Immunoexpression of PD-L1, CD4+ and CD8+ cell infiltrates and tumor-infiltrating lymphocytes (TILs) in the microenvironment of actinic cheilitis and lower lip squamous cell carcinoma

Abstract Lower lip squamous cell carcinomas (LLSCC) could be associated with a previous history of potentially malignant oral diseases (PMOD), especially actinic cheilitis (AC), with high sun exposure being a well-described risk factor. Immune evasion mechanisms, such as the PD-1/PD-L1 (programmed cell death protein 1/programmed death-ligand 1) pathway has been gaining prominence since immunotherapy with immune checkpoint inhibitors showed a positive effect on the survival of patients with different types of neoplasms. Concomitant with the characterization of the tumor microenvironment, the expression of either or both PD-1 and PD-L1 molecules may estimate mutual relations of progression or regression of the carcinoma and prognostic values of the patient. Objective: Considering the importance of tumor microenvironment characterization, this study aims to determine the immunoexpression of PD-L1 and correlate with the frequency of CD4+ and CD8+ cells in AC and LLSCC lesions and with tumor-infiltrating lymphocytes (TILs) in LLSCC and its relationship with histopathological characteristics. Methodology: This sample includes 33 cases of AC and 17 cases of LLSCC. The cases were submitted to histopathological analysis and to CD4+, CD8+, and PD-L1+ cell determination by immunohistochemistry. Results: There was a significant difference among the frequencies of CD4+, CD8+, and PD-L1+ cells between AC and LSCC cases, higher in the last group. Moreover, histopathological and atypical changes in AC and LLSCC were correlated with the frequencies of PD-L1+, CD4+, and CD8+ cells. In AC, PD-L1+ cases had a low frequency of CD4+ cells, but on the other hand, PD-L1+ cases of LLSCC had a higher frequency of CD4+ and CD8+ cells. Conclusion: Therefore, the PD-L1 molecule may be a potential escape route for the immune response in oral lesions, but the mechanisms differ between AC and LLSCC. Future studies related to immune evasion and immunotherapy in oral lesions should consider the analysis of inflammatory infiltrate and TILs.

SAMHD1 inhibits PD-L1 expression in lung adenocarcinoma cells / 国际肿瘤学杂志

Objective:To explore the correlation between SAM domain and HD domain-containing protein 1 (SAMHD1) and programmed death-ligand 1 (PD-L1) expression in lung adenocarcinoma.Methods:The expression of SAMHD1 in lung adenocarcinoma and its effect on prognosis were analyzed by online database GEPIA and Kaplan-Meier Plotter. The expression of SAMHD1 in lung adenocarcinoma cell lines was detected by quantitative real-time PCR (qPCR) and Western blotting. SAMHD1 gene was silenced in H1975, H1299 and LLC cells by small interfering RNA transfection and lentivirus infection, respectively. The mRNA and protein expression levels of PD-L1 in lung adenocarcinoma cells of control group, siSAMHD1-1 group and siSAMHD1-2 group were detected by qPCR and Western blotting. The membrane PD-L1 level was detected by flow cytometry. A mouse lung adenocarcinoma xenograft model was constructed. The PD-L1 levels in the tumor tissues of control group and shSAMHD1 group were detected by immunohistochemistry. Cell proliferation activities of the control, siSAMHD1-1 and siSAMHD1-2 groups were detected by CCK-8 assays.Results:The GEPIA database results showed that the mRNA expression of SAMHD1 in lung adenocarcinoma was lower than that in normal lung tissue (4.81±0.90 vs. 5.99±0.76, t=20.67, P<0.001) . The median overall survival time of patients with high SAMHD1 expression was significantly longer than that of patients with low SAMHD1 expression (109.0 months vs. 87.7 months, χ2=26.83, P=0.002) . The relative mRNA expression levels of SAMHD1 in A549, PC9, H1299 and H1975 cells were 1.00±0.02, 0.75±0.05, 3.49±0.19 and 7.25±0.38 ( F=589.00, P<0.001) , and the relative protein expression levels were 1.00±0.06, 0.34±0.07, 1.67±0.22 and 2.11±0.63 ( F=15.79, P=0.001) . In H1975 cells, the relative mRNA levels of PD-L1 in the control, siSAMHD1-1 and siSAMHD1-2 groups were 1.00±0.00, 1.54±0.26 and 2.89±0.13 ( F=102.30, P<0.001) , and the relative protein expression levels were 1.00±0.01, 1.50±0.10 and 1.52±0.33 ( F=6.65, P=0.030) . In H1299 cells, the relative mRNA levels of PD-L1 in the three groups were 1.00±0.08, 1.63±0.03 and 2.14±0.03 ( F=368.80, P<0.001) , and the relative protein levels of PD-L1 were 1.00±0.07, 1.88±0.35 and 2.05±0.38 ( F=10.66, P=0.011) . The expression level of PD-L1 in the siSAMHD1-1 and siSAMHD1-2 groups was higher than that in the control group (all P<0.05) . Flow cytometry results showed that in H1975 cells, the fluorescence intensity of membrane PD-L1 in the control, siSAMHD1-1 and siSAMHD1-2 groups were 246.83±27.59, 325.60±8.00 and 308.93±7.60 ( F=17.56, P=0.003) , and in H1299 cells, the fluorescence intensity of membrane PD-L1 in the three groups were 959.00±6.25, 1 084.33±7.64 and 1 085.33±21.22 ( F=86.74, P<0.001) . The fluorescence intensity of PD-L1 in the siSAMHD1-1 group and siSAMHD1-2 group was higher than that in the control group (all P<0.05) . In xenograft mouse model, the H-SCORE of PD-L1 in the shSAMHD1 group was higher than that in the control group (7.99±1.10 vs. 4.49±0.43, t=5.13, P=0.007) . The proliferative activities of H1975 cells in the control group, siSAMHD1-1 group and siSAMHD1-2 group at 72 h were 0.50±0.02, 0.75±0.05 and 0.73±0.06 ( F=25.01, P=0.001) . The proliferative activities of H1299 cells in the three groups at 72 h were 0.80±0.01, 1.00±0.04 and 0.93±0.07 ( F=13.90, P=0.006) . The cell proliferation activity in the siSAMHD1-1 group and siSAMHD1-2 group was higher than that in the control group (all P<0.05) . Conclusion:SAMHD1 silencing induces PD-L1 expression in lung adenocarcinoma.

Expression of lymphocyte-activation gene 3, programmed cell death protein 1 and its ligand 1 in decidual lymphocytes of women with preeclampsia / 中华围产医学杂志

Objective:To investigate the expression of programmed cell death protein 1 (PD-1), programmed cell death ligand 1(PD-L1), and lymphocyte-activation gene 3(LAG-3) in different subsets of lymphocytes and their relationship with the immunologic imbalance in preeclampsia.Methods:We enrolled 25 cases of singleton pregnant women with preeclampsia who were delivered by cesarean section in the Shandong Provincial Hospital Affiliated to Shandong First Medical University from May 2019 to January 2020 as the preeclampsia group. According to the allocation ratio of 1∶1 matched for the date of cesarean section and pregnancy week at delivery, another 25 healthy singleton pregnant women underwent elective cesarean section were selected as the normal group. The decidua tissue was obtained during cesarean section. The expression levels of PD-1, PD-L1, and LAG-3 on decidual T cells, natural killer (NK), and natural killer T (NKT) cells were measured by flow cytometry and compared between the two groups using two independent samples- t test. Results:(1) The expression of PD-1 on decidual T cells and NK cells of the preeclampsia group were lower than those of the normal group (37.84±3.82 vs 57.02±3.89, t=3.529, P<0.001; 3.28±0.48 vs 5.69±0.99, t=2.184, P=0.034), but did not differ significantly in the expression on decidual NKT cells ( P=0.461). PD-L1 expression on decidual NK cells of preeclampsia group was lower than that of the normal group (0.60±0.11 vs 1.32±0.19, t=3.319, P=0.002), but showed no significant difference in the expression level on T cells and NKT cells (both P>0.05). The preeclampsia group was noted for a lower expression of LAG-3 on decidual T cells and NKT cells compared with the normal group (2.32±0.36 vs 4.09±0.67, t=2.335, P=0.024; 35.40±4.97 vs 56.27±4.49, t=3.282, P=0.002), while showed no significant difference in the expression level of NK cells ( P=0.112). Conclusions:The decreased expression of PD-1, PD-L1, and LAG-3 in the decidual lymphocyte subsets may be involved in the immunologic imbalance of preeclampsia through the over-activation of immunocytes at the maternal-fetal interface.

Significance of PD-L1 expression in carbon-ion radiotherapy for uterine cervical adeno/adenosquamous carcinoma / 부인종양

OBJECTIVE: Programmed cell death-ligand 1 (PD-L1) is expressed in tumor cells and has been shown to predict clinical outcomes of several types of malignancies. The aim of this study was to investigate the effects of carbon-ion (C-ion) beam irradiation on PD-L1 expression in human uterine cervical adeno/adenosquamous carcinoma (UCAA) cells and clinical samples and to identify the prognostic factors for outcomes after C-ion radiotherapy (CIRT).METHODS: The effects of C-ion irradiation on PD-L1 expression in human UCAA and cervical squamous cell carcinoma cells were examined by flow cytometry. We examined PD-L1 expression in UCAA biopsy specimens from 33 patients before CIRT started (pre-CIRT) and after 12 Gy (relative biological effectiveness [RBE]) irradiation (post-12Gy-C) in 4 fractions of CIRT to investigate the correlation between PD-L1 status and clinical outcomes.RESULTS: The PD-L1 expression was upregulated by C-ion beam in a dose-dependent manner in HeLa and SiHa cells through phosphorylated Chk1. The overall frequencies of pre-CIRT and post-12Gy-C PD-L1 positivity were 45% (15/33) and 67% (22/33), respectively. The post-12Gy-C PD-L1 expression was significantly elevated compared to the pre-CIRT PD-L1 expression. There was no significant relationship between the pre-CIRT PD-L1 status and clinical outcomes, such as local control (LC), progression-free survival (PFS), and overall survival (OS). However, the post-12Gy-C PD-L1 expression had better correlation with PFS, but not with LC and OS.CONCLUSION: CIRT can induce PD-L1 expression in UCAA and we propose that PD-L1 expression after starting CIRT may become as a predictive prognostic marker in CIRT for UCAA.

Current status and prognostic value of PD-1/PD-l1 in treatment of colorectal cancer / 肿瘤

The death rate of colorectal cancer (CRC) is the fourth in worldwide. It is an important cause of cancer-related death and seriously affects the survival and quality of life of patients. Surgery, chemotherapy and radiotherapy are the main treatments for CRC. However, the overall survival of CRC patients has not been significantly improved. So the new treatments are urgently needed. Tumor immune escape plays a key role in tumor proliferation, recurrence and metastasis. Immune checkpoints programmed death-1 (PD-1) and programmed death-ligand 1 (PD-L1) play an important role in tumor immune escape. Anti-PD-1/PD-L1 therapy has become a hotspot in cancer research. More and more studies have showed anti-PD-1/PD-L1 immunotherapy has achieved remarkable efficacy in the treatment of microsatellite instability-high (MSI-H) CRC. Therefore, this paper summarizes the clinical application of anti-PD-1/PD-L1 therapy in the treatment of CRC and the various strategies to improve its low response rate. And the predictive value of PD-L1 expression on the surface of tumor cells in the prognosis of CRC is also reviewed.

Study on the Effects of Biyuanshu Oral Solution on mRNA Expression of IFN-γ and Immune Checkpoint B7-H1/ PD-1 of Nasal Sinus Mucosa in CRS Model Mice / 中国药房

OBJECTIVE：To investigate the effects of Biyuanshu （BYS）oral solution on IFN-γ of chronic rhinosinusitis （CRS）model mice ，and to investigate its potential mechanism on the basis of B 7-H1/PD-1 immune checkpoint. METHODS ：Male C57 mice were randomly divided into normal group ，sham operation group ，chemical medicine control group （clarithromycin，103 mg/kg），BYS low-dose ，medium-dose and high-dose groups （BYS oral solution ，3.1，6.2，12.4 mL/kg），with 20 mice in each group. Except for normal group without any treatment ，other mice were all open maxillary sinus ，sham operation group was not filled with sponge with bacteria ，while model group and administration groups were filled with sponge with bacteria to induce CRS model. Since 8th week after modeling ，normal group ，sham operation group and model group were given normal saline 0.2 mL intragastrically，administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 14 d. The nasal symptoms and general condition of mice were observed ，and the pathological changes of mice ’s nasal sinus mucosa were observed by HE staining ；qRT-PCR was used to measure the mRNA expression of IFN-γ，B7-H1 and PD- 1 in nasal sinus mucosa of mice. RESULTS：The normal group and sham operation group had no abnormal in nose ，and the epithelium and cilia of the nasal sinus mucosa were intact ；there was no significant difference f8y3j0127@163.com in the relative mRNA expression of IFN-γ，B7-H1 and PD- 1 between 2 groups（P＞0.05）. In model group ，the mice were found to have runny n ose，frequent scratching and sneezing ，a small amount of yellow secretion in the nasal cavity ，and serious depilation ；the nasal sinus mucosa was seriously damaged ，cilia was exfoliated ，and the gland in the submucosa was hyperplasia ，lymphocyte infiltration was also found ；the relative mRNA expression of IFN-γ，B7-H1 and PD- 1 were significantly increased compared with normal group （P＜0.01）. Compared with model group，the nasal symptoms ，general condition and pathological changes of the nasal sinuses in each administration group were improved in varying degrees ；the relative mRNA expression of IFN-γ and B7-H1 in chemical medicine control group ，BYS medium-dose and high-dose groups ，as well as the relative mRNA expression of PD- 1 in administration groups were decreased significantly；above indexes of BYS medium-dose and high-dose groups were significantly lower than BYS low-dose group ，while relative mRNA expression of IFN-γ in BYS high-dose group were significantly higher than BYS medium-dose group. The relative mRNA expression of IFN-γ in BYS low-dose and medium-dose groups，the relative mRNA expression of B 7-H1 in BYS low-dose group，the relative mRNA expression of PD- 1 in BYS groups were significantly higher than chemical medicine control group ； mRNA expression of IFN-γ in BYS high-dose group was significantly higher than chemical medicine control group（P＜0.05 or P＜ 0.01）. Above indexes of BYS medium-dose group were similar to those of chemical medicine control group （P＞0.05）. CONCLUSIONS：BYS oral solution can improve chronic inflammation in nasal sinus mucosa of mice ，the mechanism of which may be associated with intervening mRNA overexpression of B 7-H1/PD-1 by inhibiting mRNA expression of IFN-γ.

Alterations in PD-L1 Expression Associated with Acquisition of Resistance to ALK Inhibitors in ALK-Rearranged Lung Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The purpose of this study was to evaluate the relationships between the resistance of anaplastic lymphoma kinase (ALK)‒positive non-small cell lung cancer (NSCLC) to ALK inhibitors and the programmed cell death-1/programmed cell death–ligand 1 (PD-L1) pathway, we evaluated alterations in PD-L1 following acquisition of resistance to ALK inhibitors in ALK-positive lung cancer. MATERIALS AND METHODS: We established ALK inhibitor-resistant cell lines (H3122CR1, LR1, and CH1) by exposing the parental H3122 ALK-translocated NSCLC cell line to ALK inhibitors. Then, the double-resistant cell lines H3122CR1LR1 and CR1CH1 were developed by exposing the H3122CR1 to other ALK inhibitors. We compared the alterations in PD-L1 expression levels using western blotting, flow cytometry, and quantitative polymerase chain reaction. We also investigated gene expression using RNA sequencing. The expression of PD-L1 in the tumors from 26 ALK-positive metastatic NSCLC patients (11 ALK inhibitor-naïve and 15 ALK inhibitor-resistant patients) was assessed by immunohistochemistry and analyzed. RESULTS: PD-L1 was expressed at higher levels in ALK inhibitor-resistant cell lines than in the ALK inhibitor-naïve parental cell line at the total protein, surface protein, and mRNA levels. Furthermore, PD-L1 expression in the double-resistant cell lines was much higher than that in the single resistant cell lines. RNA sequencing demonstrated that expression of immune-related genes were largely involved in ALK inhibitor resistance. The mean value of the PD-L1 H-score was 6.5 pre-treatment and 35.0 post-treatment, and the fold difference was 5.42 (p=0.163). CONCLUSION: PD-L1 expression increased following acquisition of ALK inhibitor resistance in ALK-positive NSCLC cell lines and tumors.

The progress of immune escape-related molecule B7-H1 on malignant tumors with multidrug resistance / 医学研究生学报

Malignant tumors with multidrug resistance is one of the major reasons for the failure of tumor treatment, and it is also an important issue in the field of cancer treatment. In recent years, with the continuous research on tumor immune microenvironment, the status of tumor immune escape is deeply understood. The close relationship between the abnormal expression of related molecules B7-H1 and tumor resistance is constantly recognized that multidrug resistant tumor may be more sensitive to immune system surveillance and killing than chemotherapy-sensitive tumor. The article reviews the progress of immune escape-related molecule B7-H1 in multidrug resistance of malignant tumors.

Molecular profile of non-small cell lung cancer in northeastern Brazil / Perfil molecular do carcinoma pulmonar de células não pequenas no Nordeste brasileiro

ABSTRACT Objective: To investigate the histological subtypes and mutational profiles of non-small cell lung cancer in Brazil, looking for correlations among histological subtypes, expression of anaplastic lymphoma kinase (ALK), EGFR mutation status, and programmed death-ligand 1 (PD-L1) expression. Methods: We evaluated 173 specimens obtained from patients with lung adenocarcinoma in northeastern Brazil. Expression of PD-L1 and ALK was evaluated by immunohistochemistry; EGFR mutation status was evaluated by sequencing. We categorized the histological subtypes in accordance with the International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society classification. Results: The most common histological subtypes of lung adenocarcinoma were solid predominant (in 46.8%), acinar predominant (in 37.0%), and lepidic predominant (in 9.8%). ALK expression was detected in 10.4% of the samples, and 22.0% of the tumors harbored EGFR mutations. The most common EGFR mutation was an exon 21 L858R point mutation (in 45.5%), followed by an exon 19 deletion (in 36.3%). The tumor proportion score for PD-L1 expression was ≥ 50% in 18.2% of the samples, 1-49% in 32.7%, and 0% in 49.5%. The solid predominant subtype was significantly associated with wild-type EGFR status (p = 0.047). Positivity for PD-L1 expression was not found to be significantly associated with ALK expression or EGFR mutation status. Conclusions: Our results suggest that the molecular profile of non-small cell lung cancer in northeastern Brazil differs from those of populations in other regions of the country, with ALK positivity being higher than the other biomarkers. Further studies including clinical and genetic information are required to confirm these differences, as well as studies focusing on populations living in different areas of the country.

RESUMO Objetivo: Investigar os subtipos histológicos e perfis de mutação do carcinoma pulmonar de células não pequenas no Brasil, bem como as correlações entre os subtipos histológicos, a expressão do gene anaplastic lymphoma kinase (ALK, quinase do linfoma anaplásico), o estado de mutação do gene EGFR e a expressão de programmed death-ligand 1 (PD-L1, ligante de morte celular programada 1). Métodos: Avaliamos 173 espécimes provenientes de pacientes com adenocarcinoma pulmonar no Nordeste brasileiro. A expressão de PD-L1 e ALK foi avaliada por meio de imuno-histoquímica, ao passo que o estado de mutação do EGFR foi avaliado por meio de sequenciamento. Os subtipos histológicos foram classificados de acordo com a International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society. Resultados: Os subtipos histológicos mais comuns de adenocarcinoma pulmonar foram o predominantemente sólido (em 46,8%), o predominantemente acinar (em 37,0%) e o predominantemente lepídico (em 9,8%). A expressão de ALK foi detectada em 10,4% das amostras, e 22,0% dos tumores apresentavam mutações do gene EGFR. As mutações mais comuns do EGFR foram a mutação pontual L858R no éxon 21 (em 45,5%) e a deleção do éxon 19 (em 36,3%). O tumor proportion score relativo à expressão de PD-L1 foi ≥ 50% em 18,2% das amostras, = 1-49% em 32,7% e = 0% em 49,5%. O subtipo predominantemente sólido relacionou-se significativamente com EGFR selvagem (p = 0,047). A expressão positiva de PD-L1 não se relacionou significativamente com a expressão de ALK ou o estado de mutação do EGFR. Conclusões: Nossos resultados sugerem que o perfil molecular do carcinoma pulmonar de células não pequenas no Nordeste brasileiro difere do de populações em outras regiões do país: a expressão positiva de ALK é maior que os demais biomarcadores. Mais estudos com informações clínicas e genéticas são necessários para confirmar essas diferenças, além de estudos que se concentrem em populações em diferentes áreas do país.

PD-L1 Expression on Circulating CD34+ Hematopoietic Stem Cells Closely Correlated with T-cell Apoptosis in Chronic Hepatitis C Infected Patients

BACKGROUND AND OBJECTIVES: Lack of understanding of the interplay between hematopoietic stem cells (HSCs) and the immune system has severely hampered stem cell research. Programmed death-1 (PD-L1) has been reported on parenchymal cells in patients with chronically inflamed livers and found to play an essential role in T cell homeostasis regulation. However, the bidirectional interaction between HSCs and lymphocytes remains elusive. Here, we aimed to get more insight into circulating CD34+ HSCs PD-L1 expression and T cell apoptosis in chronic HCV infected patients. METHODS: CD34+ HSCs were isolated and purified by immunomagnetic separation. PD-L1 expression was analyzed by quantitative PCR and flow cytometry. Furthermore, co-culture experiments between CD34+ HSCs and T-lymphocytes were established. T-cell lymphocyte apoptosis in peripheral blood and in cultures was detected. RESULTS: CD34+ HSCs constitutively express low levels of PD-L1. Its expression is up-regulated in chronic HCV infected patients. Moreover, PD-L1 expression on circulating CD34+ HSCs enhanced T cell apoptosis in peripheral blood and co-culture. CONCLUSION: Our results suggest novel bidirectional interplay between HSCs and lymphocytes mediated by PD-L1 expression on CD34+ HSCs. PD-L1 expression correlated with T-cell lymphocyte apoptosis. This may contribute to immunomodulatory properties of HSCs which improves its use for allogeneic transplantation.

miR-195/miR-497 Regulate CD274 Expression of Immune Regulatory Ligands in Triple-Negative Breast Cancer / 한국유방암학회지

PURPOSE: Immune suppression is common in patients with advanced breast cancer but the mechanisms underlying this phenomenon have not been sufficiently studied. In this study, we aimed to identify B7 family members that were able to predict the immune status of patients, and which may serve as potential targets for the treatment of breast cancer. We also aimed to identify microRNAs that may regulate the expression of B7 family members. METHODS: The Cancer Genome Atlas data from 1,092 patients with breast cancer, including gene expression, microRNA expression and survival data, were used for statistical and survival analyses. Polymerase chain reaction and Western blot were used to measure messenger RNA and protein expression, respectively. Luciferase assay was used to investigate direct microRNA target. RESULTS: Bioinformatic analysis predicted that microRNA (miR)-93, miR-195, miR-497, and miR-340 are potential regulators of the immune evasion of breast cancer cells, and that they exert this function by targeting CD274, PDCD1LG2, and NCR3LG1. We chose CD274 for further investigations. We found that miR-195, miR-497, and CD274 expression levels were inversely correlated in MDA-MB-231 cells, and miR-195 and miR-497 expressions mimic inhibited CD274 expression in vitro. Mechanistic investigations demonstrated that miR-195 and miR-497 directly target CD274 3′ untranslated region. CONCLUSION: Our data indicated that the level of B7 family members can predict the prognosis of breast cancer patients, and miR-195/miR-497 regulate CD274 expression in triple negative breast cancer. This regulation may further influence tumor progression and the immune tolerance mechanism in breast cancer and may be able to predict the effect of immunotherapy on patients.

Relationships between expressions of B7-H1 and B7-H4 and Foxp3+ regulated T-cell infiltration in colorectal cancer tissue / 吉林大学学报(医学版)

Objective: To explore the expressions of B7-H1 and B7-H4 in colorectal cancer (CRC) tissue and their relationships with the Foxp3 regulated T-cell (Treg) infiltration in tumor tissue, and to provide the basis for judging the biological behaviours and prognosis of CRC. Methods: Immunohistochemistry was used to detect the expressions of B7-H1 and B7-H4 and the infiltration of Treg in 56 cases of CRC tissue samples and adjacent normal tissue samples, and their relationships with the clinicopathological features of CRC patients were analyzed. Results: The positive expression rate of B7-H1 in CRC tissue of the CRC patients was higher than that in adjacent normal tissue (χ2= 72. 34, P<0. 01). The differences of expression frequencies of B7-H1 in CRC tissues with different infiltration depths were significant (P<0. 01). The high expression frequency of B7-H1 in CRC tissue of the patients with positive lymphnode metastasis was higher than that in the patients with negative lymphnode metastasis (P<0. 01). The high expression frequency of B7-H1 in CRC tissue of the patients at Duke' s stage (C+D) was higher than that of the patients at Duke' s stage (A+B) (P<0. 01). The positive expression rate of B7-H4 in CRC tissue was higher than that in adjacent normal tissue (χ2=78. 92, P<0. 01). The differences of expression frequencies of B7-H4 in CRC tissues with different infiltration depths were significant (P<0. 05). The high expression frequency of B7-H4 in CRC tissue of the patients with positive lymphnode metastasis was higher than that of the patients with negative lymphnode metastasis (P<0. 05). The number of Treg positive expression cells in CRC tissue was significantly higher than that in adjacent normal tissue (P<0. 01), the number of Treg postive expression cells in CRC tissues with different differentiation degrees were different (P<0. 01), and the number of Treg positive expression cells in CRC tissue of the patients with positive lymphnode metastasis was higher than that of the patients with negative lymph node metastasis (P<0. 01). The expression of B7-H1 was correlated with Treg infiltration (P<0. 01, Cramer ' s = 0. 463); the expression of B7-H4 was correlated with Treg infiltration (P<0. 05, Cramer' s = 0. 320). Conclusion: B7-H1 and B7-H4 are highly expressed in human CRC tissue, and they are associated with the increasing of Treg infiltration; they can be used as the indicators to judge the prognosis of CRC.

Related research of immunosuppression in intermingled skin transplantation induced by B7-H1 expressed on auto-keratinocytes / 中国免疫学杂志

Objective:To explore the role and mechanism of the B7-H1 expressed by auto-keratinocytes in the intermingled skin grafting model in vitro(MELC). Methods:The intermingled skin grafting model(MELC) was established in vitro. Flow cytometry was performed to detect the expressions of B7-H1 in keratinocytes. The expressions of PD-1 in lymphocytes were measured at the same time. The levels of IL-10,Foxp3 and GATA-3 mRNA in lymphocytes were detected by Real-time quantitative PCR. Results: Through flow eytometry,in the MELC with auto-keratinocytes,the expression of B7-H1 in auto-keratinocytes and the PD-1 in lymphocytes were rising trend and the rising rate was in time-dependent manners(P<0. 01). RT-PCR assay indicated that the relative levels of IL-10, Foxp3,GATA-3mRNA expression were significant raised and the rising rate was in time-dependent manners (P<0. 01). Conclusion:In the intermingled skin grafting model,the auto-keratinocytes could express B7-H1 to enhance the expression of PD-1 in T cells. When B7-H1 combined with PD-1,the Th2 cells and Foxp3+Tregs were induced and suppressed the immune response in the intermingled skin grafting model.

Clinical Observation of Trastuzumab Combined with Neoadjuvant Chemotherapy for HER2 Overexpressing Breast Cancer / 中国药房

OBJECTIVE:To investigate clinical efficacy and safety of trastuzumab combined with neoadjuvant chemotherapy on breast cancer,and discuss its effects on the expression of HER2 protein,B7 homolog 1(B7-H1)and IL-2. METHODS:62 patients with breast cancer confirmed by pathological biopsy,whose test results of immunohistochemistry combined with fluorescence in situ hybridization(FISH)was overexpression of HER2,were selected as the research object. They were randomly divided into observa-tion group and control group with 31 cases in each group. Control group was given carboplatin combined with docetaxel neoadjuvant chemotherapy before surgery. Observation group was additionally given Trastuzumab for injection intravenously 4 mg/kg in the first week,2 mg/kg in second-eighteenth week,once a week,for consecutive 18 weeks. Modified radical mastectomy was conducted 2 weeks after chemotherapy. Clinical efficacy and the occurrence of ADR were observed in 2 groups. The expression of HER2 protein, B7-H1 and IL-2 in tumor tissues were compared between 2 groups before and after treatment. With 1-year follow-up,tumor recur-rence rates,metastasis rates and survival rates were recorded in 2 groups. RESULTS:After treatment,total effective rate of observa-tion group was 83.87%,which was significantly higher than that of control group (58.06%),with statistical significance (P0.05). During 1-year follow-up period,except that there was no statistically significant difference in survival rate be-tween 2 groups(P>0.05),the recurrence rate and metastasis rate of observation group were significantly lower than those of control group,with statistical significance(P<0.05). CONCLUSIONS:The application of trastuzumab combined with neoadjuvant chemo-therapy before breast cancer modified radical mastectomy can effectively improve the expression levels of HER2 protein,B7-H1 and IL-2 in lesion tissue of patients with breast cancer,promote clinical efficacy with good safety and low risk of tumor recurrence and metastasis after operation.

Effects of lung cancer cell-associated B7-H1 on T-cell proliferation in vitro and in vivo

B7 homolog 1 (B7-H1) is the most potent immunoinhibitory molecule in the B7 family. In this study, we examined the effects of tumor-associated B7-H1 on T-cell proliferation in lung cancer. The expression of B7-H1 in human adenocarcinoma A549 and mouse Lewis lung carcinoma (LLC) cells were examined by flow cytometry. To assess the in vitro effect of tumor-associated B7-H1 on T-cell proliferation, we isolated T cells from peripheral blood mononuclear cells (PBMCs) of healthy individuals, labeled them with carboxyfluorescein succinimidyl ester, and co-cultured them with A549 cells in the absence or presence of anti-B7-H1 antibody. For in vivo analysis, LLC cells were subcutaneously injected into mice treated or not with anti-B7-H1 antibody. T-cell proliferation in both in vitro and in vivo assays was analyzed by flow cytometry. In vitro, co-culturing T cells with A549 cells significantly inhibited the proliferation of the former compared with the proliferation of T cells alone (P<0.01), and the addition of B7-H1 blocking antibody dramatically reversed the inhibition of T-cell proliferation by A549 cells. Similarly, in mice bearing LLC-derived xenograft tumors, in vivo administration of anti-B7-H1 antibody significantly increased the total number of spleen and tumor T cells compared to levels in control mice that did not receive anti-B7-H1 antibody. Functionally, in vivo administration of anti-B7-H1 antibody markedly reduced tumor growth. Tumor-associated B7-H1 may facilitate immune evasion by inhibiting T-cell proliferation. Targeting of this mechanism offers a promising therapy for cancer immunotherapy.

Expressions of B7-H1 and Bcl-2 in epithelial ovarian cancer and their clinical significance / 国际肿瘤学杂志

Objective To investigate the expressions of B7-H1 and Bcl-2 proteins in epithelial ovarian cancer,and to explore the association of the expressions of B7-H1 and Bcl-2 with clinicopathological features.Methods The expressions of B7-H1 and Bcl-2 proteins were detected by immunohistochemistry in 80 cases of epithelial ovarian cancer tissues and 20 normal ovary tissues.The association of the expressions of B7-H1 and Bcl-2 with the clinicopathological features was analyzed.Results The positive expression rates of B7-H1 and Bcl-2 in epithelial ovarian cancer tissues were 77.5％ (62/80) and 58.8％ (47/80),both higher than 15.0％ (3/20) and 10.0％ (2/20) in normal ovary tissues with significant difference (x2 =27.473,P ＜ 0.05 ; x2 =15.216,P ＜ 0.05).Both of Bcl-2 and B7-H1 expressions in epithelial ovarian cancer tissues were negatively correlated with the differentiation degree of epithelial ovarian cancer (x2 =9.367,P ＜ 0.01 ; x2 =11.702,P ＜ 0.01).The Bcl-2 expression in epithelial ovarian cancer tissues was positively correlated with the FIGO stage (x2 =7.766,P ＜ 0.01).The expression of B7-H1 was positively correlated with the expression of Bcl-2 in epithelial ovarian cancer (r =0.400,P ＜0.01).Conclusion The expressions of B7-H1 and Bcl-2 are up-regulated in epithelial ovarian cancer and they correlate to each other positively.The expressions of B7-H1 and Bcl-2 are correlated with the invasion and metastasis of epithelial ovarian cancer.The detection of B7-H1 combined with Bcl-2 may have an important clinical significance in the diagnosis and treatment for patients with epithelial ovarian cancer.

Tumores renais: avaliação de fatores prognósticos de 98 casos em hospital de referência de Porto Alegre, Brasil / Renal tumors: evaluation of prognostic factors in 98 cases from a reference hospital in Porto Alegre, Brazil

Introduction: Renal cell carcinoma (RCC) is an aggressive disease worldwide. Objective: Study traditional prognostic factors associated with pathological reports and the novel markers survivin and B7-H1 by immunohistochemistry. Methods: In a reference hospital of Porto Alegre, Brazil, we conducted a cross-sectional study of RCC in patients who underwent radical nephrectomy between 2006 and 2009. We selected those who were diagnosed with the most common histologic subtypes: clear cell and papillary RCC. We retrospectively reviewed pathological data to determine traditional prognostic factors, like size, presence of coagulative necrosis, Fuhrman grade and tumor-node metastasis (TNM) system. Besides, we performed an immunohistochemistry (IHC) study with survivin and B7-H1. Results: Our sample had 98 cases, 90% of the cases were composed by clear cell histologic subtype, 73% were tumors classified as T1 and T2 in the TNM system, most were Fuhrman nuclear grade 2 or 3, and 70% were positive for necrosis. In relation to the new prognostic markers, we found 50 cases positive to survivin and 38 to B7-H1. In this investigation of traditional prognostic markers and new markers we observed that only necrosis was associated with positive results of biomarkers. < 0.001). Conclusion: This finding confirms previous studies that necrosis is an important factor to consider in the prognosis of RCC...

Introdução: O carcinoma de células renais (CCR) é uma doença de comportamento agressivo em todo o mundo. Objetivo: Estudar os fatores prognósticos tradicionais identificados no exame anatomopatológico e sua correlação com a expressão imunoistoquímica dos novos marcadores survivina e B7-H1. Materiais e métodos: Em um hospital de referência de Porto Alegre, foi realizado um estudo transversal de CCR, com pacientes que realizaram nefrectomia total, no período de 2006 a 2009. Foram selecionados aqueles com os tipos histológicos mais comuns: células claras e papilares. Fatores prognósticos tradicionais foram obtidos por meio da revisão de dados patológicos relevantes dos casos, como tamanho, necrose, tipo histológico, grau nuclear e sistema tumor-linfonodo-metástase (TNM). Também se realizou estudo imunoistoquímico (IMQ) da amostra, com o uso dos marcadores survivina e B7-H1. Resultados: Obtivemos 98 casos, com 90% deles do tipo células claras, 73% classificados como T1 e T2, a maioria com grau nuclear de Fuhrman 2 e 3 e cerca de 70% da amostra positivos para necrose. Já no estudo IMQ foi encontrada positividade em 38 casos para o B7-H1 e em 50 para survivina. Ao considerarmos a associação entre os fatores prognósticos tradicionais e a expressão dos marcadores, encontramos associação somente entre o grupo positivo para os marcadores e necrose (p < 0,001). Conclusão: Tal achado vai ao encontro dos dados da literatura que vêm realçando a importância da necrose no prognóstico dos CCR...

Inhibition effect of B7-H1 gene-modified regulatory dendritic cells on thyroid -associated ophthalmopathy in mice / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To construct adenovirus vector expressing mice B7-H1 gene, transfect dendritic cells ( DCs ) , and to study the therapeutic effect of modified DC on thyroid-associated ophthalmopathy ( TAO) in mice. METHODS: We designed and constructed B7-H1 gene adenovirus expression vector, and transfected DCs from mouse bone marrow, tested the phenotype and function of modified DCs, identificated its negative regulation to immune responses. The modified DCs were infected the sicked mice. And then the immunotherapeutic effect of modified DCs to TAO were tested. RESULTS: B7 - H1 gene adenovirus vector was constructed and transfected DCs from bone marrow. The titer of the recombinant adenovirus was 1. 8í109 PFU/mL. B7-H1 gene modified DCs characteristics of regulatory DCs, could inhibit positive immune responses. The inhibition proceeding of TAO into mice infected modified DCs, was obviously prior to the control mice. The gene modified DCs, maybe become the new immunotherapy biological agent to thy TAO. CONCLUSION: We constructed the expression of mouse B7 - H1 gene adenovirus expressed vector successfully, transfected DCs, by vector have properties of regulatory DCs, inhibiting positive immune response and the occurrence and development of thyroid eye disease. Gene modified DCs, reveal potent to the treatment of thyroid eye disease.

The expressions of B7-H1 and Bcl-2 in human colorectal cancer and their clinical significance / 肿瘤

Objective: To investigate the expressions of B7-H1 and Bcl-2 proteins in human colorectal cancer, and to explore the association of the expressions of B7-H1 and Bcl-2 with clinicopathological features. Methods: The expressions of B7-H1 and Bcl-2 proteins were detected by immunohistochemistry in colorectal cancer tissues and their corresponding para-cancerous normal tissues from 57 cases of colorectal cancer. The association of the expressions of B7-H1 and Bcl-2 with the clinicopathological features was analyzed. Results: The positive expression rates of B7-H1 and Bcl-2 in colorectal cancer tissues were both higher than those in para-cancerous normal tissues (P <0.05). Both of Bcl-2 and B7-H1 expressions in colorectal cancer tissues were negatively correlated with the degree of differentiation of colorectal cancer (P <0.01). The positive expression rate of B7-H1 was significantly elevated in patients with lymph node metastasis (Dukes' stage C-D) compared to the patients without lymph node metastasis (Dukes' stage A-B) (P <0.05). For the patients with colorectal cancer classified as well or moderately differentiated, the expression rate of Bcl-2 in patients with negative expression of B7-H1 was significantly higher than that in patients with positive expression of B7-H1 (P <0.01). Short-term followup showed that the B7-H1 was expressed in colorectal cancer tissues from the patients died of colorectal cancer. Conclusion: The expression of B7-H1 was correlated with the invasion and metastasis of colorectal cancer as well as the survival. The detection of B7-H1 combined with Bcl-2 may have an important clinical significance in the diagnosis, treatment and prognostic prediction for patients with well/moderately differentiated colorectal cancer. Copyright © 2012 by TUMOR.

Progress on B7H1/PD-1 pathway in tumors / 国际外科学杂志

B7H1 is a recently described B7 family member.Ligation of PD-1 receptors with B7hl on the surface of activated T cell inhibits T cell proliferation and cytokine production. B7H1 plays a negative regulatory role in peripheral tolerance, autoimmune diseases and chronic infections.B7H1 also has been found to be largely expressed on a broad range of cancers and is thought to contribute to immune evasion by cancers. Blockade of B7H1/PD-1 pathway may contribute to the treatment of autoimmune diseases and maligant tumors.