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Peripheral changes in immune cell populations and soluble mediators after anti-PD-1 therapy in non-small cell lung cancer and renal cell carcinoma patients

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Abstract

Patients with non-small cell lung cancer (NSCLC) and renal cell carcinoma (RCC) have shown benefit from anti-PD-1 therapies. However, not all patients experience tumor shrinkage, durable responses or prolonged survival, demonstrating the need to find response markers. In blood samples from NSCLC and RCC patients obtained before and after anti-PD-1 treatment, we studied leukocytes by complete blood cell count, lymphocyte subsets using flow cytometry and plasma concentration of nine soluble mediators, in order to find predictive biomarkers of response and to study changes produced after anti-PD-1 therapy. In baseline samples, discriminant analysis revealed a combination of four variables that helped differentiate stable disease-response (SD-R) from progressive disease (PD) patients: augmented frequency of central memory CD4+ T cells and leukocyte count was associated with response while increased percentage of PD-L1+ natural killer cells and naïve CD4+ T cells was associated with lack of response. After therapy, differential changes between responders and non-responders were found in leukocytes, T cells and TIM-3+ T cells. Patients with progressive disease showed an increase in the frequency of TIM-3 expressing CD4+ and CD8+ T cells, whereas SD-R patients showed a decrease in these subsets. Our findings indicate that a combination of immune variables from peripheral blood (PB) could be useful to distinguish response groups in NSCLC and RCC patients treated with anti-PD-1 therapy. Frequency of TIM-3+ T cells showed differential changes after treatment in PD vs SD-R patients, suggesting that it may be an interesting marker for monitoring progression during therapy.

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Abbreviations

ALC:

Absolute leukocyte count

CM:

Central memory

CBC:

Complete blood cell count

CRP:

C-reactive protein

EM:

Effector memory

FSC-A:

Forward scatter-area

FSC-H:

Forward scatter-height

NLR:

Neutrophil-to-lymphocyte ratio

PD:

Progressive disease

RCC:

Renal cell carcinoma

SSC-A:

Side scatter-area

SD-R:

Stable disease-response

TIM-3:

T cell immunoglobulin and mucin-domain containing-3

TE:

Terminal effector

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Acknowledgements

We thank Holliday Cartar for her assistance in language correction. We thank Dr. Laura Noro and all the laboratory staff from Alexander Fleming Institute for their help in this study.

Funding

This work was supported by grants from Fundación Sales, Fundación Cáncer, Fundación Pedro F. Mosoteguy, Argentina. José Mordoh and Estrella Mariel Levy are members of Consejo Nacional de Investigaciones Científicas y Técnicas-CONICET. Estefanía Paula Juliá is a fellow from Consejo Nacional de Investigaciones Científicas y Técnicas-CONICET.

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Authors and Affiliations

  1. Centro de Investigaciones Oncológicas, Fundación Cáncer-FUCA, Zabala 2836 1º Floor, C1426AOE, Ciudad Autónoma de Buenos Aires, Argentina

    Estefanía Paula Juliá, Pablo Mandó, José Mordoh & Estrella Mariel Levy

  2. Fundación Cáncer-FUCA, Ciudad Autónoma de Buenos Aires, Argentina

    Pablo Mandó, Manglio Miguel Rizzo & Romina Luca

  3. Grupo de Bioestadística Aplicada, Departamento de Ecología, Genética y Evolución, Instituto de Ecología, Genética y Evolución de Buenos Aires (IEGEBA-UBA/CONICET), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina

    Gerardo Rubén Cueto

  4. Instituto Alexander Fleming, Ciudad Autónoma de Buenos Aires, Argentina

    Florencia Tsou, Carmen Pupareli, Walter Astorino, José Mordoh & Claudio Martín

  5. Hospital Interzonal Eva Perón HIGA, San Martín, Buenos Aires, Argentina

    Alicia Inés Bravo

  6. Fundación Instituto Leloir-IIBBA, Ciudad Autónoma de Buenos Aires, Argentina

    José Mordoh

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  1. Estefanía Paula Juliá
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  11. Claudio Martín
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Contributions

EPJ collected, analyzed and interpreted the data, and wrote the manuscript. PM and EML analyzed and interpreted the data, and wrote the manuscript. MMR, FT and RL contributed with patients’ clinical data analysis. GRC contributed to the statistical analysis and performed discriminant analyses. AIB and WA performed IHC analysis. JM, CP and CM interpreted the data and revised the manuscript. All authors contributed to manuscript revision; read and approved the final manuscript version.

Corresponding author

Correspondence to Estrella Mariel Levy.

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Conflict of interest

Claudio Martín has served as speaker and advisory board member for Bristol Myers Squibb and Merck Sharp and Dohme. Carmen Pupareli has served as speaker and advisor board member for Merck Sharp and Dohme and as speaker for Bristol Myers Squibb. The authors declare that there is no other conflict of interest.

Ethical approval and ethical standards

All procedures involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The study was approved by the Comité de Ética en Investigación del Instituto Alexander Fleming CEIAF, approval number: 616, 14th June 2016.

Informed consent

All samples were taken after patients gave written informed consent approved by Comité de Ética en Investigación del Instituto Alexander Fleming CEIAF. Patients consented to the use of their specimens and data for research and for publication.

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Juliá, E.P., Mandó, P., Rizzo, M.M. et al. Peripheral changes in immune cell populations and soluble mediators after anti-PD-1 therapy in non-small cell lung cancer and renal cell carcinoma patients. Cancer Immunol Immunother 68, 1585–1596 (2019). https://doi.org/10.1007/s00262-019-02391-z

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