B7-H3 Levels Research Articles

Differential expression of PD-L1 and Th1 response of lymphocytes co-cultured with human gastric organoids infected with Helicobacter pylori strains isolates from different gastric pathologies

Abstract Helicobacter pylori (Hp) bacteria successfully establishes chronic infection leading to chronic gastritis, peptic ulcer disease (PUD), and gastric cancer (GC). Since we have previously shown that Hp hijacks expression of checkpoint immunoregulators (i.e., PD-L1), we hypothesized that strains from different gastric pathologies differ in their ability to evade the host response. We used human gastric organoids (hGOs), which recapitulate polarized epithelium, gastric gland and pit cell markers observed in the stomach. We cultured hGOs in transwell inserts which were exposed to Hp on the apical surface and cultured with naïve T cells on the basolateral side of polarized epithelial cells, thereby reproducing the interactions observed in vivo. After 7 days of culture, we recovered all cells and evaluated levels of PD-L1, B7-H3, B7-H4, and CTLA4; and in CD4+ T cells markers of Th1, Th2, Th17 and Treg. Infection of hGOs with Hp strains isolated from cases of gastritis, PUD or GC, all led to increased expression of negative immune checkpoint regulators, predominantly PD-L1, which binds PD-1 on T cells and promotes loss of effector functions, apoptosis, and reduced T cell-target cell contact. Interestingly, PUD strains are the stronger inducers of PD-L1 and GC strains were the lowest inducers of Th1 cells (Tbet+, IFNγ+), which suggest that different strains differ in their orchestration of the host response, which may contribute to the type of pathology elicited. Our data suggest that this co-culture system to evaluate expression of immune checkpoint regulators and CD4+ T cell differentiation in the context of strains comparison may provide insights in differential expression of candidate biomarkers of disease and future therapies.

MP29-19 SIPULEUCEL-T IMMUNOTHERAPY FOR CASTRATE-RESISTANT PROSTATE CANCER MODULATES SOLUBLE B7-H3: A NOVEL MECHANISM OF ACTION

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I1 Apr 2018MP29-19 SIPULEUCEL-T IMMUNOTHERAPY FOR CASTRATE-RESISTANT PROSTATE CANCER MODULATES SOLUBLE B7-H3: A NOVEL MECHANISM OF ACTION Andrew Chang, Smita Nair, Daniel George, and Brant Inman Andrew ChangAndrew Chang More articles by this author , Smita NairSmita Nair More articles by this author , Daniel GeorgeDaniel George More articles by this author , and Brant InmanBrant Inman More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.939AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES B7-H3 is an immune checkpoint molecule widely expressed by prostate cancer, that exists in soluble form in the blood, and that impairs T cell function. We examined whether a cell-based immunotherapy for castrate-resistant prostate cancer (CRPC), sipuleucel-T, might modulate B7-H3 as part of its mechanism of action. METHODS We conducted a clinical trial (NCT02036918) where men with lymph node predominant oligometastatic CRPC were randomized to undergo either (a) sipuleucel-T immunotherapy followed by salvage lymphadenectomy (n=15) or (b) salvage lymphadenectomy followed by sipuleucel-T (n=5). Serum samples were taken at baseline, prior to each sipuleucel-T leukapheresis session (x3), and two weeks after the last sipuleucel-T infusion. Serum samples were then tested in triplicate for soluble B7-H3 using a solid phase sandwich ELISA assay. The assay was calibrated against highly purified NS0-expressed recombinant human B7-H3. ELISA was performed in 90 well microplates that were read in a SpectraMax microplate reader at 450 nm with wavelength correction set to 540 nm. B7-H3 concentrations were calculated using a 4-parameter logistic model curve fit (Hill equation). Pre- and post-intervention serum B7-H3 levels were compared with a paired T test. RESULTS The mean minimum detectable dose of B7-H3 for the assay was 0.160 ng/mL. Vaccination with sipuleucel-T resulted in significant drop in serum B7-H3 levels from a mean pre-sipuleucel-T level of 29.6 ng/mL [SD = 6.7] to a mean post-sipuleucel-T level of 24.0 ng/mL [SD = 5.5] (P <0.001). In contradistinction, pre-surgery mean serum B7-H3 was 22.1 ng/mL (SD = 5.5) and post-surgery mean serum B7-H3 was 29.2 ng/mL (SD = 9.3), a difference of 7.1 ng/mL (P = 0.002), indicating that surgery caused an increase in serum B7-H3 levels (Figure). CONCLUSIONS Sipuleucel-T immunotherapy is known to prolong survival in CRPC. Our results suggest that one potential mechanism of action of this vaccine (and possibly other prostate cancer vaccines) is to reduce the production of soluble B7-H3 by prostate cancer cells, which could lead to improved immune responsiveness of the host immune system. However, the postoperative inflammatory response may abrogate some of this benefit. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e373 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Andrew Chang More articles by this author Smita Nair More articles by this author Daniel George More articles by this author Brant Inman More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

B7-H3 in tumors: friend or foe for tumor immunity?

B7-H3 is a type I transmembrane co-stimulatory molecule of the B7 family. B7-H3 mRNA is widely distributed in most tissues; however, B7-H3 protein is not constitutively expressed. Few molecules have been shown to mediate the regulation of B7-H3 expression, and their regulatory mechanisms remain unexplored. Recently, TREM-like transcript 2 (TLT-2) has been identified as a potential receptor of B7-H3. However, TLT-2 may not be the only receptor of B7-H3, as B7-H3 has many contradictory roles. As a co-stimulatory molecule, B7-H3 increases the proliferation of both CD4+ and CD8+ T-cells and enhances cytotoxic T-cell activity. However, greatly increased T-cell proliferation and IL-2 levels have been observed in the absence of B7-H3. Thus far, it has been shown that various tumors test positive for B7-H3 expression and that B7-H3 levels correlate with tumor growth, invasion, metastasis, malignant stage, and recurrence rate. Furthermore, transfection of cells with a B7-H3 plasmid and treatment with monoclonal antibodies to block B7-H3 are the main immunotherapeutic strategies for cancer treatment. Several groups have generated anti-B7-H3 antibodies and observed tumor growth suppression in vitro and in vivo. Therefore, it is likely that B7-H3 plays an important role in cancer diagnosis and treatment, aside from its role as a co-stimulatory molecule.

P2.07-021 A Checkpoint Molecule B7-H3 as a Novel Immune Therapy Target for Non-Small Cell Lung Cancer (NSCLC)

Anti-PD-1 immune therapy improved survival in NSCLC, whereas some patients were not responding to this treatment, indicating the requirement of alternative strategies for these patients. B7-H3, an immune checkpoint molecule, has known to be expressed in some cancer cells including NSCLC. In this study, we examined the therapeutic potential of targeting B7-H3 using a mouse model, also elucidated the expression levels of B7-H3 on NSCLC tumor cells.

B7-H3 Expression in NSCLC and Its Association with B7-H4, PD-L1 and Tumor-Infiltrating Lymphocytes.

Purpose: The immune checkpoint PD-1 and its receptor B7-H1 (PD-L1) are successful therapeutic targets in cancer but less is known about other B7 family members. Here, we determined the expression level of B7-H3 protein in non-small cell lung cancer (NSCLC) and evaluated its association with tumor-infiltrating lymphocytes (TIL), PD-L1, B7-H4, and major clinicopathologic characteristics is in 3 NSCLC cohorts.Experimental design: We used multiplexed automated quantitative immunofluorescence (QIF) to assess the levels of B7-H3, PD-L1, B7-H4, and TILs in 634 NSCLC cases with validated antibodies. Associations between the marker levels, major clinicopathologic variables and survival were analyzed.Results: Expression of B7-H3 protein was found in 80.4% (510/634) of the cases. High B7-H3 protein level (top 10 percentile) was associated with poor overall survival (P < 0.05). Elevated B7-H3 was consistently associated with smoking history across the 3 cohorts, but not with sex, age, clinical stage, and histology. Coexpression of B7-H3 and PD-L1 was found in 17.6% of the cases (112/634) and with B7-H4 in 10% (63/634). B7-H4 and PD-L1 were simultaneously detected only in 1.8% of NSCLCs (12/634). The expression of B7-H3 was not associated with the levels of CD3-, CD8-, and CD20-positive TILs.Conclusions: B7-H3 protein is expressed in the majority of NSCLCs and is associated with smoking history. High levels of B7-H3 protein have a negative prognostic impact in lung carcinomas. Coexpression of B7-H3 with PD-L1 and B7-H4 is relatively low, suggesting a nonredundant biological role of these targets. Clin Cancer Res; 23(17); 5202-9. ©2017 AACR.

Different expression of B7-H3 in the caput, corpus, and cauda of the epididymis in mouse

BackgroundB7-H3, a member of the B7 family of the Ig superfamily of proteins, has been detected in the epididymis, which is a storage organ related to sperm maturation. However, the characteristics of its expression in different regions of the epididymis remain unknown. Our aim was to investigate the expression of B7-H3 in the caput, corpus, and cauda of the epididymis.MethodsWe extracted epididymis specimens from adult male C57BL/6 mice. The expression of B7-H3 was then measured with immunohistochemistry, enzyme-linked immunosorbent assay (ELISA) and western blotting.ResultsB7-H3 protein was predominantly detected on the membrane and in the cytoplasm of the principal cells in the epididymis. Moreover, the level of B7-H3 in the corpus of the mouse epididymis was significantly higher than that in the caput (p < 0.05) or the cauda of the epididymis (P < 0.05). However, there was no remarkable difference in the level of B7-H3 between the caput and the cauda (p > 0.05).ConclusionsThe caput, corpus, and cauda of the mouse epididymis all expressed B7-H3 protein. However, the levels of B7-H3 were different in the three regions of the epididymis.

Early Detection of Hepatocellular Carcinoma in Patients with Hepatocirrhosis by Soluble B7-H3

Increasing evidence has demonstrated an association between increased soluble B7-H3 (sB7-H3) levels and unfavorable progression in patients with malignant tumors. In the present study, we detected sB7-H3 levels in serum to investigate the value of sB7-H3 as a tool for differential diagnosis of cirrhotic patients with or without early-stage hepatocellular carcinoma (ESHCC). We also assessed the diagnostic value of sB7-H3regarding the prediction of overall survival (OS) of cirrhotic patients with ESHCC. sB7-H3 expression was measured in 91 healthy volunteers, 149 cirrhotic patients with ESHCC, and 87 cirrhotic patients by ELISA, and correlations between DCP1a level and clinical characteristics were analyzed. SB7-H3 concentrations were significantly higher in patients with ESHCC than in cirrhotic patients (P < 0.001). Using 48.34 ng/mL as a cutoff value, the sensitivity and specificity of sB7-H3 in differentiating between cirrhotic patients and cirrhotic patients with ESHCC were 76.5 and 93.1%, respectively. Moreover, high serum sB7-H3 in cirrhotic patients with ESHCC correlated with tumor size, tumor stage, vascular invasion, and tumor differentiation. The area under the curve (AUC) value for sB7-H3 (0.898) was significantly higher than those for AFP (0.789), CA199 (0.627), and CA125 (0.545) for differentiating between cirrhotic patients with ESHCC and sex- and age-matched cirrhotic patients without ESHCC. Our data indicate that serum sB7-H3 serves as a valuable biomarker for cirrhotic patients with ESHCC and that high levels of sB7-H3 correlate with poor clinical outcomes.

B7-H3 promotes the proliferation, migration and invasiveness of cervical cancer cells and is an indicator of poor prognosis.

B7-H3 is an immune regulatory molecule whose aberrant expression in tumors is associated with adverse outcomes. Upregulation of B7-H3 may promote tumor cell proliferation and metastasis invitro, but the role of B7-H3 in cervical cancer has not yet been investigated. We measured B7-H3 expression in 90cervical cancer patient and 20non‑cervical lesion patient tissues using immunohistochemistry and in 30cervical cancer patient and 30healthy donor blood samples using ELISA. The association of B7-H3 expression and the prognosis of cervical cancer patients was investigated. B7-H3 knockdown in CaSki and SiHa cell lines was performed using small hairpin (sh)RNA lentiviral transfection and B7-H3 overexpression in CaSki and HeLa cell lines was performed using plasmid-vector lentivirus transduction. Cell proliferation, invasion and migration were then measured using MTT and Transwell assays invitro. B7-H3 expression was significantly higher in the cervical cancer tissues compared to that noted in the normal cervical tissues (mean 72.22 vs.15.00%; p<0.001). Using Kaplan‑Meier and Cox analyses, our data revealed that patients with strong intensity staining were significantly more likely to have a worse prognosis. The B7-H3 level in cervical cancer patient blood was significantly higher than that in the normal donors (13.41±6.12 vs. 9.90±3.16ng/ml; p=0.007). MTT assay revealed that high expression of B7-H3 promoted cervical cancer cell proliferation. Transwell assay data revealed that high expression of B7-H3 enhanced cervical cancer cell migration and invasion (CaSki,p=0.003; HeLa,p=0.03). In conclusion, expression of B7-H3 was significantly higher in cervical cancer tissues compared to normal cervical tissues, and this high expression was associated with worse prognosis for cervical cancer patients. In addition, B7-H3 promoted proliferation, invasion and migration of cervical cancer and may be a potential target for treating cervical cancer.

P2.01-046 Quantitative Measurement of B7-H3 Protein Expression and Its Association with B7-H4, PD-L1 and TILs in NSCLC

B7-H3 (CD276) is a type I transmembrane protein that belongs to the B7 immunoregulatory family including PD-L1 (B7-H1) and is upregulated in multiple malignancies including Non-Small Cell Lung Cancer (NSCLC). Clinical activity of monoclonal B7-H3 blocking antibodies such as Enoblituzumab are under investigation. In this study we measured the levels of B7-H3 protein in NSCLC and studied its association with major tumor infiltrating lymphocyte (TIL) subsets, levels of PD-L1, B7-H4 and clinico-pathological characteristics in three independent NSCLC cohorts. We used automated quantitative immunofluorescence (QIF) to assess the levels of B7-H3 (clone D9M2L, CST), PD-L1 (clone SP142, Spring), B7-H4 (Clone D1M8I, CST) CD3, CD8 and CD20 in 634 NSCLC cases from 3 retrospective cohorts represented in tissue microarray format. The targets were selectively measured in the tumor and stromal compartments using co-localization with cytokeratin. Associations between the marker levels, major clinic-pathological variables and survival were analyzed. Expression of B7-H3 protein was found in 80.4% (510/634) of the cases and the levels were higher in the tumor than in the stromal compartment. High B7-H3 protein expression level (top 10 percentile) was associated with poor survival in two out of three of the cohorts (p <0.05). Elevated B7-H3 was consistently associated with smoking history across the 3 cohorts, but not with sex, age, clinical stage and histology. Co-expression of B7-H3 and PD-L1 was found in 17.6% of the cases (112/634) and with B7-H4 in 10% (63/634). B7-H4 and PD-L1 were simultaneously detected only in 1.8% of NSCLCs (12/634). The expression of B7-H3 was not associated with the levels of CD3, CD8 and CD20 positive TILs. B7-H3 protein is expressed in the majority of NSCLCs and is associated with smoking history. High B7-H3 protein levels may have a prognostic effect in lung carcinomas. Elevated levels of B7-H3 are not associated with lymphocyte infiltration. Co-expression of B7-H3 with PD-L1 and B7-H4 is relatively low, suggesting a non-redundant biological role of these targets and possibilities for combination therapies using monoclonal antibodies.

B7-H3 regulates lipid metabolism of lung cancer through SREBP1-mediated expression of FASN

B7-H3 is a glycoprotein overexpressed in cancer, but its functional contribution in this setting remains poorly understood. In the present study, we identified that the overexpression of B7-H3 in lung cancer resulted in aberrant lipid metabolism via SREBP-1/FASN signaling pathway. Immunohistochemical analysis of tissue microarrays revealed that approximately 80.4% (37/46) of lung cancer tissues were positive for B7-H3 accompanying poor prognosis. Notably, Oil red O staining and total triglyceride assay exhibited that down-regulation of B7-H3 decreased lipid synthesis in lung cancer A549 and H446 cell lines. Mechanistic investigations showed that B7-H3 modulated the expression of FASN, a fatty acid synthase, specifically. Furthermore, deletion of B7-H3 down-regulated the mRNA and protein levels of SREBP-1, a transcription factor governing the expression of FASN. Finally, correlation analysis between expression levels of B7-H3 and FASN exhibited a positive correlation in clinical lung cancer tissues. Overall, we conclude that B7-H3 hijacks SREBP-1/FASN signaling mediating abnormal lipid metabolism in lung cancer. Our finding provides new insights into the function and mechanism of B7-H3 in the development of lung cancer.

Assessment of plasma B7-H3 levels in pediatric patients with different degrees of surgical stress.

BackgroundSurgical stress initiates a series of host hormone, metabolism and immune responses, which predominantly affect the homeostatic mechanism of patients with major surgery. B7-H3 is a co-stimulatory molecule and has been shown to participate in both adaptive and innate immune responses. In this study we evaluated the clinical significance of plasma B7-H3 levels in pediatric patients with different types of operation and degrees of surgical stress.MethodsA total of 48 children received pediatric general and cardiac surgery were recruited into this study. Based on the surgical stress scoring, children were divided into moderate stress (n = 14) and severe stress (n = 34) groups. Plasma B7-H3 levels were assessed at selected time points: before surgery, immediately after surgery, at day 1, day 3, and day 7 after surgery. Correlations between plasma B7-H3 levels and surgical stress scores were also examined.ResultsPlasma B7-H3 levels were significantly decreased in all 48 pediatric patients after surgery compared to the B7-H3 level before surgery (p < 0.01). Children with general surgery showed significant decreases in plasma B7-H3 immediately after surgery, and at day 3 and day 7 after surgery (p < 0.05, p < 0.01), whereas children with cardiac surgery showed reduced plasma B7-H3 immediately after surgery and at day 3 after surgery (p < 0.05). Plasma B7-H3 in cardiac surgery group was dropped much lower than that in general surgery group at day 1 (p < 0.05) and day 3 (p < 0.01) after surgery. Significantly reduced plasma B7-H3 was observed in the severe stress group, but not in the moderate stress group, immediately after surgery and at day 3 after surgery (p < 0.05), and severe stress group had significantly lower plasma B7-H3 levels than moderate stress group at day 1, day 3, and day 7 after surgery (p < 0.05). Furthermore, plasma B7-H3 levels at day 1 (p = 0.01) and day 3 (p = 0.025) after surgery correlated negatively with surgical stress scores.ConclusionsPlasma B7-H3 levels were decreased significantly in children subjected to pediatric general and cardiac surgery, which is closely associated with the severity of surgical stress. The negative correlation of plasma B7-H3 levels at day 1 and day 3 after surgery with surgical stress scoring implicates that the plasma B7-H3 level might be a useful biomarker for monitoring stress intensity during pediatric surgery.

Abstract 4133: Suppressed B7-H3 expression reduces glycolytic capacity and sensitizes tumor cells to anti-cancer agents

Abstract B7 family proteins are important immune response regulators, and can mediate oncogenicity-related signals and support cancer development. We have prevously shown that the B7-H3 protein promotes tumor cell invasion, proliferation and mestastasis, and increases resistance to chemotherapy. Now we have used human breast and melanoma cancer cell lines with different expression levels of B7-H3 to further evaluate the role of B7-H3 on the sensitivity to anticancer compounds. B7-H3 knockdown tumor cells showed enhanced chemosensitivity compared to their B7-H3 expressing counterparts, as well as increased inhibition to treatment with compounds that target proteins in the PI3K/AKT/mTOR- and MAPK-pathways (Nunes-Xavier et al. in revision). Treatment of control cells with an anti-B7-H3 monoclonal antibody resulted in similar sensitization, whereas B7-H3 overexpressing cells and xenografts were less sensitive. Of note, knockdown of B7-H3 decreased and B7-H3 overexpression increased the glycolytic capacity of the breast cancer and malignant melanoma cells. In conclusion, we have demonstrated a previously unknown relationship between B7-H3 expression and glycolysis in tumor cells, and found that B7-H3 confers resistance to also to pathway inhibiting agents. The results provide novel insights into the function of B7-H3 in cancer, and suggest that targeting of B7-H3 expression may be a promising alternative to improve current anticancer therapy. Citation Format: Caroline E. Nunes-Xavier, Karine Flem Karlsen, Tove Øyjord, Ming Tan, Øystein Fodstad. Suppressed B7-H3 expression reduces glycolytic capacity and sensitizes tumor cells to anti-cancer agents. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4133.

Increased concentrations of soluble B7-H3 and interleukin 36 in bronchoalveolar lavage fluid of Children with Mycoplasma pneumoniae pneumonia.

BackgroundThe purpose of this study is to explore the correlations of interleukin 36 (IL-36) and Soluble B7-H3 (sB7-H3) levels in bronchoalveolar lavage fluid (BALF) with clinical characteristics and laboratory findings.MethodsA total of 35 children with M. pneumnoiae pneumonia (MPP) and 15 control subjects were enrolled. BALF concentrations of sB7-H3 and IL-36 were detected using enzyme-linked immunosorbent assays and clinical profiles of children with MPP were obtained.ResultsChildren with MPP had significantly higher levels of sB7-H3 and IL-36 compared to control subjects (both P < 0.05). Meanwhile, children with pleural effusion had significantly higher levels of sB7-H3 and IL-36 compared to children without pleural effusion (both P < 0.05). BALF concentration of sB7-H3 was strongly associated with concentration of IL-36 (r = 0.796, P < 0.0001) and sB7-H3 was correlated with duration of fever (r = 0.427, P = 0.11) and length of stay (r = 0.345, P = 0.043). Both concentrations of sB7-H3 and IL-36 were significantly decreased in convalescent phase after treatment (both P < 0.05).ConclusionBoth soluble B7-H3 and IL-36 may play an important role in pathogenesis of M. pneumoniae infection and sB7-H3 could be useful as a prognostic predictor or biomarker of MPP.Electronic supplementary materialThe online version of this article (doi:10.1186/s12879-016-1555-6) contains supplementary material, which is available to authorized users.

Anti-B7-H3 monoclonal antibody ameliorates the damage of acute experimental pancreatitis by attenuating the inflammatory response

B7-H3, a recently discovered B7 family member, is documented as a regulator in the inflammatory response as well as T cell-mediated immune responses. In this paper, we find that patients with acute pancreatitis revealed overwhelming levels of serum soluble B7-H3 (sB7-H3) associated with the clinical outcomes. Furthermore, B7-H3 protein was marked increased in l-arginine-induced acute experimental pancreatitis. Anti-B7-H3 monoclonal antibody treatment attenuated the proinflammatory cytokine production, downregulated the activation of the NF-κB signaling pathway, and ameliorated the pancreas disruption in l-arginine-induced pancreatitis. In addition, although l-arginine alone failed to induce the production of proinflammatory cytokine and anti-B7-H3 mAb had no effect on the proinflammatory cytokine production of acinar cells, administration of anti-B7-H3 mAb in the coculture model of acinar cells and macrophages stimulated by l-arginine displayed the similar effects. On the whole, B7-H3 participates in the development of acute pancreatitis, and anti-B7-H3 monoclonal antibody ameliorates severity of acute experimental pancreatitis via attenuation of the inflammatory response.

B7-H3, B7-H4, Foxp3 and IL-2 expression in cervical cancer: Associations with patient outcome and clinical significance.

The aim of this study was to determine the expression of B7-H3, B7-H4, Foxp3 and IL-2 in cervical cancer tissues, and evaluate the corresponding clinical significance. The expression of B7-H3, B7-H4, Foxp3 and IL-2 in 108 cervical cancer specimens was detected using immunohistochemistry, and their relationship with clinicopathologic parameters was determined. B7-H3, B7-H4 and Foxp3 had high levels of expression in cervical cancer cells (72.22, 80.56, and 91.56%, respectively). B7-H3 levels were only significantly associated with tumor size (P=0.013), while B7-H4, Foxp3 and IL-2 levels were significantly associated with International Federation of Gynecology and Obstetrics (FIGO) stage (P=0.023, 0.014 and 0.036, respectively) and tumor size (P=0.045, 0.010 and 0.021, respectively). Their expression levels were not correlated with age, histologic type, differentiation and lymph node metastasis (all P>0.05). Cox regression multivariate analysis confirmed that B7-H3 or B7-H4 overexpression was an independent prognostic factor. In addition, there were significant positive relationships between the expression of B7-H3 and B7-H4 with Foxp3 (P<0.001). In contrast, the expression of B7-H3 and B7-H4 was negatively correlated with IL-2 (P<0.05). B7-H3, B7-H4 and Foxp3 may be useful biomarkers in patients with cervical cancer for predicting treatment.

Decreased expression of B7-H3 reduces the glycolytic capacity and sensitizes breast cancer cells to AKT/mTOR inhibitors.

B7 family proteins are important immune response regulators, and can mediate oncogenic signaling and cancer development. We have used human triple-negative breast cancer cell lines with different expression levels of B7-H3 to evaluate its effects on the sensitivity to 22 different anticancer compounds in a drug screen. API-2 (triciribidine) and everolimus (RAD-001), two inhibitors that target the PI3K/AKT/mTOR pathway, showed enhanced inhibition of cell viability and proliferation in B7-H3 knockdown tumor cells compared to their B7-H3 expressing counterparts. Similar inhibition was seen in control cells treated with an anti-B7-H3 monoclonal antibody. In B7-H3 overexpressing cells, the effects of the two drugs were reduced, supported also by in vivo experiments in which B7-H3 overexpressing xenografts were less sensitive to everolimus than control tumors. In API-2 and everolimus-treated B7-H3 overexpressing cells, phospho-mTOR levels were decreased. However, phosphorylation of p70S6K was differentially regulated in B7-H3 cells treated with API-2 or everolimus, suggesting a different B7-H3-mediated mechanism downstream of mTOR. Both API-2 and everolimus decreased the glycolysis of the cells, whereas knockdown of B7-H3 decreased and B7-H3 overexpression increased the glycolytic capacity. In conclusion, we have unveiled a previously unknown relationship between B7-H3 expression and glycolytic capacity in tumor cells, and found that B7-H3 confers resistance to API-2 and everolimus. The results provide novel insights into the function of B7-H3 in cancer, and suggest that targeting of B7-H3 may be a novel alternative to improve current anticancer therapies.

The relationship of B7H3 expression to androgen and prostate cancer outcomes in a large natural history cohort of men undergoing prostatectomy.

256 Background: B7-H3 (CD276), part of the B7 superfamily, has been shown to play an immunomodulatory role, however its regulation, receptor and mechanism of action remain unclear. Protein levels of B7-H3 have been previously shown to relate to prostate cancer outcomes and currently, humanized monoclonal antibodies are being developed for clinical use (MGA271, Macrogenics). Here we use genomic expression data to examine the relationship of B7-H3 to prostate cancer outcomes and molecular subtypes. Methods: Prostatectomy tissue from 905 patients were profiled using the Affymetrix HuEx 1.0 ST microarray. Kruskal-Wallis tests were used to identify significant associations of B7-H3 expression with clinico-pathologic variables, and survival analysis were used to evaluate the prognostic value of B7-H3. Pearson’s correlation analyses were also performed to assess the relationship of B7-H3 expression with molecular subtypes and individual transcripts. Androgen receptor (AR) occupancy of promoter regions was derived in silico from chromosomal immune-precipitation (ChIP) data. Results: B7-H3 expression was positively associated with Gleason score (p &lt; 0.01) and tumor stage (p &lt; 0.01). High B7-H3 expression also correlated with the development of metastasis and prostate cancer specific mortality (HR of 3.4 and 2.4 respectively, p &lt; 0.05 for both), but this was not significant on multi-variable analysis. B7-H3 was positively associated with ERG+ disease (n = 670, r = 0.85, p &lt; 0.05) and AR expression (n = 670, r = 0.46, p &lt; 0.001). B7-H3 was found to be one of the most correlated genes with AR (95th percentile) and ChIP analysis revealed AR binding upstream of B7-H3, suggesting potential androgen dependent regulation. Conclusions: B7-H3 expression correlates with high Gleason grade and advanced prostate cancer stage with higher quartiles of expression portending poor oncologic outcomes in two independent prostatectomy cohorts. B7-H3 expression appears to relate to the androgen receptor.

Astragaloside IV Enhances Cisplatin Chemosensitivity in Non-Small Cell Lung Cancer Cells Through Inhibition of B7-H3

Background: Chemoresistance is a major obstacle to successful chemotherapy for human non-small cell lung cancer (NSCLC). Astragaloside IV, the component of Astragalus membranaceus, has been reported to exhibit anti-inflammation, anti-cancer and immunoregulatory properties. In the present study, we investigated the role of astragaloside IV in the chemoresistance to cisplatin in NSCLC cells. Methods: We established astragaloside IV-suppressed NSCLC cell lines including A549, HCC827, and NCI-H1299 and evaluated their sensitivity to cisplatin in vitro. In addition, we examined the mRNA and protein levels of B7-H3 in response to cisplatin-based chemotherapy. Results: We showed that high doses of astragaloside IV (10, 20, 40 ng/ml) inhibited NSCLC cell growth, whereas low concentrations of astragaloside IV (1, 2.5, 5 ng/ml) had no obvious cytotoxicity on cell viability. Moreover, combined treatment with astragaloside IV significantly increased chemosensitivity to cisplatin in NSCLC cells. On the molecular level, astragaloside IV co-treatment significantly inhibited the mRNA and protein levels of B7-H3 in the presence of cisplatin. In addition, ectopic expression of B7-H3 diminished the sensitization role of astragaloside IV in cellular responses to cisplatin in NSCLC cells. Conclusion: These results demonstrate that astragaloside IV enhances chemosensitivity to cisplatin via inhibition of B7-H3 and that treatment with astragaloside IV and inhibition of B7-H3 serve as potential therapeutic approach for lung cancer patients.

The Expression and Effect of B7-H3 in Mantle Cell Lymphoma

▪Objective: To investigate the effects of B7-H3 (CD276) on oncogenesis and chemosensitivity in mantle cell lymphoma (MCL).Methods: The B7-H3 expression was detected by flow cytometry in cell lines, 20 patients with MCL and 20 volunteers. B7-H3 knockdown was performed using lentivirus transduction in the Maver and Z138 mantle cell lymphoma cell lines, respectively. The effects of B7-H3 on cell proliferation, cycle, migration and invasion were investigated by CCK-8 assay, methyl cellulose colony forming assay, PI staining, and Transwell assays in vitro. By establishing Maver and Z138 xenograft models, the effects of B7-H3 on tumourigenicity were observed, and Ki-67 and PCNA was detected through immunohistochemical. Moreover, the impacts of B7-H3 RNAi on the anti-tumor effect of chemotherapy drugs were determined with CCK-8, Annexin V-FITC/PI and Hoechst 33342 staining assays in vitro and with xenograft models in vivo.Results: The frequency of B7-H3positive expression cases was 65.0% (13/20) in MCL patients and 10.0% (2/20) in volunteers. The down-regulation of B7-H3 significantly decreased tumor proliferation in MCL in vitro and in vivo. In the B7-H3 knockdown groups of Maver and Z138 xenograft models, the mean inhibition rate of tumor growth was 59.1% and 65.0% (p = 0.010 and 0.003), and the expression of both Ki-67 and PCNA were significantly lower, respectively. After B7-H3 silencing, the cell cycles of Maver and Z138 were both arrested at G0/G1 phase, and the expression of cell cycle-related proteins Cyclin D1 and CDK4 was lower. The cell migration rates and invasion capacity were decreased, and the rates of distant metastasis in B7-H3 knockdown both Maver and Z138 xenografts were significantly declined as well. The expression of invasion-related proteins MMP-2 and MMP-9 was lower in B7-H3 knockdown cells and xenografts. The silencing of B7-H3 increased the sensitivity of Maver and Z138 cells to Rituximab and Bendamustine and enhanced the drug-induced apoptosis, respectively. The activity of caspase-3 in vitro and the expression of caspase-3 in both Maver and Z138 xenografts was significantly increased in the B7-H3 shRNA combined with chemotherapy drugs groups.Conclusions: B7-H3 levels in MCL patients were signifiantly higher than that in volunteers. Our study demonstrates for the first time that B7-H3 promotes mantle cell lymphoma progression and B7-H3 knockdown significantly enhances the chemosensitivity. This may provide a new therapeutic approach to mantle cell lymphoma. DisclosuresNo relevant conflicts of interest to declare.

B7-H3 expression in children with asthma exacerbation.

Recent studies have shown that B7-H3, a recently identified B7 family member, plays a critical role in the development of asthma. This study is to explore the expression of B7-H3 in children with asthma exacerbation. Twenty-one Chinese children with asthma exacerbation as well as 18 nonasthmatic control Chinese children were enrolled. B7-H3 level and cytokines (interferon [IFN]-γ, interleukin [IL]-4, and IL-10) determination were performed by enzyme-linked immunosorbent assay (ELISA) technique. Meanwhile, clinical parameters including laboratory findings, forced expiratory volume in one second (FEV(1)) and fractional exhaled nitric oxide were obtained. Children with asthma exacerbation had significantly higher levels of B7-H3 than controls (4.46 ± 1.33 versus 3.42 ± 1.48 ng/mL; p = 0.027). Plasma IL-4 level was significantly higher in asthma exacerbation subjects than controls (157.98 ± 21.57 versus 121.92 ± 24.37 pg/mL; p < 0.0001), and IFN-γ level was significantly lower in asthma exacerbation subjects (292.73 ± 152.47 versus 421.78 ± 145.84 pg/mL; p = 0.0107). Level of B7-H3 in asthma exacerbation subjects with inhaled corticosteroid (ICS) treatment recently was significantly lower than subjects without ICS treatment (t = 2.706; p = 0.0136). Additionally, levels of B7-H3 decreased remarkably after prednisone treatment. Level of sB7-H3 in asthma exacerbation subjects was inversely correlated with level of IFN-γ (r(p) = -0.605; p = 0.005) after adjustment. B7-H3 may play an important role in asthma exacerbation and was a useful clinical biomarker to evaluate asthma exacerbation.