Exosomes define a local and systemic communication network in healthy pancreas and pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC), a lethal disease, requires a grasp of its biology for effective therapies. Exosomes, implicated in cancer, are poorly understood in living systems. Here we use the genetically engineered mouse model (ExoBow) to map the spatiotemporal distribution of exosomes from healthy and PDAC pancreas in vivo to determine their biological significance. We show that, within the PDAC microenvironment, cancer cells establish preferential communication routes through exosomes with cancer associated fibroblasts and endothelial cells. The latter being a conserved event in the healthy pancreas. Inhibiting exosomes secretion in both scenarios enhances angiogenesis, underscoring their contribution to vascularization and to cancer. Inter-organ communication is significantly increased in PDAC with specific organs as most frequent targets of exosomes communication occurring in health with the thymus, bone-marrow, brain, and intestines, and in PDAC with the kidneys, lungs and thymus. In sum, we find that exosomes mediate an organized intra- and inter- pancreas communication network with modulatory effects in vivo.

Effectiveness of Helicobacter pylori Treatments According to Antibiotic Resistance.

INTRODUCTION: Antibiotic resistance is one of the main factors that determine the efficacy of treatments to eradicate Helicobacter pylori infection. Our aim was to evaluate the effectiveness of first-line and rescue treatments against H. pylori in Europe according to antibiotics resistance. METHODS: Prospective, multicenter, international registry on the management of H. pylori (European Registry on H. pylori Management). All infected and culture-diagnosed adult patients registered in the Spanish Association of Gastroenterology-Research Electronic Data Capture from 2013 to 2021 were included. RESULTS: A total of 2,852 naive patients with culture results were analyzed. Resistance to clarithromycin, metronidazole, and quinolones was 22%, 27%, and 18%, respectively. The most effective treatment, regardless of resistance, were the 3-in-1 single capsule with bismuth, metronidazole, and tetracycline (91%) and the quadruple with bismuth, offering optimal cure rates even in the presence of bacterial resistance to clarithromycin or metronidazole. The concomitant regimen with tinidazole achieved an eradication rate of 99% (90/91) vs 84% (90/107) with metronidazole. Triple schedules, sequential, or concomitant regimen with metronidazole did not achieve optimal results. A total of 1,118 non-naive patients were analyzed. Resistance to clarithromycin, metronidazole, and quinolones was 49%, 41%, and 24%, respectively. The 3-in-1 single capsule (87%) and the triple therapy with levofloxacin (85%) were the only ones that provided encouraging results. DISCUSSION: In regions where the antibiotic resistance rate of H. pylori is high, eradication treatment with the 3-in-1 single capsule, the quadruple with bismuth, and concomitant with tinidazole are the best options in naive patients. In non-naive patients, the 3-in-1 single capsule and the triple therapy with levofloxacin provided encouraging results.

SMC3 epigenetic silencing regulates Rab27a expression and drives pancreatic cancer progression.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is expected to soon surpass colorectal cancer as a leading cause of cancer mortality in both males and females in the US, only lagging behind lung cancer. The lethality of PDAC is driven by late diagnosis and inefficient therapies. The complex biology of PDAC involves various cellular components, including exosomes that carry molecular information between cells. Thus, recipient cells can be reprogrammed, impacting tumorigenesis. Rab27a is a GTPase responsible for the last step of exosomes biogenesis. Hence, dissecting the mechanisms that regulate the expression of Rab27a and that control exosomes biogenesis can provide fundamental insights into the molecular underpinnings regulating PDAC progression. METHODS: To assess the mechanism that regulates Rab27a expression in PDAC, we used PDAC cell lines. The biological significance of these findings was validated in PDAC genetically engineered mouse models (GEMMs) and human samples. RESULTS: In this work we demonstrate in human PDAC samples and GEMMs that Rab27a expression decreases throughout the development of the disease, and that Rab27a knockout promotes disease progression. What is more, we demonstrate that Rab27a expression is epigenetically regulated in PDAC. Treatment with demethylating agents increases Rab27a expression specifically in human PDAC cell lines. We found that SMC3, a component of the cohesin complex, regulates Rab27a expression in PDAC. SMC3 methylation is present in human PDAC specimens and treatment with demethylating agents increases SMC3 expression in human PDAC cell lines. Most importantly, high levels of SMC3 methylation are associated with a worse prognosis in PDAC. Mechanistically, we identified an enhancer region within the Rab27a gene that recruits SMC3, and modulates Rab27a expression. CONCLUSION: Overall, we dissected a mechanism that regulates Rab27a expression during PDAC progression and impacts disease prognosis.

The indirect effect of body image on distress in women with breast cancer undergoing chemotherapy.

BACKGROUND: This study assessed the indirect effect of body image in the relationship between illness duration, optimism, coping strategies and psychological distress as well as the moderator role of being or not submitted to surgery and type of surgery, in women with breast cancer undergoing chemotherapy. PARTICIPANTS AND PROCEDURE: This cross-sectional study included eighty-seven women with breast cancer undergoing chemotherapy, who answered instruments that assessed sociodemographic and clinical issues, optimism, coping, concerns with body image and psychological distress. Bayesian statistics were performed to test the indirect effect model that included also the moderator effects. RESULTS: Lower optimism, lower use of humor, and higher denial and illness duration predicted lower body image and higher distress. Longer illness duration was associated with higher distress. Body image had an indirect effect in the relationship between optimism and distress; between denial coping and distress; between humor coping and distress and between illness duration and distress. Being submitted to surgery but not the type of surgery was a moderator in the indirect effect model. CONCLUSIONS: Body image is critical to psychological distress. Future interventions for women with breast cancer should consider body image as a target, in order to promote adaptive coping strategies specially when women have had surgery.

The influence of the gastric microbiota in gastric cancer development.

Colonization of the stomach by Helicobacter pylori is the trigger for a series of gastric mucosal changes that culminate in gastric cancer. Infection with this bacterium is considered the major risk factor for this malignancy. The introduction of high-throughput sequencing technologies coupled to advanced computational pipelines offered an improved understanding of the microbiome, and it is now currently accepted that, besides H. pylori, the stomach harbours a complex microbial community. While it is well established that H. pylori plays a central role in gastric carcinogenesis, the significance of the non-H. pylori microbiota is yet to be clarified. This review will address the state of the art on the relationship between the gastric microbiota and gastric cancer development, and identify areas where additional research is needed before translating microbiome research into preventive and therapeutic strategies to reduce gastric cancer burden.

Glycans as Immune Checkpoints: Removal of Branched N-glycans Enhances Immune Recognition Preventing Cancer Progression.

Tumor growth is accompanied with dramatic changes in the cellular glycome, such as the aberrant expression of complex branched N-glycans. However, the role of this protumoral N-glycan in immune evasion and whether its removal contributes to enhancement of immune recognition and to unleashing an antitumor immune response remain elusive. We demonstrated that branched N-glycans are used by colorectal cancer cells to escape immune recognition, instructing the creation of immunosuppressive networks through inhibition of IFNγ. The removal of this "glycan-mask" exposed immunogenic mannose glycans that potentiated immune recognition by DC-SIGN-expressing immune cells, resulting in an effective antitumor immune response. We revealed a glycoimmune checkpoint in colorectal cancer, highlighting the therapeutic efficacy of its deglycosylation to potentiate immune recognition and, thus, improving cancer immunotherapy.

The Dysfunctional Immune System in Common Variable Immunodeficiency Increases the Susceptibility to Gastric Cancer.

Gastric carcinoma (GC) represents the most common cause of death in patients with common variable immunodeficiency (CVID). However, a limited number of cases have been characterised so far. In this study, we analysed the clinical features, bacterial/viral infections, detailed morphology and immune microenvironment of nine CVID patients with GC. The study of the immune microenvironment included automated digital counts of CD20+, CD4+, CD8+, FOXP3+, GATA3+ and CD138+ immune cells, as well as the evaluation of PD-L1 expression. Twenty-one GCs from non-CVID patients were used as a control group. GC in CVID patients was diagnosed mostly at early-stage (n = 6/9; 66.7%) and at younger age (median-age: 43y), when compared to non-CVID patients (p < 0.001). GC pathogenesis was closely related to Helicobacter pylori infection (n = 8/9; 88.9%), but not to Epstein-Barr virus (0.0%) or cytomegalovirus infection (0.0%). Non-neoplastic mucosa (non-NM) in CVID-patients displayed prominent lymphocytic gastritis (100%) and a dysfunctional immune microenvironment, characterised by higher rates of CD4+/CD8+/Foxp3+/GATA3+/PD-L1+ immune cells and the expected paucity of CD20+ B-lymphocytes and CD138+ plasma cells, when compared to non-CVID patients (p < 0.05). Changes in the immune microenvironment between non-NM and GC were not equivalent in CVID and non-CVID patients, reflecting the relevance of immune dysfunction for gastric carcinogenesis and GC progression in the CVID population.

Genetic Variants of the MGAT5 Gene Are Functionally Implicated in the Modulation of T Cells Glycosylation and Plasma IgG Glycome Composition in Ulcerative Colitis.

OBJECTIVES: The impact of genetic variants (single nucleotide polymorphisms [SNPs]) in the clinical heterogeneity of ulcerative colitis (UC) remains unclear. We showed that patients with UC exhibit a deficiency in MGAT5 glycogene transcription in intestinal T cells associated with a hyperimmune response. Herein, we evaluated whether MGAT5 SNPs might functionally impact on T cells glycosylation and plasma IgG glycome in patients with UC, as well as in UC clinical outcomes. METHODS: Three selected MGAT5 SNPs (rs3814022, rs4953911, and rs1257220), previously associated with severity of autoimmune disease or with plasma glycome composition in healthy individuals, were functionally evaluated in patients with UC through analysis of MGAT5 mRNA levels in colonic (n = 14) and circulating (n = 24) T cells and through profiling the plasma IgG Fc glycosylation (n = 152). MGAT5 SNPs were genotyped in 931 patients with UC from 2 European cohorts and further associated with patients' prognosis. Targeted next-generation sequencing for MGAT5 coding and regulatory regions was also performed. RESULTS: MGAT5 SNPs were shown to be functionally associated with low transcription levels of MGAT5 in colonic and circulating T cells from patients with UC and with agalactosylation of IgGs, often associated with a proinflammatory phenotype. The SNPs rs3814022 and rs4953911 were further associated with the need of biologics. Next-generation sequencing data further revealed a combination of MGAT5 SNPs that stratify patients with UC according to their severity. DISCUSSION: Our results revealed that MGAT5 SNPs have a phenotypic impact on T cells glycosylation and in plasma IgG glycome composition associated with UC pathogenesis. MGAT5 SNPs display a tendency in the association with a worse disease course in patients with UC.

Induction of apoptosis increases sensitivity to detect cancer mutations in plasma.

BACKGROUND: The study of cell-free DNA (cfDNA), namely the fraction derived from tumors (ctDNA), is a clinically relevant noninvasive biomarker for cancer management. However, the intrinsic low abundance of ctDNA in plasma limits its implementation in the clinic. AIM OF THE STUDY: In this study, the objective was to demonstrate that induction of apoptosis-the major source of ctDNA-increases ctDNA concentration, thereby increasing the sensitivity to detect clinically relevant mutations in plasma. METHODS: In vitro models were used to test the effect of docetaxel on the release levels of DNA from lung cancer cells. In vivo, Rag2-/-IL2rg-/- immunodeficient C57BL/6 xenografted mice were treated with docetaxel for 24 h or 48 h. Tumor tissue and blood were collected to evaluate the levels of apoptosis DNA release levels, respectively. RESULTS: We observed increased levels of apoptosis in H1975 cells and a consequent increase in cfDNA released into the culture medium after docetaxel treatment. In vivo, the results show increased cfDNA concentration in plasma of xenografted mice after apoptosis stimulation. Importantly, treatment increased the sensitivity of detection of relevant cancer mutations, namely 24 h after treatment. CONCLUSION: This study provides new insights regarding the importance of timing for blood collection. In our experimental model, we demonstrate that blood collection should be performed 24 h after treatment (apoptosis induction), for optimal ctDNA analysis. Translating these results into the clinical setting is likely to increase sensitivity to detect tumor-derived mutations in plasma, might help guide the therapeutic decision, and optimize current liquid biopsy procedures for situations where tissue analysis is not possible.

Systematic review: gastric microbiota in health and disease.

BACKGROUND: Helicobacter pylori is the most infamous constituent of the gastric microbiota and its presence is the strongest risk factor for gastric cancer and other gastroduodenal diseases. Although historically the healthy stomach was considered a sterile organ, we now know it is colonised with a complex microbiota. However, its role in health and disease is not well understood. AIM: To systematically explore the literature on the gastric microbiota in health and disease as well as the gut microbiota after bariatric surgery. METHODS: A systematic search of online bibliographic databases MEDLINE/EMBASE was performed between 1966 and February 2019 with screening in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Randomised controlled trials, cohort studies and observational studies were included if they reported next-generation sequencing derived microbiota analysis on gastric aspirate/tissue or stool samples (bariatric surgical outcomes). RESULTS: Sixty-five papers were eligible for inclusion. With the exception of H pylori-induced conditions, overarching gastric microbiota signatures of health or disease could not be determined. Gastric carcinogenesis induces a progressively altered microbiota with an enrichment of oral and intestinal taxa as well as significant changes in host gastric mucin expression. Proton pump inhibitors usage increases gastric microbiota richness. Bariatric surgery is associated with an increase in potentially pathogenic proteobacterial species in patient stool samples. CONCLUSION: While H pylori remains the single most important risk factor for gastric disease, its capacity to shape the collective gastric microbiota remains to be fully elucidated. Further studies are needed to explore the intricate host/microbial and microbial/microbial interplay.

Longitudinal changes on the quality of life in caregivers of type 2 diabetes amputee patients.

PURPOSE: To explore changes in the quality of life of caregivers of amputees due to type 2 diabetes ten months after amputation. METHODS: This is a longitudinal study with three moments of evaluation (T1: one month after surgery, T2: 7 months, T3: 10 months). The sample comprised 110, 101, and 84 caregivers of amputated patients with type 2 diabetes. Caregivers answered a Socio-demographic questionnaire; the Self-Assessment Caregiver; the Family Disruption from Illness Scale; and the Short Form Health Survey (SF36). RESULTS: Stress levels were not significantly reflected in changes on mental quality of life over time, except in the caregivers who presented less stress, emphasizing the adverse role of stress when experienced on a continuous basis for ten months on the caregivers' mental well-being. Caregivers presented greater number of physical symptoms at T2 that decreased at T3. CONCLUSIONS: According to the results, in order to promote caregivers' physical and mental quality of life, it would be important to evaluate stress levels especially in patients who presented somatic complaints.

Quality of life in patients with mild Alzheimer disease: the mediator role of mindfulness and spirituality.

OBJECTIVES: This study examined the mediator role of mindfulness and spirituality in the relationship between psychological morbidity, awareness of the disease, functionality, social support, family satisfaction, and quality of life (QoL) in patients with mild AD. METHOD: The sample consisted of 128 patients who answered the Cognitive and Affective Mindfulness Scale-Revised (CAMS-R), the Assessment Scale of Psychosocial Impact of the Diagnosis of Dementia (ASPIDD), the Hospital Anxiety and Depression Scales (HADS), the Satisfaction with Social Support Scale (SSSS), the Family Satisfaction Scale (FSS), the Spiritual and Religious Attitudes in Dealing with Illness (SpREUK), the Index of Barthel, and the Quality of Life-Alzheimer's Disease (QoL-AD). RESULTS: Mindfulness and spirituality mediated the relationship between functionality, awareness of the disease, family satisfaction and QoL. Psychological morbidity had a direct negative impact on QoL and was negatively associated with awareness of the disease, family satisfaction and social support. Mindfulness was negatively associated with spirituality and the latter was negatively associated with QoL. More social support was associated with greater awareness of the disease and family satisfaction. More functionality, awareness of the disease and family satisfaction contributed to more QoL and this relationship was mediated by mindfulness and spirituality. CONCLUSION: Interventions directed at the promotion of the QoL of patients with mild AD should focus on the promotion of mindfulness skills in AD patients, in addition to the reduction of psychological morbidity and the promotion of functionality, awareness of the disease, family relationships and social support.

Brief emotional screening in oncology: Specificity and sensitivity of the emotion thermometers in the Portuguese cancer population.

OBJECTIVE: This study aimed to determine the cutoff and the specificity and sensitivity of the Emotion Thermometers (ET) in a Portuguese sample of cancer patients. METHOD: A total of 147 patients (mean age = 49.2; SD = 12.6) completed the ET, the Brief Symptom Inventory (BSI), and the Subjective Experiences of Illness Suffering Inventory. Data were collected in a cancer support institution and in a major hospital in the North of Portugal. RESULT: The optimal cutoff for the Anxiety Thermometer was 5v6 (until 5 and 6 or more), which identified 74% of the BSI-anxiety cases and 70% of noncases. The Depression Thermometer cutoff was 4v5 (until 4 and 5 or more), which identified 85% of BSI-depression cases and 82% of noncases. Cutoff for the Anger Thermometer was 4v5 (until 4 and 5 or more), which identified 83% of BSI-hostility cases and 73% of noncases; for the Distress Thermometer, the optimal cutoff was 4v5 (until 4 and 5 or more), which identified 84% of the suffering cases and 73% of noncases. Finally, for the Help Thermometer, it was 3v4 (until 3 and 4 or more), which helped to identify 93% of the suffering cases and 64% of noncases. SIGNIFICANCE OF RESULTS: Results supported the Portuguese version of the ET as an important screening tool for identifying the emotional distress in cancer patients.

Differences, predictors, and moderators of therapeutic adherence in patients recently diagnosed with type 2 diabetes.

This study analyzed the differences over time in newly diagnosed type 2 diabetes patients on reported adherence. A longitudinal design with two assessment moments was used with 268 patients who were assessed on adherence to self-care behaviors and medication, beliefs about medicines, psychological distress, trust in the physician, and satisfaction with care. HbA1c and general beliefs about medicines decreased from T1 to T2 while adherence to foot care, the needs of medicines, and psychological distress increased. Beliefs about medicines, satisfaction with communication/information, and trust in physician predicted adherence. Intervention should consider these variables when promoting adherence.

The Two Faces of Tumor-Associated Macrophages and Their Clinical Significance in Colorectal Cancer.

Macrophages are one of the immune populations frequently found in colorectal tumors and high macrophage infiltration has been associated with both better and worst prognosis. Importantly, according to microenvironment stimuli, macrophages may adopt different polarization profiles, specifically the pro-inflammatory or M1 and the anti-inflammatory or M2, which display distinct functions. Therefore, concomitantly with the number of tumor-associated macrophages (TAMs), their characterization is fundamental to unravel their relevance in cancer. Here, we profiled macrophages in a series of 150 colorectal cancer (CRC) cases by immunohistochemistry, using CD68 as a macrophage lineage marker, CD80 as a marker of pro-inflammatory macrophages, and CD163 as a marker of anti-inflammatory macrophages. Quantifications were performed by computer-assisted analysis in the intratumoral region, tumor invasive front, and matched tumor adjacent normal mucosa (ANM). Macrophages, specifically the CD163+ ones, were predominantly found at the tumor invasive front, whereas CD80+ macrophages were almost exclusively located in the ANM, which suggests a predominant anti-inflammatory polarization of TAMs. Stratification according to tumor stage revealed that macrophages, specifically the CD163+ ones, are more prevalent in stage II tumors, whereas CD80+ macrophages are predominant in less invasive T1 tumors. Specifically in stage III tumors, higher CD68, and lower CD80/CD163 ratio associated with decreased overall survival. Importantly, despite the low infiltration of CD80+ cells in colorectal tumors, multivariate logistic regression revealed a protective role of these cells regarding the risk for relapse. Overall, this work supports the involvement of distinct microenvironments, present at the intra-tumor, invasive front and ANM regions, on macrophage modulation, and uncovers their prognostic value, further supporting the relevance of including macrophage profiling in clinical settings.

Review: Gastric malignancies: Basic aspects.

Gastric cancer is the third deadliest cancer in the world, and the absolute number of cases is increasing every year due to aging and growing of high-risk populations. This disease is a consequence of the complex interaction of microbial agents, with environmental and host factors, resulting in the dysregulation of multiple oncogenic and tumor-suppressing signaling pathways. Despite the advances in our understanding of carcinogenesis, there are still reduced therapeutic options for patients with gastric cancer. In recent years, genomic analyses of gastric tumors have emphasized their molecular heterogeneity. The distinction of gastric cancer molecular subtypes may be a key to identify novel therapeutic targets, to predict patient outcome and response to therapy, and to guide early diagnosis strategies. In this review, we summarize the most recent updates on the relationship between microbial agents and gastric cancer, in particular, Helicobacter pylori, the non-H pylori microbiome, and Epstein-Barr virus. We also highlight the main advances made in the past year regarding the molecular characterization of gastric cancer, especially the signatures with potential clinical utility.

Beliefs about medicines as mediators in medication adherence in type 2 diabetes.

AIMS: This study analysed whether beliefs about medicines mediated the relationship between illness representations and medication adherence. BACKGROUND: Adherence to medication is required in diabetes treatment, contributing to decreased blood glycaemic levels. The knowledge and perception of patients about diabetes as well as the beliefs about medicines are considered to be key factors for medication adherence. DESIGN: The study used a cross-sectional design that included 387 patients recently diagnosed with type 2 diabetes. METHODS: Participants were assessed, between 2010 and 2013, and answered the Medication Adherence Scale, the Beliefs about Medicines Questionnaire, and the Brief Illness Perception Questionnaire. RESULTS: The results of the path analysis showed that beliefs about medicines had a mediating role on self-report medication adherence with the exception of beliefs about specific concerns with medicines. Therefore, both general beliefs and specific needs about medicines mediated the relationship between diabetes consequences and self-report medication adherence as well as between treatment control and self-report medication adherence. Needs about medicines mediated the relationship between personal control and self-report medication adherence. CONCLUSION: Health professionals should target beliefs about medicines besides illness representations regarding medication adherence. The current study may help optimize adherence to medication in early-diagnosed type 2 diabetes patients.

Quality of life in elderly Portuguese war veterans with post-traumatic stress symptoms.

BACKGROUND: Studies show that post-traumatic stress symptoms among Portuguese veterans who participated in Colonial War (1961-75) are high, even though 43 years have gone by since the end of the war. AIMS: This study analyzed the role of family type, personality traits, and social support as predictors of post-traumatic stress symptoms and quality of life in war veterans, and whether satisfaction with social support was a mediator between neuroticism/post-traumatic stress symptoms and quality of life. METHOD: A cross-sectional study was conducted including 230 war veterans with a mean age of 60 years (SD=3.82). RESULTS: Results indicated a high prevalence of post-traumatic stress symptoms as well as high neuroticism, 16.5 (SD=4.41); 81% of veterans presented high psychological distress, suggesting emotional disturbance and 71% belonged to extreme families (families with cohesion and adaptability problems). Results showed that age (ß=-0.166, p<0.05), social support (ß=-0.184, p<0.01), and neuroticism (ß=0.325, p<0.001) predicted post-traumatic stress symptoms. Age, professional status, social support, post-traumatic stress symptoms, family type, neuroticism, and extroversion predicted different dimensions of quality of life. Finally, a path analysis showed that satisfaction with social support was a mediator in the relationship between neuroticism and quality of life (ß=-0.066; p<0.01) and between post-traumatic stress symptoms and quality of life (ß=-0.108; p<0.01). CONCLUSION: Four decades after the Colonial War have passed, there is still a high prevalence of post-traumatic stress symptoms. Screening elderly veterans who present post-traumatic stress symptoms, for the presence of neuroticism traits, and assessing family type and social support, should be a standard practice in health care services, especially in the oldest and those who are retired. Social support should be promoted in order to enhance quality of life in this population.

Family and Couple Variables Regarding Adherence in Type 2 Diabetes Patients in the Initial Stages of the Disease.

This study analyzed which family and couple variables predicted adherence to standard care treatment, in patients recently diagnosed with type 2 diabetes. The sample comprised 224 dyads assessed during the first year of diagnosis (T1) and 4 months later (T2). The results showed that family stress, dyadic adjustment, family coping, and positive support assessed by patients at T1 predicted medication adherence and glucose monitoring at T2. Positive support and dyadic adjustment, assessed by partners at T1, predicted patients' adherence to glucose monitoring and diet at T2. This study highlights the important role of the partner in patient`s adherence. Therefore, standard care in type 2 diabetes should be offered in the context of the dyad.

New insights into the inflamed tumor immune microenvironment of gastric cancer with lymphoid stroma: from morphology and digital analysis to gene expression.

BACKGROUND: Gastric cancer with lymphoid stroma (GCLS) is characterized by prominent stromal infiltration of T-lymphocytes. The aim of this study was to investigate GCLS biology through analysis of clinicopathological features, EBV infection, microsatellite instability (MSI), immune gene-expression profiling and PD-L1 status in neoplastic cells and tumor immune microenvironment. METHODS: Twenty-four GCLSs were analyzed by RNA in situ hybridization for EBV (EBER), PCR/fragment analysis for MSI, immunohistochemistry (PD-L1, cytokeratin, CD3, CD8), co-immunofluorescence (CK/PD-L1, CD68/PD-L1), NanoString gene-expression assay for immune-related genes and PD-L1 copy number alterations. CD3+ and CD8+ T-cell densities were calculated by digital analysis. Fifty-four non-GCLSs were used as control group. RESULTS: GCLSs displayed distinctive clinicopathological features, such as lower pTNM stage (p = 0.02) and better overall survival (p = 0.01). EBV+ or MSI-high phenotype was found in 66.7 and 16.7% cases, respectively. GCLSs harbored a cytotoxic T-cell-inflamed profile, particularly at the invasive front of tumors (p < 0.01) and in EBV+ cases (p = 0.01). EBV+ GCLSs, when compared to EBV- GCLSs, showed higher mRNA expression of genes related to Th1/cytotoxic and immunosuppressive biomarkers. PD-L1 protein expression, observed in neoplastic and immune stromal cells (33.3 and 91.7%, respectively), and PD-L1 amplification (18.8%) were restricted to EBV+/MSI-high tumors and correlated with high values of PD-L1 mRNA expression. CONCLUSIONS: This study shows that GCLS has a distinctive clinico-pathological and molecular profile. Furthermore, through an in-depth study of tumor immune microenvironment-by digital analysis and mRNA expression profiling-it highlights the role of EBV infection in promoting an inflamed tumor microenvironment, with putative therapeutic implications.