Tumoral immune-infiltrate (IF), PD-L1 expression and role of CD8/TIA-1 lymphocytes in localized osteosarcoma patients treated within protocol ISG-OS1

Emanuela Palmerini, Claudio Agostinelli, Piero Picci, Stefano Pileri, Teresa Marafioti, Pier-Luigi Lollini, Katia Scotlandi, Alessandra Longhi, Maria Serena Benassi, Stefano Ferrari, Emanuela Palmerini, Claudio Agostinelli, Piero Picci, Stefano Pileri, Teresa Marafioti, Pier-Luigi Lollini, Katia Scotlandi, Alessandra Longhi, Maria Serena Benassi, Stefano Ferrari

Abstract

Background: We hypothesized that immune-infiltrates were associated with superior survival, and examined a primary osteosarcoma tissue microarrays (TMAs) to test this hypothesis.

Methods: 129 patients (pts) with localized osteosarcoma treated within protocol ISG-OS1 were included in the study. Clinical characteristics, expression of CD8, CD3, FOXP3, CD20, CD68/CD163 (tumor associated macrophage, TAM), Tia-1 (cytotoxic T cell), CD303 (plasmacytoid dendritic cells: pDC), Arginase-1 (myeloid derived suppressor cells: MDSC), PD-1 on immune-cells (IC), and PD-L1 on tumoral cells (TC) and IC were analysed and correlated with outcome.

Results: Most of the cases presented tumor infiltrating lymphocytes (TILs) (CD3+ 90%; CD8+ 86%). Tia-1 was detected in 73% of the samples. PD-L1 expression was found in 14% patients in IC and 0% in TC; 22% showed PD-1 expression in IC.With a median follow-up of 8 years (range 1-13), the 5-year overall survival (5-year OS) was 74% (95% CI 64-85). Univariate analysis showed better 5-year OS for: a) pts with a good histologic response to neoadjuvant chemotherapy (p = 0.0001); b) pts with CD8/Tia1 tumoral infiltrates (p = 0.002); c) pts with normal alkaline phosphatas (sALP) (p = 0.04). After multivariate analysis, histologic response (p = 0.007) and CD8/Tia1 infiltration (p = 0.01) were independently correlated with survival. In the subset of pts with CD8+ infiltrate, worse (p 0.02) OS was observed for PD-L1(IC)+ cases.

Conclusions: Our findings support the hypothesis that CD8/Tia1 infiltrate in tumor microenvironment at diagnosis confers superior survival for pts with localized osteosarcoma, while PD-L1 expression is associated with worse survival.

Keywords: CD8; PD-1; PD-L1; osteosarcoma; tumor microenvironment.

Conflict of interest statement

CONFLICTS OF INTEREST The authors declare that they have no conflicts of interest.

Figures

Figure 1. Mechanisms for intratumoral programmed cell…
Figure 1. Mechanisms for intratumoral programmed cell death ligand-1 (PD-L1) expression
Adaptive focal expression of PD-L1 by macrophages (CD68+/CD163+) occurs at the interface of tumor cell nests with immune infiltrates secreting pro-inflammatory factors such as interferon-γ. The ligation of PD-L1 on macrophage and, in some histotypes, on tumor cells, with programmed cell death protein 1 (PD-1) molecules will down-modulate T cell function, essentially creating a negative feedback loop that reduces antitumor immunity (the so called ‘tumor shield’ effect), eventually reducing CD8 tumoricidal function.
Figure 2. Tumor microenvironment in localized osteosarcoma
Figure 2. Tumor microenvironment in localized osteosarcoma
Immunohistochemical expression of CD3, CD8, CD20 (T tumor infiltrating lymphocytes: TILs), Tia-1 (cytotoxic T cell), FOXP3 (T regulatory lymphocytes: T-regs), PD-1, PD-L1, CD68 (tumor associated macrophages: TAM), BDCA-2/CD303 (plasmacytoid dendritic cells: pDC), Arginase-1 (myeloid derived suppressor cells: MDSC) proteins.
Figure 3. Survival and immune-infiltrate
Figure 3. Survival and immune-infiltrate
(A) 5-year overall survival according to CD8/Tia1 expression in localized osteosarcoma; (B) 5-year overall survival according to PD-L1 expression in patients with CD8+ localized osteosarcoma.
Figure 4. Proposal for ‘Immune-infiltrate based treatment…
Figure 4. Proposal for ‘Immune-infiltrate based treatment algorithm’ for localized osteosarcoma

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