Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1997 Nov 1;100(9):2254–2262. doi: 10.1172/JCI119763

Pretreatment with antibody to eosinophil major basic protein prevents hyperresponsiveness by protecting neuronal M2 muscarinic receptors in antigen-challenged guinea pigs.

C M Evans 1, A D Fryer 1, D B Jacoby 1, G J Gleich 1, R W Costello 1
PMCID: PMC508421  PMID: 9410903

Abstract

In antigen-challenged guinea pigs there is recruitment of eosinophils into the lungs and to airway nerves, decreased function of inhibitory M2 muscarinic autoreceptors on parasympathetic nerves in the lungs, and airway hyperresponsiveness. A rabbit antibody to guinea pig eosinophil major basic protein was used to determine whether M2 muscarinic receptor dysfunction, and the subsequent hyperresponsiveness, are due to antagonism of the M2 receptor by eosinophil major basic protein. Guinea pigs were sensitized, challenged with ovalbumin and hyperresponsiveness, and M2 receptor function tested 24 h later with the muscarinic agonist pilocarpine. Antigen-challenged guinea pigs were hyperresponsive to electrical stimulation of the vagus nerves compared with controls. Likewise, loss of M2 receptor function was demonstrated since the agonist pilocarpine inhibited vagally-induced bronchoconstriction in control but not challenged animals. Pretreatment with rabbit antibody to guinea pig eosinophil major basic protein prevented hyperresponsiveness, and protected M2 receptor function in the antigen-challenged animals without inhibiting eosinophil accumulation in the lungs or around the nerves. Thus, hyperresponsiveness is a result of inhibition of neuronal M2 muscarinic receptor function by eosinophil major basic protein in antigen-challenged guinea pigs.

Full Text

The Full Text of this article is available as a PDF (322.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abu-Ghazaleh R. I., Fujisawa T., Mestecky J., Kyle R. A., Gleich G. J. IgA-induced eosinophil degranulation. J Immunol. 1989 Apr 1;142(7):2393–2400. [PubMed] [Google Scholar]
  2. Ayala L. E., Ahmed T. Is there loss of protective muscarinic receptor mechanism in asthma? Chest. 1989 Dec;96(6):1285–1291. doi: 10.1378/chest.96.6.1285. [DOI] [PubMed] [Google Scholar]
  3. Barker R. L., Gundel R. H., Gleich G. J., Checkel J. L., Loegering D. A., Pease L. R., Hamann K. J. Acidic polyamino acids inhibit human eosinophil granule major basic protein toxicity. Evidence of a functional role for ProMBP. J Clin Invest. 1991 Sep;88(3):798–805. doi: 10.1172/JCI115379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barnes P. J. Modulation of neurotransmission in airways. Physiol Rev. 1992 Jul;72(3):699–729. doi: 10.1152/physrev.1992.72.3.699. [DOI] [PubMed] [Google Scholar]
  5. Blaber L. C., Fryer A. D., Maclagan J. Neuronal muscarinic receptors attenuate vagally-induced contraction of feline bronchial smooth muscle. Br J Pharmacol. 1985 Nov;86(3):723–728. doi: 10.1111/j.1476-5381.1985.tb08951.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coyle A. J., Ackerman S. J., Burch R., Proud D., Irvin C. G. Human eosinophil-granule major basic protein and synthetic polycations induce airway hyperresponsiveness in vivo dependent on bradykinin generation. J Clin Invest. 1995 Apr;95(4):1735–1740. doi: 10.1172/JCI117850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Monchy J. G., Kauffman H. F., Venge P., Koëter G. H., Jansen H. M., Sluiter H. J., De Vries K. Bronchoalveolar eosinophilia during allergen-induced late asthmatic reactions. Am Rev Respir Dis. 1985 Mar;131(3):373–376. doi: 10.1164/arrd.1985.131.3.373. [DOI] [PubMed] [Google Scholar]
  8. Diamant Z., Timmers M. C., van der Veen H., Page C. P., van der Meer F. J., Sterk P. J. Effect of inhaled heparin on allergen-induced early and late asthmatic responses in patients with atopic asthma. Am J Respir Crit Care Med. 1996 Jun;153(6 Pt 1):1790–1795. doi: 10.1164/ajrccm.153.6.8665036. [DOI] [PubMed] [Google Scholar]
  9. Diaz P., Gonzalez M. C., Galleguillos F. R., Ancic P., Cromwell O., Shepherd D., Durham S. R., Gleich G. J., Kay A. B. Leukocytes and mediators in bronchoalveolar lavage during allergen-induced late-phase asthmatic reactions. Am Rev Respir Dis. 1989 Jun;139(6):1383–1389. doi: 10.1164/ajrccm/139.6.1383. [DOI] [PubMed] [Google Scholar]
  10. Dixon W. E. Contributions to the physiology of the lungs: Part I. The bronchial muscles, their innervation, and the action of drugs upon them. J Physiol. 1903 Mar 16;29(2):97–173. doi: 10.1113/jphysiol.1903.sp000947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Elbon C. L., Jacoby D. B., Fryer A. D. Pretreatment with an antibody to interleukin-5 prevents loss of pulmonary M2 muscarinic receptor function in antigen-challenged guinea pigs. Am J Respir Cell Mol Biol. 1995 Mar;12(3):320–328. doi: 10.1165/ajrcmb.12.3.7873198. [DOI] [PubMed] [Google Scholar]
  12. Frigas E., Loegering D. A., Gleich G. J. Cytotoxic effects of the guinea pig eosinophil major basic protein on tracheal epithelium. Lab Invest. 1980 Jan;42(1):35–43. [PubMed] [Google Scholar]
  13. Frigas E., Loegering D. A., Solley G. O., Farrow G. M., Gleich G. J. Elevated levels of the eosinophil granule major basic protein in the sputum of patients with bronchial asthma. Mayo Clin Proc. 1981 Jun;56(6):345–353. [PubMed] [Google Scholar]
  14. Fryer A. D., Costello R. W., Yost B. L., Lobb R. R., Tedder T. F., Steeber D. A., Bochner B. S. Antibody to VLA-4, but not to L-selectin, protects neuronal M2 muscarinic receptors in antigen-challenged guinea pig airways. J Clin Invest. 1997 Apr 15;99(8):2036–2044. doi: 10.1172/JCI119372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fryer A. D., Elbon C. L., Kim A. L., Xiao H. Q., Levey A. I., Jacoby D. B. Cultures of airway parasympathetic nerves express functional M2 muscarinic receptors. Am J Respir Cell Mol Biol. 1996 Dec;15(6):716–725. doi: 10.1165/ajrcmb.15.6.8969265. [DOI] [PubMed] [Google Scholar]
  16. Fryer A. D., Jacoby D. B. Function of pulmonary M2 muscarinic receptors in antigen-challenged guinea pigs is restored by heparin and poly-L-glutamate. J Clin Invest. 1992 Dec;90(6):2292–2298. doi: 10.1172/JCI116116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fryer A. D., Jacoby D. B. Parainfluenza virus infection damages inhibitory M2 muscarinic receptors on pulmonary parasympathetic nerves in the guinea-pig. Br J Pharmacol. 1991 Jan;102(1):267–271. doi: 10.1111/j.1476-5381.1991.tb12164.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fryer A. D., Maclagan J. Muscarinic inhibitory receptors in pulmonary parasympathetic nerves in the guinea-pig. Br J Pharmacol. 1984 Dec;83(4):973–978. doi: 10.1111/j.1476-5381.1984.tb16539.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Fryer A. D., Wills-Karp M. Dysfunction of M2-muscarinic receptors in pulmonary parasympathetic nerves after antigen challenge. J Appl Physiol (1985) 1991 Dec;71(6):2255–2261. doi: 10.1152/jappl.1991.71.6.2255. [DOI] [PubMed] [Google Scholar]
  20. Gleich G. J., Loegering D. A., Maldonado J. E. Identification of a major basic protein in guinea pig eosinophil granules. J Exp Med. 1973 Jun 1;137(6):1459–1471. doi: 10.1084/jem.137.6.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gross N. J., Skorodin M. S. Anticholinergic, antimuscarinic bronchodilators. Am Rev Respir Dis. 1984 May;129(5):856–870. doi: 10.1164/arrd.1984.129.5.856. [DOI] [PubMed] [Google Scholar]
  22. Ito Y., Yoshitomi T. Autoregulation of acetylcholine release from vagus nerve terminals through activation of muscarinic receptors in the dog trachea. Br J Pharmacol. 1988 Mar;93(3):636–646. doi: 10.1111/j.1476-5381.1988.tb10321.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jacoby D. B., Gleich G. J., Fryer A. D. Human eosinophil major basic protein is an endogenous allosteric antagonist at the inhibitory muscarinic M2 receptor. J Clin Invest. 1993 Apr;91(4):1314–1318. doi: 10.1172/JCI116331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jolobe O. M. Impaired atropine responsiveness in asthma: role of atopy. Respiration. 1983;44(2):97–102. doi: 10.1159/000194534. [DOI] [PubMed] [Google Scholar]
  25. Kita H., Weiler D. A., Abu-Ghazaleh R., Sanderson C. J., Gleich G. J. Release of granule proteins from eosinophils cultured with IL-5. J Immunol. 1992 Jul 15;149(2):629–635. [PubMed] [Google Scholar]
  26. Larsen G. L., Fame T. M., Renz H., Loader J. E., Graves J., Hill M., Gelfand E. W. Increased acetylcholine release in tracheas from allergen-exposed IgE-immune mice. Am J Physiol. 1994 Mar;266(3 Pt 1):L263–L270. doi: 10.1152/ajplung.1994.266.3.L263. [DOI] [PubMed] [Google Scholar]
  27. Lefort J., Nahori M. A., Ruffie C., Vargaftig B. B., Pretolani M. In vivo neutralization of eosinophil-derived major basic protein inhibits antigen-induced bronchial hyperreactivity in sensitized guinea pigs. J Clin Invest. 1996 Feb 15;97(4):1117–1121. doi: 10.1172/JCI118505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lewis D. M., Loegering D. A., Gleich G. J. Antiserum to the major basic protein of guinea pig eosinophil granules. Immunochemistry. 1976 Sep;13(9):743–746. doi: 10.1016/0019-2791(76)90194-4. [DOI] [PubMed] [Google Scholar]
  29. Maclagan J., Barnes P. J. Muscarinic pharmacology of the airways. Trends Pharmacol Sci. 1989 Dec;Suppl:88–92. [PubMed] [Google Scholar]
  30. Mauser P. J., Pitman A. M., Fernandez X., Foran S. K., Adams G. K., 3rd, Kreutner W., Egan R. W., Chapman R. W. Effects of an antibody to interleukin-5 in a monkey model of asthma. Am J Respir Crit Care Med. 1995 Aug;152(2):467–472. doi: 10.1164/ajrccm.152.2.7633694. [DOI] [PubMed] [Google Scholar]
  31. McCaig D. J. Comparison of autonomic responses in the trachea isolated from normal and albumin-sensitive guinea-pigs. Br J Pharmacol. 1987 Dec;92(4):809–816. doi: 10.1111/j.1476-5381.1987.tb11385.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Minette P. A., Barnes P. J. Prejunctional inhibitory muscarinic receptors on cholinergic nerves in human and guinea pig airways. J Appl Physiol (1985) 1988 Jun;64(6):2532–2537. doi: 10.1152/jappl.1988.64.6.2532. [DOI] [PubMed] [Google Scholar]
  33. Minette P. A., Lammers J. W., Dixon C. M., McCusker M. T., Barnes P. J. A muscarinic agonist inhibits reflex bronchoconstriction in normal but not in asthmatic subjects. J Appl Physiol (1985) 1989 Dec;67(6):2461–2465. doi: 10.1152/jappl.1989.67.6.2461. [DOI] [PubMed] [Google Scholar]
  34. Minette P. A., Lammers J. W., Dixon C. M., McCusker M. T., Barnes P. J. A muscarinic agonist inhibits reflex bronchoconstriction in normal but not in asthmatic subjects. J Appl Physiol (1985) 1989 Dec;67(6):2461–2465. doi: 10.1152/jappl.1989.67.6.2461. [DOI] [PubMed] [Google Scholar]
  35. Nadel J. A., Cabezas G. A., Austin J. H. In vivo roentgenographic examination of parasympathetic innervation of small airways. Use of powdered tantalum and a fine focal spot x-ray tube. Invest Radiol. 1971 Jan-Feb;6(1):9–17. doi: 10.1097/00004424-197101000-00002. [DOI] [PubMed] [Google Scholar]
  36. Roffel A. F., Elzinga C. R., Zaagsma J. Muscarinic M3 receptors mediate contraction of human central and peripheral airway smooth muscle. Pulm Pharmacol. 1990;3(1):47–51. doi: 10.1016/0952-0600(90)90009-8. [DOI] [PubMed] [Google Scholar]
  37. Rossi G. A., Crimi E., Lantero S., Gianiorio P., Oddera S., Crimi P., Brusasco V. Late-phase asthmatic reaction to inhaled allergen is associated with early recruitment of eosinophils in the airways. Am Rev Respir Dis. 1991 Aug;144(2):379–383. doi: 10.1164/ajrccm/144.2.379. [DOI] [PubMed] [Google Scholar]
  38. Schultheis A. H., Bassett D. J., Fryer A. D. Ozone-induced airway hyperresponsiveness and loss of neuronal M2 muscarinic receptor function. J Appl Physiol (1985) 1994 Mar;76(3):1088–1097. doi: 10.1152/jappl.1994.76.3.1088. [DOI] [PubMed] [Google Scholar]
  39. Sedgwick J. B., Calhoun W. J., Gleich G. J., Kita H., Abrams J. S., Schwartz L. B., Volovitz B., Ben-Yaakov M., Busse W. W. Immediate and late airway response of allergic rhinitis patients to segmental antigen challenge. Characterization of eosinophil and mast cell mediators. Am Rev Respir Dis. 1991 Dec;144(6):1274–1281. doi: 10.1164/ajrccm/144.6.1274. [DOI] [PubMed] [Google Scholar]
  40. Sedgwick J. B., Calhoun W. J., Vrtis R. F., Bates M. E., McAllister P. K., Busse W. W. Comparison of airway and blood eosinophil function after in vivo antigen challenge. J Immunol. 1992 Dec 1;149(11):3710–3718. [PubMed] [Google Scholar]
  41. Van Oosterhout A. J., Ladenius A. R., Savelkoul H. F., Van Ark I., Delsman K. C., Nijkamp F. P. Effect of anti-IL-5 and IL-5 on airway hyperreactivity and eosinophils in guinea pigs. Am Rev Respir Dis. 1993 Mar;147(3):548–552. doi: 10.1164/ajrccm/147.3.548. [DOI] [PubMed] [Google Scholar]
  42. Walters E. H., O'Byrne P. M., Graf P. D., Fabbri L. M., Nadel J. A. The responsiveness of airway smooth muscle in vitro from dogs with airway hyper-responsiveness in vivo. Clin Sci (Lond) 1986 Nov;71(5):605–611. doi: 10.1042/cs0710605. [DOI] [PubMed] [Google Scholar]
  43. Wassom D. L., Loegering D. A., Gleich G. J. Measurement of guinea pig eosinophil major basic protein by radioimmunoassay. Mol Immunol. 1979 Sep;16(9):711–719. doi: 10.1016/0161-5890(79)90012-9. [DOI] [PubMed] [Google Scholar]
  44. Wilson P. O., Barber P. C., Hamid Q. A., Power B. F., Dhillon A. P., Rode J., Day I. N., Thompson R. J., Polak J. M. The immunolocalization of protein gene product 9.5 using rabbit polyclonal and mouse monoclonal antibodies. Br J Exp Pathol. 1988 Feb;69(1):91–104. [PMC free article] [PubMed] [Google Scholar]
  45. Yu D. Y., Galant S. P., Gold W. M. Inhibition of antigen-induced bronchoconstriction by atropine in asthmatic patients. J Appl Physiol. 1972 Jun;32(6):823–828. doi: 10.1152/jappl.1972.32.6.823. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES