BACKGROUND: Immune checkpoint inhibitors (ICIs), including those targeting PD-1, are currently used in a wide range of tumors, but only 20-40% of patients achieve clinical benefit. The objective of our study was to find predictive peripheral blood-based biomarkers for ICI treatment. METHODS: In 41 patients with advanced malignant melanoma (MM) and NSCLC treated with PD-1 inhibitors, we analyzed peripheral blood-based immune subsets by flow cytometry before treatment initialization and the second therapy dose. Specifically, we assessed basic blood differential count, overall T cells and their subgroups, B cells, and myeloid-derived suppressor cells (MDSC). In detail, CD4 + and CD8 + T cells were assessed according to their subtypes, such as central memory T cells (TCM), effector memory T cells (TEM), and naïve T cells (TN). Furthermore, we also evaluated the predictive value of CD28 and ICOS/CD278 co-expression on T cells. RESULTS: Patients who achieved disease control on ICIs had a significantly lower baseline proportion of CD4 + TEM (p = 0.013) and tended to have a higher baseline proportion of CD4 + TCM (p = 0.059). ICI therapy-induced increase in Treg count (p = 0.012) and the proportion of CD4 + TN (p = 0.008) and CD28 + ICOS- T cells (p = 0.012) was associated with disease control. Patients with a high baseline proportion of CD4 + TCM and a low baseline proportion of CD4 + TEM showed significantly longer PFS (p = 0.011, HR 2.6 and p ˂ 0.001, HR 0.23, respectively) and longer OS (p = 0.002, HR 3.75 and p ˂ 0.001, HR 0.15, respectively). Before the second dose, the high proportion of CD28 + ICOS- T cells after ICI therapy initiation was significantly associated with prolonged PFS (p = 0.017, HR 2.51) and OS (p = 0.030, HR 2.69). Also, a high Treg count after 2 weeks of ICI treatment was associated with significantly prolonged PFS (p = 0.016, HR 2.33). CONCLUSION: In summary, our findings suggest that CD4 + TEM and TCM baselines and an early increase in the Treg count induced by PD-1 inhibitors and the proportion of CD28 + ICOS- T cells may be useful in predicting the response in NSCLC and MM patients.

Department of Comprehensive Cancer Care Faculty of Medicine Masaryk University Brno Czech Republic

Department of Comprehensive Cancer Care Masaryk Memorial Cancer Institute Brno Czech Republic

Department of Laboratory Medicine Department of Laboratory Methods Faculty of Medicine and University Hospital Brno Masaryk University Brno Czech Republic

Department of Laboratory Medicine Masaryk Memorial Cancer Institute Brno Czech Republic

Department of Pharmacology Faculty of Medicine Masaryk University Brno Czech Republic

Faculty of Medicine Masaryk University Brno Czech Republic

Research Center for Applied Molecular Oncology Masaryk Memorial Cancer Institute Brno Czech Republic

Zobrazit více v PubMed

Larkin J, Chiarion-Sileni V, Gonzalez R, Grob J-J, Rutkowski P, Lao CD, et al. Five-year survival with combined Nivolumab and Ipilimumab in Advanced Melanoma. N Engl J Med. 2019;381:1535–46. PubMed

Espinosa E, Márquez-Rodas I, Soria A, Berrocal A, Manzano JL, Gonzalez-Cao M, et al. Predictive factors of response to immunotherapy—a review from the Spanish Melanoma Group (GEM). Ann Transl Med. 2017;5:389. PubMed PMC

Hodi FS, O’Day SJ, McDermott DF, Weber RW, Sosman JA, Haanen JB, et al. Improved survival with Ipilimumab in patients with metastatic melanoma. N Engl J Med. 2010;363:711–23. PubMed PMC

Topalian SL, Hodi FS, Brahmer JR, Gettinger SN, Smith DC, McDermott DF, et al. Safety, Activity, and Immune correlates of Anti–PD-1 antibody in Cancer. N Engl J Med. 2012;366:2443–54. PubMed PMC

Wang Y, Tong Z, Zhang W, Zhang W, Buzdin A, Mu X, et al. FDA-Approved and emerging next generation predictive biomarkers for Immune checkpoint inhibitors in Cancer patients. Front Oncol. 2021;11:683419. PubMed PMC

Ashktorab H, Ahuja S, Kannan L, Llor X, Ellis NA, Xicola RM, et al. A meta-analysis of MSI frequency and race in colorectal cancer. Oncotarget. 2016;7:34546. PubMed PMC

An HJ, Chon HJ, Kim C. Peripheral blood-based biomarkers for Immune Checkpoint inhibitors. Int J Mol Sci. 2021;22:9414. PubMed PMC

Kamphorst AO, Wieland A, Nasti T, Yang S, Zhang R, Barber DL, et al. Rescue of exhausted CD8 T cells by PD-1-targeted therapies is CD28-dependent. Science. 2017;355:1423–7. PubMed PMC

R Core Team. (2023). R: A Language and Environment for Statistical Computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org/

Reck M, Rodríguez-Abreu D, Robinson AG, Hui R, Csőszi T, Fülöp A, et al. Five-year outcomes with Pembrolizumab Versus Chemotherapy for metastatic non-small-cell lung Cancer with PD-L1 tumor proportion score ≥ 50. J Clin Oncol. 2021;39:2339–49. PubMed PMC

Rizvi NA, Hellmann MD, Snyder A, Kvistborg P, Makarov V, Havel JJ, et al. Mutational landscape determines sensitivity to PD-1 blockade in non–small cell lung cancer. Volume 348. New York, NY: Science; 2015. p. 124. PubMed PMC

Katayama Y, Yamada T, Chihara Y, Tanaka S, Tanimura K, Okura N, et al. Significance of inflammatory indexes in atezolizumab monotherapy outcomes in previously treated non-small-cell lung cancer patients. Sci Rep. 2020;10:17495. PubMed PMC

Nakaya A, Kurata T, Yoshioka H, Takeyasu Y, Niki M, Kibata K, et al. Neutrophil-to-lymphocyte ratio as an early marker of outcomes in patients with advanced non-small-cell lung cancer treated with nivolumab. Int J Clin Oncol. 2018;23:634–40. PubMed PMC

Ferrucci PF, Gandini S, Battaglia A, Alfieri S, Di Giacomo AM, Giannarelli D, et al. Baseline neutrophil-to-lymphocyte ratio is associated with outcome of ipilimumab-treated metastatic melanoma patients. Br J Cancer. 2015;112:1904–10. PubMed PMC

Dusselier M, Deluche E, Delacourt N, Ballouhey J, Egenod T, Melloni B, et al. Neutrophil-to-lymphocyte ratio evolution is an independent predictor of early progression of second-line nivolumab-treated patients with advanced non-small-cell lung cancers. PLoS ONE. 2019;14:e0219060. PubMed PMC

Ohashi H, Takeuchi S, Miyagaki T, Kadono T. Increase of lymphocytes and eosinophils, and decrease of neutrophils at an early stage of anti-PD-1 antibody treatment is a favorable sign for advanced malignant melanoma. Drug Discoveries Ther. 2020;14:117–21. PubMed

Ueda K, Suekane S, Kurose H, Ogasawara N, Hiroshige T, Chikui K, et al. Absolute lymphocyte count is an independent predictor of survival in patients with metastatic renal cell carcinoma treated with nivolumab. Jpn J Clin Oncol. 2022;52:179–86. PubMed

Rossi S, Toschi L, Finocchiaro G, Santoro A. Neutrophil and lymphocyte blood count as potential predictive indicators of nivolumab efficacy in metastatic non-small-cell lung cancer. Immunotherapy. 2020;12:715–24. PubMed

Rebuzzi SE, Signori A, Banna GL, Maruzzo M, De Giorgi U, Pedrazzoli P, et al. Inflammatory indices and clinical factors in metastatic renal cell carcinoma patients treated with nivolumab: the development of a novel prognostic score (Meet-URO 15 study). Ther Adv Med Oncol. 2021;13:17588359211019642. PubMed PMC

Zhang N, Bevan MJ. CD8 + T cells: Foot soldiers of the Immune System. Immunity. 2011;35:161–8. PubMed PMC

Garrido F, Cabrera T, Aptsiauri N. Hard and soft lesions underlying the HLA class I alterations in cancer cells: implications for immunotherapy. Int J Cancer. 2010;127:249–56. PubMed

Borst J, Ahrends T, Bąbała N, Melief CJM, Kastenmüller W. CD4 + T cell help in cancer immunology and immunotherapy. Nat Rev Immunol. 2018;18:635–47. PubMed

Marty Pyke R, Thompson WK, Salem RM, Font-Burgada J, Zanetti M, Carter H. Evolutionary pressure against MHC class II binding Cancer mutations. Cell. 2018;175:1991. PubMed

Sallusto F, Lenig D, Förster R, Lipp M, Lanzavecchia A. Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature. 1999;401:708–12. PubMed

Zhang G, Liu A, Yang Y, Xia Y, Li W, Liu Y, et al. Clinical predictive value of naïve and memory T cells in advanced NSCLC. Front Immunol. 2022;13:996348. PubMed PMC

Manjarrez-Orduño N, Menard LC, Kansal S, Fischer P, Kakrecha B, Jiang C, et al. Circulating T cell subpopulations correlate with Immune responses at the Tumor Site and clinical response to PD1 inhibition in Non-small Cell Lung Cancer. Front Immunol. 2018;9:1613. PubMed PMC

Klenerman P, Hill A. T cells and viral persistence: lessons from diverse infections. Nat Immunol. 2005;6:873–9. PubMed

Caserta S, Borger JG, Zamoyska R. Central and effector memory CD4 and CD8 T-cell responses to tumor-associated antigens. Crit Rev Immunol. 2012;32:97–126. PubMed

Kim HR, Park S-M, Seo S-U, Jung I, Yoon HI, Gabrilovich DI, et al. The Ratio of Peripheral Regulatory T Cells to Lox-1 + polymorphonuclear myeloid-derived suppressor cells predicts the early response to Anti–PD-1 therapy in patients with non–small cell Lung Cancer. Am J Respir Crit Care Med. 2019;199:243. PubMed PMC

Araujo B, de Lima V, Hansen M, Spanggaard I, Rohrberg K, Reker Hadrup S, Lassen U et al. Immune Cell profiling of peripheral blood as signature for response during checkpoint inhibition Across Cancer types. Front Oncol. 2021;11. PubMed PMC

Sojka DK, Huang Y-H, Fowell DJ. Mechanisms of regulatory T-cell suppression – a diverse arsenal for a moving target. Immunology. 2008;124:13–22. PubMed PMC

Zuazo M, Arasanz H, Fernández-Hinojal G, García‐Granda MJ, Gato M, Bocanegra A, et al. Functional systemic CD4 immunity is required for clinical responses to PD‐L1/PD‐1 blockade therapy. EMBO Mol Med. 2019;11:e10293. PubMed PMC

Luckheeram RV, Zhou R, Verma AD, Xia B. CD4 + T cells: differentiation and functions. Clin Dev Immunol. 2012;2012:925135. PubMed PMC

Xia L, Wang H, Sun M, Yang Y, Yao C, He S, et al. Peripheral CD4 + T cell signatures in predicting the responses to anti-PD-1/PD-L1 monotherapy for Chinese advanced non-small cell lung cancer. Sci China Life Sci. 2021;64:1590–601. PubMed

Lerrer S, Tocheva AS, Bukhari S, Adam K, Mor A. PD-1-stimulated T cell subsets are transcriptionally and functionally distinct. iScience. 2021;24:103020. PubMed PMC

Garcia-Bates T, Palma M, Shen C, Gambotto A, Macatangay B, Ferris R et al. Contrasting roles of the PD-1 signaling pathway in dendritic cell-mediated induction and regulation of HIV-1-Specific effector T cell functions. J Virol. 2019;93. PubMed PMC

Riley JL, Blair PJ, Musser JT, Abe R, Tezuka K, Tsuji T, et al. ICOS Costimulation requires IL-2 and can be prevented by CTLA-4 Engagement1. J Immunol. 2001;166:4943–8. PubMed

Marinelli O, Nabissi M, Morelli MB, Torquati L, Amantini C, Santoni G. ICOS-L as a potential therapeutic target for Cancer Immunotherapy. Curr Protein Pept Sci. 2018;19:1107–13. PubMed

Liakou CI, Kamat A, Tang DN, Chen H, Sun J, Troncoso P, et al. CTLA-4 blockade increases IFNγ-producing CD4 + ICOShi cells to shift the ratio of effector to regulatory T cells in cancer patients. Proc Natl Acad Sci U S A. 2008;105:14987–92. PubMed PMC

Dart SJ, Cook AM, Millward MJ, McDonnell AM, Chin WL, Hakeem MU, et al. Changes in expression of PD-L1 on peripheral T cells in patients with melanoma and lung cancer treated with PD-1 inhibitors. Sci Rep. 2021;11:15312. PubMed PMC

Hui E, Cheung J, Zhu J, Su X, Taylor MJ, Wallweber HA, et al. T cell costimulatory receptor CD28 is a primary target for PD-1–mediated inhibition. Science. 2017;355:1428–33. PubMed PMC