Analysis of immune responses directed toward a recombinant early secretory antigenic target six-kilodalton protein-culture filtrate protein 10 fusion protein in Mycobacterium bovis-infected cattle

Alexander C Maue; W Ray Waters; William C Davis; Mitchell V Palmer; F Chris Minion; D Mark Estes

doi:10.1128/IAI.73.10.6659-6667.2005

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Analysis of immune responses directed toward a recombinant early secretory antigenic target six-kilodalton protein-culture filtrate protein 10 fusion protein in Mycobacterium bovis-infected cattle

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Analysis of immune responses directed toward a recombinant early secretory antigenic target six-kilodalton protein-culture filtrate protein 10 fusion protein in Mycobacterium bovis-infected cattle

Alexander C Maue, W Ray Waters, William C Davis, Mitchell V Palmer, F Chris Minion and D Mark Estes

Infection and immunity, Vol.73(10), pp.6659-6667

10/2005

DOI: https://doi.org/10.1128/IAI.73.10.6659-6667.2005

Handle:

https://hdl.handle.net/2376/113969

PMCID: PMC1230946

PMID: 16177342

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https://doi.org/10.1128/IAI.73.10.6659-6667.2005View

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Abstract

Recombinant Fusion Proteins - immunology

Up-Regulation

Dipeptidyl Peptidase 4 - metabolism

Cell Proliferation

Leukocyte Common Antigens - metabolism

Mycobacterium bovis - immunology

Tuberculosis, Bovine - immunology

Down-Regulation

Antigens, Bacterial - immunology

Bacterial Proteins - genetics

Receptors, Interleukin-2 - metabolism

Antigens, Bacterial - genetics

CD4-Positive T-Lymphocytes - immunology

Lymphocyte Activation - immunology

Animals

Leukocyte Common Antigens - analysis

Cattle

Bacterial Proteins - immunology

CD8-Positive T-Lymphocytes - immunology

Dipeptidyl Peptidase 4 - analysis

Receptors, Interleukin-2 - analysis

Cell-mediated immune responses are critical for protective immunity to mycobacterial infections. Recent progress in defining mycobacterial antigens has determined that region of difference 1 (RD1) gene products induce strong T-cell responses, particularly the early secretory antigenic target 6-kDa (ESAT-6) protein and culture filtrate protein 10 (CFP10). However, comprehensive analysis of the immune response towards these antigens is incompletely characterized. To evaluate recall responses to ESAT-6 and CFP10, peripheral blood mononuclear cells from M. bovis-infected cattle were stimulated in vitro with a recombinant ESAT-6 (rESAT-6)-CFP10 fusion protein and compared to responses induced by M. bovis-derived purified protein derivative. Following antigenic stimulation, activation marker expression was evaluated. Significant proliferative responses (P < 0.05) were evident in CD4(+), CD8(+), immunoglobulin M-positive, and CD172a(+) cell fractions after 6 days of culture. Expression of CD25 and CD26 was increased (P < 0.05) on CD4(+), CD8(+), and gammadelta T-cell-receptor-positive cells. CD4(+) and CD8(+) cells also exhibited significant changes (P < 0.05) in expression of CD45 isoforms. Using a flow cytometry-based proliferation assay, it was determined that CD45R expression is downregulated (P < 0.05) and that CD45RO expression is upregulated (P < 0.05) on proliferating (i.e., activated) CD4(+) cells. Collectively, data indicate that recall immune responses directed toward the rESAT-6-CFP10 fusion protein or purified protein derivative are comparable and that recall to mycobacterial antigens correlates with a CD45RO(+) phenotype.

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Title: Analysis of immune responses directed toward a recombinant early secretory antigenic target six-kilodalton protein-culture filtrate protein 10 fusion protein in Mycobacterium bovis-infected cattle
Creators: Alexander C Maue - Department of Molecular Microbiology and Immunology, University of Missouri, Columbia, 65211, USA
W Ray Waters
William C Davis
Mitchell V Palmer
F Chris Minion
D Mark Estes
Publication Details: Infection and immunity, Vol.73(10), pp.6659-6667
Academic Unit: Veterinary Microbiology and Pathology, Department of
Publisher: United States
Identifiers: 99900548241601842
Language: English
Resource Type: Journal article