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. 1984 Oct;46(1):87–93. doi: 10.1128/iai.46.1.87-93.1984

Blastomyces dermatitidis chemotactic factor: kinetics of production and biological characterization evaluated by a modified neutrophil chemotaxis assay.

L M Thurmond, T G Mitchell
PMCID: PMC261425  PMID: 6480118

Abstract

Chemotactic activity for human polymorphonuclear neutrophils (PMNs) was detectable in culture filtrates (CFs) of Blastomyces dermatitidis and may have influenced the pathogenesis of blastomycosis. Production of this chemotaxin depended upon culture age and medium; peak levels were achieved after incubation for 17 days or more in minimal essential medium. This factor was also chemotactic for human monocytes. CF was temperature stable even after treatment at 100 degrees C for 60 min. The activity was stable under alkaline conditions but was destroyed below pH 7. Dialyzed, chemotactically active CF contained approximately 60 micrograms of carbohydrate per ml; total protein was estimated to be less than 0.8 micrograms/ml. Preincubation of PMNs with CF or N-formyl-L-methionyl-L-leucyl-L-phenylalanine deactivated their chemotactic response to each agent, whereas the chemotactic response to zymosan-activated serum was not affected. In addition, deactivation with N-formyl-L-methionyl-L-leucyl-L-phenylalanine reduced the response to CF. Pretreatment of CF with PMNs decreased chemotactic activity, which may reflect binding of the chemotaxin molecules to PMN receptors. A modified chemotaxis assay was developed in which commercial, disposable multiwell plates are used. This method was rapid, efficient, and inexpensive and permitted the assay of larger numbers of samples than was previously feasible with conventional chemotaxis methods.

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Selected References

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

  1. Axelsson L. G., Nilsson G., Björkstén B. Statistical aspects of cell motility determinations with a modified chemotaxis assembly for multiwell filter assays. J Immunol Methods. 1981;46(2):251–258. doi: 10.1016/0022-1759(81)90141-1. [DOI] [PubMed] [Google Scholar]
  2. Baker R. D. Tissue Reactions in Human Blastomycosis: An Analysis of Tissue from Twenty-Three Cases. Am J Pathol. 1942 May;18(3):479–497. [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Brass C., Volkmann C. M., Klein H. P., Halde C. J., Archibald R. W., Stevens D. A. Pathogen factors and host factors in murine pulmonary blastomycosis. Mycopathologia. 1982 Jun 18;78(3):129–140. doi: 10.1007/BF00466066. [DOI] [PubMed] [Google Scholar]
  5. Brummer E., Stevens D. A. Enhancing effect of murine polymorphonuclear neutrophils (PMN) on the multiplication of Blastomyces dermatitidis in vitro and in vivo. Clin Exp Immunol. 1983 Nov;54(2):587–594. [PMC free article] [PubMed] [Google Scholar]
  6. Brummer E., Stevens D. A. Opposite effects of human monocytes, macrophages, and polymorphonuclear neutrophils on replication of Blastomyces dermatitidis in vitro. Infect Immun. 1982 Apr;36(1):297–303. doi: 10.1128/iai.36.1.297-303.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cutler J. E. Chemotactic factor produced by Candida albicans. Infect Immun. 1977 Dec;18(3):568–573. doi: 10.1128/iai.18.3.568-573.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Donabedian H., Gallin J. I. Deactivation of human neutrophil chemotaxis by chemoattractants: effect on receptors for the chemotactic factor f-Met-Leu-Phe. J Immunol. 1981 Sep;127(3):839–844. [PubMed] [Google Scholar]
  9. Harvath L., Falk W., Leonard E. J. Rapid quantitation of neutrophil chemotaxis: use of a polyvinylpyrrolidone-free polycarbonate membrane in a multiwell assembly. J Immunol Methods. 1980;37(1):39–45. doi: 10.1016/0022-1759(80)90179-9. [DOI] [PubMed] [Google Scholar]
  10. Mitchell T. G., Turner S. R. Effects of culture filtrates of Blastomyces dermatitidis on neutrophil locomotion. Sabouraudia. 1984;22(2):145–153. [PubMed] [Google Scholar]
  11. Nelson R. D., McCormack R. T., Fiegel V. D., Herron M., Simmons R. L., Quie P. G. Chemotactic deactivation of human neutrophils: possible relationship to stimulation of oxidative metabolism. Infect Immun. 1979 Feb;23(2):282–286. doi: 10.1128/iai.23.2.282-286.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Neuman R. E., Ainsworth S. K. A simplified procedure for the assay of leukocyte chemotaxis. J Reticuloendothel Soc. 1980 Oct;28(4):305–312. [PubMed] [Google Scholar]
  13. O'Flaherty J. T., Kreutzer D. L., Showell H. J., Vitkauskas G., Becker E. L., Ward P. A. Selective neutrophil desensitization to chemotactic factors. J Cell Biol. 1979 Mar;80(3):564–572. doi: 10.1083/jcb.80.3.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Repine J. E., Clawson C. C., Rasp F. L., Jr, Sarosi G. A., Roidal J. R. Defective neutrophil locomotion in human blastomycosis: evidence for a serum inhibitor. Am Rev Respir Dis. 1978 Aug;118(2):325–334. doi: 10.1164/arrd.1978.118.2.325. [DOI] [PubMed] [Google Scholar]
  15. Sixbey J. W., Fields B. T., Sun C. N., Clark R. A., Nolan C. M. Interactions between human granulocytes and Blastomyces dermatitidis. Infect Immun. 1979 Jan;23(1):41–44. doi: 10.1128/iai.23.1.41-44.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sugar A. M., Chahal R. S., Brummer E., Stevens D. A. Susceptibility of Blastomyces dermatitidis strains to products of oxidative metabolism. Infect Immun. 1983 Sep;41(3):908–912. doi: 10.1128/iai.41.3.908-912.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Turner S. R. ACDAS: an automated chemotaxis data acquisition system. J Immunol Methods. 1979;28(3-4):355–360. doi: 10.1016/0022-1759(79)90200-x. [DOI] [PubMed] [Google Scholar]
  18. Zigmond S. H., Hirsch J. G. Leukocyte locomotion and chemotaxis. New methods for evaluation, and demonstration of a cell-derived chemotactic factor. J Exp Med. 1973 Feb 1;137(2):387–410. doi: 10.1084/jem.137.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]

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