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Clinical and Diagnostic Laboratory Immunology, March 2001, p. 460-464, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.460-464.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Use of Hoechst 33342 Staining To Detect Apoptotic Changes in Bovine Mononuclear Phagocytes Infected with Mycobacterium avium subsp. paratuberculosis

S. Allen,1,2 J. Sotos,2 M. J. Sylte,2 and C. J. Czuprynski2,*

Department of Pathobiological Sciences, School of Veterinary Medicine,2 and Department of Bacteriology, College of Agriculture and Life Sciences,1 University of Wisconsin---Madison, Madison, Wisconsin 53706

Received 6 April 2000/Returned for modification 20 June 2000/Accepted 12 December 2000

Mycobacterium avium subsp. paratuberculosis is an intracellular pathogen of macrophages that causes a chronic enteritis (Johne's disease) in ruminants. The purpose of this study was to determine whether M. avium subsp. paratuberculosis infection causes apoptosis in bovine monocytes. Using Hoechst 33342 staining, we observed increased numbers of apoptotic monocytes within 6 h of infection with M. avium subsp. paratuberculosis, and these numbers increased further at 24 and 48 h. This effect appeared to require viable bacilli, because monocytes infected with heat-killed M. avium subsp. paratuberculosis did not exhibit a significant increase in apoptosis. Preincubation of monocytes with bovine growth hormone prior to infection with M. avium subsp. paratuberculosis did not significantly alter the number of apoptotic cells.

* Corresponding author. Mailing address: Department of Pathological Sciences, University of Wisconsin, 2015 Linden Dr. West, Madison, WI 53706. Phone and fax: (608) 262-8102. E-mail: czuprync{at}svm.vetmed.wisc.edu.

Clinical and Diagnostic Laboratory Immunology, March 2001, p. 460-464, Vol. 8, No. 2
1071-412X/01/$04.00+0   DOI: 10.1128/CDLI.8.2.460-464.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.