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[cat] TREX2 (three-prime repair exonuclease 2) és una exonucleasa no processiva 3'-5', expressada específicament en la pell que pot estar implicada en el manteniment de la integritat del genoma editant els extrems 3' del DNA en diversos processos, des de la replicació a la reparació del dany passant per la degradació en l'apoptosi. L'estudi de TREX2 mitjançant immunohistofluorescència de mostres humanes de lesions precanceroses (queratosis actínica) i tumors derivats de queratinòcits, incloent cSCC (cutaneous squamous cell carcinomas) i HNSCCs (head and neck SCC), revela que en les carcinogènesis escamoses l'expressió de TREX2 pateix es troba alterada. La deleció de Trex2 en ratolins provoca un augment de la susceptibilitat de tumors induïts en pell provocats per radiació ultraviolada B (UVB). TREX2 promou l'eliminació del DNA danyat i l'apoptosi dels queratinòcits en les pells de ratolí irradiades amb UVB. Les lesions al DNA induïdes per UVB (CPD: cyclobutane pyrimidine_dimers), són més persistents en les pells dels ratolins Trex21- respecte els wt, paral• lelament també es dóna una menor activació de la caspasa 3, indicador d'apoptosi, en les pells Trex21-. Els resultat obtinguts en queratinòcits wt i deficients en TREX2 in vitxo reflexen una multifuncionalitat de la proteïna TREX2 intervenint en la reparació del dany al DNA en resposta a la radiació UV i promovent l'apoptosi de forma irreversible, essent aquesta última funció la que preval. TREX2 pot ser reclutada diferencialment a diferents localitzacions subnuclears en funció de l'estat de la cromatina, com per exemple els micronuclis i les regions de cromatina poc condensada. Els estudis de co-immunoprecipitació demostren que TREX2 interacciona amb yH2AX, així TREX2 pot accedir als extrems lliures del DNA interaccionant amb la proteïna H2AX fosforilada, preferentment en la cromatina destinada a ser degradada. L'estudi histopatològic de les pells de ratolins irradiats amb UVB, revela que els ratolins Trex21- són més resistents que els wt a la inflamació de la pell induïda per UVB. Addicionalment, en les pells irradiades repetidament del ratolí Trex2-/, l'augment en el nombre de capes cel• lulars en l'epidermis com a mesura de protecció de la pell contra la irradiació, és menor que en el wt. En resum, les nostres dades subratllen la rellevància de TREX2 a l'hora de promoure la resposta inflamatòria contra la UVB. Per altra part, a lesions psoriàsiques es dóna un fort increment i canvis en el patró d'expressió de TREX2 respecte la pell sana. La deficiència de TREX2 a ratolins disminueix el fenotip psoriàsic induït per l'Imiquimod. Concretament, TREX2 afavoreix la mort cel• lular i la resposta inflamatòria inicial. A més a més, TREX2 està implicada en la degradació del DNA durant la diferenciació terminal dels corneòcits, disminuint la paraqueratosi. Així, TREX2 té un paper actiu i rellevant en el desenvolupament de la patogènesi associada a la psoriasi., [eng] TREX2 is a 3 '-DNA exonuclease specifically expressed in keratinocytes that acts as tumor suppressor in skin carcinogenesis, but the importance in human pathologies like carcinogenesis and psoriasis and the underlying mechanisms remained unknown. Here, we investigated the relevance and molecular events of TREX2 in the DNA damage skin response. TREX2 expression was up-regulated by chronic exposure to ultraviolet (UV) radiation, whereas was de-regulated or lost in human squamous cell carcinomas (SCCs). TREX2 deficiency in mice enhanced the susceptibility to UVB-induced skin carcinogenesis, which was associated with alterations in DNA damage removal and degradation, cell death and inflammation, indicating multifaceted functions of TREX2. Notably, TREX2 deficiency diminished the up-regulation of key cytokines related to DNA repair and antitumor immunity. Interestingly, TREX2 was recruited to low-density chromatin regions in nuclei that were beginning to condensate chromatin and to micronuclei, interacting with phosphorylated H2AX histone (yH2AX). Altogether, our data establish cell and non-cell autonomous functions of TREX2 in the defense against UV-induced skin carcinogenesis. Moreover TREX2 is overexpressed in psoriasic lesions. To elucidate the importance of this exonuclease in the development of the disease, we induced a skin inflammation in mice by topical application of Imiquimod. The Trex24- mice were more resistant to the Imiquimod-induced skin inflammation than wt. Thus, clinical parameters like skin thickness, erythema and scaling were lower in the Trex24- mice compared to the wt, whereas parakeratosis was more abundant in the Trex24- skins. Altogether, our results show thatTREX2 contribute to the pathologic features of psoriasis by enhancing the keratinocyte apoptosis and the inflammation in response to Imiquimod application.

Background: The clinical benefit of immune response is largely unknown. We systematically explored the correlation of immune response with patient outcome in human cancers. Results: The global immune gene signature was primarily located on the plasma membrane with a high gene density at 6p21 and 1q23-1q24. Immune responses varied with a wide range in human cancers. A total of 11 cancer types exhibited significant correlation of immune response with overall survival. Higher immune response was significantly associated with longer overall survival in 7 types and with shorter overall survival in 4 types. In addition, 11 cancer types exhibited significant correlation of immune response with progression-free interval. Higher immune response was significantly associated with longer progression-free interval in 7 types and with shorter progression-free interval in 4 types. Methods: The Ingenuity Knowledge Base and human genome assembly GRCh38 were used to annotate the immune gene signature by cellular components and genomic coordinates, respectively. We devised an mRNA-based metric of pre-existing immune conditions by using the gene signature, and calculated the metric for 10,062 The Cancer Genome Atlas tumor samples across 32 different cancer types. The Kaplan-Meier method was used to evaluate the overall survival and progression-free interval differences between dichotomic groups stratified by the median metric for each cancer type. Conclusions: Immune responses have different impacts on patient outcome in different human cancers. Prospective verification is needed before the findings can be applied for clinical trial development.

In this study, erianin was found to reduce the viability of cancer cells, inhibit their proliferation and migration, induce G2/M phase arrest, enhance cancer cell apoptosis, promote an increase in levels of intracellular reactive oxygen species and a decrease in mitochondrial membrane potential, and regulate the expression levels of anti- and pro-apoptosis-related proteins in HepG2 and SMMC-7721 cells. Erianin inhibited tumor growth in HepG2- and SMMC-7721-xenograft tumor nude mouse models, reduced the expression levels of anti-apoptosis proteins and enhanced the expression levels of pro-apoptosis proteins in tumor tissues. Erianin inhibited tumor growth in immunosuppressed BALB/c mice bearing heterotopic tumors. Among 111 types of cytokines detected in proteome profiling of tumor tissues, erianin substantially influenced levels of 38 types of cytokines in HepG2-xenografted tumors and of 15 types of cytokines in SMMC-7721-xenografted tumors, most of which are related to immune functions. Erianin strongly affected the serum levels of cytokines, and regulated the activation of nuclear factor-kappa B (NF-κB), and the expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream proteins in spleen. The anti-liver cancer properties of erianin were found to be related mostly to its modulation of oxidative stress-mediated mitochondrial apoptosis and immune response.

// Yuexin Liu 1 1 Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA Correspondence to: Yuexin Liu, email: yliu8@mdanderson.org Keywords: clinicopathological characteristics; immune response; immunogenicity; cancer Received: August 01, 2019 Accepted: September 24, 2019 Published: October 08, 2019 ABSTRACT Background: The clinicopathologic association of tumor immune response is largely unknown. We systematically investigated this matter in human cancers. Results: Different cancer types exhibited distinct immune gene profiling. Four cancer types exhibited a significant and positive correlation of immune response with patient age. Significant but inconsistent correlation of immune response was observed with gender, surgical stage, and TNM stage in a small number of cancer types. In contrast, the histological grade appears to have much stronger and more consistent association with immune response as compared to the other clinicopathologic factors. Specifically, patients with high grade had significantly higher immune responses than those with low grade in 5 out of 12 analyzed cancer types. In addition, both histological and molecular classifications had a significant and strong association with tumor immune response. Methods: t-distributed stochastic neighbor embedding was used to assess similarity of immune gene profiling in human cancers. The Mann-Whitney or Kruskal-Wallis test was, respectively, used to compare the tumor immune response in two or more groups that were stratified by patient clinicopathological characteristics, such as gender, grade, stage (including surgical and TNM stage), histology, and molecular subtypes. Spearman correlation with student’s t -test was used to examine the association of patient age with immune response. Multiple tests with the Benjamini-Hochberg correction also were performed. Conclusions: Tumor grade should be taken into account in selection of patient candidates for immunotherapy. Prospective verification is needed before use of the findings for clinical practice.

Background OGN could modify tissue inflammation and immune response via local and circulating innate immune cells, which was suggestive of a reciprocal relationship between OGN and T cell infiltration in cancer. Hence, we aim to measure the OGN expression patterns and immune cells response in colorectal cancer(CRC). Methods This study enrolled three independent sets of patients from TCGA and the Fudan University Shanghai Cancer Center(FUSCC). The effect of OGN on T cell infiltration and the mechanism were examined in vitro and in vivo. Findings Tumor OGN expression levels were positively associated with CD3, CD8, and PTPRC expressions in the training and testing sets from TCGA, respectively. In validation set from FUSCC, OGN expression level also paralleled positively with CD8+ cell density in colorectal cancer tissue (p, Highlights • OGN is positively associated with T cell density in CRC • OGN inhibited the activation of HIF-1α, then significantly impeded the expression of VEGF.

// Julien Broseus 1,2,* , Samia Mourah 3,4,5,* , Gerard Ramstein 6 , Sophie Bernard 7 , Nicolas Mounier 8 , Wendy Cuccuini 9 , Philippe Gaulard 10,11 , Christian Gisselbrecht 7,12 , Josette Briere 13 , Remi Houlgatte 1,14,** and Catherine Thieblemont 3,7 1 Inserm U954, Faculty of Medicine, University of Lorraine, Nancy, France 2 University Hospital of Nancy, Hematology Laboratory, Nancy, France 3 Paris Diderot University, Sorbonne Paris Cite, Paris, France 4 APHP, Saint Louis University Hospital, Pharmacology-Biologic Laboratory, Paris, France 5 Inserm UMRS 976, France 6 LS2N - DUKe, University of Nantes, Nantes, France 7 APHP, Saint-Louis University Hospital, Hemato-Oncology, Paris, France 8 University Hospital of L’archet, Nice, France 9 APHP, Saint-Louis University Hospital, Hematology Laboratory, Paris, France 10 Department of Pathology, APHP, Henri Mondor University Hospital, Creteil, France 11 Inserm U955, University Paris-Est, Creteil, France 12 Lymphoma Study Association, Pierre-Benite, France 13 Department of Pathology, APHP, Saint-Louis University Hospital, Paris, France 14 University Hospital of Nancy, DRCI, Nancy, France * Julien Broseus and Samia Mourah have contributed equally to this work ** Catherine Thieblemont and Remi Houlgatte have contributed equally to this work Correspondence to: Catherine Thieblemont, email: // Remi Houlgatte, email: // Keywords : ABC, like DLBCL, angiogenesis, immune response, cancer Received : June 12, 2017 Accepted : July 03, 2017 Published : July 19, 2017 Abstract Tumor microenvironment including endothelial and immune cells plays a crucial role in tumor progression and has been shown to dramatically influence cancer survival. In this study, we investigated the clinical relevance of the gene expression of key mediators of angiogenesis, VEGF isoforms 121, 165, and 189, and their receptors (VEGFR-1 and R-2) in a cohort of patients ( n = 37) with relapsed/refractory diffuse large B-cell lymphoma (DLBCL) from the Collaborative Trial in Relapsed Aggressive Lymphoma (CORAL). In patients with ABC-like DLBCL, but not in patients with GCB-like DLBCL, low VEGF 121 expression was associated with a significantly better survival than in those with high VEGF 121 level: 4-year overall survival at 100% vs 36% ( p = .011), respectively. A specific gene signature including 57 genes was correlated to VEGF 121 expression level and was analyzed using a discovery process in 1,842 GSE datasets of public microarray studies. This gene signature was significantly expressed in other cancer datasets and was associated with immune response. In conclusion, low VEGF 121 expression level was significantly associated with a good prognosis in relapsed/refractory ABC-like DLBCL, and with a well-conserved gene-expression profiling signature related to immune response. These findings pave the way for rationalization of drugs targeting immune response in refractory/relapsed ABC-like DLBCL.

// Yaxin Wang 2 , Di Wang 1 , Gengchen Xie 1 , Yuping Yin 1 , Ende Zhao 1 , Kaixiong Tao 1 , Ruidong Li 1 1 Department of Gastrointestinal Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China 2 Department of Critical Care Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China Correspondence to: Kaixiong Tao, email: tao_kaixiongg@yahoo.com Ruidong Li, email: liruidonghust@163.com Keywords: gastric cancer, microRNA-152, B7-H1, immune response Received: December 19, 2016 Accepted: February 20, 2017 Published: March 06, 2017 ABSTRACT MiR-152 has been reported may be involved in carcinogenesis in gastric cancer. However, its role has not been comprehensively investigated in gastric cancer. We found miR-152 in human gastric cancer tissues were significantly lower than that in matched adjacent normal tissues. Meanwhile, lower miR-152 was also found in gastric cancer cell lines. The stage, tumor size and lymph node metastasis rate were significant higher in low–miR-152 group in clinical patients. Furthermore, there was a marked correlation between the levels of miR-152 and B7-H1 mRNA in gastric cancer tissues. Mechanistically, miR-152 directly bind to B7-H1 3′ untranslated region in gastric cancer cell and inhibited B7-H1 expression. Functional study demonstrated that elevation of miR-152 enhanced T cells proliferation and effector cytokines production via inhibiting B7-H1/PD-1 pathway. In conclusion, our work identified a novel mechanism by which immune response is increased by expression of miR-152 via targeting B7-H1. MiR-152 may be a potential therapeutic approach for gastric cancer.

Extranodal natural killer/T-cell lymphoma, nasal type (ENKTL) is a rare subtype of lymphoma that is often associated with poor clinical prognosis. Several studies have shown that hepatitis B virus (HBV) infection may be associated with increased risk of B-cell non-Hodgkin lymphoma; however, because of the rarity of ENKTL, little is known about its association with HBV. Our study aimed to assess whether HBV infection was associated with increased odds of ENKTL. We conducted a hospital-based case-control study including 417 ENKTL cases and 488 age- and sex-matched subjects with nonmalignant diseases unrelated to HBV infection. Multivariable unconditional logistic regression analyses were performed to estimate adjusted odds ratios [AOR] and their corresponding 95% confidence intervals (CI). The results of the multivariable analysis showed that after adjustment for a set of known risk factors, patients previously infected with HBV (HBsAg-seronegative/anti-HBc-seropositive) and naturally immune to HBV (anti-HBs-seropositive/anti-HBc-seropositive) were at significantly greater odds of being diagnosed with ENKTL (AOR, 1.497; 95% CI 1.098-2.042, P=0.033 and AOR, 1.871; 95% CI 1.302-2.689, P=0.001, respectively). After adjusting for other factors, significantly greater odds of being diagnosed with ENKTL were observed among cases who reported ever drinking alcohol (AOR, 1.675; 95% CI 1.054-2.660, P=0.029). The odds of ENKTL diagnosis were not significantly associated with ABO blood type, cigarette smoking status or family history of cancer. The results of our study suggest that patients previously infected with HBV and naturally immune to HBV were at greater odds of being diagnosed with ENKTL.

Cancer gene therapy has attracted increasing attention for its advantages over conventional therapy in specific killing of tumor cells. Here, we attempt to prove a novel therapeutic approach that targets tumors by harnessing the blood antigen immune response system, which is inherently present in patients with breast cancers. Breast cancer MDA-MB-231 cells expressed blood group H antigen precursor. After ectopic expression of blood group A glycosyltransferase, we found that the H precursor was converted into the group A antigen, appearing on the surface of tumor cells. Incubation with group B plasma from breast cancer patients activated the antigen-antibody-complement cascade and triggered tumor cell killing. Interestingly, expression of blood A antigen also reduced tumorigenesis in breast cancer cells by inhibiting cell proliferation, migration, and tumor sphere formation. Cell cycle analysis revealed that cancer cells were paused at S phase due to the activation of cell cycle regulatory genes. Furthermore, pro-apoptotic genes were unregulated by the A antigen, including BAX, P21, and P53, while the anti-apoptotic BCL2 was down regulated. Importantly, we showed that extracellular HMGB1 and ATP, two critical components of the immunogenic cell death pathway, were significantly increased in the blood A antigen-expressing tumor cells. Collectively, these data suggest that blood antigen therapy induces specific cancer cell killing by activating the apoptosis and immunogenic cell death pathways. Further translational studies are thereby warranted to apply this approach in cancer immuno-gene therapy.

// Xiaoxin Zhang 1 , Qiaoqiao Xu 1 , Lirun Yang 1 , Yueyang Lai 1 , Zhuangzhuang Zhang 1 , Chao Han 1 , Chizhou Jiang 1 , Jiahuang Li 1 , Yixin Shi 2 , Zi-Chun Hua 1, 3, 4, 5 1 The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science and School of Stomatology, Affiliated Stomatological Hospital, Nanjing University, Nanjing, 210023, Jiangsu, China 2 The School of Life Sciences and the Center for Infectious Diseases and Vaccinology at the Biodesign Institute, Arizona State University, Tempe, Arizona, 85287-4501, USA 3 Changzhou High-Tech Research Institute of Nanjing University and Jiangsu TargetPharma Laboratories Inc., Changzhou, 213164, Jiangsu, China 4 The State Key Lab of Natural Medicines, China Pharmaceutical University, Nanjing 210017, China 5 The State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210008, China Correspondence to: Zi-Chun Hua, email: zchua@nju.edu.cn Yixin Shi, email: yixin.shi@asu.edu Keywords: VNP20009, cancer, bacteria, tumor-targeting, immune response Received: April 29, 2016 Accepted: October 14, 2016 Published: November 08, 2016 ABSTRACT VNP20009 is a very effective anti-cancer agent and can specifically target tumors and inhibit tumor growth. It was assumed that the tumor targeting ability of VNP20009 correlated to its anticancer capacity. However, our observation contradicted to this assumption. Three VNP20009 mutant strains ( ΔslyA , ΔSTM3120 and ΔhtrA ) with reduced fitness in normal tissues and unchanged fitness in tumors partially or completely lost their anti-cancer capacities. The genes slyA , STM3120 and htrA were required for survival within macrophages and were indispensable for tumor microenvironment remodeling by VNP20009. The infiltration of immune cells occurred less in the tumors of mice infected with the mutant strains. In addition, the mRNA levels of TNF-α and IL-1β were significantly decreased in the tumors of mice treated with the mutant strains. Our results indicate that the immune responses elicited by bacteria rather than the bacterial titer in tumors play a “decisive” role in VNP20009-mediated bacterial cancer therapy, which provides a novel perspective for the underlying mechanism of bacterial cancer therapy.

Reactive oxygen species (ROS) modulator 1 (Romo1) is a mitochondrial membrane protein that is essential for the regulation of mitochondrial ROS production and redox sensing. Although the physiological functions of Romo1 have been studied for the past few years, the role of Romo1 in cancer remained unclear. In this study, we found that the high expression of Romo1 is associated with the poor prognosis of glioblastoma patients. Further study revealed that Romo1 is highly expressed in macrophages, indicating that the overexpression of Romo1 may participate in the function of macrophages and contribute to the progression of glioblastoma. Through the glioblastoma mouse model, we found that the overexpression of Romo1 in bone marrow cells significantly inhibited the immune response within tumor microenvironment, and that the overexpression of Romo1 resulted in the M2 polarization of bone marrow derived macrophages (BMDMs) through mTORC1 signaling pathway. In addition, the inhibition of Romo1 combining with anti-PD-1 immunotherapy significantly improved the survival outcome of glioblastoma in mouse model. Collectively, our study highlights the important role of Romo1 in immune response especially the function of macrophages, and implicates it as a potential target of immunotherapy for glioblastoma.

International audience; Pancreatic cancer is one of the most aggressive human cancers. PD1/PDL1-inhibitors recently showed promising results in different cancers with correlation between PDL1 tumor expression and responses. Expression of programmed cell death receptor ligand 1 (PDL1) has been scarcely studied in pancreatic cancer. In this retrospective study, we analyzed PDL1 mRNA expression in 453 clinical pancreatic cancer samples profiled using DNA microarrays and RNASeq. Compared to normal pancreatic samples, PDL1 expression was upregulated in 19% of cancer samples. Upregulation was not associated with clinicopathological features such as patients' age and sex, pathological type, tumor size, lymph node status, and grade, but was associated with shorter disease-free survival and overall survival in multivariate analyses. Analysis of correlations with biological parameters showed that PDL1 upregulation was associated with some degree of lymphocyte infiltration and signs of anti-tumor T-cell response, but to a lesser extent than what has been reported in breast cancer and GIST. PDL1-up pancreatic cancers displayed profiles of lymphocyte exhaustion, were more enriched in inhibitory molecules and pro-tumor populations (Tregs with upregulation of FOXP3 and IL10, myeloid-derived suppressor cells with upregulation of CD33 and S100A8/A9), and demonstrated a down-modulation of most MHC class I members (HLA-A/B/C, HLA-E/F/G) suggestive of a defect in antigen processing and presentation. In conclusion, our results suggest that PDL1 expression might refine the prediction of metastatic relapse in operated pancreatic cancer, and that PD1/PDL1 inhibitors might reactivate inhibited T-cells to increase the anti-tumor immune response in PDL1-upregulated tumors.

Tumor-associated macrophages (TAMs) polarized to the M2 phenotype play key roles in tumor progression in different cancer types, including lung cancer. MUC1 expression in various types of cancer is an indicator of poorer prognosis. Elevated MUC1 expression has been reported in inflammatory lung macrophages and is associated with lung cancer development. Here, we investigated the role of M2-polarized TAMs (M2-TAMs) in the generation of lung cancer stem cells (LCSCs) and tested pterostilbene, a small-molecule agent that modulates MUC1 expression in lung cancer cells, with the goal of subverting the microenvironment toward a favorable anti-tumor impact. We found that MUC1 was overexpressed in lung cancer patients, which was associated with poor survival rates. M2-TAMs and cancer cell lines were co-cultured in an experimental tumor microenvironment model. The expression levels of MUC1 and cancer stemness genes significantly increased in lung cancer cells in the presence of the M2-TAM cells. Intriguingly, pterostilbene dose-dependently suppressed self-renewal ability in M2-TAMs-co-cultured lung cancer cells, and this suppression was accompanied by downregulation of MUC1, NF-κB, CD133, β-catenin, and Sox2 expression. Moreover, MUC1-silenced M2-TAMs exhibited a significantly lower ability to promote LCSC generation and decreased levels of NF-κB, CD133, and Sox2. The results suggest that MUC1 plays an important role in TAM-induced LCSC progression. Pterostilbene may have therapeutic potential for modulating the unfavorable effects of TAMs in lung cancer progression.

// Josef Haik 1,2,3 , Gil Nardini 1 , Noga Goldman 4 , Gilli Galore-Haskel 5 , Moti Harats 1 , Isaac Zilinsky 1 , Oren Weissman 1 , Jacob Schachter 3,5 , Eyal Winkler 1,3 and Gal Markel 2,3,5 1 Department of Plastic Surgery, Tel Aviv University, Tel Aviv, Israel 2 Talpiot Medical Leadership Program, Tel Aviv University, Tel Aviv, Israel 3 Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel 4 Department of General Surgery C, Tel Aviv University, Tel Aviv, Israel 5 Ella Institute of Melanoma, Sheba Medical Center, Ramat Gan, Israel Correspondence to: Gal Markel, email: // Keywords : burns, immune suppression, natural killer, lysis receptors, MICB, Immunology and Microbiology Section, Immune response, Immunity Received : September 26, 2015 Accepted : December 23, 2015 Published : December 29, 2015 Abstract Immune suppression following major thermal injury directly impacts the recovery potential. Limited data from past reports indicate that natural killer cells might be suppressed due to a putative soluble factor that has remained elusive up to date. Here we comparatively study cohorts of patients with Major and Non-Major Burns as well as healthy donors. MICB and ULBP1 are stress ligands of NKG2D that can be induced by heat stress. Remarkably, serum concentration levels of MICB and ULBP1 are increased by 3-fold and 20-fold, respectively, already within 24h post major thermal injury, and are maintained high for 28 days. In contrast, milder thermal injuries do not similarly enhance the serum levels of MICB and ULBP1. This kinetics coincides with a significant downregulation of NKG2D expression among peripheral blood NK cells. Downregulation of NKG2D by high concentration of soluble MICB occurs in cancer patients and during normal pregnancy due to over production by cancer cells or extravillous trophoblasts, respectively, as an active immune-evasion mechanism. In burn patients this seems an incidental outcome of extensive thermal injury, leading to reduced NKG2D expression. Enhanced susceptibility of these patients to opportunistic viral infections, particularly herpes viruses, could be explained by the reduced NKG2D expression. Further studies are warranted for translation into innovative diagnostic or therapeutic technologies.

MAGE-A4 antigen is a cancer-testis antigen that is frequently expressed in tumor tissues. Cholesteryl pullulan (CHP) is a novel antigen delivery system for cancer vaccines. This study evaluated the safety, immune responses and clinical outcomes of patients who received a CHP-MAGE-A4 vaccine. Twenty-two patients with advanced or metastatic cancer were enrolled, and were subcutaneously vaccinated with either 100 μg or 300 μg of CHP-MAGE-A4. Seven and 15 patients, respectively, were repeatedly vaccinated with 100 μg or 300 μg of CHP-MAGE-A4; patients in both groups received a median of 7 doses. No serious adverse events related to the vaccine were observed. Of 7 patients receiving the 100 μg dose, 2 (29%) showed immune responses, compared with 3 of the 14 (21%) patients who received the 300 μg dose. In total, MAGE-A4-specific antibody responses were induced in 5 of 21 (24%) patients. No differences in survival were seen between patients receiving the 100 μg and 300 μg doses, or between immune responders and non-responders. Eleven (50%) patients had pre-existing antibodies to NY-ESO-1. In 16 patients with esophageal or head/neck squamous cell carcinoma, the survival time was significantly shorter in those who had NY-ESO-1-co-expressing tumors. Patients with high pre-existing antibody responses to NY-ESO-1 displayed worse prognosis than those with no pre-existing response. Therefore, in planning clinical trials of MAGE-A4 vaccine, enrolling NY-ESO-1-expressing tumor or not would be a critical issue to be discussed. Combination vaccines of MAGE-A4 and NY-ESO-1 antigens would be one of the strategies to overcome the poor prognosis.

// Christian D. Fankhauser 1 , Peter J. Schuffler 2, 3 , Silke Gillessen 4 , Aurelius Omlin 4 , Niels J. Rupp 5 , Jan H. Rueschoff 5 , Thomas Hermanns 1 , Cedric Poyet 1 , Tullio Sulser 1 , Holger Moch 5 and Peter J. Wild 5 1 Department of Urology, University Hospital Zurich, University of Zurich, Zurich, Switzerland 2 Department of Computer Science, ETH Zurich, Zurich, Switzerland 3 Department of Medical Physics, Memorial Sloan Kettering Cancer Center, New York, NY, USA 4 Department of Medical Oncology and Hematology, Cantonal Hospital, St. Gallen, Switzerland 5 Department of Pathology and Molecular Pathology, University Hospital Zurich, University of Zurich, Zurich, Switzerland Correspondence to: Peter J. Wild, email: peter.wild@usz.ch Keywords: prostate cancer, PD-L1, immunotherapy, immune response Received: September 30, 2016 Accepted: November 15, 2017 Published: December 04, 2017 ABSTRACT Background: We aimed to analyze the frequency and distribution of PD-L1 expression in specimens from prostate cancer (PC) patients using two different anti-PD-L1 antibodies (E1L3N, SP263). Materials and Methods: PD-L1 immunohistochemistry was performed in a tissue microarray consisting of 82 castration-resistant prostate cancer (CRPC) specimens, 70 benign prostate hyperplasia (BPH) specimens, 96 localized PC cases, and 3 PC cell lines, using two different antibodies (clones E1L3N, and SP263). Staining images for CD4, CD8, PD-L1, and PanCK of a single PD-L1 positive case were compared, using a newly developed dot-wise correlation method for digital images to objectively test for co-expression. Results: Depending on the antibody used, in tumor cells (TC) only five (E1L3N: 6%) and three (SP263: 3.7%) samples were positive. In infiltrating immune cells (IC) 12 (SP263: 14.6%) and 8 (E1L3N: 9.9%) specimens showed PD-L1 expression. Two PC cell lines (PC3, LnCaP) also displayed membranous immunoreactivity. All localized PCs or BPH samples tested were negative. Dot-wise digital correlation of expression patterns revealed a moderate positive correlation between PD-L1 and PanCK expression, whereas both PanCK and PD-L1 showed a weak negative Pearson correlation coefficient between CD4 and CD8. Conclusions: PD-L1 was not expressed in localized PC or BPH, and was only found in a minority of CRPC tumors and infiltrating immune cells. Protein expression maps and systematic dot-wise comparison could be a useful objective way to describe the relationship between immuno- and tumor-related proteins in the future, without the need to develop multiplex staining methods.

Background Melanoma is one of the fastest-rising types of cancer in North American. Accumulating evidence suggests that anti-tumor immune tolerance plays a critical role in tumor development. Methods B16 melanoma cells were injected into wild type and miR-17 overexpressing transgenic mice. Tumor growth was monitored and tumor bearing mice were sacrificed by the end of the forth week. Peripheral blood and spleen cells were subject to flow cytometry analysis and tumor samples were subject to immunohistochemistry staining. Meanwhile, Jurkat cells transfected with mock-control or miR-17 overexpressing plasmid were co-cultured with B16 cells. The influence of miR-17 on cell cycle, proliferation and survival was evaluated. Results The melanoma tumors formed in mice overexpressing miR-17 were less than that in wild type mice. In addition, the miR-17 tumors were less invasive and less angiogenic. The percentage of CD8+ T cells was suppressed in miR-17 transgenic mice before melanoma cell injection. Its level was significantly increased upon tumor grafting. More tumor infiltrating CD8+ cytotoxic T lymphocyte could be found in transgenic mice with tumor formation. Luciferase assay and protein analysis indicated that STAT3 was the target of miR-17. Decreased levels of STAT3 were associated with miR-17 over-expression. Down-regulation of STAT3 in Jurkat cells promoted cell proliferation and mitosis. Conclusions MiR-17 inhibits melanoma growth by stimulating CD8+ T cells mediated host immune response, which is due to its regulation of STAT3.

// Kelli A. Connolly 1 , Brian A. Belt 2 , Nathania M. Figueroa 2 , Aditi Murthy 1 , Ankit Patel 2 , Minsoo Kim 1 , Edith M. Lord 1 , David C. Linehan 2 , Scott A. Gerber 1, 2 1 Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY 14642, USA 2 Department of Surgery, University of Rochester Medical Center, Rochester, NY 14642, USA Correspondence to: Scott A. Gerber, email: scott_gerber@urmc.rochester.edu Keywords: radiotherapy, myeloid cells, chemokines, immunotherapy, immune response Received: August 02, 2016 Accepted: October 29, 2016 Published: November 11, 2016 ABSTRACT Although radiotherapy (RT) is widely used to control tumor growth across many cancer types, there is a relatively high incidence of RT failure exhibited by tumor recurrence, therefore a clear need exists to achieve improved effectiveness of RT. The RT-elicited immune response largely impacts the efficacy of RT and includes immune cells that kill tumor cells, but also immunosuppressive cells, which dampen anti-tumor immunity. Using murine models in which syngeneic tumor cell lines (Colon38, Glioma261, Line1) are grown intramuscularly and treated with 15 Gy local RT, we assessed the effects of RT on both the systemic and intratumoral immune response. Here we demonstrate that RT stimulates increased production of two chemokines, CCL2 and CCL5, at the tumor site. Further, that this leads to increased CCR2+ CCR5+ monocytes in circulation and subsequently alters the intratumoral immune infiltrate favoring the largely immunosuppressive CCR2+ CCR5+ monocytes. Importantly, a CCR2/CCR5 antagonist administered daily (15 mg/kg subcutaneously) starting two days prior to RT reduces both circulating and intratumoral monocytes resulting in increased efficacy of RT in radioresponsive tumors. Overall, these data have important implications for the mechanism of RT and present a means to improve RT efficacy across many cancer types.

// Julie N. Graff 1,2 , Joshi J. Alumkal 1 , Charles G. Drake 3 , George V. Thomas 4 , William L. Redmond 5 , Mohammad Farhad 5,6 , Jeremy P. Cetnar 1 , Frederick S. Ey 1 , Raymond C. Bergan 1 , Rachel Slottke 1 and Tomasz M. Beer 1 1 Division of Hematology/Oncology, Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA 2 VA Portland Health Care System, Portland, OR, USA 3 Sidney Kimmel Comprehensive Cancer Center and the Brady Urological Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA 4 Pathology and Laboratory Medicine, Oregon Health and Science University, Portland, OR, USA 5 Robert W. Franz Cancer Research Center, Earle A. Chiles Research Institute, Providence Portland Medical Center, Portland, OR, USA 6 Cell, Developmental, and Cancer Biology Department, Oregon Health and Science University, Portland, OR, USA Correspondence to: Julie N. Graff, email: // Keywords : prostate cancer, PD-1, immunotherapy, enzalutamide, Immunology and Microbiology Section, Immune response, Immunity Received : May 31, 2016 Accepted : June 17, 2016 Published : July 12, 2016 Abstract While programmed cell death 1 (PD-1) inhibitors have shown clear anti-tumor efficacy in several solid tumors, prior results in men with metastatic castration resistant prostate cancer (mCRPC) showed no evidence of activity. Here we report unexpected antitumor activity seen in mCRPC patients treated with the anti-PD-1 antibody pembrolizumab. Patients with evidence of progression on enzalutamide were treated with pembrolizumab 200 mg IV every 3 weeks for 4 doses; pembrolizumab was added to standard dose enzalutamide. Three of the first ten patients enrolled in this ongoing phase II trial experienced rapid prostate specific antigen (PSA) reductions to ≤ 0.2 ng/ml. Two of these three patients had measurable disease upon study entry; both achieved a partial response. There were three patients with significant immune-related adverse events. One had grade 2 myositis, one had grade 3 hypothyroidism, and one had grade 2 hypothyroidism. None of these patients had a response. Two of the three responders had a baseline tumor biopsy. Immunohistochemistry from those biopsies showed the presence of CD3 + , CD8 + , and CD163 + leukocyte infiltrates and PD-L1 expression. Genetic analysis of the two responders revealed markers of microsatellite instability in one. The surprising and robust responses seen in this study should lead to re-examination of PD-1 inhibition in prostate cancer.

Vascular endothelial growth factor receptor-2 (VEGFR-2) is an attractive target in oncology due to its crucial role in angiogenesis. In this study a DNA vaccine coding for human VEGFR-2 was evaluated in healthy mice and dogs, administered by intradermal injection and electroporation. In mice, three doses and vaccination schedules were evaluated. Cellular immune responses were measured by intracellular IFN-gamma staining and a cytotoxicity assay and antibodies by ELISA. Safety was assessed by measuring regulatory T cells and myeloid derived suppressor cells and a wound healing assay. The vaccine was subsequently evaluated in dogs, which were vaccinated three times with 100 mu g. Cellular immune responses were measured by intracellular IFN-gamma staining and antibodies by a flow cytometric assay. In mice, maximal cellular responses were observed after two vaccinations with 5 mu g. Humoral responses continued to increase with higher dose and number of vaccinations. No abnormalities in the measured safety parameters were observed. The vaccine was also capable of eliciting a cellular and humoral immune response in dogs. No adverse effects were observed, but tolerability of the electroporation was poor. This study will facilitate the evaluation of the vaccine in tumor bearing animals, ranging from rodent models to dogs with spontaneous tumors.

Foxp3(+) regulatory T cells (Tregs) are often highly enriched within the tumor-infiltrating T cell pool. Using a well-characterised model of carcinogen-induced fibrosarcomas we show that the enriched tumor-infiltrating Treg population comprises largely of CXCR3(+) T-bet(+) 'TH1-like' Tregs which are thymus-derived Helios(+) cells. Whilst IL-2 maintains homeostatic ratios of Tregs in lymphoid organs, we found that the perturbation in Treg frequencies in tumors is IL-2 independent. Moreover, we show that the TH1 phenotype of tumor-infiltrating Tregs is dispensable for their ability to influence tumor progression. We did however find that unlike Tconvs, the majority of intra-tumoral Tregs express the activation markers CD69, CD25, ICOS, CD103 and CTLA4 and are significantly more proliferative than Tconvs. Moreover, we have found that CD69(+) Tregs are more suppressive than their CD69- counterparts. Collectively, these data indicate superior activation of Tregs in the tumor microenvironment, promoting their suppressive ability and selective proliferation at this site.

// Bo Jiang 1 , Ting Zhang 2 , Fen Liu 2 , Zhangzhang Sun 3 , Hanping Shi 1 , Dong Hua 3 and Chen Yang 4 1 Department of Medical Oncology, Beijing Institute of Translational Medicine, Chinese Academy of Sciences/Cancer Center, Aviation General Hospital, Beijing, China 2 Institute of Cancer, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China 3 Department of Medical Oncology, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China 4 Department of Nuclear-Medicine, Suzhou Hospital Affiliated to Nanjing Medical University, Suzhou, Jiangsu, China Correspondence to: Bo Jiang, email: // Chen Yang, email: // Keywords : B7-H3, colorectal cancer, epithelial-mesenchymal transition, co-stimulatory molecule, metastasis, Immunology and Microbiology Section, Immune response, Immunity Received : July 17, 2015 Accepted : March 31, 2016 Published : April 27, 2016 Abstract B7-H3, first recognized as a co-stimulatory molecule, is abnormally expressed in cancer tissues and is associated with cancer metastasis and a poor prognosis. However, as an initial event of metastasis, the relationship between the Epithelial–Mesenchymal Transition (EMT ) in cancer cells and B7-H3 has still not been investigated. In this study, we first analyzed B7-H3 expression by immunohistochemistry in colorectal cancer tissues. B7-H3 was expressed in the cancer cell membrane and was associated with the T stage of colorectal cancer; it also showed a positive correlation with MMP2 and MMP9 expression in cancer tissues. Over-expression of B7-H3 in SW480 cells allowed cancer cells to invade and metastasize more than the control cells, whereas invasion and metastasis capabilities were decreased after B7-H3 was knocked down in Caco-2 cells. We further showed that B7-H3 down-regulated the expression of E-cadherin and β-catenin and up-regulated N-cadherin and Vimentin expression, implying that B7-H3 promoted the EMT in colorectal cancer cells. We also checked another character of the EMT, the stemness of cancer cells. CD133, CD44 and Oct4 were significantly elevated after the SW480 cells were transfected with B7-H3 and reduced in Caco-2 cells after B7-H3 was inhibited. In subsequent studies, we proved that B7-H3 upregulated the expression of Smad1 via PI3K-Akt. In conclusion, B7-H3 promotes the EMT in colorectal cancer cells by activating the PI3K-Akt pathway and upregulating the expression of Smad1.

Chromosomal instability (CIN) is a hallmark of cancer and has been implicated in cancer initiation, progression and the development of resistance to traditional cancer therapy. Here we identify a new property of CIN cells, showing that inducing CIN in proliferating Drosophila larval tissue leads to the activation of innate immune signalling in CIN cells. Manipulation of this immune pathway strongly affects the survival of CIN cells, primarily via JNK, which responds to both Toll and TNFα/Eiger. This pathway also activates Mmp1, which recruits hemocytes to the CIN tissue to provide local amplification of the immune response that is needed for effective elimination of CIN cells. Refereed/Peer-reviewed

// A. Marijne Heeren 1, 2 , Eline de Boer 1 , Maaike C.G. Bleeker 3 , Rene J.P. Musters 4 , Marrije R. Buist 5 , Gemma G. Kenter 1, 5, 6 , Tanja D. de Gruijl 2 , Ekaterina S. Jordanova 1 1 Center Gynecological Oncology Amsterdam (CGOA), Department of Obstetrics and Gynecology, VU University Medical Center, 1081 HV Amsterdam, The Netherlands 2 Department of Medical Oncology, VU University Medical Center-Cancer Center Amsterdam, 1081 HV Amsterdam, The Netherlands 3 Department of Pathology, VU University Medical Center, 1081 HV Amsterdam, The Netherlands 4 Laboratory for Physiology, Institute for Cardiovascular Research, VU University Medical Center, 1081 BT Amsterdam, The Netherlands 5 Center Gynecological Oncology Amsterdam (CGOA), Department of Obstetrics and Gynecology, Academic Medical Center, 1105 AZ Amsterdam, The Netherlands 6 Center Gynecological Oncology Amsterdam (CGOA), Department of Gynecology, Netherlands Cancer Institute - Antoni van Leeuwenhoek, 1006 BE Amsterdam, The Netherlands Correspondence to: Ekaterina S. Jordanova, e-mail: e.jordanova@vumc.nl Keywords: Immunology and Microbiology Section, Immune response, Immunity, cervical cancer, tumor-draining lymph nodes, metastatic niche, tregs, PD-L1-myeloid cells Received: August 04, 2015 Accepted: September 16, 2015 Published: September 28, 2015 ABSTRACT In cervical cancer, high frequencies of regulatory T cells (Tregs) and immunosuppressive PD-L1 + CD14 + antigen-presenting cells dominate the microenvironment of tumor-positive lymph nodes (LN+). It is unknown whether this is restricted to LN+ or precedes metastasis, emanating from the primary tumor and spreading through tumor-draining lymph nodes (TDLNs). To investigate immunosuppression in the lymphatic basin of cervical tumors, all dissected TDLNs of five cervical cancer patients (in total 9 LN+ and 74 tumor-negative lymph nodes (LN-)) were analyzed for FoxP3 + Tregs, CD8 + T cells, HLA-DR + - and PD-L1 + myeloid cells by immunohistochemistry. Tregs and PD-L1 + cells were found to form an immunosuppressive cordon around metastatic tumor cells. Importantly, whereas high HLA-DR + - and PD-L1 + cell rates were strongly associated with LN+, elevated Treg levels and decreased CD8 + T cell/Treg ratios were found similar in LN+ and adjacent LN-, as compared to LN- at more distant anatomical localizations. These data suggest that delineated fields of Treg-associated immune suppression in anatomically co-localized TDLNs enable metastasis by creating metastatic niches. This may be of importance for decision-making regarding (surgical) intervention in cervical cancer. Future efforts should include the implementation of immunotherapeutic regimens to overcome this immune suppression, establish loco-regional control and halt systemic tumor spread.

// Barbara Chirullo 1 , Serena Ammendola 2 , Leonardo Leonardi 3 , Roberto Falcini 4 , Paola Petrucci 1 , Claudia Pistoia 1 , Silvia Vendetti 5 , Andrea Battistoni 2 , Paolo Pasquali 1 1 Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanita, Rome 00161, Italy 2 Department of Biology, University of Rome Tor Vergata, Rome 00133, Italy 3 Universita degli Studi di Perugia, Department of Veterinary Medicine, Perugia 06126, Italy 4 Veterinary Clinic, Rieti 02043, Italy 5 Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanita, Rome 00161, Italy Correspondence to: Barbara Chirullo, e-mail: barbara.chirullo@iss.it Paolo Pasquali, e-mail: paolo.pasquali@iss.it Keywords: bacterial therapy, cancer therapy, antitumor efficacy, attenuated-Salmonella, immune response Received: February 09, 2015 Accepted: April 24, 2015 Published: May 07, 2015 ABSTRACT Salmonella Typhimurium has been shown to be highly effective as antitumor agent. The aim of this study was to investigate the tumor targeting efficacy and the mechanism of action of a specific attenuated mutant strain of Salmonella Typhimurium (STM) devoid of the whole operon coding for the high-affinity zinc transporter ZnuABC, which is required for bacterial growth in environments poor in zinc and for conferring full virulence to different Gram-negative pathogens. We showed that STM is able to penetrate and replicate into tumor cells in in vitro and in vivo models. The subcutaneous administration of STM in mammary adenocarcinoma mouse model led to both reduction of tumor growth and increase in life expectancy of STM treated mice. Moreover, investigating the potential mechanism behind the favorable clinical outcomes, we provide evidence that STM stimulates a potent inflammatory response and a specific immune pattern, recruiting a large number of innate and adaptive immune cells capable to contrast the immunosuppressive environment generated by tumors.