Upregulation of B7-H4 Is Involved in and Related to the Severity of Acute Pancreatitis.

The expression and clinical significance of co-stimulator B7-H4 in acute pancreatitis (AP) is still unclear. In vitro study showed that the expression of soluble B7-H4 (sB7-H4) and proportions of membrane B7-H4-positive CD14+ cells in the peripheral blood mononuclear cells were upregulated in response to stimulation with plasma from AP patients, lipopolysaccharides, or tumor necrosis factor α (TNF-α). sB7-H4 in the plasma of AP patients were positively correlated with interleukin (IL)-6, IL-10, IL-17A, TNF-α, and interferon-γ The areas under the curves (AUCs) of receiver operating characteristic (ROC) curves of plasma sB7-H4 to distinguish the AP patients from healthy donors, the mild AP (MAP) from the moderately severe acute pancreatitis (MSAP)+severe acute pancreatitis (SAP) or the SAP from the MAP+MSAP were 0.78 (P < 0.001) or 0.773 (P < 0.001) or 0.764 (P < 0.001). sB7-H4 in the plasma of patients were positively correlated with the RANSON scores, Bedside Index of Severity of Acute Pancreatitis scores, Marshall scores, and Acute Physiology And Chronic Health Evaluation II scores; and the AUCs of ROC curves of plasma sB7-H4 in the prediction of local complications was 0.726 (P = 0.001). In conclusion, the co-stimulator B7-H4 is involved in the immune response in AP.

Mifepristone inhibited the expression of B7-H2, B7-H3, B7-H4 and PD-L2 in adenomyosis.

BACKGROUND: The immune mechanism was shown to be involved in the development of adenomyosis. The aim of the current study was to evaluate the expression of the immune checkpoints B7-H2, B7-H3, B7-H4 and PD-L2 in adenomyosis and to explore the effect of mifepristone on the expression of these immune checkpoints. METHODS: The expression of B7-H2, B7-H3, B7-H4 and PD-L2 in normal endometria and adenomyosis patient samples treated with or without mifepristone was determined by immunohistochemistry analysis. RESULTS: In adenomyosis patient samples, the expression of B7-H2, B7-H3 and B7-H4 was increased in the eutopic and ectopic endometria compared with normal endometria, both in the proliferative and secretory phases. Moreover, the expression of B7-H2 and B7-H3 was higher in adenomyotic lesions than in the corresponding eutopic endometria, both in the proliferative and secretory phases. The expression of PD-L2 was higher in adenomyotic lesions than in normal endometria in both the proliferative and secretory phases. In the secretory phase but not the proliferative phase, the expression of B7-H4 and PD-L2 in adenomyotic lesions was significantly higher than that in the corresponding eutopic endometria. In normal endometria and eutopic endometria, the expression of B7-H4 was elevated in the proliferative phase compared with that in the secretory phase, while in the ectopic endometria, B7-H4 expression was decreased in the proliferative phase compared with the secretory phase. In addition, the expression of B7-H2, B7-H3, B7-H4 and PD-L2 was significantly decreased in adenomyosis tissues after treatment with mifepristone. CONCLUSIONS: The expression of the immune checkpoint proteins B7-H2, B7-H3, B7-H4 and PD-L2 is upregulated in adenomyosis tissues and is downregulated with mifepristone treatment. The data suggest that B7 immunomodulatory molecules are involved in the pathophysiology of adenomyosis.

Enhanced B7-H4 expression in gliomas with low PD-L1 expression identifies super-cold tumors.

BACKGROUND: Characterizing expression profiles of different immune checkpoint molecules are promising for personalized checkpoint inhibitory immunotherapy. Gliomas have been shown as potential targets for immune checkpoint inhibitors recently. Our study was performed to determine coexpression levels of two major B7 immune regulatory molecules programmed death ligand 1 (PD-L1) and B7-H4, both of which have been demonstrated to inhibit antitumor host immunity in gliomas. METHODS: We assessed tumor tissues from stage II-IV primary gliomas (n=505) by immunohistochemistry (IHC) for protein levels of both PD-L1 and B7-H4. Gene coexpression analysis assessing clusters based on extent of PD-L1/B7-H4 classifier genes expression were investigated in two transcriptome datasets (The Cancer Genome Atlas and Chinese Glioma Genome Atlas). In addition, levels of immune cell infiltrates were estimated with IHC and RNA-seq data for assessing the tumor immune microenvironment of PD-L1/B7-H4 subgroups. RESULTS: High expression of PD-L1 and B7-H4 in gliomas was 23% and 20%, respectively, whereas coexpression of two proteins at high levels was limited to 2% of the cases. Comparable results were seen in RNA-seq datasets where PD-L1 mRNA expression levels negatively correlated with that of B7-H4. Gene coexpression modules clustered within each grade of gliomas demonstrated lack of double-high modules (cluster with high expression of both PD-L1 and B7-H4 classifier genes). B7-H4 mRNA expression levels showed negative correlation with extent of immune cell infiltration and High-B7-H4 module gliomas (high B7-H4 but low PD-L1 classifier genes expression) had less tumor-infiltrating lymphocytes (TILs) and tumor-associated macrophages (TAMs). IHC assessment also showed few TILs and TAMs in High-B7-H4 subgroup gliomas. CONCLUSIONS: The majority of gliomas express PD-L1 or B7-H4, however, coexpression of both at high levels is minimal. The high-B7-H4 patients could be considered as 'super-cold' gliomas with significantly deficient in TILs, suggesting that B7-H4 might inhibit T-cell trafficking into the central nervous system. This study demonstrated that PD-L1 and B7-H4 may serve as mutually compensatory immune checkpoint molecules in gliomas for immune targeted or active-specific immunotherapy. The distinct B7-H4 pathways modulating T-cell function and immune evasion in glioma patients deserved to be further explored in the future during immunotherapy.

High expression of BTLA and B7-H4 on the surface of myeloid dendritic cells has a negative regulatory effect on their anti-tuberculosis immunity activity in pleural tuberculosis patients.

To investigate the effects of the surface markers B- and T-lymphocyte attenuator (BTLA) and B7 homologous body 4 (B7-H4) on expression of CD83, and Human Leukocyte Antigen-DR isotype (HLA-DR) that can activate dendritic cells (DCs). Flow cytometry was used to detect the co-expression of BTLA and B7-H4 on myeloid DCs (mDCs) in peripheral blood (PB) and pleural effusions (PE) in 15 volunteers and 20 tuberculous pleurisy (TP) patients. Co-expression of BTLA and B7-H4 (double positive (DP)) mDCs in PB and PE of TP patients were enhanced. The proportion of DP mDC in PB decreased markedly after 2 weeks treatment, but was still greater than in controls. Expression of CD83 and HLA-DR on DP mDCs was higher than on BTLA and B7-H4 double negative (DN) expressing mDCs in PB of different TP groups. Expression of CD83 on DP mDCs in PB and PE of TP patients was greater than that of controls. Expression of HLA-DR on DP mDCs in TP patient PB was lower than in TP PE and controls. In pleural tuberculosis (TB) patients, high expression of BTLA and B7-H4 promoted a high level of CD83 and HLA-DR, which had a negative regulatory effect on mDCs on anti-TB immunity.

Does B7-H4 expression correlate with clinicopathologic characteristics and survival in ovarian cancer?: A systematic review and PRISMA-compliant meta-analysis.

BACKGROUND: Several studies have shown that B7-H4 expression is significantly increased in ovarian cancer. However, the role of B7-H4 expression in ovarian cancer remains unclear, and some studies reporting conflicting results. A systematic review of the literature and meta-analysis were conducted to assess the clinicopathologic characteristics and prognostic significance of B7-H4 in ovarian cancer. METHODS: Eligible studies were searched in the PubMed, MEDLINE, Cochrane Library, and the China National Knowledge Infrastructure databases. The included studies assessed the relationship between B7-H4 expression and clinicopathologic features or prognosis in patients with ovarian cancer through September 2017. A total of 1045 patients in 10 studies were included in the meta-analysis. Stata software version 12.0 was used to analyze the data. We used an odds ratio (OR) or hazard ratio (HR) with a 95% confidence interval (CI) to assess the risk or hazard association. RESULTS: B7-H4 expression in ovarian cancer patients was significantly increased (OR: 4.20, 95% CI: 2.85-6.18, Z = 6.91, P < .05), and heterogeneity was low between studies (I = 8.2%, P = .366). With respect to the clinicopathologic features, no relation was detected between B7-H4 expression and International Federation of Gynaecology and Obstetricsstages stages (OR: 0.81, 95% CI: 0.64-1.03, Z = 1.70, P = .09), pathologic grade (OR: 0.91, 95% CI: 0.72-1.16, Z = 0.76, P = .45), tumor metastasis (OR: 1.25, 95% CI: 0.90-1.74, Z = 1.34, P = .18), or histologic type (OR: 1.17, 95% CI: 0.85-1.60, Z = 0.96, P = .34) in ovarian cancer. Furthermore, B7-H4 expression was significantly associated with a worse progression-free survival (PFS) (HR: 1.30, 95% CI: 1.17-1.45, Z = 4.79, P < .05). CONCLUSION: B7-H4 expression was related to ovarian cancer, but not to patients' clinicopathologic characteristics. High B7-H4 expression was negatively correlated with survival outcome, suggesting that B7-H4 plays an essential role in poor prognosis in ovarian cancer patients.

B7-Homolog 4 Promotes Epithelial-Mesenchymal Transition and Invasion of Bladder Cancer Cells via Activation of Nuclear Factor-κB.

B7-homolog 4 (B7-H4), a member of the B7 family of costimulatory molecules, has been reported to be upregulated in urothelial cell carcinoma. This study was conducted to explore the biological role of B7-H4 in the aggressiveness of bladder cancer and the associated molecular mechanism. We found that the mRNA and protein levels of B7-H4 were significantly greater in bladder cancer cell lines than in SV-HUC-1 (normal human urothelial cells). Overexpression of B7-H4 significantly promoted bladder cancer cell migration and invasion, whereas knockdown of B7-H4 exerted an opposite effect. However, the growth of bladder cancer cells was not altered by B7-H4 overexpression or knockdown. Overexpression of B7-H4 promoted epithelial-mesenchymal transition (EMT), as evidenced by decreased E-cadherin and increased vimentin expression. The EMT inducers Twist1 and Snail were upregulated by B7-H4 overexpression and downregulated by B7-H4 silencing. Mechanistically, overexpression of B7-H4 induced the activation of NF-κB signaling. Pharmacological inhibition of NF-κB partially prevented B7-H4-mediated bladder cancer cell invasion. Taken together, B7-H4/NF-κB signaling is involved in the EMT and invasion of bladder cancer cells and represents a new candidate target for the treatment of bladder cancer.

Diagnosis value of serum soluble B7-H4 expression in non-small cell lung cancer.

INTRODUCTION: B7-H4, a member of the inhibitory B7 family, can restrain T cell proliferation, activation, cytokine secretion, and may be involved in immune evasion in cancer patients. OBJECTIVES: This aim of the study was to determine the expression level of soluble B7-H4 (sB7-H4) in circulation and to subsequently evaluate the clinical significance of circulating sB7-H4 in patients with non-small cell lung cancer (NSCLC). METHODS: Serum specimens from 128 patients with NSCLC, 100 healthy volunteers (HV), and 80 patients with benign lung diseases (BLD) were collected. The concentrations of sB7-H4 were measured by sandwich enzyme-linked immunosorbent assay. RESULTS: Serum sB7-H4 levels in patients with NSCLC were significantly higher than those in patients with BLD (P < 0.05), or those in HV (P < 0.05). Using a cutoff of 27.8 ng/mL, the sensitivity and specificity of sB7-H4 in differentiating between patients with NSCLC and patients with BLD, and between patients with NSCLC and HV was, 46.9% and 92.5%, and 54.7% and 95.0%, respectively. An area under the curve (AUC) for NSCLC resulting from sB7-H4 (0.863), which was significantly better than any other tumour markers tested including CA125 (0.763), and CEA (0.775). CONCLUSION: In conclusion, assessment of serum sB7-H4 levels could be considered as a diagnostic biomarker for NSCLC.

TGF-ß1 promotes colorectal cancer immune escape by elevating B7-H3 and B7-H4 via the miR-155/miR-143 axis.

Transforming growth factor-beta 1 (TGF-ß1) suppresses T cell function, promoting tumor immune escape. Yet, whether the depression of TGF-ß1 on T cell function is mediated by co-inhibitory molecules B7-H3 and B7-H4 remains largely unclear. Here, we demonstrated that TGF-ß1 elevated the expression of miR-155 in colorectal cancer cells through SMAD3 and SMAD4. The upregulated miR-155 attenuated miR-143 by inhibiting its direct target, the transcription factor CEBPB. Consequently, the direct target genes of miR-143, B7-H3 and B7-H4, were augmented in the cytoplasm and membrane of tumor cells. Over-expression of B7-H3 and B7-H4 in HCT-116 cells induced T cells to secrete TGF-ß1 and the immunosuppressive cytokines IL-2, IL-6, and IL-17. Restoration of miR-143 inhibited the growth of HCT-116 xenograft tumors in mice, and also repressed the expression of B7-H3 and B7-H4 in the tumors. Thus, this study reveals the mechanism by which TGF-ß1 leads to T cell-mediated tumor evasion through an increase in B7-H3 and B7-H4 expression.

Unstable B7-H4 cell surface expression and T-cell redirection as a means of cancer therapy.

Tumor immune regulation has been demonstrated in clinical studies using antibodies targeted to the B7/CD28 family. B7 homolog 4 (B7-H4) negatively regulates immune responses and is overexpressed in many types of human cancer, indicating that B7-H4 may be a potential target of cancer therapy. B7-H4 expression is affected by the microenvironment, and its presence has been reported in cancer tissues and immune cells. We found an upregulation of B7-H4 expression using comprehensive whole exome sequencing and gene expression profiling (project HOPE) launched by the Shizuoka Cancer Center based on tumor tissue samples from 1,058 cancer patients. We were successful in producing monoclonal antibodies for B7-H4 and demonstrated B7-H4 dimerization and rapid cell surface disappearance by antibody cross-linking in breast cancer cells, even under typical conditions. These observations may explain why antibody-dependent cellular cytotoxicity (ADCC) did not function in vivo on the B7-H4-expressing tumor cells. Unstable cell surface antigens are not suitable as targets for ADCC, and we therefore performed an indirect ADCC-redirecting T-cell cytotoxicity assay to study B7-H4 using polyclonal anti-mouse IgG antibody-mediated linking. Our results showed the possibility of targeting the B7-H4 molecule as a means of treating cancer.

B7-H4 expression indicates poor prognosis of oral squamous cell carcinoma.

Checkpoint blockade therapy utilizing monoclonal antibodies to reactivate T cells and recover their antitumor activity makes an epoch in cancer immunotherapy. The role of B7-H4, a novel negative immune checkpoint, in oral squamous cell carcinoma (OSCC) has still not been elucidated. In this study, tissue samples from human OSCC, which contains 165 primary OSCC, 48 oral epithelial dysplasia and 43 normal oral mucosa specimens, and Tgfbr1/Pten 2cKO mice OSCC model were stained with B7-H4 antibody to analyze the correlations between B7-H4 expression and clinicopathological characteristics. Kaplan-Meier analysis was used to compare the survival of patients with high B7-H4 expression and patients with low B7-H4 expression. We found B7-H4 is highly expressed in human OSCC tissue, and the B7-H4 expression level was associated with the clinicopathological parameters containing pathological grade and lymph node status. Moreover, we confirmed that B7-H4 was overexpressed in Tgfbr1/Pten 2cKO mice OSCC model. Our data also indicated that patients with high B7-H4 expression had poor overall survival compared with those with low B7-H4 expression. Furthermore, this study demonstrated that B7-H4 was positively associated with PD-L1, CD11b, CD33, PI3Kα p110, and p-S6 (S235/236). Taken together, these findings suggest B7-H4 is a potential target in the treatment of OSCC.

B7-H4 facilitates proliferation of esophageal squamous cell carcinoma cells through promoting interleukin-6/signal transducer and activator of transcription 3 pathway activation.

B7-H4, one of the costimulatory molecules of the B7 family, has been found to be widely expressed in many kinds of tumor tissues and to play an important part in tumor progression and poor prognosis. However, the role of B7-H4 in esophageal squamous cell carcinoma (ESCC) cells has not been elucidated. In this study, we found that, compared with normal esophageal tissue, B7-H4 was highly expressed in three ESCC cell lines, Eca109, TE1, and TE13. B7-H4 silenced cells suppressed cellular proliferation and colony formation. Additionally, compared with control cells, B7-H4 silenced cells showed higher apoptosis rates, Bcl-2 and Survivin upregulation, and BAX downregulation. Further study revealed that B7-H4 silenced cells also showed reduction in interleukin-6 (IL-6) secretion, signal transducer and activator of transcription 3 (STAT3) activation, and p-STAT3 translocation from cytoplasm to nucleus. Moreover, B7-H4 depletion inhibited the IL-6 secretion of control cells but not JAK2/STAT3 inhibitor FLLL32-treated cells. Interleukin-6 receptor antagonist tocilizumab did not block the p-JAK2 or p-STAT3 downregulation induced by B7-H4 silence. It was suggested that B7-H4 silence suppressed IL-6 secretion through JAK2/STAT3 inactivation. Furthermore, cell proliferation and colony formation were downregulated by tocilizumab in control cells but not in B7-H4 silenced cells, indicating that IL-6 upregulation induced by B7-H4 was necessary for cell growth. On the other hand, B7-H4 expression was downregulated by tocilizumab. In all, our study provided the first evidence that B7-H4 facilitated ESCC cell proliferation through promoting IL-6/STAT3 positive loopback pathway activation.

Effect of VTCN1 on progression and metastasis of ovarian carcinoma in vitro and vivo.

BACKGROUND AND PURPOSES: Through reducing immune response, VTCN1 could promote carcinoma indirectly. However, the direct effect of VTCN1 on carcinoma was not studied clearly, especially on ovarian carcinoma. In this paper, we verified the potential effect and mechanism of VTCN1 on ovarian carcinoma. METHODS: The influence of high or low VTCN1 expression on the viability of ovarian cancer was detected by CKK-8 and annexin V-PI kit. The orthotopicxenograft tumor model was performed to evaluate the effect of VTCN1 on the promotion of tumor in vivo. Western blot was used to verify the signaling pathways predicted by bioinformatics analysis. RESULTS: Low expression of VTCN1 could inhibit the viability and metastasis of ovarian carcinoma directly in vitro and vivo; Information analysis demonstrated that cell cycle and JAK2/STAT were involved in the regulation of VTCN1. The CDK2/4 and CDC25C expression and phosphorylation of JAK2/STAT had a direct relationship with the reduction of VTCN1. CONCLUSIONS: VTCN1 could affect the viability and metastasis of ovarian carcinoma by reducing the expression of CDK2/4 and CDC25C and phosphorylation of JAK2/STAT. It indicated that VTCN1 was a potential target for treating ovarian carcinoma.

Role of B7-H4 siRNA in Proliferation, Migration, and Invasion of LOVO Colorectal Carcinoma Cell Line.

OBJECTIVES: Colorectal cancer is one of the most common malignancies. Recent studies investigated that B7-H4 is highly expressed in various cancers. We aimed at exploring the effect of B7-H4 siRNA on proliferation, invasion, and migration of LOVO cells which expressed B7-H4 notably. DESIGN AND METHODS: Colon adenocarcinoma dataset was downloaded from The Cancer Genome Atlas. 35 colorectal cancer patients admitted to Shanghai Tongren Hospital were enrolled in this study. Cell proliferation and cell cycle distribution were identified by CCK8 and flow cytometry, respectively. Transwell assay was performed to detect the invasion and migration of LOVO cells. CXCL12/CXCR4 expression and JAK2/STAT3 phosphorylation were determined by real-time PCR and western blot. RESULTS: B7-H4 expressed is elevated in colorectal cancer tissues than in the adjacent normal tissues. B7-H4 siRNA effectively inhibited the proliferation at 24 h and 48 h, arrested cell cycle at G0/G1, and suppressed cell invasion and migration. Gene set enrichment analysis showed that CXCL12/CXCR4 and JAK/STAT were correlative with the B7-H4 expression. Additionally, CXCL12/CXCR4 expression and JAK2/STAT3 phosphorylation were reduced. CONCLUSIONS: B7-H4 siRNA can effectively inhibit proliferation, invasion, and migration of LOVO cells by targeting CXCL12/CXCR4 and JAK2/STAT3 signaling, which can serve as a new target for colorectal carcinoma treatment.

B7-H4 expression is associated with tumor progression and prognosis in patients with osteosarcoma.

Increasing evidences have demonstrated that B7-H4 is associated with tumor development and prognosis. However, the clinical significance of B7-H4 expression in human osteosarcoma (OS) remains unclear. The aim of present study was to examine the B7-H4 expression and to explore its contribution in OS. B7-H4 expression in OS tissues was examined by immunohistochemistry. Soluble B7-H4 (sB7-H4) levels in blood were examined by ELISA. The association of B7-H4 expression with clinicopathological factors or prognosis was statistically analyzed. Our findings demonstrated that B7-H4 expression in OS tissues was significantly higher than those in paired normal bone tissues (P < 0.001). sB7-H4 level in OS serum samples was significantly higher than that in healthy controls (P = 0.005). High B7-H4 expression in tissues and sB7-H4 level were both correlated with advanced tumor stage (P < 0.001, P = 0.017, resp.) and distant metastasis (P = 0.034, P = 0.021, resp.). Additionally, high B7-H4 expression or serum sB7-H4 levels were significantly related to poor overall survival (P = 0.028, P = 0.005, resp.). B7-H4 in tissues and serum samples were an independent factor for affecting the survival time of OS patients (P = 0.004, P = 0.041, resp.). Collectively, our data suggest that the evaluation of B7-H4 expression in tissues and blood is a useful tool for predicting the progression of osteosarcoma and prognosis.

Cancer cell-associated cytoplasmic B7-H4 is induced by hypoxia through hypoxia-inducible factor-1α and promotes cancer cell proliferation.

Aberrant B7-H4 expression in cancer tissues serves as a novel prognostic biomarker for poor survival in patients with cancer. However, the factor(s) that induce cancer cell-associated B7-H4 remain to be fully elucidated. We herein demonstrate that hypoxia upregulates B7-H4 transcription in primary CD138(+) multiple myeloma cells and cancer cell lines. In support of this finding, analysis of the Multiple Myeloma Genomics Portal (MMGP) data set revealed a positive correlation between the mRNA expression levels of B7-H4 and the endogenous hypoxia marker carbonic anhydrogenase 9. Hypoxia-induced B7-H4 expression was detected in the cytoplasm, but not in cancer cell membranes. Chromatin immunoprecipitation analysis demonstrated binding of hypoxia-inducible factor-1α (HIF-1α) to proximal hypoxia-response element (HRE) sites within the B7-H4 promoter. Knockdown of HIF-1α and pharmacological inhibition of HIF-1α diminished B7-H4 expression. Furthermore, knockdown of cytoplasmic B7-H4 in MCF-7 decreased the S-phase cell population under hypoxia. Finally, MMGP analysis revealed a positive correlation between the transcript levels of B7-H4 and proliferation-related genes including MKI67, CCNA1, and Myc in several patients with multiple myeloma. Our results provide insight into the mechanisms underlying B7-H4 upregulation and its role in cancer cell proliferation in a hypoxic tumor microenvironment.

Prognostic value of soluble H7-B4 in pleural effusion associated with lung cancer.

B7-H4, a member of the inhibitory B7 family, can restrain T cell proliferation, activation, and cytokine secretion and may be involved in immune evasion in cancer patients. This study aimed to evaluate the diagnostic and prognostic value of pleural effusion levels of soluble B7-H4 (sB7-H4) in lung cancer patients with malignant pleural effusion (MPE). Pleural effusion samples were collected from 98 lung cancer patients with malignant effusion and from 60 patients with nonmalignant pleural effusion. Pleural effusion concentrations of sB7-H4 were measured using sandwich enzyme-linked immunosorbent assay. Malignant effusion exhibited higher sB7-H4 levels than those in nonmalignant effusion (P < 0.01). Lung cancer patients with pleural effusion sB7-H4 levels below 35.8 ng/ml had a longer overall survival than those with higher levels (P < 0.05). By multivariate analysis, pleural effusion sB7-H4 was an independent prognostic factor in patients with MPE. In conclusion, measurement of sB7-H4 might be a useful diagnostic and prognostic value for MPE patients.

B7-H4 overexpression impairs the immune response of T cells in human cervical carcinomas.

OBJECTIVE: To investigate B7-H4 expression and its correlation with the number of infiltrating T lymphocytes and cytokine production by those lymphocytes in human cervical cancer and to determine the effect of recombinant B7-H4 on the active peripheral blood T cells of the patients in vitro. METHODS: B7-H4 expression was detected in 67 cases of cervical cancer using immunohistochemical staining. Tumor-infiltrating CD8(+)T, CD4(+)T, and FOXP3(+) (Forkhead Box P3) T lymphocytes and their levels of IFN-γ and TGF-ß1 production were determined by immunofluorescent double-staining. After the peripheral blood T lymphocytes of patients were co-cultured with B7-H4, proliferation, apoptosis, and cell subtypes were analyzed using flow cytometry. Cytokines in the supernatant were detected by ELISA. RESULTS: B7-H4 was expressed in 46% (31/67) of the cases of cervical cancer. The number of infiltrating CD8(+)T lymphocytes and their IFN-γ production in positive B7-H4 expression cervical cancers was significantly lower than in negative B7-H4 cases (P<0.01, P<0.05), but there was no significant difference between cases positive and negative for B7-H4 with respect to infiltrating FOXP3(+)T and CD4(+)T cells or TGF-ß1 production. After co-culture with B7-H4 for 48 h, the patients' activated T lymphocytes were arrested at G1/G2 phase. The Ki67 positive rates of CD4(+)T and CD8(+)T cells were 2.13 ± 0.13% and 1.03 ± 1.33%, and they were lower than in the blank group. The proportion of CD4(+)T and CD8(+)T cells decreased, but CD4(+)T/CD8(+)T and the proportion of CD4(+)CD25(+)Foxp3(+)T cells increased. In addition, concentrations of IL-10 and TGF-ß1 in the supernatant of co-cultured T cells increased significantly (P<0.05, P<0.05), but that of IFN-γ decreased. B7-H4 had no significant effect on apoptosis of the T cells. CONCLUSION: B7-H4 is overexpressed in human cervical cancers, and it is associated with lower numbers of tumor-infiltrating CD8(+)T lymphocytes and therefore less IFN-γ production. In vitro, B7-H4 inhibits the proliferation of CD4(+)T and CD8(+)T but promotes the proliferation of Tregs and the secretion of IL-10 and TGF-ß1. B7-H4 plays an important role in depressing the anti-tumor immunity of CD8(+)T cell in microenvironments of cervical cancer.

B7-H4 as a potential target for immunotherapy for gynecologic cancers: a closer look.

B7-H4 is a transmembrane protein that binds an unknown receptor on activated T cells resulting in inhibition of T-cell effector function via cell cycle arrest, decreased proliferation, and reduced IL-2 production. B7-H4 is up-regulated on the surface of cancer cells and immunosuppressive tumor-associated macrophages (TAMs) in a variety of human cancers. Notably, B7-H4 expression levels inversely correlate with patient survival in ovarian cancer, making B7-H4 an attractive candidate for therapeutic intervention. Here, we summarize the experimental data and methodologies that have revealed B7-H4's mRNA and protein expression and function in both mice and humans since its discovery in 2003, with a specific focus on B7-H4's role in ovarian cancer. We also underscore the discrepancies in published data due to high variability in methodology and use of different antibodies, most of which are not commercially available. Finally, since B7-H4 is expressed on tumor cells and TAMs in various cancer types, directing therapeutics against B7-H4 could have tremendous synergistic outcomes in favorably altering the tumor micro-environment and eliminating cancer cells. We highlight the therapeutic potential of targeting B7-H4, both by comparing other negative immune modulators such as PD-1 and CTLA-4 and by identifying novel methods to target B7-H4 directly or indirectly to overcome B7-H4-mediated T-cell inhibition.

The expressions of co-stimulatory molecules are altered on putative antigen-presenting cells in cord blood.

PROBLEM: The aim of our study was to estimate the expressions of both B7-H1 and B7-H4 as well as CD200 and CD200R co-stimulatory molecules on immature myeloid and lymphoid dendritic cells, B CD19(+) lymphocytes and monocytes in umbilical cord blood of healthy neonates and in peripheral blood of healthy adults. METHOD OF STUDY: Thirty-nine healthy full-term neonates from physiological single pregnancies and 27 healthy adults were included in the study. The expressions of B7-H1, B7-H4, CD200, CD200R antigens were estimated using flow cytometry. Statistical analysis was performed using a non-parametric Mann-Whitney U-test and parametric Wilcoxon's test. RESULTS: The expressions of B7-H1 and B7-H4 molecules on immature BDCA-1(+) myeloid dendritic cells were significantly lower in umbilical cord blood of healthy neonates when compared with those cells in peripheral blood of healthy adults (P < 0.0001). Furthermore, the suppression of B7-H4 molecule on BDCA-2(+) lymphoid dendritic cells was observed in cord blood of healthy neonates when compared with peripheral blood of healthy adults (P < 0.02). The expressions of CD200 antigen on BDCA-1(+) cells, CD200R antigen on BDCA-2(+) cells and CD200R antigen on B CD19(+) cells were significantly lower in cord blood of healthy neonates. On the other hand, the expressions of CD200 and CD200R as well as B7-H4 co-stimulatory molecules on CD14(+) cells were significantly higher in cord blood when compared with peripheral blood. CONCLUSION: The increased percentages of CD14(+) monocytes with the expressions of CD200 and CD200R as well as B7-H4 co-stimulatory molecules can suggest the increased immunomodulatory properties of neonatal monocytes in cord blood.

B7x in the periphery abrogates pancreas-specific damage mediated by self-reactive CD8 T cells.

B7x (B7-H4 or B7S1) is the seventh member of the B7 family, and its in vivo function remains largely unknown. Despite new genetic data linking the B7x gene with autoimmune diseases, how exactly it contributes to peripheral tolerance and autoimmunity is unclear. In this study, we showed that B7x protein was not detected on APCs or T cells in both human and mice, which is unique in the B7 family. Because B7x protein is expressed in some peripheral cells such as pancreatic ß cells, we used a CD8 T cell-mediated diabetes model (AI4αß) in which CD8 T cells recognize an endogenous self-Ag, and found that mice lacking B7x developed more severe diabetes than control AI4αß mice. Conversely, mice overexpressing B7x in the ß cells (Rip-B7xAI4αß) were diabetes free. Furthermore, adoptive transfer of effector AI4αß CD8 T cells induced diabetes in control mice, but not in Rip-B7xAI4αß mice. Mechanistic studies revealed that pathogenic effector CD8 T cells were capable of migrating to the pancreas but failed to robustly destroy tissue when encountering local B7x in Rip-B7xAI4αß mice. Although AI4αß CD8 T cells in Rip-B7xAI4αß and AI4αß mice showed similar cytotoxic function, cell death, and global gene expression profiles, these cells had greater proliferation in AI4αß mice than in RIP-B7xAI4αß mice. These results suggest that B7x in nonlymphoid organs prevents peripheral autoimmunity partially through inhibiting proliferation of tissue-specific CD8 T cells, and that local overexpression of B7x on pancreatic ß cells is sufficient to abolish CD8 T cell-induced diabetes.