Limits...
Distinct differences in the expansion and phenotype of TB10.4 specific CD8 and CD4 T cells after infection with Mycobacterium tuberculosis.

Hoang TT, Nansen A, Roy S, Billeskov R, Aagaard C, Elvang T, Dietrich J, Andersen P - PLoS ONE (2009)

Bottom Line: Recently we and others have identified CD8 and CD4 T cell epitopes within the highly expressed M. tuberculosis protein TB10.4.We focused on T cells directed to two epitopes in TB10.4; the MHC class I restricted epitope TB10.4 (3-11) (CD8/10.4 T cells) and the MHC class II restricted epitope TB10.4 (74-88) (CD4/10.4 T cells).In addition, the observed strong expansion of CD8 T cells in the late stages of infection could have implications for the development of post exposure vaccines against latent TB.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Disease Immunology, Statens Serum Institut, Copenhagen, Denmark.

ABSTRACT

Background: Recently we and others have identified CD8 and CD4 T cell epitopes within the highly expressed M. tuberculosis protein TB10.4. This has enabled, for the first time, a comparative study of the dynamics and function of CD4 and CD8 T cells specific for epitopes within the same protein in various stages of TB infection.

Methods and findings: We focused on T cells directed to two epitopes in TB10.4; the MHC class I restricted epitope TB10.4 (3-11) (CD8/10.4 T cells) and the MHC class II restricted epitope TB10.4 (74-88) (CD4/10.4 T cells). CD4/10.4 and CD8/10.4 T cells displayed marked differences in terms of expansion and contraction in a mouse TB model. CD4/10.4 T cells dominated in the early phase of infection whereas CD8/10.4 T cells were expanded after week 16 and reached 5-8 fold higher numbers in the late phase of infection. In the early phase of infection both CD4/10.4 and CD8/10.4 T cells were characterized by 20-25% polyfunctional cells (IL-2(+), IFN-gamma(+), TNF-alpha(+)), but whereas the majority of CD4/10.4 T cells were maintained as polyfunctional T cells throughout infection, CD8/10.4 T cells differentiated almost exclusively into effector cells (IFN-gamma(+), TNF-alpha(+)). Both CD4/10.4 and CD8/10.4 T cells exhibited cytotoxicity in vivo in the early phase of infection, but whereas CD4/10.4 cell mediated cytotoxicity waned during the infection, CD8/10.4 T cells exhibited increasing cytotoxic potential throughout the infection.

Conclusions/significance: Our results show that CD4 and CD8 T cells directed to epitopes in the same antigen differ both in their kinetics and functional characteristics throughout an infection with M. tuberculosis. In addition, the observed strong expansion of CD8 T cells in the late stages of infection could have implications for the development of post exposure vaccines against latent TB.

Show MeSH

Related in: MedlinePlus

Changes in cytokine profiles of CD4/10.4 and CD8/10.4 T cells during a chronic infection.Cells from infected lungs were stimulated with TB10.4 3–11 or TB10.4 74–88 prior to staining with anti-CD4, -CD8, -IFN-γ, -TNF-α and –IL-2. (A) Cytokine profiles of CD4/10.4 was determined by first dividing the CD4 T cells into IFN-γ positive (+) or IFN-γ negative (−) cells. Both the IFN-γ+ and IFN-γ- cells were analyzed with respect to the production of TNF-α and IL-2. The numbers in the quadrant gates of the plots denominates each distinct population based on their cytokine production and is color coded as shown. To illustrate the gating sequence two examples from a late infection stage are shown. (B and C). The pie-charts illustrate the relative contribution of the different cytokine T cell populations to the total CD4/10.4 population and data is depicted for both an early and a late time point, for 3 mice at each time point. Background (>0.5%) has been deducted. (D) The proportion of IFN-γ+ TNF-α+ IL-2+ within the CD4/10.4 or CD8/10.4 T cells populations in the early stage compared to the late stage of infection.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2691482&req=5

pone-0005928-g005: Changes in cytokine profiles of CD4/10.4 and CD8/10.4 T cells during a chronic infection.Cells from infected lungs were stimulated with TB10.4 3–11 or TB10.4 74–88 prior to staining with anti-CD4, -CD8, -IFN-γ, -TNF-α and –IL-2. (A) Cytokine profiles of CD4/10.4 was determined by first dividing the CD4 T cells into IFN-γ positive (+) or IFN-γ negative (−) cells. Both the IFN-γ+ and IFN-γ- cells were analyzed with respect to the production of TNF-α and IL-2. The numbers in the quadrant gates of the plots denominates each distinct population based on their cytokine production and is color coded as shown. To illustrate the gating sequence two examples from a late infection stage are shown. (B and C). The pie-charts illustrate the relative contribution of the different cytokine T cell populations to the total CD4/10.4 population and data is depicted for both an early and a late time point, for 3 mice at each time point. Background (>0.5%) has been deducted. (D) The proportion of IFN-γ+ TNF-α+ IL-2+ within the CD4/10.4 or CD8/10.4 T cells populations in the early stage compared to the late stage of infection.

Mentions: We next looked at the co-expression of multiple cytokines in CD8/10.4 and CD4/10.4 T cells. Cells from infected lungs, taken at week 6 or week 43 post infection, were stimulated with TB10.4 3–11 or TB10.4 74–88 prior to staining with anti-CD4, -CD8, -IFN-γ, -TNF-α and –IL-2. The results showed that TNF-α/IFN-γ effector cells constituted the major population observed among both CD4 and CD8/10.4 T cells (Fig. 5A–C), reflecting an ongoing stimulation in the M.tb infected animals, and in agreement with the results shown in figure 4. However, in terms of polyfunctionality, i.e. the co-expression of multiple cytokines such as TNF-α, IFN-γ and IL-2, we observed a difference between the two T cell populations as the infection progressed. Thus, at the late time point there was still a significant proportion of the CD4/10.4 cells that were polyfunctional in contrast to the CD8/10.4 cells (Fig. 5B and C). For the CD8/10.4 T cell population the proportion of TNF-α, IFN-γ and IL-2 co-expressing cells decreased from around 25% during the early timepoint to 0.5% during the late timepoint. However, the proportion of polyfunctional CD4/10.4 T cells did not change significantly despite the ongoing infection (Fig. 5D). Taken together, these results showed that in contrast to the CD8/10.4 T cells, a considerable proportion of the CD4/10.4 T cells were maintained as polyfunctional T cells.


Distinct differences in the expansion and phenotype of TB10.4 specific CD8 and CD4 T cells after infection with Mycobacterium tuberculosis.

Hoang TT, Nansen A, Roy S, Billeskov R, Aagaard C, Elvang T, Dietrich J, Andersen P - PLoS ONE (2009)

Changes in cytokine profiles of CD4/10.4 and CD8/10.4 T cells during a chronic infection.Cells from infected lungs were stimulated with TB10.4 3–11 or TB10.4 74–88 prior to staining with anti-CD4, -CD8, -IFN-γ, -TNF-α and –IL-2. (A) Cytokine profiles of CD4/10.4 was determined by first dividing the CD4 T cells into IFN-γ positive (+) or IFN-γ negative (−) cells. Both the IFN-γ+ and IFN-γ- cells were analyzed with respect to the production of TNF-α and IL-2. The numbers in the quadrant gates of the plots denominates each distinct population based on their cytokine production and is color coded as shown. To illustrate the gating sequence two examples from a late infection stage are shown. (B and C). The pie-charts illustrate the relative contribution of the different cytokine T cell populations to the total CD4/10.4 population and data is depicted for both an early and a late time point, for 3 mice at each time point. Background (>0.5%) has been deducted. (D) The proportion of IFN-γ+ TNF-α+ IL-2+ within the CD4/10.4 or CD8/10.4 T cells populations in the early stage compared to the late stage of infection.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2691482&req=5

pone-0005928-g005: Changes in cytokine profiles of CD4/10.4 and CD8/10.4 T cells during a chronic infection.Cells from infected lungs were stimulated with TB10.4 3–11 or TB10.4 74–88 prior to staining with anti-CD4, -CD8, -IFN-γ, -TNF-α and –IL-2. (A) Cytokine profiles of CD4/10.4 was determined by first dividing the CD4 T cells into IFN-γ positive (+) or IFN-γ negative (−) cells. Both the IFN-γ+ and IFN-γ- cells were analyzed with respect to the production of TNF-α and IL-2. The numbers in the quadrant gates of the plots denominates each distinct population based on their cytokine production and is color coded as shown. To illustrate the gating sequence two examples from a late infection stage are shown. (B and C). The pie-charts illustrate the relative contribution of the different cytokine T cell populations to the total CD4/10.4 population and data is depicted for both an early and a late time point, for 3 mice at each time point. Background (>0.5%) has been deducted. (D) The proportion of IFN-γ+ TNF-α+ IL-2+ within the CD4/10.4 or CD8/10.4 T cells populations in the early stage compared to the late stage of infection.
Mentions: We next looked at the co-expression of multiple cytokines in CD8/10.4 and CD4/10.4 T cells. Cells from infected lungs, taken at week 6 or week 43 post infection, were stimulated with TB10.4 3–11 or TB10.4 74–88 prior to staining with anti-CD4, -CD8, -IFN-γ, -TNF-α and –IL-2. The results showed that TNF-α/IFN-γ effector cells constituted the major population observed among both CD4 and CD8/10.4 T cells (Fig. 5A–C), reflecting an ongoing stimulation in the M.tb infected animals, and in agreement with the results shown in figure 4. However, in terms of polyfunctionality, i.e. the co-expression of multiple cytokines such as TNF-α, IFN-γ and IL-2, we observed a difference between the two T cell populations as the infection progressed. Thus, at the late time point there was still a significant proportion of the CD4/10.4 cells that were polyfunctional in contrast to the CD8/10.4 cells (Fig. 5B and C). For the CD8/10.4 T cell population the proportion of TNF-α, IFN-γ and IL-2 co-expressing cells decreased from around 25% during the early timepoint to 0.5% during the late timepoint. However, the proportion of polyfunctional CD4/10.4 T cells did not change significantly despite the ongoing infection (Fig. 5D). Taken together, these results showed that in contrast to the CD8/10.4 T cells, a considerable proportion of the CD4/10.4 T cells were maintained as polyfunctional T cells.

Bottom Line: Recently we and others have identified CD8 and CD4 T cell epitopes within the highly expressed M. tuberculosis protein TB10.4.We focused on T cells directed to two epitopes in TB10.4; the MHC class I restricted epitope TB10.4 (3-11) (CD8/10.4 T cells) and the MHC class II restricted epitope TB10.4 (74-88) (CD4/10.4 T cells).In addition, the observed strong expansion of CD8 T cells in the late stages of infection could have implications for the development of post exposure vaccines against latent TB.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious Disease Immunology, Statens Serum Institut, Copenhagen, Denmark.

ABSTRACT

Background: Recently we and others have identified CD8 and CD4 T cell epitopes within the highly expressed M. tuberculosis protein TB10.4. This has enabled, for the first time, a comparative study of the dynamics and function of CD4 and CD8 T cells specific for epitopes within the same protein in various stages of TB infection.

Methods and findings: We focused on T cells directed to two epitopes in TB10.4; the MHC class I restricted epitope TB10.4 (3-11) (CD8/10.4 T cells) and the MHC class II restricted epitope TB10.4 (74-88) (CD4/10.4 T cells). CD4/10.4 and CD8/10.4 T cells displayed marked differences in terms of expansion and contraction in a mouse TB model. CD4/10.4 T cells dominated in the early phase of infection whereas CD8/10.4 T cells were expanded after week 16 and reached 5-8 fold higher numbers in the late phase of infection. In the early phase of infection both CD4/10.4 and CD8/10.4 T cells were characterized by 20-25% polyfunctional cells (IL-2(+), IFN-gamma(+), TNF-alpha(+)), but whereas the majority of CD4/10.4 T cells were maintained as polyfunctional T cells throughout infection, CD8/10.4 T cells differentiated almost exclusively into effector cells (IFN-gamma(+), TNF-alpha(+)). Both CD4/10.4 and CD8/10.4 T cells exhibited cytotoxicity in vivo in the early phase of infection, but whereas CD4/10.4 cell mediated cytotoxicity waned during the infection, CD8/10.4 T cells exhibited increasing cytotoxic potential throughout the infection.

Conclusions/significance: Our results show that CD4 and CD8 T cells directed to epitopes in the same antigen differ both in their kinetics and functional characteristics throughout an infection with M. tuberculosis. In addition, the observed strong expansion of CD8 T cells in the late stages of infection could have implications for the development of post exposure vaccines against latent TB.

Show MeSH
Related in: MedlinePlus