Expression of Immune Checkpoint Regulators, Cytotoxic T-Lymphocyte Antigen-4, and Programmed Death-Ligand 1 in Epstein-Barr Virus-associated Nasopharyngeal Carcinoma

Abstract

Background: Nasopharyngeal carcinoma (NPC) is the most common cancer arising from the nasopharynx with a poor prognosis. Targeting immune checkpoint is one of the new promising lines in cancer treatment. Cytotoxic T-lymphocyte antigen-4 (CTLA-4) and programmed death-ligand 1 (PD-L1) are immune checkpoints that regulate T-cell immune function.Aim: This work aimed to assess the immunohistochemical expression of PD-L1 and CTLA-4 in NPC and their ability to predict survival and response therapy and to check their validity as immunotherapy targets. Twenty-six cases of NPC were studied by immunohistochemistry for PD-L1 and CTLA-4 and by nested polymerase chain reaction followed by DNA sequencing for the presence of EBNA-1 gene of Epstein-Barr virus (EBV). All investigated cases were diagnosed and treated in the Zagazig University Hospital in the period from August 2015 to July 2018. EBNA-1 gene was identified in 84.6% of the cases. Whereas the expression of PD-L1 was noted in 46.2% of all cases studied, 54.6% of EBV-associated NPCs were found to express PD-L1. There was a significant association between PD-L1 expression and the advanced stage of the tumor (P < 0.001). CTLA-4 expression was observed in 88.4% of all NPC cases as cytoplasmic staining in both tumor cells and tumor-infiltrating lymphocytes. CTLA-4 expression in lymphocytes was associated with the presence of EBV. A significant association was detected between CTLA-4 and tumor-infiltrating lymphocyte expression on one side and the stage of the tumor on the other. High expression of CTLA-4 was significantly associated with disease progression and worse overall survival.Conclusion: PD-L1 and CTLA-4 are adverse prognostic markers in NPC. The authors propose that targeted therapy against PD-L1 and CTLA-4 will be a hopeful therapy for cases of NPC with resistance to concurrent chemoradiation treatment in Egypt, especially EBV-associated cases.

Combined Association of Tumoral PD-L1 Expression and Pretreatment Presence of Epstein-Barr Virus DNA With Risk Stratification and Prognosis of Patients With Nasopharyngeal Carcinoma

“…Therefore, PD-L1 may be an effective target for immunotherapy as well as a useful biomarker for predicting metastasis and survival. Many studies have investigated the role of PD-L1 expression and EBV in the prognosis and risk stratification of NPC, as well as other EBV-associated malignancies, but most of them did not combine tumor PD-L1 expression with EBV-DNA to further stratify NPC patients ( 30 , 33 , 47 – 51 ) Hu and colleagues stratified NPC patients into three groups based on the combination of EBV DNA load and PD-L1 expression on tumor infiltrating lymphocytes (TILs). The results indicated that a high log EBV-DNA load with low TIL PD-L1 was a poor prognostic factor for PFS ( 52 ).…”

The Prognostic Value of Cancer Stem Cell Marker, Cluster of Differentiation 133 Expression in Stage III Colorectal Carcinoma

“…Using IGg antibody against CD133 (CD133/Prominin-1 (PROM1) Concentrated, Code: RM0202, Clone: MD49R, Source: Rabbit, The EnVision TM FLEX + system (DAKO, North America Inc, CA, USA) was used for immunohistochemical staining, which was a two-step polymer-enhanced IHC detection method. Recommended 3in-1 specimen preparation procedure using pretreatment procedure: Deparaffinization, rehydration, and heat-induced epitope recovery were performed in paraffin-embedded tissue sections (7,8) . The staining steps with the recommended incubation time were reprogrammed in the Auto Stainer Link 48 software.…”

Role of Neoadjuvant Chemotherapy Followed by Interval Cyto-Reductive Surgery in Reducing Progression and Recurrence of Patients with Advanced Endometrial Cancer

“…Representative samples from all specimens were immunohistochemically stained with the PAX8, CK7, p16, p53, HNF-1B and, Napsin A antibodies. The immunohistochemistry technique was performed as fellow; initially, heating was used to extract antigens from tissue sections (16)(17)(18) . The sections were treated and chilled overnight with PAX8, CK7, p16, p53, HNF-1B and Napsin A rabbit monoclonal primary antibodies.…”