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Blockade of PD-1 and TIM-3 immune checkpoints fails to restore the function of exhausted CD8+ T cells in early clinical stages of chronic lymphocytic leukemia

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Abstract

Blocking antibodies targeting immune checkpoint molecules achieved invaluable success in tumor therapy and amazing clinical responses in a variety of cancers. Although common treatment protocols have improved overall survival in patients with chronic lymphocytic leukemia (CLL), they continue to relapse and progress. In the present in vitro study, the application of anti-PD-1 and anti-TIM-3 blocking antibodies was studied to restore the function of exhausted CD8+ T cells in CLL. CD8+ T cells were isolated from peripheral blood of 20 patients with CLL, treated with blocking antibodies, and cocultured with mitomycin-frozen non-CD8+ T cell fraction as target cells. Cultures were stimulated with anti-CD3/CD28 antibodies to assess the proliferation of CD8+ T cells by MTT and stimulated with PMA/ionomycin to measure the levels of CD107a expression and cytokine production by flow cytometry and ELISA, respectively. Our results showed that the blockade of PD-1 and TIM-3 does not improve the proliferation of CD8+ T cells in CLL patients. No significant difference was found between control and blocked groups in terms of degranulation properties and production of IFN-γ, TNF-α, IL-2, and IL-10 by CD8+ T cells. We observed that pre-treatment of CD8+ T cells with blocking antibodies in CLL patients at early clinical stages had no effects on restoring their functional properties. Further in vitro and in vivo complementary studies are required to more explore the utility of checkpoint inhibitors for CLL patients.

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Acknowledgments

The authors thank the patients and their families for their support, cooperation, and patience. We would like to thank the staff of the departments associated with the care and management of the patients.

Funding

This study was financially supported by grants from the National Institute for Medical Research Development (NIMAD, grant number: 962337) and Mazandaran University of Medical Sciences (grant number: 3160).

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Authors and Affiliations

  1. Department of Immunology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

    Hadiseh Rezazadeh, Mojgan Astaneh, Mohsen Tehrani, Hadi Hossein-Nataj & Hossein Asgarian-Omran

  2. Gastrointestinal Cancer Research Center, Non-Communicable Diseases Institute, Mazandaran University of Medical Sciences, Sari, Iran

    Mohsen Tehrani, Ehsan Zaboli, Ramin Shekarriz & Hossein Asgarian-Omran

  3. Department of Hematology and Oncology, Imam Khomeini Hospital, Mazandaran University of Medical Sciences, Sari, Iran

    Ehsan Zaboli & Ramin Shekarriz

  4. Immunogenetics Research Center, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran

    Hossein Asgarian-Omran

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  1. Hadiseh Rezazadeh
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Contributions

Hadiseh Rezazadeh performed the experiments and wrote the manuscript. Mojgan Astaneh contributed to the MACS isolation and cell culture. Mohsen Tehrani consulted in the optimization of the experiments. Hadi Hossein-Nataj contributed to the performing and analyzing of the flow cytometry experiments. Ehsan Zaboli and Ramin Shekarriz consulted as clinicians to obtain samples. Hossein Asgarian-Omran conceived the original idea, supervised the project, and edited the final manuscript.

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Correspondence to Hossein Asgarian-Omran.

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Written informed consent letters were obtained from all participants in accordance with the Ethical Committee of Mazandaran University of Medical Sciences.

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The authors state that written informed consent was obtained from all participants and the study has been approved by the Ethical Committee of Mazandaran University of Medical Sciences according to the Declaration of Helsinki.

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Rezazadeh, H., Astaneh, M., Tehrani, M. et al. Blockade of PD-1 and TIM-3 immune checkpoints fails to restore the function of exhausted CD8+ T cells in early clinical stages of chronic lymphocytic leukemia. Immunol Res 68, 269–279 (2020). https://doi.org/10.1007/s12026-020-09146-4

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