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Cellular Immune Responses to 35 kDa Recombinant Antigen of Mycobacterium avium paratuberculosis

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Abstract

Mycobacterium avium paratuberculosis is the causative agent of Johne disease, a chronic ulcerative intestinal condition in ruminant animals. Owing to the predominance of cellular response in subclinical forms of the infection, identification of M. a. paratuberculosis antigens eliciting host cell-mediated immune (CMI) reaction is crucial for early control of the disease. A 35 kDa protein of M. a. paratuberculosis was studied for its ability to elicit CMI responses using a mouse model. Lymphoproliferation and IFN-γ response were used to measure the CMI response. Recombinant 35 kDa protein (P35) stimulated proliferation of mouse mononuclear splenocytes sensitized with M. a. paratuberculosis. The P35 elicited increased nitrite production from mononuclear splenocytes from M. a. paratuberculosis-sensitized mice. In addition, RT-PCR-based semiquantitative IFN-γ measurement showed that stimulation with P35 is associated with significant expression of IFN-γ mRNA in M. a. paratuberculosis-sensitized mouse splenocytes. The results indicate that the 35 kDa protein of M. a. paratuberculosis is associated with CMI response in the host.

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Abbreviations

CMI:

cell-mediated immune response

PBS:

1 × phosphate buffered saline, pH 7.4

P35:

recombinant 35 kDa protein of M. a. paratuberculosis

RT-PCR:

reverse transcription–polymerase chain reaction

IFN-γ:

interferon gamma

ConA:

concanavalin A

IFA:

incomplete Freund's adjuvant

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

ST:

stimulation index

TNF-α:

tumour necrosis factor α

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Author notes

  1. S. H. Basagoudanavar

    Present address: Pennsylvania State University College of Medicine, Hershey, PA, USA

Authors and Affiliations

  1. National Biotechnology Centre, Indian Veterinary Research Institute (IVRI), Izatnagar, 243 122, India

    S. H. Basagoudanavar, P. P. Goswami & V. Tiwari

Authors
  1. S. H. Basagoudanavar
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  2. P. P. Goswami
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  3. V. Tiwari
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Corresponding author

Correspondence to P. P. Goswami.

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Basagoudanavar, S.H., Goswami, P.P. & Tiwari, V. Cellular Immune Responses to 35 kDa Recombinant Antigen of Mycobacterium avium paratuberculosis. Vet Res Commun 30, 357–367 (2006). https://doi.org/10.1007/s11259-006-3253-0

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  • DOI: https://doi.org/10.1007/s11259-006-3253-0

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