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Characterization of effector and memory T cell subsets in the immune response to bovine tuberculosis in cattle.

Maggioli MF, Palmer MV, Thacker TC, Vordermeier HM, Waters WR - PLoS ONE (2015)

Bottom Line: The objective of the present study was to characterize the relative contribution of Tcm (CCR7+, CD62Lhi, CD45RO+), T effector memory (Tem, defined as: CCR7-, CD62Llow/int, CD45RO+), and T effector cells (CCR7-, CD62L-/low, CD45RO-), in the immune response to Mycobacterium bovis.These findings suggest that proliferative responses of Tcm cells to some extent occurs simultaneously with reversion to effector phenotypes (mostly Tem).The present study characterizes Tcm cells of cattle and their participation in the response to M. bovis infection.

View Article: PubMed Central - PubMed

Affiliation: Infectious Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, Ames, IA, United States of America; Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Ames, IA, United States of America.

ABSTRACT
Cultured IFN-γ ELISPOT assays are primarily a measure of central memory T cell (Tcm) responses with humans; however, this important subset of lymphocytes is poorly characterized in cattle. Vaccine-elicited cultured IFN-γ ELISPOT responses correlate with protection against bovine tuberculosis in cattle. However, whether this assay measures cattle Tcm responses or not is uncertain. The objective of the present study was to characterize the relative contribution of Tcm (CCR7+, CD62Lhi, CD45RO+), T effector memory (Tem, defined as: CCR7-, CD62Llow/int, CD45RO+), and T effector cells (CCR7-, CD62L-/low, CD45RO-), in the immune response to Mycobacterium bovis. Peripheral blood mononuclear cells (PBMC) from infected cattle were stimulated with a cocktail of M. bovis purified protein derivative, rTb10.4 and rAg85A for 13 days with periodic addition of fresh media and rIL-2. On day 13, cultured PBMC were re-stimulated with medium alone, rESAT-6:CFP10 or PPDb with fresh autologous adherent cells for antigen presentation. Cultured cells (13 days) or fresh PBMCs (ex vivo response) from the same calves were analyzed for IFN-γ production, proliferation, and CD4, CD45RO, CD62L, CD44, and CCR7 expression via flow cytometry after overnight stimulation. In response to mycobacterial antigens, ~75% of CD4+ IFN-γ+ cells in long-term cultures expressed a Tcm phenotype while less than 10% of the ex vivo response consisted of Tcm cells. Upon re-exposure to antigen, long-term cultured cells were highly proliferative, a distinctive characteristic of Tcm, and the predominant phenotype within the long-term cultures switched from Tcm to Tem. These findings suggest that proliferative responses of Tcm cells to some extent occurs simultaneously with reversion to effector phenotypes (mostly Tem). The present study characterizes Tcm cells of cattle and their participation in the response to M. bovis infection.

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CD62L and CD44 expression by Tcm, Tem and effector CD4+ cells in long-term (14 day) cultures.Analysis of long-term cultured PBMCs was performed ~ 8 weeks after aerosol challenge with virulent M. bovis. Cells were stimulated with a cocktail of rAg85A (1 μg/ml), rTB10.4 (1 μg/ml), and rESAT-6:CFP10 (1 μg/ml), as well as PPDb (5 μg/ml) for 13 days followed by transfer to 96 well round bottom plates with APCs and addition of PPDb or rESAT-6:CFP10. Data are presented as mean fluorescence intensity (MFI, y-axis, ± SEM) of CD62L or CD44 by CD4+ cells of the various effector / memory subsets (x-axis). (A) CD62L expression by Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. (B) CD44 expression on Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. Tcm, Tem and effector cell phenotypes were as defined in Fig 2 and S3 Fig Paired Student's t-tests (n = 8).
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pone.0122571.g004: CD62L and CD44 expression by Tcm, Tem and effector CD4+ cells in long-term (14 day) cultures.Analysis of long-term cultured PBMCs was performed ~ 8 weeks after aerosol challenge with virulent M. bovis. Cells were stimulated with a cocktail of rAg85A (1 μg/ml), rTB10.4 (1 μg/ml), and rESAT-6:CFP10 (1 μg/ml), as well as PPDb (5 μg/ml) for 13 days followed by transfer to 96 well round bottom plates with APCs and addition of PPDb or rESAT-6:CFP10. Data are presented as mean fluorescence intensity (MFI, y-axis, ± SEM) of CD62L or CD44 by CD4+ cells of the various effector / memory subsets (x-axis). (A) CD62L expression by Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. (B) CD44 expression on Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. Tcm, Tem and effector cell phenotypes were as defined in Fig 2 and S3 Fig Paired Student's t-tests (n = 8).

Mentions: Tcm cells highly expressed (P < 0.05) CD62L and CD44 in response to either rESAT-6:CFP10 or PPDb stimulation (Fig 4). Expression of CD62L was intermediate with Tem and low to non-existent with effector cells (Fig 4A). CD44 expression was low in both Tem and effector cells (Fig 4B).


Characterization of effector and memory T cell subsets in the immune response to bovine tuberculosis in cattle.

Maggioli MF, Palmer MV, Thacker TC, Vordermeier HM, Waters WR - PLoS ONE (2015)

CD62L and CD44 expression by Tcm, Tem and effector CD4+ cells in long-term (14 day) cultures.Analysis of long-term cultured PBMCs was performed ~ 8 weeks after aerosol challenge with virulent M. bovis. Cells were stimulated with a cocktail of rAg85A (1 μg/ml), rTB10.4 (1 μg/ml), and rESAT-6:CFP10 (1 μg/ml), as well as PPDb (5 μg/ml) for 13 days followed by transfer to 96 well round bottom plates with APCs and addition of PPDb or rESAT-6:CFP10. Data are presented as mean fluorescence intensity (MFI, y-axis, ± SEM) of CD62L or CD44 by CD4+ cells of the various effector / memory subsets (x-axis). (A) CD62L expression by Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. (B) CD44 expression on Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. Tcm, Tem and effector cell phenotypes were as defined in Fig 2 and S3 Fig Paired Student's t-tests (n = 8).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4400046&req=5

pone.0122571.g004: CD62L and CD44 expression by Tcm, Tem and effector CD4+ cells in long-term (14 day) cultures.Analysis of long-term cultured PBMCs was performed ~ 8 weeks after aerosol challenge with virulent M. bovis. Cells were stimulated with a cocktail of rAg85A (1 μg/ml), rTB10.4 (1 μg/ml), and rESAT-6:CFP10 (1 μg/ml), as well as PPDb (5 μg/ml) for 13 days followed by transfer to 96 well round bottom plates with APCs and addition of PPDb or rESAT-6:CFP10. Data are presented as mean fluorescence intensity (MFI, y-axis, ± SEM) of CD62L or CD44 by CD4+ cells of the various effector / memory subsets (x-axis). (A) CD62L expression by Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. (B) CD44 expression on Tcm, Tem and effector cells in response to PPDb and rESAT-6:CFP10. Tcm, Tem and effector cell phenotypes were as defined in Fig 2 and S3 Fig Paired Student's t-tests (n = 8).
Mentions: Tcm cells highly expressed (P < 0.05) CD62L and CD44 in response to either rESAT-6:CFP10 or PPDb stimulation (Fig 4). Expression of CD62L was intermediate with Tem and low to non-existent with effector cells (Fig 4A). CD44 expression was low in both Tem and effector cells (Fig 4B).

Bottom Line: The objective of the present study was to characterize the relative contribution of Tcm (CCR7+, CD62Lhi, CD45RO+), T effector memory (Tem, defined as: CCR7-, CD62Llow/int, CD45RO+), and T effector cells (CCR7-, CD62L-/low, CD45RO-), in the immune response to Mycobacterium bovis.These findings suggest that proliferative responses of Tcm cells to some extent occurs simultaneously with reversion to effector phenotypes (mostly Tem).The present study characterizes Tcm cells of cattle and their participation in the response to M. bovis infection.

View Article: PubMed Central - PubMed

Affiliation: Infectious Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, Ames, IA, United States of America; Department of Veterinary Pathology, College of Veterinary Medicine, Iowa State University, Ames, IA, United States of America.

ABSTRACT
Cultured IFN-γ ELISPOT assays are primarily a measure of central memory T cell (Tcm) responses with humans; however, this important subset of lymphocytes is poorly characterized in cattle. Vaccine-elicited cultured IFN-γ ELISPOT responses correlate with protection against bovine tuberculosis in cattle. However, whether this assay measures cattle Tcm responses or not is uncertain. The objective of the present study was to characterize the relative contribution of Tcm (CCR7+, CD62Lhi, CD45RO+), T effector memory (Tem, defined as: CCR7-, CD62Llow/int, CD45RO+), and T effector cells (CCR7-, CD62L-/low, CD45RO-), in the immune response to Mycobacterium bovis. Peripheral blood mononuclear cells (PBMC) from infected cattle were stimulated with a cocktail of M. bovis purified protein derivative, rTb10.4 and rAg85A for 13 days with periodic addition of fresh media and rIL-2. On day 13, cultured PBMC were re-stimulated with medium alone, rESAT-6:CFP10 or PPDb with fresh autologous adherent cells for antigen presentation. Cultured cells (13 days) or fresh PBMCs (ex vivo response) from the same calves were analyzed for IFN-γ production, proliferation, and CD4, CD45RO, CD62L, CD44, and CCR7 expression via flow cytometry after overnight stimulation. In response to mycobacterial antigens, ~75% of CD4+ IFN-γ+ cells in long-term cultures expressed a Tcm phenotype while less than 10% of the ex vivo response consisted of Tcm cells. Upon re-exposure to antigen, long-term cultured cells were highly proliferative, a distinctive characteristic of Tcm, and the predominant phenotype within the long-term cultures switched from Tcm to Tem. These findings suggest that proliferative responses of Tcm cells to some extent occurs simultaneously with reversion to effector phenotypes (mostly Tem). The present study characterizes Tcm cells of cattle and their participation in the response to M. bovis infection.

Show MeSH
Related in: MedlinePlus