PD-L1 and PD-L2 expression in colorectal cancer

Context: The programmed death-1 (PD-1) is an immune checkpoint molecule that suppresses T-cell response. The binding of PD-1 to PD-L1/PD-L2 results cytokine production, and T-cell proliferation are reduced. Tumors expressing PD-L1 and PD-L2 escape from cytotoxic T-cells and are exposed to tumor progression. For this reason, immunotherapy has become a new option in the treatment of cancer. Aims: In this study, we examined the PD-L1 and PD-L2 expression in colorectal carcinoma (CRC), and evaluated the relationship between clinicopathological parameters and CD8+ T cells. Methods and Material: We evaluated CD8 expression in tumor-infiltrating lymphocytes and surrounding tumor lymphocytes with PD-L1, PD-L2 staining in tumor cells and immune cells formalin-fixed paraffin embedded samples of 124 patient diagnosed with CRC. Statistical Analysis Used: Pearson Chi-Square, Fisher Exact Chi-Square, and Pearson Exact Chi-Square analyses were used in the analysis of the cross tables. Survival distributions predicted Kaplan--Meier method and it was evaluated using log-rank statistics. Results: In our study, a significant correlation was found between PD-L1 expression and female sex and tumors with medullary morphology. No expression of PD-L2 was observed in tumors containing medullary morphology, and a statistically inverse relationship was observed between PD-L2 and the medullary component. PD-L1 positive tumor-infiltrating lymphocytes were determined to be an important predictor for recurrence-free survival. Conclusions: We believe that the evaluation of these parameters may be useful in the selection of patients who will benefit from immunotherapy in CRC cases.

Desbalances genómicos del locus 9p24.1 en pacientes argentinos con linfoma de Hodgkin clásico / Genomic imbalances at 9p24.1 locus in Argentine patients with classical Hodgkin lymphoma

Resumen Introducción: El linfoma de Hodgkin clásico presenta escasas células de Reed Sternberg/Hodgkin inmersas en un abundante microambiente tumoral. Los desbalances genómicos del locus 9p24.1 han sido asociados con alteraciones en la expresión de los genes del ligando de muerte celular 1 y 2, ambos reguladores de la respuesta inmune. Objetivo: Evaluar desbalances genómicos del locus 9p24.1 en células de Reed Sternberg/Hodgkin y del microambiente tumoral en biopsias de pacientes con linfoma de Hodgkin clásico y correlacionarlo con la expresión del ligando de muerte celular 1 y la presentación de la enfermedad. Materiales y Métodos: Se efectuó hibridación in situ en biopsias de 22 pacientes con linfoma de Hodgkin clásico dirigida a los genes del ligando de muerte celular 1 y 2. Las alteraciones se clasificaron en: amplificación, ganancia y polisomía. La expresión se evaluó mediante inmunohistoquímica. Resultados: Todos los pacientes mostraron alteraciones del número de copias. Se diferenciaron dos grupos: con amplificación (32%) y sin amplificación (68%); este último subdividido en: rico en ganancia (53%) y rico en polisomías (47%). El grupo rico en polisomías mostró mayor edad (p=0,027). El 40% de los pacientes con amplificación y rico en ganancias no presentó masa bulky. La expresión proteica mostró score +3 sólo en estos últimos. El title% de los casos ricos en polisomías presentaron monosomía del cromosoma 9 en los linfocitos circundantes respecto al 36,4% de los otros dos grupos. Conclusiones: Nuestros datos constituyen un aporte a la caracterización biológica del LHC, de interés en el marco de las nuevas modalidades terapéuticas. MÉD.UIS.2020;34(1):35-44.

Abstract Background: Classical Hodgkin lymphoma shows scarce tumor Reed-Sternberg/Hodgkin cells surrounded by a dense immune microenvironment. Genetic alterations at the 9p24.1 locus result in genomic imbalances in the copy number of PD-L1/PD-L2 genes, both of them being immune response regulators. Aim: To characterize genomic imbalances at the 9p24.1 locus in Reed-Sternberg/Hodgkin cells and immune microenvironment in biopsies of patients with Classical Hodgkin lymphoma and correlate it with PD-L1 protein expression and disease presentation. Material and Methods: Paraffin embedded biopsies of 22 patients with CHL were retrospectively evaluated by fluorescence in situ hybridization using SPEC CD274/PDCD1LG2/CEN9 DNA probe. The frequency of 9p24.1 alterations, amplification, copy gain and polysomy, were determined taking into account the number of gene copies with respect to the centromere. PD-L1 protein expression was evaluated by immunohistochemistry. Results: All cases presented alterations in the number of copies of PD-L1 / PD-L2, which are differentiated in two groups: with amplification (32%) and without amplification (68%). The latter was subdivided into rich in gains (RG) (53%) and rich in polysomies (RP) (47%). Groups with amplification and RG were younger than the RP group (p = 0.027). The latter was not associated with bulky disease, a fact observed in 40% of patients with amplification and RG. Protein expression showed score +3 only in the latter. All RP cases presented chromosome 9 monosomy in the surrounding lymphocytes, compared to 36.4% of the other two groups. Conclusions: Our data contributes to the biologic characterization of CHL, of interest in the context of new therapeutic modalities. MÉD.UIS.2020;34(1):35-44

Application of PD-L1, PD-L2, CD30, CD23 and BCL-6 in Diagnosis and Prognostic Evaluation of Primary Mediastinal Large B-cell Lymphoma / 肿瘤防治研究

Objective To explore the application value of treatment-related markers PD-L1, PD-L2, CD30, CD23, BCL-2, BCL-6, MUM1 and GATA3 in the diagnosis and prognostic evaluation of primary mediastinal B-cell lymphoma(PMBL). Methods A retrospective study was conducted on 34 patients diagnosed with PMBL, and 31 patients with DLBCL-NOS which was not primary in the mediastinum were taken as control group. The expressions of 8 proteins were detected by IHC staining. Results The median percentages of tumor cells with PD-L1, PD-L2 and CD30 expression in PMBL group were 70% (30%, 90%), 25% (0, 70%) and 17.5% (0, 60%) respectively, which were significantly higher than those in the DLBCL-NOS group (P < 0.05). The positive rates of CD30 and CD23 in PMBL group were 61.76% (21/34) and 76.47% (26/34) respectively, significantly different with those in the DLBCL-NOS group (P=0.000). The survival curve of PMBL patients with CD30 or BCL-6 expression showed a trend of poor prognosis, despite the P value was > 0.05. Conclusion The high expression levels of PD-L1, PD-L2 and CD30 in PMBL are helpful to accurately identify more patients who may respond to immune or targeted therapy. Immunohistochemical staining of PD-L1, PD-L2, CD30 and CD23 is helpful for the differential diagnosis of PMBL and DLBCL-NOS. As candidate prognostic indicators of PMBL, CD30 and BCL-6 should be further studied in a larger number of samples.

Research Progress of PD-1/PD-L1/2 Pathway in Multiple Myeloma / 肿瘤防治研究

Multiple myeloma is a type of hematological malignancy caused by clonal plasma cell proliferation. In recent years, the application of proteasome inhibitors and immune drugs (IMiDs) in multiple myeloma has prolonged the survival period of patients and improved the quality of life. However, due to the heterogeneity, recurrence and drug resistance of the disease, MM is still incurable. Relatively good response rates of immune checkpoint inhibitors such as Pembrolizumab, Nivolumab, Pidilizumab, Atezolizumab and Durvalumab offer MM a new hope. This article reviews the relevant literature in recent years, introduces the immune pathogenesis of MM, PD-1/PD-L1/2-related immune pathway and the current status of immunotherapy, to provide reference for the immunotherapy of MM.

Protocol for a prospective multicenter intervention study (URivo study) assessing biomarkers in patients with previously treated advanced clear cell renal cell carcinoma receiving nivolumab

Background:In recent years, immune checkpoint inhibitors have been introduced into routine clinical practice for treating patients with a wide variety of malignant tumors, including advanced renal cell carcinoma (aRCC), resulting in the significant improvement of the prognosis of these patients. However, a reliable biomarker prediciting the clinical course in patients receiving nivolumab has not yet been developed; accordingly, the URivo study was planned to investigate the significance of various candidate biomarkers for aRCC patients treated with nivolumab.Methods:This was designed as a prospective multicenter intervention study, and will include a total of 200 aRCC patients who are scheduled to receive nivolumab followed by treatment with either 1 or 2 tyrosine kinase inhibitors (TKIs). Using resected tumor tissues and serum samples prior to the introduction of nivolumab, the following assessments will be conducted: programmed death ligand-1 (PD-L1) and PD-L2 gene copy number gains by fluorescence in situ hybridization, serum concentrations of PD-L1 and PD-L2 by enzyme-linked immunosorbent assay, and expression ofseveral proteins involved in apoptosis, epithelial-mesenchymal transition, signal transduction and immune reaction by immunohistochemical staining. The outcomes of these assays will be evaluated focusing on the association with the response to nivolumab, overall survival, progression-free survival and disease-specific survival.Conclusions: The significance of various types of candidate biomarker, particularly PD-L1 and PD-L2, will be intensively investigated in this study, and this may offer unique information to determine the optimal indication of nivolumab for aRCC patients following the failure of TKIs.Trial Registration:UMIN000030400; registered April 1, 2018

Expression and clinical significance of PD-1/PD-Ls in EBV-positive T/NK lymphoprolif-erative disorders / 中国肿瘤临床

Objective: To investigate the expression and clinical significance of programmed death-ligand 1 (PD-L1), programmed death-ligand 2 (PD-L2), and their receptor programmed cell death protein 1 (PD-1) in EBV-positive T/NK lymphoproliferative disease [Epstein-Barr virus-positive T/natural killer (NK)-cell lymphoproliferative disease, EBV(+)-T/NK-LPD]. Methods: The pathological paraffin-embedded tissues of 17 patients with EBV(+)-T/NK-LPD from the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2017 were collected. These patients include 12 males and 5 females, aged 10-82 years old, the average age being 29 years, 4 people in gradeⅠ, 7 in gradeⅡ, 3 in gradeⅢ, and 3 people with hydroa vacciniforme-like lymphoproliferative disorders. Immunohistochemical SP method was used to detect the expression of PD-1, PD-L1, and PD-L2 in human EBV(+)-T/NK-LPD tissues. The relationship between PD-1, PD-L1, PD-L2 expression, and clinicopathological parameters, pathological grades and prognosis were analyzed by Fisher's exact probabilities and Spearman rank correlation. Result: After statistical analysis, the results showed that in 17 cases of tissue samples, there were 12 cases with positive PD-1 expression, 6 cases with positive PD-L1 expression and 5 cases with positive PD-L2 expression. There was no significant correlation between PD-1 and PD-L2 expression and prognosis (P>0.05). PD-L1 expression showed a positive correlation with prognosis (P<0.05). There was no significant correlation between the expression of PD-L1 and PD-L2 with age, sex, as well as LDH and Ki-67 levels (P>0.05). Moreover, there was no significant correlation of PD-1 and PD-L2 expression with pathological grade (r=0.141, r=-0.149, both P>0.05). However, there was a negative correlation between the PD-L1 expression and pathological grade (r=-0.563), and the correlation between the PD-L1 ex-pression and pathological grade was statistically significant (P<0.05). Conclusions: PD-1, PD-L1, and PD-L2 are abnormally expressed in the pathological tissues of EBV(+)-T/NK-LPD. Although there was no significant correlation between the expression of PD-1 and prognosis or pathological grade, it was significantly higher in EBV+T/NK-LPD. PD-1/PD-Ls associated signaling pathway is expected to be a potential new target for EBV(+)-T/NK-LPD immunotherapy.

PD-L1 and PD-L2 expressions in human breast cancer and their significance / 临床与实验病理学杂志

Purpose To investigate the expressions of costimulation molecules PD-L1 and PD-L2 in human breast cancers and their association with clinical and pathologic characteristics of the patients.Methods The expressions of PD-L1,PD-L2 in 10 cases of normal human breast tissue,18 cases of ductal carcinoma in situ,and 40 cases of breast cancer were detected by immunohistochemistry SP two-step staining,and the relationship between their expressions and clinical and pathologic characteristics was evaluated.Results There was no PD-L1 or PD-L2 expression in normal breast tissue,faint expressions of PD-L1 and PD-L2 were determined in ductal carcinoma in situ.45％ (18/40) PD-L1 and 35％ (14/40) PD-L2 were detected in breast cancers.PD-L1 and PD-L2 expression in cancer cells were related to the lymph node metastasis,ER and PR (P ＜ 0.05).High expression of PD-L1 and PD-L2 in breast cancer patients were correlated with negative ER,PR and HER-2 (P ＜ 0.05).Conclusion PD-L1 and PD-L2 express aberrantly in breast cancer and are closely associated with the lymph node metastasis of breast cancer.PD-L1 and PD-L2 may be the potential immunotherapy targets of breast cancer.

Analysis and experimental evidence on interaction networks among PD-1 and other proteins / 国际检验医学杂志

Objective An interaction network among PD-1-ligands and other proteins were built to add deeper understanding of PD-1 signal pathway and to supply the theoretical data on the clinical application study of PD-1 and its ligands.Methods Searching the literature about PD-1 and its ligands in the PubMed,the result of which would be used to summarize the proteins that had been reported to have interactions with PD-1 and its ligands,and then,quadratic search would be performed on these proteins.Finally,the software Cytoscape would be used to build and analyze the interaction network and verified by PCR.Results There were 122 and 126 nodes in the PD-1/PD-L1 network and PD-1/PD-L2 network respectively.These proteins were involved in TCR signal path-way,cell adhesion,JAK-STAT signal pathway and interaction between cytokines.Meanwhile,a perspective that PD-L1 may influ-ence on CXCR4 through JAK-STAT pathway based on the network which had been supported by qPCR.Conclusion The bioinfor-matics suggested that there are differences between the mechanism of PD-1/PD-L1 and PD-1/PD-L2.Additionally,the method, which was based on the literature mining and bioinformatics,is good to put the biological literature to rational utilization and to pro-vide guidance for experiments.

Expression and clinical significance of PD-L2 molecule on monocytes of peripher-al blood in systemic lupus erythematosus / 中国免疫学杂志

Objective:To analyze PD-L2 expression on monocytes of peripheral blood cells in systemic lupus erythematosus ( SLE) and it′s correlation with the degree of disease activity .Methods:Peripheral blood of 26 cases of SLE patiens and 38 cases of healthy controls were collected .Peripheral blood mononuclear cells ( PBMC) were isolated and realtime PCR was carried on to analyze the PD-L2 gene expression.At the same time flow cytometry was performed to analyze the CD 14 and PD-L2 expression.Results:PD-L2 was significantly up-regulated on monocytes in RA patients than in healthy controls and had correlation with the disease activity and the SLEAI score.Conclusion:These findings help to clarify the function of PD-L2,including its potential role as a biomarker for SLE .

Progress in Clinical Research on Novel PD-1 inhibitor Nivolumab / 中国药师

Objective:To introduce a programmed death-1 (PD-1) inhibitor nivolumab used as a new antitumor agent. Methods:According to the literatures, the action mechanism of nivolumab and the clinical trial results on the main indications approved or being investigated in phase III trials were reviewed and evaluated. Results:Nivolumab could restore the antitumor activity of T cells through binding with PD-1 and consequently blocking its interaction with the key ligands of PD-L1 and PD-L2. Several completed and ongoing clinical trials showed that nivolumab used alone or combined with chemotherapy or CTLA-4 inhibitor ipilimumab exhibited better effica-cy when compared with current clinical used chemotherapy drugs in the treatment of melanoma, non-small cell lung cancer and renal cell carcinoma. Nivolumab was well tolerated during the treatment with such main grade 3-4 adverse events as immune-mediated pneu-monia, abnormal liver functions and fatigue. Conclusion:Through its anti-tumor immune response, nivolumab can improve the clinical efficacy in the treatment of various tumors including melanoma, non-small cell lung cancer and renal cell carcinoma.

The expression of PD-L2 in human cervical carcinomas and the influence to peripheral blood T lymphocytes apoptosis / 重庆医学

Objective To determine PD-L2 expression in human cervical squamous cell carcinomas and analyze its association with the clinical and pathologic characteristics of these cases ;observe the role of recombinant PD-L2 protein on apoptosis of active peripheral blood T lymphocytes of cervical carcinoma patients .Methods PD-L2 expression in cervical squamous cell carcinomas was determined by immunohistochemistry staining ,and the association between PD-L2 expression with the clinical and pathologic characteristics of these cases was analyzed .In vitro ,the peripheral blood T lymphocytes of cervical carcinoma patients were divided into blank group ,PD-L2 group and PD-L2+anti-PD-1 group respectively .After these active T cells were cultured 72 h ,their apop-totic rates were detected by flow cytometry .Results PD-L2 expressed in 53 .3% (32/60)cervical squamous cell carcinomas ,and it′s expression associated with lymph node metastasis of these cases (P< 0 .05) .In vitro ,PD-L2 promoted apoptosis of CD4+ T and CD8+ T lymphocytes ,the apoptotic rates were 17 .0% and 22 .4% respectively ,which were higher than 9 .0% and 16 .2% of blank group;however ,the apoptotic rates dropped to 11 .1% and 17 .5% in PD-L2+anti-PD-1 group .Conclusion PD-L2 aberrantly ex-presses in cervical squamous cell carcinomas and is associated with their lymph node metastasis .PD-L2 promotes apoptosis of T lymphocytes and depresses the anti-tumor immunity of cervical microenvironment ,and promotes lymph node metastasis of these cancers .PD-L2/PD-1 pathway may be a potential immunotherapy target of cervical squamous cell carcinomas .