Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1966 Jan 1;123(1):173–190. doi: 10.1084/jem.123.1.173

ISOLATION OF RABBIT IGA ANTIHAPTEN ANTIBODY AND DEMONSTRATION OF SKIN-SENSITIZING ACTIVITY IN HOMOLOGOUS SKIN

Kaoru Onoue 1, Yasuo Yagi 1, David Pressman 1
PMCID: PMC2138123  PMID: 4159250

Abstract

Multiple antibody components of rabbit antisera against p-azobenzenearsonate (Rp) were studied with respect to their globulin nature and skin-sensitizing activity. IgA antibody was characterized by isolating two IgA-rich fractions from a specifically purified antibody preparation. Examination of these fractions showed that IgA antibodies existed in two molecular forms, one with a sedimentation constant of 7S and the other 9S. Skin-sensitizing activity was examined by a P-K type test and a PCA test with Rp-rabbit serum albumin in homologous (rabbit) species. Only the 7S but not 9S IgA antibody sensitized rabbit skin. IgM antibody showed no activity and IgG antibody showed very low activity. In contrast, only IgG antibody was active in the P-K type test to sensitize a heterologous species (guinea pig). None of the antibodies of other classes showed sensitizing activity in heterologous skin. The 7S IgA antibody lost its sensitizing activity upon reduction and alkylation, although no change in its molecular size could be observed. The loss of sensitizing activity was not due to the destruction of antigen-binding activity since the treated 7S IgA antibody retained this activity as shown by radioimmunoelectrophoresis and by binding to the specific immunoadsorbent. The 9S IgA antibody was more resistant to these treatments than the IgM antibody and showed no indication of dissociation. The treated 9S IgA also retained antigen-binding activity. Both the P-K type and PCA reactions were considerably stronger when the interval between injections of antibody and antigen was 24 hr rather than 4 to 5 hr.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. ANDERSEN B. R., VANNIER W. E. THE SEDIMENTATION PROPERTIES OF THE SKIN-SENSITIZING ANTIBODIES OF RAGWEED-SENSITIVE PATIENTS. J Exp Med. 1964 Jul 1;120:31–43. doi: 10.1084/jem.120.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BIOZZI G., HALPERN B. N., BINAGHI R. The competitive effect of normal serum proteins from various animal species on antibody fixation in passive cutaneous anaphylaxis in the guinea pig. J Immunol. 1959 Mar;82(3):215–218. [PubMed] [Google Scholar]
  3. BLOCH K. J., KOURILSKY F. M., OVARY Z., BENACERRAF B. Properties of guinea pig 7S antibodies. III. Identification of antibodies involved in complement fixation and hemolysis. J Exp Med. 1963 Jun 1;117:965–981. doi: 10.1084/jem.117.6.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barth W. F., Fahey J. L. Heterologous and homologous skin-sensitizing activity of mouse 7S gamma 1- and 7S gamma 2-globulins. Nature. 1965 May 15;206(985):730–731. doi: 10.1038/206730a0. [DOI] [PubMed] [Google Scholar]
  5. DEUTSCH H. F. MOLECULAR TRANSFORMATIONS OF A GAMMA1-GLOBULIN OF HUMAN SERUM. J Mol Biol. 1963 Dec;7:662–671. doi: 10.1016/s0022-2836(63)80114-x. [DOI] [PubMed] [Google Scholar]
  6. FAHEY J. L., MCKELVEY E. M. QUANTITATIVE DETERMINATION OF SERUM IMMUNOGLOBULINS IN ANTIBODY-AGAR PLATES. J Immunol. 1965 Jan;94:84–90. [PubMed] [Google Scholar]
  7. FIREMAN P., VANNIER W. E., GOODMAN H. C. The association of skin-sensitizing antibody with the beta 2A-globulins in sera from ragweed-sensitive patients. J Exp Med. 1963 Apr 1;117:603–620. doi: 10.1084/jem.117.4.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. FISHER J. P., COOKE R. A. Passive cutaneous anaphylaxis (PCA) in the guinea pig; an immunologic and pathologic study. J Allergy. 1957 Mar;28(2):150–169. doi: 10.1016/0021-8707(57)90021-7. [DOI] [PubMed] [Google Scholar]
  9. FRANKLIN E. C., OVARY Z. ON THE SENSITIZING PROPERTIES OF SOME NORMAL AND PATHOLOGIC HUMAN IMMUNE GLOBULINS AND FRAGMENTS OBTAINED BY PAPAIN OF PEPSIN DIGESTION. Immunology. 1963 Sep;6:434–438. [PMC free article] [PubMed] [Google Scholar]
  10. FRANKLIN E. C. Two types of gamma-1A-globulin in sera from normals and patients with multiple myeloma. Nature. 1962 Jul 28;195:393–394. doi: 10.1038/195393a0. [DOI] [PubMed] [Google Scholar]
  11. Fahey J. L. HETEROGENEITY OF MYELOMA PROTEINS. J Clin Invest. 1963 Jan;42(1):111–123. doi: 10.1172/JCI104688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. GELOTTE B., FLODIN P., KILLANDER J. Fractionation of human plasma proteins by gel filtration and zone electrophoresis or ion-exchange chromatography. Arch Biochem Biophys. 1962 Sep;Suppl 1:319–326. [PubMed] [Google Scholar]
  13. GROSSBERG A. L., RADZIMSKI G., PRESSMAN D. Effect of iodination on the active site of several antihapten antibodies. Biochemistry. 1962 May 25;1:391–401. doi: 10.1021/bi00909a004. [DOI] [PubMed] [Google Scholar]
  14. HJERTEN S. Agarose as an anticonvection agent in zone electrophoresis. Biochim Biophys Acta. 1961 Nov 11;53:514–517. doi: 10.1016/0006-3002(61)90210-4. [DOI] [PubMed] [Google Scholar]
  15. ISHIZAKA K., DENNIS E. G., HORNBROOK M. PRESENCE OF REAGIN AND GAMMA-GLOBULIN IN SALIVA. J Allergy. 1964 Mar-Apr;35:143–148. doi: 10.1016/0021-8707(64)90028-0. [DOI] [PubMed] [Google Scholar]
  16. ISHIZAKA K., ISHIZAKA T., HORNBROOK M. M. BLOCKING OF PRAUSNITZ-KUESTNER SENSITIZATION WITH REAGIN BY NORMAL HUMAN BETA-2A GLOBULIN. J Allergy. 1963 Sep-Oct;34:395–403. doi: 10.1016/0021-8707(63)90002-9. [DOI] [PubMed] [Google Scholar]
  17. LEDDY J. P., FREEMAN G. L., LUZ A., TODD R. H. Inactivation of the skin-sensitizing antibodies of human allergy by thiols. Proc Soc Exp Biol Med. 1962 Oct;111:7–12. doi: 10.3181/00379727-111-27690. [DOI] [PubMed] [Google Scholar]
  18. NUSSENZWEIG R. S., MERRYMAN C., BENACERRAF B. ELECTROPHORETIC SEPARATION AND PROPERTIES OF MOUSE ANTIHAPTEN ANTIBODIES INVOLVED IN PASSIVE CUTANEOUS ANAPHYLAXIS AND PASSIVE HEMOLYSIS. J Exp Med. 1964 Aug 1;120:315–328. doi: 10.1084/jem.120.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. ONOUE K., YAGI Y., PRESSMAN D. MULTIPLICITY OF ANTIBODY PROTEINS IN RABBIT ANTI-P-AZOBENZENEARSONATE SERA. J Immunol. 1964 Feb;92:173–184. [PubMed] [Google Scholar]
  20. OVARY Z., BARTH W. F., FAHEY J. L. THE IMMUNOGLOBULINS OF MICE. 3. SKIN SENSITIZING ACTIVITY OF MOUSE IMMUNOGLOBULINS. J Immunol. 1965 Mar;94:410–415. [PubMed] [Google Scholar]
  21. OVARY Z., BENACERRAF B., BLOCH K. J. Properties of guinea pig 7S antibodies. II. Identification of antibodies involved in passive cutaneous and systemic anaphylaxis. J Exp Med. 1963 Jun 1;117:951–964. doi: 10.1084/jem.117.6.951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. OVARY Z., BIER O. G. Action empêchante du sérum normal de lapin sur l'anaphylaxie cutanée passive du cobaye. Ann Inst Pasteur (Paris) 1953 Feb;84(2):443–445. [PubMed] [Google Scholar]
  23. OVARY Z., FUDENBERG H., KUNKEL H. G. Anuphylactic reactions in the skin of the guinea pig with high and low molecular weight antibodies and gamma globulins. J Exp Med. 1960 Nov 1;112:953–961. doi: 10.1084/jem.112.5.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. OVARY Z. Immediate reactions in the skin of experimental animals provoked by antibody-antigen interaction. Prog Allergy. 1958;5:459–508. [PubMed] [Google Scholar]
  25. Onoue K., Tanigaki N., Yagi Y., Pressman D. IgM and IgG anti-hapten antibody: hemolytic, hemagglutinating and precipitating activity. Proc Soc Exp Biol Med. 1965 Nov;120(2):340–346. doi: 10.3181/00379727-120-30531. [DOI] [PubMed] [Google Scholar]
  26. Onoue K., Yagi Y., Pressman D. Immunoadsorbents with high capacity. Immunochemistry. 1965 Jun;2(2):181–194. doi: 10.1016/0019-2791(65)90020-0. [DOI] [PubMed] [Google Scholar]
  27. TABACHNICK M., SOBOTKA H. Azoproteins. II. A spectrophotometric study of the coupling of diazotized arsanilic acid with proteins. J Biol Chem. 1960 Apr;235:1051–1054. [PubMed] [Google Scholar]
  28. TERR A. I., BENTZ J. D. DENSITY-GRADIENT SEDIMENTATION OF SKIN-SENSITIZING ANTIBODY AND BETA-2A-GLOBULIN IN SERUM OF ALLERGIC INDIVIDUALS. Proc Soc Exp Biol Med. 1964 Mar;115:721–723. doi: 10.3181/00379727-115-29018. [DOI] [PubMed] [Google Scholar]
  29. TERR A. I., BENTZ J. D. GEL FILTRATION OF HUMAN SKIN-SENSITIZING ANTIBODY AND BETA 2A-GLOBULIN. J Allergy. 1964 May-Jun;35:206–214. doi: 10.1016/0021-8707(64)90003-6. [DOI] [PubMed] [Google Scholar]
  30. VAERMAN J. P., EPSTEIN W., FUDENBERG H., ISHIZAKA K. DIRECT DEMONSTRATION OF REAGIN ACTIVITY IN PURIFIED GAMMA 1A GLOBULIN. Nature. 1964 Sep 5;203:1046–1048. doi: 10.1038/2031046a0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES