anatomy and Physiology, Tulane University School of Medicine New Orleans, Louisiana, 70112

Abstract

Two novel polypeptides known as pituitary adenylate cyclase activating polypeptide with 38 residues (PACAP38) and a shorter form of the peptide corresponding to the N-terminal 27 residues (PACAP27) were isolated from ovine hypothalamus. The N-terminal 28 residues of PACAP show 68% homology with vasoactive intestinal peptide (VIP). VIP has been reported to have specific binding sites in lymphocytes and inhibit mitogen-stimulated lymphocyte proliferation through a receptor- mediated stimulation of cAMP-dependent protein kinase. Using concanavalin A-induced proliferation of murine splenocytes as a model system, we now report that both PACAP38 and PACAP 27 can inhibit the proliferation of these cells in the same dose-dependent manner as VIP. The minimal effective concentration of the PACAPs was 10^-10-10^-9 M. However, neither PACAP affected lipopolysaccharide-induced proliferation of murine splenocytes. The binding of [¹²⁵I]PACAP27 to these splenocytes was rapid, time dependent, reversible, and proportional to the numbers of murine splenocytes. Scatchard analysis of displacement of the bound tracer by unlabeled PACAP27 indicated the existence of two classes of binding sites. The dissociation constant (K_d) was 0.86±0.24 nM and the maximal binding capacity (B_max) was 1.13±0.39 fmol/10⁶ cells for the high affinity binding site. The low affinity binding site had a K_d of 0.13 ± 0.03 MM with a Bmaj[ of 73.5 ± 9.5 fmol/10⁶ cells. PACAP38 and VIP displaced the binding of [¹²⁵I]PACAP27 in the same manner as PACAP27 and Scatchard analyses indicated the presence of two classes of binding sites with K_d and B_max similar to those for PACAP27. Furthermore, when [12SI]VIP was used as a radiolabeled ligand, PACAP27 and PACAP38 displaced the [¹²⁵I] VIP binding to the same degree as unlabeled VIP. Scatchard analysis indicated that there was no significant difference of the K_d or B_max between PACAP and VIP. Taken together, these data suggest that PACAPs bind to a site similar or identical to that used by VIP which inhibit the proliferation of murine splenocytes induced by concanavalin A. (Endocrinology 128: 728–734, 1991)

Footnotes

^* This study was supported in part by a research grant from Takeda Chemical Industries, Ltd. and NIH Grant DK-09094.

Received August 3, 1990.

This article has been cited by other articles:

				C. Martinez, Y. Juarranz, C. Abad, A. Arranz, B. G. Miguel, F. Rosignoli, J. Leceta, and R. P. Gomariz Analysis of the role of the PAC1 receptor in neutrophil recruitment, acute-phase response, and nitric oxide production in septic shock J. Leukoc. Biol., May 1, 2005; 77(5): 729 - 738. [Abstract] [Full Text] [PDF]

				M. Delgado, D. Pozo, and D. Ganea The Significance of Vasoactive Intestinal Peptide in Immunomodulation Pharmacol. Rev., June 1, 2004; 56(2): 249 - 290. [Abstract] [Full Text] [PDF]

				F. S. Lee, R. Rajagopal, A. H. Kim, P. C. Chang, and M. V. Chao Activation of Trk Neurotrophin Receptor Signaling by Pituitary Adenylate Cyclase-activating Polypeptides J. Biol. Chem., March 8, 2002; 277(11): 9096 - 9102. [Abstract] [Full Text] [PDF]

				C. Abad, C. Martinez, J. Leceta, R. P. Gomariz, and M. Delgado Pituitary Adenylate Cyclase-Activating Polypeptide Inhibits Collagen-Induced Arthritis: An Experimental Immunomodulatory Therapy J. Immunol., September 15, 2001; 167(6): 3182 - 3189. [Abstract] [Full Text] [PDF]

				N. M. Erhardt, E. A. Fradinger, L. A. Cervini, J. E. Rivier, and N. M. Sherwood Early Expression of Pituitary Adenylate Cyclase-Activating Polypeptide and Activation of its Receptor in Chick Neuroblasts Endocrinology, April 1, 2001; 142(4): 1616 - 1625. [Abstract] [Full Text]

				N. M. Sherwood, S. L. Krueckl, and J. E. McRory The Origin and Function of the Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP)/Glucagon Superfamily Endocr. Rev., December 1, 2000; 21(6): 619 - 670. [Abstract] [Full Text]

				F. El-Gehani, M. Tena-Sempere, and I. Huhtaniemi Evidence That Pituitary Adenylate Cyclase-Activating Polypeptide Is a Potent Regulator of Fetal Rat Testicular Steroidogenesis Biol Reprod, November 1, 2000; 63(5): 1482 - 1489. [Abstract] [Full Text]

				D. Vaudry, B. J. Gonzalez, M. Basille, L. Yon, A. Fournier, and H. Vaudry Pituitary Adenylate Cyclase-Activating Polypeptide and Its Receptors: From Structure to Functions Pharmacol. Rev., June 1, 2000; 52(2): 269 - 324. [Abstract] [Full Text] [PDF]

				M. Delgado, D. Pozo, C. Martinez, J. Leceta, J. R. Calvo, D. Ganea, and R. P. Gomariz Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide Inhibit Endotoxin-Induced TNF-{alpha} Production by Macrophages: In Vitro and In Vivo Studies J. Immunol., February 15, 1999; 162(4): 2358 - 2367. [Abstract] [Full Text] [PDF]

				M. B. P. Soares, R. G. Titus, C. B. Shoemaker, J. R. David, and M. Bozza The Vasoactive Peptide Maxadilan from Sand Fly Saliva Inhibits TNF-{alpha} and Induces IL-6 by Mouse Macrophages Through Interaction with the Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) Receptor J. Immunol., February 15, 1998; 160(4): 1811 - 1816. [Abstract] [Full Text] [PDF]