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. 1983 May;17(5):819–823. doi: 10.1128/jcm.17.5.819-823.1983

Agar microdroplet assay for delayed hypersensitivity to Legionella pneumophila serogroup 1.

R Widen, T Klein, I Lee, H Friedman
PMCID: PMC272747  PMID: 6863501

Abstract

An agarose microdroplet technique was utilized to assess the cellular immunity of guinea pig lymphoid cells to Legionella pneumophila antigen in vitro. Both direct and indirect migration inhibition procedures were shown to be capable of detecting sensitization of guinea pigs to L. pneumophila antigens. Animals injected with adjuvant alone or unrelated antigens did not yield spleen cells responsive to L. pneumophila, indicating the specificity of the response. Migration inhibition factor induction by Legionella antigen in vitro correlated well with skin test responses in vivo. The positive reaction detected by migration inhibition occurred at times similar to that of skin reactivity but later than that of the earliest serum antibody titers. The assay appears to be useful for monitoring sensitization to Legionella and may be applicable to the study of cell-mediated immunity to this bacterium in infected individuals.

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Selected References

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

  1. Bloom B. R., Bennett B. Mechanism of a reaction in vitro associated with delayed-type hypersensitivity. Science. 1966 Jul 1;153(3731):80–82. doi: 10.1126/science.153.3731.80. [DOI] [PubMed] [Google Scholar]
  2. Cherry W. B., Pittman B., Harris P. P., Hebert G. A., Thomason B. M., Thacker L., Weaver R. E. Detection of Legionnaires disease bacteria by direct immunofluorescent staining. J Clin Microbiol. 1978 Sep;8(3):329–338. doi: 10.1128/jcm.8.3.329-338.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. David J. R. Delayed hypersensitivity in vitro: its mediation by cell-free substances formed by lymphoid cell-antigen interaction. Proc Natl Acad Sci U S A. 1966 Jul;56(1):72–77. doi: 10.1073/pnas.56.1.72. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fraser D. W., Tsai T. R., Orenstein W., Parkin W. E., Beecham H. J., Sharrar R. G., Harris J., Mallison G. F., Martin S. M., McDade J. E. Legionnaires' disease: description of an epidemic of pneumonia. N Engl J Med. 1977 Dec 1;297(22):1189–1197. doi: 10.1056/NEJM197712012972201. [DOI] [PubMed] [Google Scholar]
  5. Freedman A. P., Katz S. M. The prevalence of serum antibodies to Legionella pneumophila in patients with chronic pulmonary disease. Am Rev Respir Dis. 1981 Feb;123(2):238–239. doi: 10.1164/arrd.1981.123.2.238. [DOI] [PubMed] [Google Scholar]
  6. Holliday M. G. The diagnosis of Legionnaires' disease by counterimmunoelectrophoresis. J Clin Pathol. 1980 Dec;33(12):1174–1178. doi: 10.1136/jcp.33.12.1174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Horwitz M. A., Silverstein S. C. Activated human monocytes inhibit the intracellular multiplication of Legionnaires' disease bacteria. J Exp Med. 1981 Nov 1;154(5):1618–1635. doi: 10.1084/jem.154.5.1618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Horwitz M. A., Silverstein S. C. Interaction of the legionnaires' disease bacterium (Legionella pneumophila) with human phagocytes. II. Antibody promotes binding of L. pneumophila to monocytes but does not inhibit intracellular multiplication. J Exp Med. 1981 Feb 1;153(2):398–406. doi: 10.1084/jem.153.2.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Horwitz M. A., Silverstein S. C. Legionnaires' disease bacterium (Legionella pneumophila) multiples intracellularly in human monocytes. J Clin Invest. 1980 Sep;66(3):441–450. doi: 10.1172/JCI109874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kishimoto R. A., Kastello M. D., White J. D., Shirey F. G., McGann V. G., Larson E. W., Hedlund K. W. In vitro interaction between normal cynolmolgus monkey alveolar macrophages and Legionnaires disease bacteria. Infect Immun. 1979 Aug;25(2):761–763. doi: 10.1128/iai.25.2.761-763.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kishimoto R. A., White J. D., Shirey F. G., McGann V. G., Berendt R. F., Larson E. W., Hedlund K. W. In vitro responses of guinea pig peritoneal macrophages to Legionella pneumophila. Infect Immun. 1981 Mar;31(3):1209–1213. doi: 10.1128/iai.31.3.1209-1213.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Klein G. C., Jones W. L., Feeley J. C. Upper limit of normal titer for detection of antibodies to Legionella pneumophila by the microagglutination test. J Clin Microbiol. 1979 Nov;10(5):754–755. doi: 10.1128/jcm.10.5.754-755.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pine L., George J. R., Reeves M. W., Harrell W. K. Development of a chemically defined liquid medium for growth of Legionella pneumophila. J Clin Microbiol. 1979 May;9(5):615–626. doi: 10.1128/jcm.9.5.615-626.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Plouffe J. F., Baird I. M. Lymphocyte transformation to Legionella pneumophila. J Clin Lab Immunol. 1981 May;5(3):149–152. [PubMed] [Google Scholar]
  15. Rocklin R. E., Bendtzen K., Greineder D. Mediators of immunity: lymphokines and monokines. Adv Immunol. 1980;29:55–136. doi: 10.1016/s0065-2776(08)60043-7. [DOI] [PubMed] [Google Scholar]
  16. Saravolatz L. D., Russell G., Cvitkovich D. Direct immunofluorescence in the diagnosis of Legionnaires' disease. Chest. 1981 May;79(5):566–570. doi: 10.1378/chest.79.5.566. [DOI] [PubMed] [Google Scholar]
  17. Vogel F. R., Klein T. W., Specter S. C., Hitchings M., Friedman H. Detection of antibodies to Legionella pneumophila in immune guinea pig serum by solid-phase immunofluorescence. J Clin Microbiol. 1981 Apr;13(4):726–729. doi: 10.1128/jcm.13.4.726-729.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wilkinson H. W., Cruce D. D., Broome C. V. Validation of Legionella pneumophila indirect immunofluorescence assay with epidemic sera. J Clin Microbiol. 1981 Jan;13(1):139–146. doi: 10.1128/jcm.13.1.139-146.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Winn W. C., Jr, Cherry W. B., Frank R. O., Casey C. A., Broome C. V. Direct immunofluorescent detection of Legionella pneumophila in respiratory specimens. J Clin Microbiol. 1980 Jan;11(1):59–64. doi: 10.1128/jcm.11.1.59-64.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Winn W. C., Jr, Glavin F. L., Perl D. P., Keller J. L., Andres T. L., Brown T. M., Coffin C. M., Sensecqua J. E., Roman L. N., Craighead J. E. The pathology of Legionnaires' disease. Fourteen fatal cases from the 1977 outbreak in Vermont. Arch Pathol Lab Med. 1978 Jul;102(7):344–350. [PubMed] [Google Scholar]

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