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PD-1(hi)TIM-3(+) T cells associate with and predict leukemia relapse in AML patients post allogeneic stem cell transplantation.

Kong Y, Zhang J, Claxton DF, Ehmann WC, Rybka WB, Zhu L, Zeng H, Schell TD, Zheng H - Blood Cancer J (2015)

Bottom Line: T-cell exhaustion is a state of T-cell dysfunction mediated by inhibitory molecules including programmed cell death protein 1 (PD-1) and T-cell immunoglobulin domain and mucin domain 3 (TIM-3).Consistent with exhaustion, PD-1(hi)TIM-3(+) T cells are functionally deficient manifested by reduced production of interleukin 2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ).In addition, these cells demonstrate a phenotype consistent with exhausted antigen-experienced T cells by losing TN and TEMRA subsets.

View Article: PubMed Central - PubMed

Affiliation: 1] Penn State Hershey Cancer Institute, Penn State University College of Medicine, Hershey, PA, USA [2] Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing Key Laboratory of Emerging Infectious Diseases, Beijing, China.

ABSTRACT
Prognosis of leukemia relapse post allogeneic stem cell transplantation (alloSCT) is poor and effective new treatments are urgently needed. T cells are pivotal in eradicating leukemia through a graft versus leukemia (GVL) effect and leukemia relapse is considered a failure of GVL. T-cell exhaustion is a state of T-cell dysfunction mediated by inhibitory molecules including programmed cell death protein 1 (PD-1) and T-cell immunoglobulin domain and mucin domain 3 (TIM-3). To evaluate whether T-cell exhaustion and inhibitory pathways are involved in leukemia relapse post alloSCT, we performed phenotypic and functional studies on T cells from peripheral blood of acute myeloid leukemia patients receiving alloSCT. Here we report that PD-1(hi)TIM-3(+) cells are strongly associated with leukemia relapse post transplantation. Consistent with exhaustion, PD-1(hi)TIM-3(+) T cells are functionally deficient manifested by reduced production of interleukin 2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). In addition, these cells demonstrate a phenotype consistent with exhausted antigen-experienced T cells by losing TN and TEMRA subsets. Importantly, increase of PD-1(hi)TIM-3(+) cells occurs before clinical diagnosis of leukemia relapse, suggesting their predictive value. Results of our study provide an early diagnostic approach and a therapeutic target for leukemia relapse post transplantation.

No MeSH data available.


Related in: MedlinePlus

PD-1hiTIM3+ cells produce less TNF-a and IL-2 upon PMA/ionomycin stimulation. Production of TNF-α, IFN-γ and IL-2 upon in vitro PMA/ionomycin stimulation. Shown is cytokine release from CD4+ (a) or CD8+ (b) T cells gated on each fraction of cells based on PD-1 and TIM-3 expression. Representative plots from a single relapse patient (09) are shown.
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fig3: PD-1hiTIM3+ cells produce less TNF-a and IL-2 upon PMA/ionomycin stimulation. Production of TNF-α, IFN-γ and IL-2 upon in vitro PMA/ionomycin stimulation. Shown is cytokine release from CD4+ (a) or CD8+ (b) T cells gated on each fraction of cells based on PD-1 and TIM-3 expression. Representative plots from a single relapse patient (09) are shown.

Mentions: To evaluate the functional status of PD-1hiTIM-3+ T cells, we performed functional assays to test cytokine release by T cells derived from AML patients with leukemia relapse post transplantation. T cells were stimulated in vitro before flow cytometry analysis for intracellular IL-2, TNF-α and IFN-γ production. The cells were costained with PD-1 and TIM-3, allowing us to dissect the function of each T-cell fraction based on PD-1 and TIM-3 expression. We first used PMA/ionomycin as the in vitro stimulator for T cells. Figure 3 shows the cytokine release of T cells from a patient with AML relapse post transplantation (patient 09). The PD-1hiTIM-3+ population in CD4+ T cells produced much lower TNF-α and IL-2 compared with that from fractions I, II, IV and V (19.0% vs 37–58.7% of TNF-α release, and 16.2% vs 37.3–41% of IL-2 release; Figure 3a). Fraction III showed moderate loss of TNF-α (28.5%) and IL-2 (35.2%). A similar pattern was observed for CD8+ T cells, in that PD-1hiTIM-3+CD8+ T cells produced extremely low TNF-α and IL-2 (Figure 3b). Interestingly, in both CD4+ and CD8+ T cells, the PD-1hiTIM-3+ population had increased production of IFN-γ (Figures 3a and b). PMA/ionomycin stimulates T-cell activation through a strong but non-T-cell receptor (TCR)-dependent pathway.32 To dissect the T-cell functional status by mimicking the physiological signal, we performed a separate study of cytokine release using anti-CD3 and anti-CD28 as in vitro stimulators, in which the activation of T cells is TCR dependent. As expected, the overall levels of cytokine release were reduced following CD3/CD28 stimulation compared with that obtained with PMA/ionomycin stimulation (Figure 4). Consistent with results obtained using PMA/ionomycin stimulation, PD-1hiTIM-3+ CD4+ T cells had much lower intracellular TNF-α and IL-2. Importantly, IFN-γ production was also decreased in this population (Figure 4a). CD8+ T cells followed the same pattern (Figure 4b) in that PD-1hiTIM-3+CD8+ T cells produced low TNF-α and IFN-γ. As expected, CD8+ T cells did not produce much IL-2. These results indicate that the PD-1hiTIM-3+ T cells that developed in leukemia relapse are dysfunctional, consistent with development of exhaustion.


PD-1(hi)TIM-3(+) T cells associate with and predict leukemia relapse in AML patients post allogeneic stem cell transplantation.

Kong Y, Zhang J, Claxton DF, Ehmann WC, Rybka WB, Zhu L, Zeng H, Schell TD, Zheng H - Blood Cancer J (2015)

PD-1hiTIM3+ cells produce less TNF-a and IL-2 upon PMA/ionomycin stimulation. Production of TNF-α, IFN-γ and IL-2 upon in vitro PMA/ionomycin stimulation. Shown is cytokine release from CD4+ (a) or CD8+ (b) T cells gated on each fraction of cells based on PD-1 and TIM-3 expression. Representative plots from a single relapse patient (09) are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4526784&req=5

fig3: PD-1hiTIM3+ cells produce less TNF-a and IL-2 upon PMA/ionomycin stimulation. Production of TNF-α, IFN-γ and IL-2 upon in vitro PMA/ionomycin stimulation. Shown is cytokine release from CD4+ (a) or CD8+ (b) T cells gated on each fraction of cells based on PD-1 and TIM-3 expression. Representative plots from a single relapse patient (09) are shown.
Mentions: To evaluate the functional status of PD-1hiTIM-3+ T cells, we performed functional assays to test cytokine release by T cells derived from AML patients with leukemia relapse post transplantation. T cells were stimulated in vitro before flow cytometry analysis for intracellular IL-2, TNF-α and IFN-γ production. The cells were costained with PD-1 and TIM-3, allowing us to dissect the function of each T-cell fraction based on PD-1 and TIM-3 expression. We first used PMA/ionomycin as the in vitro stimulator for T cells. Figure 3 shows the cytokine release of T cells from a patient with AML relapse post transplantation (patient 09). The PD-1hiTIM-3+ population in CD4+ T cells produced much lower TNF-α and IL-2 compared with that from fractions I, II, IV and V (19.0% vs 37–58.7% of TNF-α release, and 16.2% vs 37.3–41% of IL-2 release; Figure 3a). Fraction III showed moderate loss of TNF-α (28.5%) and IL-2 (35.2%). A similar pattern was observed for CD8+ T cells, in that PD-1hiTIM-3+CD8+ T cells produced extremely low TNF-α and IL-2 (Figure 3b). Interestingly, in both CD4+ and CD8+ T cells, the PD-1hiTIM-3+ population had increased production of IFN-γ (Figures 3a and b). PMA/ionomycin stimulates T-cell activation through a strong but non-T-cell receptor (TCR)-dependent pathway.32 To dissect the T-cell functional status by mimicking the physiological signal, we performed a separate study of cytokine release using anti-CD3 and anti-CD28 as in vitro stimulators, in which the activation of T cells is TCR dependent. As expected, the overall levels of cytokine release were reduced following CD3/CD28 stimulation compared with that obtained with PMA/ionomycin stimulation (Figure 4). Consistent with results obtained using PMA/ionomycin stimulation, PD-1hiTIM-3+ CD4+ T cells had much lower intracellular TNF-α and IL-2. Importantly, IFN-γ production was also decreased in this population (Figure 4a). CD8+ T cells followed the same pattern (Figure 4b) in that PD-1hiTIM-3+CD8+ T cells produced low TNF-α and IFN-γ. As expected, CD8+ T cells did not produce much IL-2. These results indicate that the PD-1hiTIM-3+ T cells that developed in leukemia relapse are dysfunctional, consistent with development of exhaustion.

Bottom Line: T-cell exhaustion is a state of T-cell dysfunction mediated by inhibitory molecules including programmed cell death protein 1 (PD-1) and T-cell immunoglobulin domain and mucin domain 3 (TIM-3).Consistent with exhaustion, PD-1(hi)TIM-3(+) T cells are functionally deficient manifested by reduced production of interleukin 2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ).In addition, these cells demonstrate a phenotype consistent with exhausted antigen-experienced T cells by losing TN and TEMRA subsets.

View Article: PubMed Central - PubMed

Affiliation: 1] Penn State Hershey Cancer Institute, Penn State University College of Medicine, Hershey, PA, USA [2] Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing Key Laboratory of Emerging Infectious Diseases, Beijing, China.

ABSTRACT
Prognosis of leukemia relapse post allogeneic stem cell transplantation (alloSCT) is poor and effective new treatments are urgently needed. T cells are pivotal in eradicating leukemia through a graft versus leukemia (GVL) effect and leukemia relapse is considered a failure of GVL. T-cell exhaustion is a state of T-cell dysfunction mediated by inhibitory molecules including programmed cell death protein 1 (PD-1) and T-cell immunoglobulin domain and mucin domain 3 (TIM-3). To evaluate whether T-cell exhaustion and inhibitory pathways are involved in leukemia relapse post alloSCT, we performed phenotypic and functional studies on T cells from peripheral blood of acute myeloid leukemia patients receiving alloSCT. Here we report that PD-1(hi)TIM-3(+) cells are strongly associated with leukemia relapse post transplantation. Consistent with exhaustion, PD-1(hi)TIM-3(+) T cells are functionally deficient manifested by reduced production of interleukin 2 (IL-2), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). In addition, these cells demonstrate a phenotype consistent with exhausted antigen-experienced T cells by losing TN and TEMRA subsets. Importantly, increase of PD-1(hi)TIM-3(+) cells occurs before clinical diagnosis of leukemia relapse, suggesting their predictive value. Results of our study provide an early diagnostic approach and a therapeutic target for leukemia relapse post transplantation.

No MeSH data available.


Related in: MedlinePlus