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. 1974 Nov;18(3):439–448.

Technical study of the leucocyte migration inhibition test in agarose. Application to PPD and to hepatitis B antigen

P Erard
PMCID: PMC1537987  PMID: 4619978

Abstract

A modification of the leucocyte migration inhibition test, in which cells migrate in an agarose layer, was proposed by Clausen (1971). In the present study, this method was applied to PPD and hepatitis B antigen (HBAg), and its advantages were reassessed.

From a technical standpoint, the migration in a monolayer facilitated a study of the morphology of the migrating cells. The proportion of the different kinds of migrating leucocytes was calculated and expressed as a function of the distance of migration. On the whole, 92% of the migrating leucocytes were polymorphonuclears and 8% lymphomonocytes. Three to 10% of the incubated cells were shown to migrate in the agarose layer. The surface area of the controls (migration without inhibition) was sufficiently constant and large (on the average 13 mm2) to allow a clear assessment of inhibition when antigen was added. In addition, the small requirement for antigen facilitated the use of this test in experimentation with purified HBAg.

Application of the technique to clinical problems revealed that, with PPD as antigen, a highly significant inhibition was obtained in PPD skin test-positive normal individuals (0·0025<P<0·005). But this inhibition was not significant (0·05<P<0·1) in PPD skin test-positive patients during the course of acute hepatitis B. With HBAg as antigen, a significant inhibition was obtained in individuals having recovered from acute hepatitis B. Such an inhibition was not demonstrable in individuals without a history of hepatitis, nor in patients during the course of the disease.

These results demonstrate the suitability of the leucocyte migration inhibition test in agarose for routine detection of cell-mediated immunity in man. They further suggest the development of cell-mediated immunity specific for HBAg after recovery from acute hepatitis B and impairment of this type of immune response to PPD during the acute phase of hepatitis.

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Selected References

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

  1. Clausen J. E. Migration inhibitory effect of cell-free supernatants from mixed human lymphocyte cultures. J Immunol. 1972 Feb;108(2):453–459. [PubMed] [Google Scholar]
  2. Clausen J. E. Migration inhibitory effect of cell-free supernatants from tuberculin-stimulated cultures of human mononuclear leukocytes demonstrated by two-step MIF agarose assay. J Immunol. 1973 Feb;110(2):546–551. [PubMed] [Google Scholar]
  3. Clausen J. E. Tuberculin-induced migration inhibition of human peripheral leucocytes in agarose medium. Acta Allergol. 1971 Feb;26(1):56–80. doi: 10.1111/j.1398-9995.1971.tb01399.x. [DOI] [PubMed] [Google Scholar]
  4. Dudley F. J., Giustino V., Sherlock S. Cell-mediated immunity in patients positive for hepatitis-associated antigen. Br Med J. 1972 Dec 30;4(5843):754–756. doi: 10.1136/bmj.4.5843.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Egeberg J., Bendixen G., Nerup J. Morphologic investigations on the leucocyte migration test (LMT). Scand J Immunol. 1972;1(3):205–216. doi: 10.1111/j.1365-3083.1972.tb01812.x. [DOI] [PubMed] [Google Scholar]
  6. Frei P. C., Erard P., Zinkernagel R. Cell-mediated immunity to hepatitis-associated antigen (HAA) demonstrated by leucocyte migration test during and after acute B hepatitis. Biomedicine. 1973 Sep 20;19(9):379–383. [PubMed] [Google Scholar]
  7. Haider S., Coutinho M. de L., Emond R. T., Sutton R. N. Tuberculin anergy and infectious mononucleosis. Lancet. 1973 Jul 14;2(7820):74–74. doi: 10.1016/s0140-6736(73)93265-0. [DOI] [PubMed] [Google Scholar]
  8. Hall C. B., Kantor F. S. Depression of established delayed hepersensitivity by mumps virus. J Immunol. 1972 Jan;108(1):81–85. [PubMed] [Google Scholar]
  9. In vitro methods in cell-mediated immunity: a progress report. Cell Immunol. 1973 Mar;6(3):331–347. doi: 10.1016/0008-8749(73)90034-8. [DOI] [PubMed] [Google Scholar]
  10. Maini R. N., Roffe L. M., Magrath I. T., Dumonde D. C. Standardization of the leucocyte migration test. Int Arch Allergy Appl Immunol. 1973;45(1):308–321. doi: 10.1159/000231048. [DOI] [PubMed] [Google Scholar]
  11. Olson G. B., Dent P. B., Rawls W. E., South M. A., Montgomery J. R., Melnick J. L., Good R. A. Abnormalities of in vitro lymphocyte responses during rubella virus infections. J Exp Med. 1968 Jul 1;128(1):47–68. doi: 10.1084/jem.128.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Reed W. D., Mitchell C. G., Eddleston A. L., Lee W. M., Williams R., Zuckerman A. J. Exposure and immunity to hepatitis-B virus in a liver unit. Lancet. 1974 Apr 6;1(7858):581–583. doi: 10.1016/s0140-6736(74)92646-4. [DOI] [PubMed] [Google Scholar]
  13. Soborg M., Bendixen G. Human lymphocyte migration as a parameter of hypersensitivity. Acta Med Scand. 1967 Feb;181(2):247–256. doi: 10.1111/j.0954-6820.1967.tb07255.x. [DOI] [PubMed] [Google Scholar]

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