Expression of PD-1/PD-L1 and PD-L2 in peripheral T-cells from non-small cell lung cancer patients

Oscar Arrieta, Edgar Montes-Servín, Juan-Manuel Hernandez-Martinez, Andrés F Cardona, Eibar Casas-Ruiz, José C Crispín, Daniel Motola, Diana Flores-Estrada, Lourdes Barrera, Oscar Arrieta, Edgar Montes-Servín, Juan-Manuel Hernandez-Martinez, Andrés F Cardona, Eibar Casas-Ruiz, José C Crispín, Daniel Motola, Diana Flores-Estrada, Lourdes Barrera

Abstract

Binding of programmed death-1 (PD-1) with its ligands (PD-L1/2) transmits a co-inhibitory signal in activated T-cells that promotes T-cell exhaustion, leading to tumor immune evasion. The efficacy of antibodies targeting PD-1 and PD-L1 has led to a paradigm shift in lung cancer treatment but the prognostic and predictive value of tumor PD-L1 expression remains controversial. Evaluating PD-1, PD-L1/2 expression in peripheral blood cells may serve as a potential biomarker for prognosis and response to therapy. In this prospective observational study, plasma cytokine levels and PD-1, PD-L1 and PD-L2 expression was evaluated in circulating CD3+, CD3+CD4+ and CD3+CD8+ cells from 70 treatment-naïve patients with advanced NSCLC (Stage IIIB and IV) and from 10 healthy donors. The primary objective was to assess OS according to PD-1, PD-L1, PD-L2 expression status on PBMCs and lymphocyte subsets. Our results indicate that the percentage of PD-L1+CD3+, PD-L1+CD3+CD8+ PD-L2+PBMCs, PD-L2+CD3+, PD-L2+CD3+CD4+ cells was higher in patients than in healthy donors. Survival was decreased among patients with a high percentage of either PD-1+PBMCs, PD-1+CD3+, PD-L1+CD3+, PD-L1+CD3+CD8+, PD-L2+CD3+, PD-L2+CD3+CD4+, or PD-L2+CD3+CD8+ cells. IL-2 and TNF-α showed the strongest association with PD-L1 and PD-L2 expression on specific subsets of T-lymphocytes. Our findings suggest that increased PD-1/PD-L1/PDL-2 expression in PBMCs, particularly in T-cells, may be an additional mechanism leading to tumor escape from immune control. This study is registered with ClinicalTrials.gov, number NCT02758314.

Keywords: checkpoint inhibitors; circulating lymphocytes; immunotherapy; lung adenocarcinoma; prognosis.

Conflict of interest statement

CONFLICTS OF INTEREST Dr. Oscar Arrieta received research funding from Bristol-Myers Squibb (Mexico). However, in this study no Bristol-Myers or competitor molecules were tested and data analyses was carried out independently. All other authors declare no conflicts of interest.

Figures

Figure 1. Immune cell subsets in NSCLC…
Figure 1. Immune cell subsets in NSCLC patients and healthy subjects
Scatter plot showing the percentage of (A) CD3 +, (B) CD3 +CD4 +, (C) CD3 +CD8 + cells in the PBMC fraction. Panel (D, E) & (F) show the percentage of PBMCs positive for PD-1, PD-L1 & PD-L2, respectively. Panel (G, H) & (I) show the percentage of PD-1 +CD3 +, PD-L1 +CD3 +, and PD-L2 +CD3 + cells. Panel (J, K) & (L) show the percentage of PD-1 +CD3 +CD4 +, PD-L1 +CD3 +CD4 +, and PD-L2 +CD3 +CD4 + cells. Panel (M, N) & (O) show the percentage of PD-1 +CD3 +CD8 +, PD-L1 +CD3 +CD8 +, and PD-L2 +CD3 +CD8 + cells.
Figure 2. Kaplan-Meier curves of OS by…
Figure 2. Kaplan-Meier curves of OS by PD-1, PD-L1, PD-L2 expression status
(A) PD-1 cutoff point ≤1.25% in PBMCs and (B) ≤1.1% in CD3 + cells; (C) PD-L1 cutoff point ≤3.6% in CD3 + cells and (D) ≤1.5% in CD3 +CD8 + cells; (E) PD-L2 cutoff point ≤0.985% in CD3 + cells, (F) ≤0.5% in CD3 +CD4 + cells, and (G) ≤0.625% in CD3 +CD8 + cells.

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