Tumor-infiltrating Mononuclear Cells Research Articles

Association between PD-L1 Expression and the Prognosis and Clinicopathologic Features of Non-Clear Cell Renal Cell Carcinoma.

PD-L1 is one of the two programmed cell death 1 (PD-1) ligands and a part of an immune checkpoint system (PD-1/PD-L1) with widespread clinical application. The aim of this study was to investigate PD-L1 expression and its association with clinicopathological and prognostic significance in non-clear cell renal cell carcinoma (non-ccRCC) patients. A total of 41 papillary (pRCC) and 20 chromophobe (chRCC) RCC tumors were examined for PD-L1 expression by immunohistochemistry in the cancer cells and tumor-infiltrating mononuclear cells (TIMCs). PD-L1 positivity was detected in 36.6% pRCC and 85.0% chRCC cancer cells, while PD-L1 positivity was observed in 73.2% pRCC and 50.0% chRCC TIMCs. PD-L1 positivity in both pRCC and chRCC tumor cells was not correlated with any of the examined clinicopathological features, while PD-L1 positivity in TIMCs was associated with the age of patients with pRCC. During follow-up, the death was documented among 6 patients with pRCC. Papillary RCC patients with PD-L1-positive tumor cells were significantly associated with an increased risk of death compared with patients with PD-L1-negative cancer cells. A similar trend was observed when comparing PD-L1 expression in TIMCs. However, no differences in overall survival for PD-L1-positive pRCC patients with compared to PD-L1-negative patients were observed in tumor cells or TIMCs.

The effects of immune checkpoint modulators on the clinical course of patients with resectable hepatocellular carcinoma.

Immune checkpoint proteins regulating T-cell mediated anti-tumor immunity have been reported to affect clinical outcomes in multiple malignancies. This study aimed to investigate the prognostic effect of histological expression of immune checkpoint proteins in patients with resected hepatocellular carcinoma (HCC). A total of 221 patients with HCC who underwent curative resection were included. Expression of programmed-cell death ligand-1 (PD-L1) in tumor cells (tPD-L1) and tumor infiltrating mononuclear cells (TIMCs) (iPD-L1), programmed-cell death-1 in TIMCs (iPD-1), and cytotoxic T lymphocyte antigen-4 in TIMCs (iCTLA-4) were measured immunohistochemically. Histo-positivity for iCTLA-4, iPD-1, iPD-L1, and tPD-L1 was 32.1%, 42.5%, 35.3%, and 14.9%, respectively. Multivariate logistic analyses revealed that male sex and tumor >5 cm were variables related to iCTLA-4 positivity (odds ratio [OR], 0.46 and 1.94, respectively; P<0.05). Poor differentiation was related to PD-L1 expression in both tumor cells and TIMCs (OR, 2.88 and 3.46, respectively; P<0.05). Microvascular invasion was significantly associated only with iPD-L1 (OR, 2.24; P<0.05). In time-dependent outcome analyses, expression of immune checkpoint proteins in TIMCs (i.e., iCTLA-4, iPD-1, and iPD-L1) was significantly related to longer overall survival and non-cancer-related survival (all P<0.05), but not to time-to-recurrence or cancer-specific deaths. Concurrent activation of the PD-1:PD-L1 and CTLA-4 pathways predicted improved outcomes in terms of overall survival and non-cancer related survival (P=0.06 and P=0.03, respectively). Immune checkpoint proteins upregulated in TIMCs in HCC tissues have individual and additive effects in prolonging the survival of patients, specifically in terms of survival not related to cancer recurrence.

Expression of programmed death ligand 1 (PD-L1) in urothelial bladder carcinoma (immunohistochemical and histopathological study)

The aim of this work is to evaluate the immunohistochemical expression of PD-L1 in tumor cells of urothelial bladder carcinoma. PD-L1 immunoexpression was assessed on 20 cases of radical cystectomy specimens using monoclonal rabbit anti- PD-L1 antibody. Extent of membranous PD-L1 expression was assigned in each case. Tumor cells showing > 5% expression were considered positive. PD-L1was seen in 6 (30%) cases [1 case score +2 and 5 cases score +3], while 14(70%) cases were negative [7 cases score 0 and 7 cases score +1]. PD-L1 expression in tumor cells was insignificantly correlated with different available clinicopathological parameters such as sex, pathological stage, associatedbilharzial infestation, necrosis, perineural invasion and associated tumor infiltrating mononuclear cells (TIMCs), although PD-L1 positivity tend to predominate in advanced pathological stage of tumor (T3 and T4). These findings may have clinical implications for the management of patients with PD-L1 positive urothelial bladder carcinoma.

Abstract PO001: Results from a preclinical study to evaluate the efficacy of polyvalent immunoglobulins on the imbalance of (anti-)inflammatory immune cell responses in clinical colorectal cancer

Abstract Introduction: The purpose of this study was to determine the efficacy of an immunoglobulin concentrate on dysregulated inflammatory responses in patients with colorectal cancer (CRC). Recent findings have shown that CRC is associated with dysbiosis of the gut microbiome and frequently a compromised gut barrier, resulting in prolonged and smoldering endotoxin leakage into the circulation. Subclinical local and systemic microinflammation is suggested to facilitate metastasis formation. Previous studies have pointed to the capacity of polyvalent immunoglobulins to neutralize endotoxins and pathogens on gut epithelial surfaces to abrogate inflammatory responses and thereby tumor recurrence. The effects of KMP01D, a colostrum preparation with a multitude of immunoglobulins against gram-negative/positive bacteria and viruses, on pro-/anti-inflammatory cytokine responses and apoptosis were determined ex vivo. Methods: Peripheral blood and tumor-derived mononuclear cells (MNC) from CRC patients at UICC stages I–IV (n=48) operated at our Center for Operative Medicine/Comprehensive Cancer Center and treated under current guidelines for (neo-)adjuvant therapy were further investigated. Expression of CD14, CD68, Toll-like receptor(TLR) 4, and insulin-like growth factor (IGF)-1 with/without co-incubation of KMP01D as well as immune cell apoptosis was analyzed by flow cytometry, RT PCR, immunohistochemistry, and Elisa in each patient. Results: KMP01D increased interleukin (IL)-10 and IL-13 anti-inflammatory cytokine expression in peripheral blood MNCs (UICC I-III). More interestingly, it likewise decreased secretion of IL-1β, IL-6, IFN-γ, TNF-α, and IL-12 driven inflammation as well as IGF-1. Moreover, CD14/CD68 and TLR4 expression critically involved in endotoxin signaling was downregulated in PBMCs and tumor-infiltrating MNCs. CD14+Fas+/TUNEL+ PBMCs and tumor-derived MNCs indicating apoptosis were likewise enhanced under KMP01D co-incubation. Addition of vitamin D3 known as a regulatory factor in immune responses demonstrated additive anti-inflammatory effects in vitro. Conclusions: Inflammatory immune responses are significantly and stage-dependently upregulated which negatively impacts anti-tumor-specific immune reactivity, immune cell apoptosis and prognosis in CRC patients despite modern adjuvant treatment protocols. KMP01D produced from natural bovine sources and composed of a multitude of immunoglobulin activity (IgG1/2 and Lactoferrin) demonstrates highly ex vivo efficacy on downregulation of inflammatory cytokine responses in PBMCs and normalized immune cell apoptosis in CRC patients. Together with previous clinical data our results strongly suggest that KMP01-mediated endotoxin and pathogen neutralization and thereby mitigated chronic inflammation can be a highly valuable add-on instrument in the treatment of CRC patients to downregulate stage-dependently their local and systemic inflammation and risk for metastasis formation. Citation Format: Martin Gasser, Reinhard Lissner, Li-Li Hsiao, Ana Maria Waaga-Gasser. Results from a preclinical study to evaluate the efficacy of polyvalent immunoglobulins on the imbalance of (anti-)inflammatory immune cell responses in clinical colorectal cancer [abstract]. In: Abstracts: AACR Virtual Special Conference: Tumor Immunology and Immunotherapy; 2020 Oct 19-20. Philadelphia (PA): AACR; Cancer Immunol Res 2021;9(2 Suppl):Abstract nr PO001.

Expression of Programmed Death-Ligand 1 and Programmed Death-1 in Patients with Extramammary Paget's Disease.

Background:Extramammary Paget's disease (EMPD) is a rare skin cancer and sometimes has fatal prognosis. For progressive cases, therapeutic options are limited. In recent years, treatment with an anti-programmed death-1 (PD-1) antibody has improved the prognosis of various malignancies. In addition, correlations between PD-ligand 1 (PD-L1) expression in tumor cells and favorable responses to anti-PD-1 therapy have been reported for several cancers. There have been a few case series of analysis of PD-L1 expression in patients with EMPD.Aims:The purpose of this study was to investigate the relationship between the EMPD and PD-L1/PD-1 expression in Japanese EMPD patients.Materials and Methods:We investigated 39 patients with EMPD by immunohistochemical staining of PD-L1 and PD-1. We counted the number of tumor cells that were positive for PD-L1 and the number of tumor-infiltrating mononuclear cells (TIMCs) that were positive for PD-L1 and PD-1. We also analyzed correlations between the expression of PD-L1 and PD-1 in EMPD and patients’ characteristics.Results:We found that none of the Paget's cells expressed PD-L1. All of the specimens contained TIMCs, and some of the TIMCs expressed PD-L1 and PD-1. However, there was no correlation between the expression of PD-L1/PD-1 in TIMCs and patients’ characteristics.Conclusion:Although tumor cells did not express PD-L1 and the expression of PD-L1/PD-1 in TIMCs did not correlate with patients’ characteristics, future clinical studies should be carried out to explore another immune escape pathway in EMPD.

Association between Genetic and Immunological Background of Hepatocellular Carcinoma and Expression of Programmed Cell Death-1

Background and Aim: Immune checkpoint inhibitors are promising agents for the treatment of hepatocellular carcinomas (HCC) refractory to conventional therapies. To enhance the efficacy of this treatment, immunological and molecular characteristics of HCC with programmed cell death ligand 1 (PD-L1) should be explored. Methods: Clinical backgrounds, PD-L1 expression, and the amount of CD8+ tumor-infiltrating mononuclear cells (TIMCs) were analyzed in 154 HCCs. The expression of 3 stem cell markers and co-inhibitory receptors on tumor cells and TIMCs, respectively, were examined by immunohistochemical analysis. Somatic mutations in the 409 cancer-associated genes and TERT promoter were determined; HCCs were classified based on the presence of gene alterations affecting the 8 oncogenic pathways. The results were validated using the dataset from the Cancer Genome Atlas. Results: The expression of PD-L1 in the HCCs was positively correlated with progressive tumor features, the presence of cytokeratin 19 (CK19), Sal-like protein 4 (SALL4), and the mutations of genes involving the phosphatidyl inositol 3-kinase (PI3K)-Akt pathway. Although CD8+ cells were densely infiltrated in PD-L1-positive tumors, these TIMCs frequently expressed multiple co-inhibitory receptors. However, a subset of PD-L1-positive tumors characterized by activating mutations of the PI3K-Akt pathway showed a low degree of TIMCs. Conversely, PD-L1-negative HCCs were associated with mutations in the β-catenin pathway and a small number of TIMCs, although the expression of co-inhibitory receptors was rare. Conclusions: PD-L1-positive HCCs frequently showed an inflamed phenotype with stem cell features; a subset of PD-L1-positive HCCs with mutations in the PI3K-Akt pathway showed a non-inflamed phenotype. In HCCs with dense infiltration of TIMCs, CD8+ cells expressed multiple co-inhibitory receptors, suggesting T cell exhaustion. On the other hand, PD-L1-negative HCCs showed mutations leading to β-catenin activation and exhibited a non-inflamed background. These characteristics should be taken into consideration for developing novel combination therapies using immune checkpoint inhibitors.

Expression of PD-1 and CTLA-4 Are Negative Prognostic Markers in Renal Cell Carcinoma.

Immuno-oncological therapy with checkpoint inhibition (CI) has become a new standard treatment in metastatic renal cell carcinoma (RCC), but the prognostic value of the expression of CI therapy target molecules is still controversial. 342 unselected consecutive RCC tumor samples were analyzed regarding their PD-1, PD-L1, and CTLA-4 expression by immunohistochemistry (IHC). The prognostic values for cancer-specific survival (CSS) and overall survival (OS) were analyzed for those not exposed to CI therapy. The expression of PD-1 in tumor-infiltrating mononuclear cells (TIMC) and PD-L1 in tumor cells was detected in 9.4% and 12.3%, respectively (Immune reactive score (IRS) > 0). Furthermore, PD-L1 expression in TIMC (IRS > 0) and CTLA-4 expression in TIMC (>1% positive cells) was detected in 4.8% and 6.3%. PD-1 expression and CTLA-4 expression were significantly associated with a worse OS and CSS in log rank survival analysis and univariate Cox regression analysis. CTLA-4 expression is a prognostic marker that is independently associated with a worse outcome in multivariate Cox regression analysis in the whole cohort (OS: p = 0.013; CSS: p = 0.048) as well as in a non-metastatic subgroup analysis (OS: p = 0.028; CSS: p = 0.022). Patients with combined CTLA-4 expression and PD-1-expression are at highest risk in OS and CSS. In RCC patients, PD-1 expression in TIMC and CTLA-4 expression in TIMC are associated with a worse OS and CSS. The combination of PD-1 expression in TIMC and CTLA-4 expression in TIMC might identify high risk patients. This is, to our knowledge, the first description of CTLA-4 expression to be a prognostic marker in RCC.

Abstract B082: PD-1 to CD8 ratio on tumor-infiltrating lymphocytes and peripheral blood mononuclear cells as a predictor for determining response of glioblastoma patients to radiation-induced ICD-based DC vaccine therapy

Abstract Glioblastoma multiforme (GBM) is the most common and aggressive glioma within the central nervous system in adults. Radiation-induced ICD-based DC vaccine (RICD-DC vaccine) therapy plus conventional multi-modal regimen for GBM has been demonstrated with promising outcomes in clinical trials. However, some GBM patients received RICD-DC vaccine therapy did not increased survival rates than other GBM patients who only received conventional therapy. To investigate this issue, we conducted a retrospective study to analyze clinical and laboratory data to evaluate the factors which is critical for affecting the response rate of RICD-DC vaccine treatment. Patients with de novo GBM were enrolled (n=47) and divided into two subgroups: the first subgroup received post-surgical adjuvant immunotherapy with autologous RICD-DC vaccine (n=27) and the second received conventional treatment without immunotherapy (control, n=20). Quantitative immunohistochemistry for CD45, CD4, CD8, programmed death-1 (PD-1) and programmed death ligand 1 (PD-L1) was performed on patient tumor samples and peripheral blood mononuclear cells (PBMCs) at initial resection/biopsy before treatment. Pearson’s correlation, Cox proportional hazard model, and Kaplan-Meier analyses were performed to examine the correlations between these biomarkers expression and survival rates. In the RICD-DC vaccine group, patients with a lower PD-1+/CD8+ ratio (≤0.21) on tumor infiltrating lymphocytes (TILs) had longer overall survival (OS) (median 60.97 months, P &amp;lt; 0.001) than patients with a higher PD-1+/CD8+ ratio (median 20.07 months, , P &amp;lt; 0.001); 80% of the patients with a lower PD-1+/CD8+ ratio survived longer than 2 years whereas only 20% of patients with a higher PD-1+/CD8+ ratio. Similar results were observed in progression-free survival (PFS). Consistently, lymphocyte counts with lower PD-1+/CD8+ ratio (≤0.197) had much longer OS and PFS in patients’ PBMCs. In addition, there was a significant correlation of PD-1+/CD8+ ratio between TILs and PBMCs (Pearson’s correlation R2 = 0.6002, P &amp;lt; 0.001). By contrast, OS and PFS did not be impacted by CD4−, CD8−, but PD-1+, CD45+ TILs (P = 0.073 and P = 0.249, respectively). Moreover, the significant prognostic factors of OS and PFS also contained younger age (&amp;lt;57 years), gross total resection, and conventional treatment of concomitant chemoradiotherapy (CCRT) and PD-1+ lymphocyte counts in the RICD-DC vaccine group. In contrast to other factors, gender, counts of CD45+, CD4+ or CD8+ lymphocytes, tumoral PD-L1 expression, isocitrate dehydrogenase 1 mutation, and methylation status of O6 methylguanine-DNA methyltransferase promoter have no impact in both groups. Taken together, we found that TILs or PBMCs with lower PD-1+/CD8+ ratio in GBM patients with younger age, with gross tumor resection may be the desirable candidates who can be benefited from conventional multimodal regimen plus RICD-DC vaccine immunotherapy. Citation Format: Fang-Yu Lin, Chia-Ing Jan, Wan-Chen Tsai, Horng-Jyh Harn, Hsin-Man Lu, Ming-Chao Liu, Shao-Chih Chiu, Der-Yang Cho. PD-1 to CD8 ratio on tumor-infiltrating lymphocytes and peripheral blood mononuclear cells as a predictor for determining response of glioblastoma patients to radiation-induced ICD-based DC vaccine therapy [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B082.

Correction to: The Immunoscore is a Superior Prognostic Tool in Stages II and III Colorectal Cancer and is Significantly Correlated with Programmed Death-Ligand 1 (PD-L1) Expression on Tumor-Infiltrating Mononuclear Cells.

In the original article, Akihiko Kawahara's first name is incorrect. It is correct as reflected here.

The Immunoscore is a Superior Prognostic Tool in Stages II and III Colorectal Cancer and is Significantly Correlated with Programmed Death-Ligand 1 (PD-L1) Expression on Tumor-Infiltrating Mononuclear Cells.

In colorectal cancer (CRC), the indication for immune checkpoint inhibitors is determined by the microsatellite instability status of the tumors. However, an optimal biomarker for their indication has not been fully identified to date. This study aimed to establish the clinicopathologic importance of the Immunoscore (IS) in CRC and to clarify the relationships between the IS, programmed death-ligand 1 (PD-L1) expression, and tumor-associated macrophages. In 132 cases, CRC was diagnosed and surgically treated in our department from 2009 to 2010. Immunohistochemical staining using primary antibodies PD-L1, CD3, CD8, CD68, and CD163 was performed. The IS was determined according to the proposal of an international task force. Statistical analyses were performed to investigate the correlation between the IS, clinicopathologic variables, and expression of immune checkpoint molecules. The overall survival (OS) and relapse-free survival (RFS) in the high-IS group (I3-4) were significantly better than in the low-IS group (I0-2) (OS: P = 0.0420; RFS: P = 0.0226). The positivity rate for PD-L1 on tumor cells (tPD-L1) was only 0.8%, whereas that for PD-L1 on interstitial tumor-infiltrating mononuclear cells (iPD-L1) was 18.2%. The iPD-L1-positive group showed significantly better survival in terms of both OS and RFS than the iPD-L1-negative group (OS: P = 0.0278; RFS: P = 0.0253). The findings showed significant correlation between the IS and iPD-L1 expression (P < 0.0001). The study found that a high IS was a good indicator of a better prognosis and significantly correlated with iPD-L1 expression in CRC.

Checkpoint molecule PD-1-assisted CD8+ T lymphocyte count in tumor microenvironment predicts overall survival of patients with metastatic renal cell carcinoma treated with tyrosine kinase inhibitors.

PurposeThe aim of this study was to determine whether CD8+ T lymphocyte and its checkpoint-associated module programmed cell death protein 1 (PD-1)/main ligand of PD-1 (PD-L1) pathway impact overall survival (OS) in patients with metastatic renal cell carcinoma (mRCC) treated with tyrosine kinase inhibitors (TKIs).Materials and methodsA total of 231 mRCC patients, from 2007 to 2017, treated with sunitinib or sorafenib in Zhongshan Hospital, Fudan University were included in the study analyses. CD8, PD-1, and PD-L1 was assessed by immunohistochemistry on continuous paraffin-embedded slides. Kaplan–Meier method and COX regression model were applied in the survival analyses.ResultsBaseline characteristics were comparable between the training (n=118) and validation (n=113) sets. Patients with high CD8+ T lymphocytes infiltration and low PD-1 expression had longer survival in both sets (P=0.0106 and P=0.0047 in training set, P=0.0291 and P=0.0011 in validation set, respectively). However, survival stratified by PD-L1 was only insignificant or marginally significant. Multivariable analyses verified that CD8+ T lymphocytes, together with PD-1, but not tumor infiltrating mononuclear cells or tumor cells PD-L1, were independent prognostic factors (training set [HR 3.202, 95% CI 1.433–7.153, P=0.011] and validation set [HR 4.012, 95% CI 2.354–6.838, P<0.001]). Subsequent analysis revealed that the PD-1 high/CD8 low group had shorter survival (16 months) than PD-1 low/CD8 high group (51 months, P<0.0001). Combining the International Metastatic Renal Cancer Database Consortium system with the PD-1/CD8 model exhibited much better accuracy for the prediction of OS.ConclusionOur findings suggest that abundant CD8+ T cells are significantly associated with longer OS in mRCC patients treated with TKIs. The most influential checkpoint-associated molecule, PD-1, assisted CD8+ T cell-stratified patients and could be used as a better predictive and prognostic factor for the mRCC patients.

Abstract 720: Anti-OX-40 agonist antibody significantly increases therapeutic efficacy of TVGV-1 vaccine for HPV-positive cancers

Abstract Background: Human papillomavirus 16 (HPV16) E7 protein is a potential target antigen for HPV-positive cancers due to its interaction with pRb tumor suppressor protein. The clinical-stage therapeutic vaccine, TVGV-1, comprises of the PE-E7-K3 fusion protein and the adjuvant GPI-0100, has shown therapeutic effect preclinically by stimulating T cell response targeting HPV16 E7 protein. TVGV-1 was well tolerated clinically in more than 30 HPV+ cervical intraepithelial neoplasia (CIN) patients. Agonistic antibodies targeting OX40 (CD134) have shown remarkable single agent anti-tumor effect by modulating T cell response surrounding tumor environment, as well as the ability to combine with other immunotherapies in several mouse tumor models. Here we evaluated the synergistic therapeutic potential of TVGV-1 vaccine combined with anti-OX-40 agonist antibody for HPV-positive cancer therapy. Methods: Therapeutic and immune efficacy of TVGV-1 in combination with anti-OX-40 agonist antibody was tested in E7 antigen expressing TC-1 tumor mouse model. Mice with various treatments were assessed for tumor growth and survival. Peripheral and tumor-infiltrating immune cell profiles with various treatments were also assessed by flow cytometry. Results: Our results revealed that both the HPV16 E7-specific humoral and cell-mediated immunities solely elicited by TVGV-1 vaccination was synergistically enhanced more than two-fold by concomitant anti-OX-40 agonist antibody treatment. In a TC-1 tumor model, tumor growth and overall survival was not protected by OX-40 antibody treatment alone, and mice was protected with prolonged survival benefit when treated in combination of TVGV-1 and OX-40 antibody. In TC-1 tumor bearing mice, about 10% decrease of peripheral CD3+ T cell was observed upon TC-1 tumor development, and restoration was only observed when mice were treated with combined TVGV-1 and OX-40 antibody. When tumor infiltrating lymphocytes recognizing HPV E7 antigen were prepared from these mice, the amount of tumor-infiltrating mononuclear cells and CD3+ T cells were dramatically increased with combination treatment when compared to control group or to treatment alone groups. Conclusions: We demonstrated efficacy with combination approach between TVGV-1 vaccine and anti-OX-40 agonistic antibody in a pre-clinically mouse model. This enhanced activity is likely due to modulation of T-cell population in tumor and in periphery. This result will guide to a better design to perform clinical studies in order to obtain better outcome with TVGV-1 cancer vaccine intended for treating HPV E7 bearing tumors. Citation Format: Yin-Ching Lin, Yi-Tsui Chiu, Yi-Chia Lin, Jiun-Ming Wu, Chien-Hung Chen, Wen-Fang Cheng, Chia-Mao Wu. Anti-OX-40 agonist antibody significantly increases therapeutic efficacy of TVGV-1 vaccine for HPV-positive cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 720.

CD73 expression and clinical significance in human metastatic melanoma.

BackgroundCD73 is an ectoenzyme involved in the production of adenosine. It exerts immunosuppressive and protumoral roles and has emerged as a potential immuno-oncology target.ResultsCD73 expression was detected in TC in 54% of melanoma metastases, involving < 50% TC in the majority of the cases, with variable intensity. CD73 expression was significantly associated with a lower Breslow's depth of the primary lesion and was more frequent in patients having received prior non-surgical therapies. In an adjusted analysis, CD73 expression in TC (H-score > 37.5 or intensity > 1) significantly correlated to decreased overall survival (OS) from biopsy. Of the samples containing TIMC, 35% presented CD73+ TIMC. Highly infiltrated tumors were more likely to contain CD73+ TIMC. CD73 expression in TIMC (percentage ≥1%) significantly correlated with improved OS from biopsy.ConclusionsImmunohistochemistry detected CD73 expression in more than half of metastatic melanomas. While CD73 expression in TC significantly correlated with decreased OS, CD73 expression in TIMC significantly associated with improved OS. These results encourage the study of anti-CD73 therapies for metastatic melanoma patients.MethodsCD73 expression was assessed by immunohistochemistry in metastatic melanomas from 114 patients. Immunostainings were evaluated in tumor cells (TC) (percentage, intensity (1–3) and H-score) and in tumor-infiltrating mononuclear cells (TIMC) (percentage).

The expression of programed death ligand‐1 could be related with unfavorable prognosis in salivary duct carcinoma

Salivary duct carcinoma (SDC) is a rare tumor occurring in the salivary gland. SDC is a highly aggressive tumor and its prognosis is extremely poor. Effective treatments in advanced SDC have not yet been established. Recently, immune checkpoint inhibitors have paved the way for the treatment of various malignancies. We examined the expressions of programed death ligand (PD-L) 1/PD-L2 and programed death (PD-1), and the correlation of clinicopathological findings. We examined 18 cases of SDC and conducted immunohistochemical staining using formalin-fixed paraffin-embedded full-face sections. The expression of PD-L1 and PD-L2 in tumor cells was observed in nine cases (50%) and 14 cases (78%), respectively. Cases with a high expression of PD-L1 and PD-L2 were found in four (22%) and seven cases (39%), respectively. The cases with a high expression of PD-L1 showed significantly shorter overall survival compared to those with low PD-L1 expression and null expression. We also examined the expression of PD-L1/PD-L2 and PD-1 of tumor-infiltrating mononuclear cells (TIMC) in stroma. The expressions of PD-L1 in tumor cells and stroma had a significant correlation. Association between the expressions of PD-L1 in tumor cells and those of PD-1 in stroma was significant. However, PD-L2 expression in the tumor had no significant correlation with expression in TIMCs. PD-L1, PD-L2 and PD-1 expressions in stroma were not associated with patient prognosis. High PD-L1 expression in SDC was strongly associated with unfavorable prognosis, indicating that PD-1/PD-L1 inhibitors could be effective in SDC.

A comparison of the local immune status between the primary and metastatic tumor in colorectal cancer: a retrospective study

BackgroundThe anticancer immune response has been reported to correlate with cancer progression. Tumor-infiltrating lymphocytes (TILs), which are one of the indicators of host immunity, affect the tumor growth, metastasis and chemoresistance. Both TILs in the primary tumor and those in the metastatic tumor have been reported to be a useful predictor of the survival and therapeutic outcome. However, the correlation between the density of TILs in the primary and metastatic tumor is unclear. The aim of this study was to elucidate the correlation between the density of TILs in the primary and metastatic tumor.MethodsA total of 24 patients with stage IV colorectal cancer who underwent concurrent resection of the primary tumor and liver metastasis were enrolled in order to assess the correlation between the density of TILs in the primary tumor and that in the metastatic tumor. Hematoxylin and eosin (HE)-stained tumor sections were used for the evaluation of TILs. The density of TILs was assessed by the measurement of the area occupied by mononuclear inflammatory cells over the total stromal area at the invasive margin. In addition, to evaluate TIL subsets and the activation/suppression status of the lymphocytes, immunohistochemistry for CD4, CD8, Forkhead boxprotein P3 (FOXP3), programmed cell death 1 (PD-1), cytotoxic T-lymphocyte-associated protein 4 (CTLA4), inducible T-cell co-stimulator (ICOS), Glucocorticoid induced tumor necrosis factor receptor related protein (GITR), Human Leukocyte Antigen - antigen D Related (HLA-DR) and Granzyme B was performed, and the number of immunoreactive lymphocytes was counted.ResultsAccording to the evaluation using the HE-stained sections, the density of tumor-infiltrating mononuclear inflammatory cells in the primary tumor was significantly associated with that in the metastatic tumor. In addition, according to the immunohistochemistry evaluation, the density of CD4+, CD8+ and FOXP3+ TILs in the primary tumor and that in the metastatic tumor were significantly correlated with that in the metastatic tumor. Furthermore, the activation/suppression marker values of the lymphocytes (i.e., such as PD-1, ICOS, Granzyme B and the PD-1/CD8 ratio) in the primary tumor were correlated with values in the metastatic tumor.ConclusionsThe local immune status of the primary tumor was revealed to be similar to that of the metastatic tumor. This suggests that the evaluation of the local immunity of the primary tumor may be a substitute for the evaluation of the local immunity of the metastatic lesion. Therefore, information on the primary tumor may be useful when considering treatment strategies for metastatic lesions.

PD-L1 expression in malignant salivary gland tumors

BackgroundProgrammed death-1 ligand-1 (PD-L1) an important cancer biomarker that can suppress the immune system and its high expression is often reported to be related with increased tumor aggressiveness in some cancers. Here, we examined and evaluated PD-L1 expression in patients with malignant salivary gland tumor. Moreover, the relationship between PD-L1 immunolocalization and clinical pathological features, as well as the prognosis of malignant salivary gland tumors was investigated.MethodsWe examined PD-L1expression in 47 patients with malignant salivary gland tumor by immunohistochemical staining. PD-L1 positivity was defined as ≥5% in tumor cell membrane and evaluated according to three categories (0% = 0, < 5% = 1, ≥5% = 2) in tumor-infiltrating mononuclear cells (TIMCs). Fisher’s exact test was used to compare between PD-L1 expression and clinico-pathological features, and Kaplan–Meier method was used to estimate the distribution of OS by PD-L1 positivity.ResultsPD-L1 expression was detected in 51.1% of malignant salivary gland tumor tissues. No association was observed between PD-L1 immunolocalization in tumor and patient gender, or age. However, PD-L1 immunodetection of tumor cell membranes was significantly associated to stage, recurrence or metastasis after surgery, and patient outcome. On the other hand, PD-L1 immunodetection of tumor-infiltrating mononuclear cells (TIMCs) was significantly associated to recurrence or metastasis after surgery, and patient outcome. PD-L1 positivity in both tumor cell membrane and TIMCs was associated with shorter overall survival (OS) (p = 0.002 and p = 0.016, respectively).ConclusionThese findings suggested that patients with PD-L1 positive tumors or TIMCs appear to have poor clinical outcomes in malignant salivary gland tumors.

Expression of PD-1 and CTLA-4 Are Negative Prognostic Markers in Renal Cell Carcinoma

Background: Immun-oncological therapy with checkpoint inhibition (CI) has become a new standard treatment in metastatic renal cell carcinoma (RCC) but the prognostic value of the expression of CI therapy target molecules is still controversial. Material and Methods: 342 unselected consecutive RCC tumor samples were analyzed regarding their PD-1, PD-L1 and CTLA-4 expression by immunohistochemistry (IHC). The prognostic value for cancer-specific (CSS) and overall survival (OS) was analyzed. Results: Expression of PD-1 in tumor-infiltrating mononuclear cells (TIMC) and PDL1 in tumor cells was detected in 9·4% and 12·3% respectively (Immune reactive score (IRS) >0). Furthermore, PD-L1 expression in TIMC (IRS>0) and CTLA-4 expression in tumor tissue (>1% positive cells) was detected in 4·8% and 6·3%. PD-1 expression and CTLA-4 expression were significantly associated with worse OS and CSS in log rank survival analysis and univariate Cox's regression analysis. CTLA-4 expression is a prognostic marker, independently associated with worse outcome in multivariate Cox's regression analysis in the whole cohort (OS: p=0·013; CSS: p=0·048) as well as in a non-metastatic subgroup analysis (OS: p=0·028; CSS: p=0·022). Patients with combined CTLA-4 expression and PD1-expression are at highest risk in OS and CSS. Conclusion: In RCC patients, PD-1 expression in TIMC and CTLA-4 expression in cells in tumor tissue are associated with worse OS and CSS. The combination of PD-1 expression in TIMC and CTLA-4 expression in cells in tumor tissue might identify high risk patients. This is to our knowledge the first description of CTLA-4 expression to be a prognostic marker in RCC. Funding Statement: All investigations were financed from clinical funds of the participating institutes. All funding sources had no involvement in data collection, analysis, interpretation, writing, and the decision to submit the paper for publication. Declaration of Interests: AK reports personal fees and non-financial support from Bayer, personal fees and non-financial support from Pfizer, non-financial support from Novartis, non-financial support from Sanofi Avensis, non-financial support from BMS, and non-financial support from Ipsen; all outside the submitted work. AH reports grants, personal fees and non-financial support from Roche, personal fees from Astra Zeneca, personal fees and non-financial support from BMS, personal fees and non-financial support from MSD, grants and personal fees from Cepheid, grants and personal fees from Biontech, grants and personal fees from Qiagen, personal fees from Abbvie, grants and personal fees from Novartis, personal fees from Boehringer Ingelheim, and grants and personal fees from Janssen; all outside the submitted work. PJG reports personal fees and non-financial support from BMS, personal fees and non-financial support from Roche, personal fees and non-financial support from Novartis, personal fees and non-financial support from Bayer, personal fees and non-financial support from AstraZeneka, personal fees and non-financial support from Janssen, personal fees and non-financial support from Ipsen, and personal fees and non-financial support from Eisal; all outside the submitted work. BW reports personal fees from Astellas and personal fees from Jansssen-Cilag; all outside the submitted work. CGS, SW, HT, and FE have nothing to disclose. Ethics Approval Statement: The study was carried out according to the latest version of the Declaration of Helsinki and is approved by the institutional ethics committee.

Germline polymorphism of interferon-lambda3 is clinically associated with progression of renal cell carcinoma.

Renal cell carcinoma (RCC) is an immunogenic tumor that shows a metabolic shift to aerobic glycolysis. The immune system can have opposing host-protective and tumor-promoting effects, and aerobic glycolysis suppresses antitumor immunity. In addition to immunostimulatory effect, increasing numbers of studies have revealed that interferon (IFN) is also involved in promoting immunosuppression. Since various single nucleotide polymorphisms (SNPs) can influence the outcome of anticancer therapy, we investigated SNPs for IFN-lambda3, a new member of IFN family, in 53 patients with metastatic RCC who underwent cytoreductive nephrectomy. The 16 patients who were heterozygous/homozygous for the minor alleles of SNPs for IFN-lambda3 had a significantly worse response to sequential vascular endothelial growth factor-targeting therapy (P = 0.0029) and shorter survival (P = 0.0033) compared with the 37 patients possessing the major alleles of SNPs for IFN-lambda3. In these 16 patients, the primary tumor showed elevated glucose uptake on positron emission tomography with [18F] fluorodeoxyglucose (P = 0.0160) and increased expression of programmed cell death 1 (PD-1)-ligand 1 (PD-L1) and phosphorylated serine/threonine kinase Akt (P = 0.0006 and P = 0.0043, respectively) compared to the tumors of the patients without these alleles. Since IFN-induced PD-L1 expression on either tumor cells or tumor-infiltrating mononuclear cells can trigger immunosuppression due to crosstalk between cancer cells and T cells, IFN-lambda3 polymorphism might be linked to the immunosuppressive effects of IFNs in cancer. Although this retrospective study lacks mechanistic insight, our findings suggest that IFN-lambda3 polymorphism might be relevant to the progression of RCC.

Programmed death-ligand 1 expression correlates with diminished CD8+ T cell infiltration and predicts poor prognosis in anal squamous cell carcinoma patients

ObjectiveIncreased expression of programmed death-ligand 1 (PD-L1) on tumor cells can be found in various malignancies; however, very limited information is known about its role in anal squamous cell carcinoma (ASCC). This study explored PD-L1 expression in ASCC patients and its association with patients’ clinicopathological features, CD8+ T cell infiltration, and prognosis.MethodsFormalin-fixed paraffin-embedded tumor samples from 26 patients with ASCC were retrieved. The levels of PD-L1 expression on the membrane of both tumor cells and tumor-infiltrating mononuclear cells (TIMCs) were evaluated by immunohistochemistry. CD8+ T cell densities, both within tumors and at the tumor–stromal interface, were also analyzed. Baseline clinicopathological characteristics, human papilloma virus (HPV) status, and outcome data correlated with PD-L1-positive staining.ResultsPD-L1 expression on tumor cells and TIMCs was observed in 46% and 50% of patients, respectively. Nineteen patients (73%) were HPV positive, with 7 showing PD-L1-positive staining on tumor cells and 9 showing PD-L1-positive staining on TIMCs. Increasing CD8+ density within tumors, but not immune stroma, was significantly associated with decreased PD-L1 expression by both tumor cells and TIMCs (P=0.0043 and P=0.0007). Patients with negative PD-L1 expression had significantly better progression-free survival (P=0.038 and P=0.0443) and a non-statistically significant trend toward longer overall survival (P=0.0882 and P=0.1222) compared with patients with positive PD-L1 expression.ConclusionPD-L1 is widely expressed on the membrane of tumor cells and TIMCs in ASCCs. Its negative impact on prognosis may be due to the diminished CD8+ T cell infiltration within tumors.

Reply by Authors

No AccessJournal of UrologyLetter to the Editor/Errata1 Mar 2018Reply by Authors View All Author Informationhttps://doi.org/10.1016/j.juro.2017.09.159AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "Reply by Authors." The Journal of Urology, 199(3), pp. 855–856 References 1 : Association of PD-L1 expression on tumor-infiltrating mononuclear cells and overall survival in patients with urothelial carcinoma. Ann Oncol2015; 26: 812. Google Scholar 2 : Safety and efficacy of durvalumab (MEDI4736), an anti-programmed cell death ligand-1 immune checkpoint inhibitor, in patients with advanced urothelial bladder cancer. J Clin Oncol2016; 34: 3119. Google Scholar 3 : Atezolizumab in patients with locally advanced and metastatic urothelial carcinoma who have progressed following treatment with platinum-based chemotherapy: a single-arm, multicentre, phase 2 trial. Lancet2016; 387: 1909. Google Scholar 4 : Nivolumab monotherapy in recurrent metastatic urothelial carcinoma (CheckMate 032): a multicentre, open-label, two-stage, multi-arm, phase 1/2 trial. Lancet Oncol2016; 17: 1590. Google Scholar 5 : PD-L1 expression in melanoma: a quantitative immunohistochemical antibody comparison. Clin Cancer Res2017; 23: 4938. Google Scholar 6 : Comparison of different antibody clones for immunohistochemistry detection of programmed cell death ligand 1 (PD-L1) on non-small cell lung carcinoma. Appl Immunohistochem Mol Morphol2017; 10.1097/PAI.0000000000000531. Crossref, Google Scholar 7 : PD-L1 immunohistochemistry assays for lung cancer: results from phase 1 of the Blueprint PD-L1 IHC Assay Comparison Project. J Thorac Oncol2017; 12: 208. Google Scholar © 2018 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 199Issue 3March 2018Page: 855-856 Advertisement Copyright & Permissions© 2018 by American Urological Association Education and Research, Inc.MetricsAuthor Information Expand All Advertisement PDF downloadLoading ...