The structure of N-glycans on specific proteins can regulate innate and adaptive immunity via sensing environmental signals. Meanwhile, the structural diversity of N-glycans poses analytical challenges that limit the exploration of specific glycosylation functions. In this work, we used THP-1-derived macrophages as examples to show the vast potential of a N-glycan structural interpretation tool StrucGP in N-glycoproteomic analysis. The intact glycopeptides of macrophages were enriched and analyzed using mass spectrometry (MS)-based glycoproteomic approaches, followed by the large-scale mapping of site-specific glycan structures via StrucGP. Results revealed that bisected GlcNAc, core fucosylated, and sialylated glycans (e.g., HexNAc₄Hex₅Fuc₁Neu5Ac₁, N₄H₅F₁S₁) were increased in M1 and M2 macrophages, especially in the latter. The findings indicated that these structures may be closely related to macrophage polarization. In addition, a high level of glycosylated PD-L1 was observed in M1 macrophages, and the LacNAc moiety was detected at Asn-192 and Asn-200 of PD-L1, and Asn-200 contained Lewis epitopes. The precision structural interpretation of site-specific glycans and subsequent intervention of target glycoproteins and related glycosyltransferases are of great value for the development of new diagnostic and therapeutic approaches for different diseases.
Humans ; B7-H1 Antigen ; Glycosylation ; Polysaccharides/metabolism*

Humans ; B7-H1 Antigen ; Neoplasms/genetics*

B7-H1 Antigen ; Biomarkers, Tumor ; Consensus ; Humans ; Immunohistochemistry ; Neoplasms

Objective: The advent of immunotherapy has significantly changed the treatment and management of patients with advanced non-small cell lung cancer (NSCLC). Prior to initiation of immunotherapy, evaluation of programmed death ligand 1 (PD-L1) expression is required. One factor that affects PD-L1 expression in NSCLC is the presence of oncogenic driver mutations; however, little data on its association is available, especially in the Philippine setting. The study aims to determine the prevalence of PD-L1 expression and its association with driver mutations among patients with non-small cell lung cancer in a private tertiary care hospital in the Philippines. Methodology: The study was undertaken for a period of two years from July 2017-July 2019 at St. Luke’s Medical Center and included 446 NSCLC samples. PD-L1 was evaluated by immunohistochemistry using 22C3 anti-PD-L1 antibody clone, EnVision FLEX visualization system on Autostainer Link 48. Patient demographics and data on driver mutation testing were recorded. Statistical analysis was performed using logistic regression. Results: PD-L1 expression was observed in 273 (61.20%) of 446 cases, 119 (61.20%) of which demonstrated high PD-L1 expression while 154 (34.50%) had low PD-L1 expression. There was no significant association between PD-L1 expression and EGFR mutation, ALK mutation, age, and gender. For histologic type, high PD-L1 expression was significantly associated with adenocarcinoma and non- small cell carcinoma, NOS. Conclusion The overall prevalence of PD-L1 expression in non-small cell lung carcinoma is 61.20% based on the cases included. Although we did not find an association between PD-L1 expression and EGFR and ALK mutation, our study observed that ALK-mutated NSCLCs have 4.7 odds of having high PD-L1 expression, however, a higher sample size is warranted to truly determine significant association. The outcome of this study may provide help in the stratification of patients and predict those who will benefit from immunotherapy.
Carcinoma, Non-Small-Cell Lung ; B7-H1 Antigen

Bladder urothelial carcinoma(BUC)is a common malignant tumor in the urinary system.Pt-based chemotherapy has long been a standard therapeutic method for resectable or metastatic BUC,but with poor outcomes.Immune checkpoint inhibitors specific to programmed death 1(PD-1)/programmed death-ligand 1(PD-L1)and cytotoxic T lymphocyte-associated protein 4(CTLA-4)pathways have shown significant antitumor activities,safety,and enduring reactivity in clinical trials,thus creating a new epoch for the treatment of advanced-stage BUC.This article reviews the relationships of BUC with PD-1/PD-L1 and CTLA-4 pathways,as demonstrated in clinical trials.In particular,the authors elucidate the clinical studies on the application of immune checkpoint inhibitors in different BUC stages and their optimal combining strategies,with an attempt to improve the clinical use of immune inhibitors for BUC treatment.
Apoptosis ; B7-H1 Antigen ; CTLA-4 Antigen ; Immunotherapy ; Programmed Cell Death 1 Receptor ; Signal Transduction

Objective: To investigate the expression of VISTA and PD-L1 in triple-negative breast cancer (TNBC) and to explore its relationship with clinicopathologic features and prognosis. Methods: Ninety TNBC patients who underwent surgical resections between 2016 to 2018 in Jiangsu Province Hospital were selected. The expression of VISTA and PD-L1 in both tumor cells and immune cells was evaluated by immunohistochemistry, and the relationship with clinicopathologic parameters and prognosis was analyzed. Results: VISTA was expressed in 17.8% (16/90) of the tumors. The expression of VISTA in tumor cells was related to a higher Ki-67 proliferation index (P=0.02) and higher number of tumor-infiltrating lymphocytes (TIL, P<0.01). VISTA was expressed in 71.1% (64/90) of the immune cells and the expression correlated with smaller tumor size (P=0.02), lower T stage (P=0.04), higher number of TIL (P<0.01), higher number of CD8⁺T cells (P=0.03) and higher Ki-67 proliferation index (P=0.02). PD-L1 was expressed in 17.8% (16/90) of the immune cells and the expression correlated with higher histologic grade (P=0.04), higher Ki-67 proliferation index (P=0.02) and higher number of TIL (P<0.01). VISTA expression was higher in immune cells within TNBC patients than PD-L1 (P<0.01). Among 90 TNBC patients, complete follow-up was obtained in 85 patients, 8 of whom had recurrences or metastasis after surgery, and two patients cases died of recurrences or metastasis. Conclusions: The expression rate of VISTA is higher than that of PD-L1 in TNBC. The expression of VISTA in immune cells predicts a lower T stage. VISTA may act as an effective immunotherapy target.
B7-H1 Antigen/metabolism* ; Humans ; Ki-67 Antigen ; Prognosis ; Recurrence ; Triple Negative Breast Neoplasms/surgery*

OBJECTIVETo investigate the relationship of PD-L1 expression with the clinical features and prognosis of patients with primary acute leukemia.

METHODSSixty patients with primary acute leukemia(AL) from January 2013 to May 2015 in our hospital were selected as AL group, while 40 healthy persons were selected as control group. The flow cytometry was used to detect the PD-L1 expression in serum, the clinical data were collected and the treatment and lethality in AL patients were traced; the therapeutic efficacy in AL patients with different expression level of PD-L1 was compared.

RESULTSThe expression rate of PD-L1(+) in primary AL patients was significantly higher than that in controls(P<0.05). There was no significant difference in age, sex, WBC count, Plt count, Hb level and blast ratio in bone marrow between PD-L1(+) and PD-L1(-) patients(>0.05). The complete remission rate in PD-L1(+) patients was all lower than that of the negative patients after 1 and 2 course of treatment,the difference was statistically significant (P<0.05); the death rate of PD-L1(+) patients was higher than that of PD-L1(-) patients after treatment, while the survival rate of PD-L1(+) patients was lower than that of PD-L1(-) patients, the difference between 2 groups was statistically significant (P<0.05); the expression rate of PD-L1(+) in patients with ALL was not significantly different from that in AML (>0.05). The short-term remission rate and long-term survival rate of PD-L1(+) patients were significantly lower than those of PD-L1(-) patients, while the long-term mortality was statistically significant higher than that of PD-L1(-) patients in ALL and AML patients(P<0.05).

CONCLUSIONThe positive expression rate of PD-L1 in the patients with acute leukemia is higher, providing an indication for the evaluation of treatment efficacy and prognosis of patients.

OBJECTIVETo investigate the role of B7-H1 expression in IL-10 production, the B7-H1 and IL-10 expression levels in pancreatic carcinoma tissues and to analyze the correlation between B7-H1 expression and IL-10 level.

METHODSThe mRNA and protein levels expressions of B7-H1 and IL-10 in 35 cases of pancreatic cancer and corresponding paracarcinoma tissues and 5 cases of normal pancreas tissues were detected by RT-PCR, Western blot and immunohistochemistry respectively.

RESULTSThe findings for the first time provided the evidences that there was a clear trend for B7-H1 and IL-10 expressions to be most highly expressed in carcinoma tissue, intermediately expressed in paracarcinoma tissue, and expressed at the lowest level in normal pancreatic tissue at mRNA and protein levels. Moreover, there were statistically significant differences in B7-H1 and IL-10 expression between pancreatic carcinoma tissues, corresponding paracarcinoma tissues and normal pancreatic tissues at mRNA and protein levels (P < 0.05). Furthermore, the immunohistochemistry indicated that there were high expression levels of B7-H1 (60.5% +/- 12.7%) and IL-10 (65.3% +/- 16.2%) in pancreatic carcinoma tissues while there were no significant expressions in normal pancreatic tissues. Meanwhile, correlation analysis revealed that B7-H1 expression was significant associated with IL-10 level in tumor tissues at mRNA (P = 0.008, r = 0.841) and protein levels (P = 0.007, r = 0.838).

CONCLUSIONSOver-expression of B7-H1 may be responsible for the increasing IL-10 production in pancreatic cancer, which caused reduced immune response to tumor cells and contributed to pancreatic carcinoma escape from immune attack.

OBJECTIVETo investigate whether programmed death ligand 1 (PD-L1) expressed in the periodontal tissue of chronic periodontitis and the correlativity of PD-L1 and different degrees of chronic periodontitis, provide experience for immunoregulation mechanism, clinical treatment and prognosis of chronic periodontitis.

METHODSGingiva and periodontal tissue of healthy people and chronic periodontitis patients were collected. Based on clinical probing, periodontal tissue were classified into three groups: periodontal tissues of healthy people, periodontal tissue of mild chronic periodontitis, periodontal tissue of severe chronic periodontitis. Fluorescent quantitation polymerase chain reaction was applied to explore the expression of PD-L1 mRNA in the periodontal tissue of the different groups. Western blot and immunohistochemistry method were utilized to test the expression of PD-L1 protein in the periodontal tissue of the different groups. Combining with clinical image data, the relationship between differentially expressions of PD-L1 and different degrees of chronic periodontitis was analyzed.

RESULTSThe relative expression quantity of PD-L1 in the periodontal tissue of the mild chronic periodontitis was significantly higher that of the severe chronic periodontitis (P<0.01). The relative expression quantity of PD-L1 in the periodontal tissue of healthy subjects and severe chronic periodontitis had no statistical significance (P>0.05).

CONCLUSIONThe expression of PD-L1 in the periodontal tissue negativelv regulates inflammatory periodontal tissue damage.

OBJECTIVETo explore the expression of PD-L1 in acute leukemia patients, and to analyze the relationship of PD-L1 expression with the patients' clinical characteristics and prognosis.

METHODSThe expression of PD-L1 in leukemia cells of 75 patients including 59 de novo patients and 16 relapse/refractory patients with acute leukemia was detected by the flow cytometry, the clinical information was collected, and the therapeutic efficacy of de novo patients was analyzed.

RESULTSThe PD-L1 was expressed in human acute leukemia cells with total expression rate 32% (24/75), and its expression level in AML-M5 was higher than that in other leukemias [56.3% (9/16) vs 25.4% (15/59)], there was statistical significance (P = 0.019). The PD-L1 possitive rate in relapse/refractory group was higher than that in de novo patient group [(56.3% (9/16) vs 25.4% (15/59)], and there was statistical significance (P = 0.019). In 59 de novo patients, the CR rate of PD-L1 positive group after 1 course of chemotherapy was lower than that in PD-L1 negative group (66.7% vs 71.4%), the CR rate of PD-L1 positive group after 2 courses of chemotherapy was also lower than that in PD-L1 negative group (70% vs 88.6%). The relapse rate and the proportion of refractory patients in PD-L1 possitive group were higher than those in PD-L1 negative group. The expression of PD-L1 did not correlated with the clinical parameters, such as sex, age, extramedullary infiltration, percentage of blast cells in bone marrow, counts of WBC, RBC and platelet, as well as molecular biological features and cytogenetical characteristics.

CONCLUSIONPD-L1 is expressed in human acute leukemia cells, and may be involved in the immune escape and primary resistant mechanisms, PD-L1 may be used as an indicator for evaluation of the the patients' prognosis and reocurrence.