Analysis of the antigen-specific IFN-γ producing T-cell subsets in cattle experimentally infected with Mycobacterium bovis

K. Walravens, V. Wellemans, V. Weynants, F. Boelaert, Vinciane De Brouchoven De Bergeyck, J. J. Letesson, K. Huygen, J. Godfroid

Research output: Contribution to journalArticle

Abstract

Three 10 months old cattle were infected by the intratracheal route with 106cfu of a field strain of Mycobacterium bovis. Blood samples were regularly collected for in vitro IFN-γ production after antigenic stimulation. Peripheral blood cells of infected animals produced IFN-γ in response to crude M. bovis antigens (live and heat-inactivated BCG and protein-purified derivative (PPD)) 3-4 weeks after infection. The ratio of the response to bovine PPD versus avian PPD indicated a specific sensitisation for M. bovis antigens. Three months post-infection (PI), animals were culled and M. bovis was cultured from tubercle lesions. At different time points, the frequency of specific M. bovis IFN-γ producing CD4+, CD8+ and WC1+ T-cells in the peripheral blood was examined by flow cytometry. Two colour immunofluorescence staining of intracellular IFN-γ and bovine cell surface molecules showed that both CD4+ and CD8+, but not WC1+, T-cells produced IFN-γ following stimulation with PPD, live or killed BCG.In two animals analysed, the relative percentage of circulating IFN-γ producing CD8+ cells decreased between week 5 and week 9 PI. The same evolution was not observed for IFN-γ secreting CD4+ cells. Magnetic positive selection of T-cells from infected animals showed that CD4+ T-cells produced specific IFN-γ only in the presence of antigen presenting cells (APCs). Positively selected CD8+ T-cells secreted IFN-γ only in the presence of recombinant human IL-2 and APCs. In vitro depletion of the CD4+ T-cells, but not the depletion of CD8+ or WC1+ T-cells, resulted in abrogation of the specific IFN-γ production showing the key role of this cell population for the specific IFN-γ production. Copyright © 2002 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)29-41
Number of pages13
JournalVeterinary immunology and immunopathology
Volume84
Issue number1-2
DOIs
Publication statusPublished - 2002

Fingerprint

Mycobacterium bovis
T-Lymphocyte Subsets
T-lymphocytes
antigens
T-Lymphocytes
Antigens
cattle
chemical derivatives
antigen-presenting cells
Antigen-Presenting Cells
animals
proteins
Avian Proteins
Infection
infection
cells
Proteins
blood
blood cells
interleukin-2

Keywords

  • Cattle
  • Flow cytometry
  • IFN-γ
  • Mycobacterium bovis
  • T-cell subsets
  • Tuberculosis

Cite this

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title = "Analysis of the antigen-specific IFN-γ producing T-cell subsets in cattle experimentally infected with Mycobacterium bovis",
abstract = "Three 10 months old cattle were infected by the intratracheal route with 106cfu of a field strain of Mycobacterium bovis. Blood samples were regularly collected for in vitro IFN-γ production after antigenic stimulation. Peripheral blood cells of infected animals produced IFN-γ in response to crude M. bovis antigens (live and heat-inactivated BCG and protein-purified derivative (PPD)) 3-4 weeks after infection. The ratio of the response to bovine PPD versus avian PPD indicated a specific sensitisation for M. bovis antigens. Three months post-infection (PI), animals were culled and M. bovis was cultured from tubercle lesions. At different time points, the frequency of specific M. bovis IFN-γ producing CD4+, CD8+ and WC1+ T-cells in the peripheral blood was examined by flow cytometry. Two colour immunofluorescence staining of intracellular IFN-γ and bovine cell surface molecules showed that both CD4+ and CD8+, but not WC1+, T-cells produced IFN-γ following stimulation with PPD, live or killed BCG.In two animals analysed, the relative percentage of circulating IFN-γ producing CD8+ cells decreased between week 5 and week 9 PI. The same evolution was not observed for IFN-γ secreting CD4+ cells. Magnetic positive selection of T-cells from infected animals showed that CD4+ T-cells produced specific IFN-γ only in the presence of antigen presenting cells (APCs). Positively selected CD8+ T-cells secreted IFN-γ only in the presence of recombinant human IL-2 and APCs. In vitro depletion of the CD4+ T-cells, but not the depletion of CD8+ or WC1+ T-cells, resulted in abrogation of the specific IFN-γ production showing the key role of this cell population for the specific IFN-γ production. Copyright {\circledC} 2002 Elsevier Science B.V.",
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Analysis of the antigen-specific IFN-γ producing T-cell subsets in cattle experimentally infected with Mycobacterium bovis. / Walravens, K.; Wellemans, V.; Weynants, V.; Boelaert, F.; De Brouchoven De Bergeyck, Vinciane; Letesson, J. J.; Huygen, K.; Godfroid, J.

In: Veterinary immunology and immunopathology, Vol. 84, No. 1-2, 2002, p. 29-41.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis of the antigen-specific IFN-γ producing T-cell subsets in cattle experimentally infected with Mycobacterium bovis

AU - Walravens, K.

AU - Wellemans, V.

AU - Weynants, V.

AU - Boelaert, F.

AU - De Brouchoven De Bergeyck, Vinciane

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AU - Huygen, K.

AU - Godfroid, J.

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AB - Three 10 months old cattle were infected by the intratracheal route with 106cfu of a field strain of Mycobacterium bovis. Blood samples were regularly collected for in vitro IFN-γ production after antigenic stimulation. Peripheral blood cells of infected animals produced IFN-γ in response to crude M. bovis antigens (live and heat-inactivated BCG and protein-purified derivative (PPD)) 3-4 weeks after infection. The ratio of the response to bovine PPD versus avian PPD indicated a specific sensitisation for M. bovis antigens. Three months post-infection (PI), animals were culled and M. bovis was cultured from tubercle lesions. At different time points, the frequency of specific M. bovis IFN-γ producing CD4+, CD8+ and WC1+ T-cells in the peripheral blood was examined by flow cytometry. Two colour immunofluorescence staining of intracellular IFN-γ and bovine cell surface molecules showed that both CD4+ and CD8+, but not WC1+, T-cells produced IFN-γ following stimulation with PPD, live or killed BCG.In two animals analysed, the relative percentage of circulating IFN-γ producing CD8+ cells decreased between week 5 and week 9 PI. The same evolution was not observed for IFN-γ secreting CD4+ cells. Magnetic positive selection of T-cells from infected animals showed that CD4+ T-cells produced specific IFN-γ only in the presence of antigen presenting cells (APCs). Positively selected CD8+ T-cells secreted IFN-γ only in the presence of recombinant human IL-2 and APCs. In vitro depletion of the CD4+ T-cells, but not the depletion of CD8+ or WC1+ T-cells, resulted in abrogation of the specific IFN-γ production showing the key role of this cell population for the specific IFN-γ production. Copyright © 2002 Elsevier Science B.V.

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