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

  • Estefanía Paula Juliá
  • Pablo Mandó
  • Manglio Miguel Rizzo
  • Gerardo Rubén Cueto
  • Florencia Tsou
  • Romina Luca
  • Carmen Pupareli
  • Alicia Inés Bravo
  • Walter Astorino
  • José Mordoh
  • Claudio Martín
  • Estrella Mariel LevyEmail author
Original Article


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.


Anti-PD-1 therapy Nivolumab Pembrolizumab NSCLC Renal cell carcinoma TIM-3 



Absolute leukocyte count


Central memory


Complete blood cell count


C-reactive protein


Effector memory


Forward scatter-area


Forward scatter-height


Neutrophil-to-lymphocyte ratio


Progressive disease


Renal cell carcinoma


Side scatter-area


Stable disease-response


T cell immunoglobulin and mucin-domain containing-3


Terminal effector



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.

Author 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.


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.

Compliance with ethical standards

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.

Supplementary material

262_2019_2391_MOESM1_ESM.pdf (810 kb)
Supplementary material 1 (PDF 811 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Estefanía Paula Juliá
    • 1
  • Pablo Mandó
    • 1
    • 2
  • Manglio Miguel Rizzo
    • 2
  • Gerardo Rubén Cueto
    • 3
  • Florencia Tsou
    • 4
  • Romina Luca
    • 2
  • Carmen Pupareli
    • 4
  • Alicia Inés Bravo
    • 5
  • Walter Astorino
    • 4
  • José Mordoh
    • 1
    • 4
    • 6
  • Claudio Martín
    • 4
  • Estrella Mariel Levy
    • 1
    Email author
  1. 1.Centro de Investigaciones OncológicasFundación Cáncer-FUCACiudad Autónoma de Buenos AiresArgentina
  2. 2.Fundación Cáncer-FUCACiudad Autónoma de Buenos AiresArgentina
  3. 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 NaturalesUniversidad de Buenos AiresCiudad Autónoma de Buenos AiresArgentina
  4. 4.Instituto Alexander FlemingCiudad Autónoma de Buenos AiresArgentina
  5. 5.Hospital Interzonal Eva Perón HIGA, San MartínBuenos AiresArgentina
  6. 6.Fundación Instituto Leloir-IIBBACiudad Autónoma de Buenos AiresArgentina

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