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Infection and Immunity, December 2004, p. 6870-6883, Vol. 72, No. 12
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.12.6870-6883.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Analysis of the Immune Response to Mycobacterium avium subsp. paratuberculosis in Experimentally Infected Calves

Hye Cheong Koo,1 Yong Ho Park,1 Mary Jo Hamilton,2 George M. Barrington,3 Christopher J. Davies,2 Jong Bae Kim,4 John L. Dahl,5 W. Ray Waters,6 and William C. Davis2*

Department of Veterinary Microbiology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul,1 Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Republic of Korea,4 Department of Veterinary Microbiology and Pathology,2 Department of Veterinary Clinical Sciences, College of Veterinary Medicine,3 School of Molecular Biosciences, Washington State University, Pullman, Washington,5 National Animal Disease Center, USDA Agricultural Research Service, Ames, Iowa6

Received 8 May 2004/ Returned for modification 23 July 2004/ Accepted 14 August 2004

Johne's disease of cattle is widespread and causes significant economic loss to producers. Control has been hindered by limited understanding of the immune response to the causative agent, Mycobacterium avium subsp. paratuberculosis, and lack of an effective vaccine and sensitive specific diagnostic assays. The present study was conducted to gain insight into factors affecting the immune response to M. avium subsp. paratuberculosis. A persistent proliferative response to M. avium subsp. paratuberculosis purified protein derivative and soluble M. avium subsp. paratuberculosis antigens was detected in orally infected neonatal calves 6 months postinfection (p.i.) by flow cytometry (FC). CD4+ T cells with a memory phenotype (CD45R0+) expressing CD25 and CD26 were the predominant cell type responding to antigens. Few CD8+ T cells proliferated in response to antigens until 18 months p.i. {gamma}{delta} T cells did not appear to respond to antigen until 18 months p.i. The majority of WC1+ CD2– and a few WC1– CD2+ {gamma}{delta} T cells expressed CD25 at time zero. By 18 months, however, subsets of {gamma}{delta} T cells from both control and infected animals showed an increase in expression of CD25, ACT2, and CD26 in the presence of the antigens. Two populations of CD3– non-T non-B null cells, CD2+ and CD2–, proliferated in cell cultures from some control and infected animals during the study, with and without antigen. The studies clearly show multicolor FC offers a consistent reliable way to monitor the evolution and changes in the immune response to M. avium subsp. paratuberculosis that occur during disease progression.

* Corresponding author. Mailing address: Department of Veterinary Microbiology and Pathology, CVM, Washington State University, Pullman, WA 99164-7040. Phone: (509) 335-6051. Fax: (509) 335-8328. E-mail: davisw{at}vetmed.wsu.edu.

Editor: J. D. Clements

Infection and Immunity, December 2004, p. 6870-6883, Vol. 72, No. 12
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.12.6870-6883.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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