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Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells
Published on: February 8, 2018
Higher PD-1/Tim-3 expression on IFN-γ+ T cells is associated with poor prognosis in patients with acute myeloid
Shuxin Huang1, Yujie Zhao1, Wenpu Lai1
1Key Laboratory for Regenerative Medicine of Ministry of Education, Institute of Hematology, School of Medicine, Jinan University, Guangzhou, China.
Abstract:
With the success of immune checkpoint inhibitors (ICI), such as anti- programmed death-1 (PD-1) antibody for solid tumors and lymphoma immunotherapy, a number of clinical trials with ICIs have been attempted for acute myeloid leukemia (AML) immunotherapy; however, limited clinical efficacy has been reported. This may be due to the heterogeneity of immune microenvironments and various degrees of T cell exhaustion in patients and may be involved in the IFN-γ pathway. In this study, we first characterized the percentage of PD-1+ and T cell immunoglobulin mucin-domain-containing-3 (Tim-3) +IFN-γ+ T cells in peripheral blood (PB) in AML compared with healthy individuals (HIs) by flow cytometry and further discussed the possibility of the reversal of T cell exhaustion to restore the secretion capacity of cytokines in T cells in AML based on blockade of PD-1 or Tim-3 (anti-PD-1 and anti-Tim-3 antibody) in vitro using a cytokine protein chip. A significantly increased percentage of PD-1+, Tim-3+, and PD-1+Tim-3+ IFN-γ+ T cells was observed in PB from patients with AML in comparison with HIs. Moreover, higher PD-1+IFN-γ+CD3+/CD8+ T cell levels were associated with poor overall survival in AML patients. Regarding leukemia cells, the percentage of Tim-3 in CD117+CD34+ AML cells was positively correlated with PD-1 in IFN-γ+CD4+ T cells. Furthermore, blocking PD-1 and Tim-3 may involve multiple cytokines and helper T cell subsets, mainly Th1 and Treg cells. Blockade of PD-1 or Tim-3 tends to restore cytokine secretion to a certain extent, a synergistic effect shown by the co-blockade of PD-1 and Tim-3. However, we also demonstrated the heterogeneity of secretory cytokines in ICI-treated T cells in AML patients.
Insights
Immune checkpoint inhibitors show limited efficacy in acute myeloid leukemia (AML) due to T cell exhaustion. Blocking PD-1 and Tim-3 pathways may restore T cell function and cytokine secretion in AML immunotherapy.
Area of Science:
- Immunology
- Oncology
- Cancer Immunotherapy
Background:
- Immune checkpoint inhibitors (ICIs) show promise in solid tumors but have limited efficacy in acute myeloid leukemia (AML).
- T cell exhaustion and immune microenvironment heterogeneity contribute to the limited clinical success of ICIs in AML.
- The interferon-gamma (IFN-γ) pathway is implicated in T cell exhaustion within the AML immune landscape.
Purpose of the Study:
- To characterize the expression of PD-1 and Tim-3 on IFN-γ-producing T cells in AML patients compared to healthy individuals.
- To investigate the potential of blocking PD-1 and/or Tim-3 to reverse T cell exhaustion and restore cytokine secretion in AML.
- To explore the correlation between T cell exhaustion markers and clinical outcomes in AML patients.
Main Methods:
- Flow cytometry was used to quantify PD-1 and Tim-3 expression on T cells in peripheral blood from AML patients and healthy individuals.
- In vitro experiments using cytokine protein chips assessed the effects of anti-PD-1 and anti-Tim-3 antibody blockade on T cell cytokine secretion.
- Correlations between T cell marker expression, leukemia cell markers, and overall survival were analyzed.
Main Results:
- AML patients exhibited significantly higher percentages of PD-1+, Tim-3+, and PD-1+Tim-3+ IFN-γ+ T cells in peripheral blood compared to healthy individuals.
- Elevated levels of PD-1+IFN-γ+CD3+/CD8+ T cells were associated with poorer overall survival in AML patients.
- Blocking PD-1 and Tim-3 individually or in combination showed potential to restore T cell cytokine secretion, with synergistic effects observed upon co-blockade, though heterogeneity in cytokine response was noted.
Conclusions:
- Increased PD-1 and Tim-3 expression on T cells in AML contributes to T cell exhaustion and is linked to poor prognosis.
- Combined blockade of PD-1 and Tim-3 offers a potential therapeutic strategy to restore T cell function and enhance anti-leukemia immunity in AML.
- The heterogeneity of cytokine secretion in response to ICI treatment highlights the complexity of AML immunotherapy and warrants further investigation.
