Immunonutrition Changes Inflammatory Response in Colorectal Cancer: Results from a Pilot Randomized Clinical Trial

Introduction: Surgery is the first choice of treatment for colorectal cancer. Nutritional support in the form of oral nutritional supplements (ONSs) in the preoperative period is widely accepted for reducing the incidence of perioperative complications, and immunonutrition is generally recommended. However, there is little clinical data regarding the impact of such treatment on tumor biology. Material and Methods: In this study, tumor tissue and blood samples were collected from 26 patients during preoperative colonoscopy at the time of clinical diagnosis (sample A). Group 1 received standard ONSs (3× Nutricia Nutridrink Protein per day) for 2 weeks before surgery. In group 2, immune ONSs (2× Nestle Impact Oral) were administered for the same duration. Tumor tissue (sample B) was then retrieved from the tumor after resection. Changes in the expression levels of inflammatory cytokines (TNF-α, interleukin 8 or chemokine (C-X-C motif) ligand (CXCL8), stromal cell-derived factor 1 (SDF1a), chemokine (C-X-C motif) ligand 6 (CXCL6), chemokine (C-X-C motif) ligand (CXCL2), myeloperoxidase (MPO), and CXCL1) were assessed during the perioperative course. Results: TNF-α expression differed after intervention between the two groups (immune group 31.63 ± 13.28; control group 21.54 ± 6.84; p = 0.049) and prior to and after intervention in the control group (prior to intervention 35.68 ± 24.41; after intervention 21.54 ± 6.84; p = 0.038). Changes in CXCL8 expression in the control group occurred prior to and after intervention (prior to intervention 2975.93 ± 1484.04; after intervention 1584.85 ± 1659.84; p = 0.041). CXCL1 expression was increased in the immune group and decreased in the control group (immune group 2698.27 (1538.14–5124.70); control group 953.75 (457.85–1534.60); p = 0.032). In both groups, a decrease in superficial neutrophil infiltration was observed, but this was only statistically significant in the immune group. There was no impact of the observed differences between the two groups on surgical outcomes (morbidity, length of stay, readmissions). Conclusions: Immunonutrition in the preoperative period compared with standard nutritional support may influence inflammatory cytokine expression and leukocyte infiltration in patients with colorectal cancer.

Identification and Verification on Prognostic Index of Lower-Grade Glioma Immune-Related LncRNAs

Previous studies have shown that the prognosis of patients with lower-grade glioma (LGG) is closely related to the infiltration of immune cells and the expression of long non-coding RNAs (lncRNAs). In this paper, we applied single-sample gene set enrichment analysis (ssGSEA) algorithm to evaluate the expression level of immune genes from tumor tissues in The Cancer Genome Atlas (TCGA) database, and divided patients into the high immune group and the low immune group, which were separately analyzed for differential expression. Venn analysis was taken to select 36 immune-related lncRNAs. To construct a prognostic model of LGG based on immune-related lncRNAs, we divided patients into a training set and a verification set at a ratio of 2:1. Univariate Cox regression and the Least Absolute Shrinkage and Selection Operator (LASSO) regression were performed to select 11 immune-related lncRNAs associated with the prognosis of LGG, and based on these selected lncRNAs, the risk scoring model was constructed. Through Kaplan-Meier analysis, the overall survival (OS) of patients in the high-risk group was significantly lower than that of the low-risk group. Then, established a nomogram including age, gender, neoplasm histologic grade, and risk score. Meanwhile, the predictive performance of the model was evaluated by calculating the C-index, drawing the calibration chart, the clinical decision curve as well as the Receiver Operating Characteristic (ROC) curve. Similar results were obtained by utilizing the validation data to verify the above consequences. Based on the TIMER database, the correlation analysis showed that the 11 immune-related lncRNAs risk score of LGG were in connection with the infiltration of the subtypes of immune cells. Subsequently, we performed enrichment analysis, whose results showed that these immune-related lncRNAs played important roles in the progress of LGG. In conclusion, these 11 immune-related lncRNAs have the potential to predict the prognosis of patients with LGG, which may play a key role in the development of LGG.

Adolescent Drug Use in Connecticut Private High Schools: Zero Tolerance, Contextual Peer Influence, and Deterrence Effectiveness

After the 1994 Gun-Free School Act, schools expanded the use of zero-tolerance policies with all Connecticut private high schools implementing punitive drug and alcohol policies. Based on the criminological theory of deterrence, zero-tolerance policies deliver severe and certain punishments designed to deter rational actors from engaging in problem behaviors. Existing research suggests that adolescents perceive rewards more strongly around peers and lack impulse control, raising the possibility that peer pressure may override rational deterrence in an adolescents’ decision-making process. An “immune group” of adolescents predisposed to ignore punitive deterrents may play a sizable role in inducing peer drug use. If peer influence supersedes deterrence in a significant number of cases, adolescents who are affected both by deterrents and peer pressure may be at a higher risk of following the example of the “immune group.” This study raises the question of whether Connecticut private high school students’ drug use is correlated with perceptions of punishment mandated by school policy and contextual peer influences. A questionnaire that measured students’ drug use on a scale of 1 to 4, perceived severity and likelihood of punishment from 0 to 10, and interaction with drug using peers from 0 to 10, was completed by 50 respondents. The study found no correlations between student drug use and perception of punishment likelihood and severity but found contextual peer influences to be positively associated with expected student drug use in the future. While the results of this study are limited to Connecticut private high school students, the observed tendency in students to disregard risks and pursue peer-involved drug use may be generalized in adolescents. Even in places where school discipline is not a wide issue, the impact of contextual factors like peer influence must be reconceptualized in thinking about school drug policies.

Lung metastases share common immune features regardless of primary tumor origin

BackgroundOnly certain disseminated cells are able to grow in secondary organs to create a metastatic tumor. Under the hypothesis that the immune microenvironment of the host tissue may play an important role in this process, we have categorized metastatic samples based on their immune features.MethodsGene expression data of metastatic samples (n=374) from four secondary sites (brain, bone, liver and lung) were used to characterize samples based on their immune and stromal infiltration using gene signatures and cell quantification tools. A clustering analysis was done that separated metastatic samples into three different immune categories: high, medium and low.ResultsSignificant differences were found between the immune profiles of samples metastasizing in distinct organs. Metastases in lung showed a higher immunogenic score than metastases in brain, liver or bone, regardless of their primary site of origin. Also, they preferentially clustered in the high immune group. Samples in this cluster exhibited a clear inflammatory phenotype, higher levels of immune infiltrate, overexpression of programmed death-ligand 1 (PD-L1) and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) pathways and upregulation of genes predicting clinical response to programmed cell death protein 1 (PD-1) blockade (T-cell inflammatory signature). A decision tree algorithm was used to select CD74 as a biomarker that identify samples belonging to this high-immune subtype of metastases, having specificity of 0.96 and sensitivity of 1.ConclusionsWe have found a group of lung-enriched metastases showing an inflammatory phenotype susceptible to be treated with immunotherapy.

Immunoscore classifier:A gene based prognostic tool in early non-small cell lung cancer.

e21030 Background: The clinical benefits of immunotherapy in patients with stage I non-small cell lung cancer (NSCLC) is still controversial. Immune status plays critical role in the development and progression of NSCLC, and is associated with the patient survival outcomes. The analysis of immune features is thus valuable for the determination of immunotherapy. However, one single immune feature cannot reflect the complex immune status, and its prognostic value is extremely limited. In this study, we aimed to construct an immunoscore classifier based on multiple immuno-genes to predict the prognosis of patients with early NSCLC. Methods: A total of 522 patients with stage I NSCLC were included in this study. All patients' follow-up records and gene expression data were completely preserved. A least absolute shrinkage and selection operator (LASSO) algorithm was used to screen immune-related genes, and a COX proportional hazard regression model was used to construct the immunoscore classifier based on multiple immune-genes. Besides, the net reclassification improvement (NRI) calculation and concordance index (C-index) were applied to quantify the improvement of usefulness added by the immunoscore classifier compared to TNM staging system. Results: The immunoscore classifier including CCL5, CD8A, CXCL9, HLA-DQA1, LAG3, STAT1, and CD276 was significantly correlated with OS (HR: 2.785 CI: 1.809-4.289 P < 0.001) in patients with stage I NSCLC. With the optimal cut-off value of 4.32, all patients can be divided into a low-risk immune group and a high-risk immune group. The 10-year survival rates of the two groups were 36.8% and 12.3%, respectively. Besides, the immunoscore classifier was superior to the traditional TNM staging system in terms of distinguishing ability (C-index improvement by 0.075) and net reclassification ability (NRI improvement by 11.29%), indicating that the immunoscore classifier plays an important role in improving prognostic value. Conclusions: Multiple immune-genes based immunoscore classifiers can effectively predict the prognosis of patients with stage I NSCLC, and is significantly superior to the traditional TNM staging system in terms of prediction effectiveness and accuracy. As a new assessment tool, the immunoscore classifier may be helpful for determining the immune status of patients with stage I NSCLC and screening patients suitable for subsequent immunotherapy.

Recombinant tandem epitope vaccination provides cross protection against Actinobacillus pleuropneumoniae challenge in mice

Actinobacillus pleuropneumoniae (A. pleuropneumoniae/APP) is the pathogen that causes porcine contagious pleuropneumonia. Actinobacillus pleuropneumoniae is divided into 18 serovars, and the cross protection efficacy of epitopes is debatable, which has resulted in the slow development of a vaccine. Consequently, epitope-based vaccines conferring Actinobacillus pleuropneumoniae cross protection have rarely been reported. In this study, B cell epitopes in the head domain of trimeric autotransporter adhesin were predicted, and 6 epitopes were selected. Then, the predicted epitopes (Ba1, Bb5, C1, PH1 and PH2) were connected by linkers to construct a recombinant tandem antigen (rta) gene. The RTA protein encoded by the recombinant rta gene was expressed, and finally the ICR mice were immunized with the RTA protein with or without inactivated Actinobacillus pleuropneumoniae (serovars 1 and 5b) and challenged with Actinobacillus pleuropneumoniae to evaluate the protective effect of the epitope-based vaccine and combined vaccine.The mice in the RTA-immunized group and RTA plus inactivated Actinobacillus pleuropneumoniae vaccine group had a significant improvement in clinical symptoms and a higher level of antibody in the serum than those in the control group. The RTA immune group had a 40% survival rate after Actinobacillus pleuropneumoniae infection, whereas the combination of RTA and inactivated Actinobacillus pleuropneumoniae produced very strong cross immune protection in mice, at least 50% (RTA IB1+ C5) and at most 100% (RTA IB5 + C1), whereas no cross immunoprotection was found in the solo Actinobacillus pleuropneumoniae immune group. Overall, the combination of the RTA protein and inactivated bacteria significantly enhanced the cross protection effects. This implies that RTA protein in combination with a suitable inactivated Actinobacillus pleuropneumoniae strain could be a candidate vaccine for porcine contagious pleuropneumonia.

Membranoproliferative Glomerulonephritis- A Disease or a Pattern of Injury?

Introduction:Membranoproliferative Glomerulonephritis (MPGN) is a pattern of glomerular injury resulting from wide range of diseases sharing a common pathogenesis and thus, presented with large spectrum of clinical presentation starting from slowly progressive subnephrotic range proteinuria to nephritic syndrome and Rapidly Progressive Glomerulonephritis (RPGN). Aim: The aim of the study was to do reanalysis of the aetiological classification of MPGN and its prognostic outcomes. Materials and Methods: A retrospective, cohort study was conducted including the patients of all age groups with MPGN pattern of injury, from February, 2018 to August, 2018 at a Tertiary Healthcare Centre of West Bengal, India. All specimens were examined by two nephropathologists at the institute using both light microscope and immunofluorescence microscope. Haematoxylin and Eosin (H&E), Periodic Acid-Schiff (PAS), silver methanamine, Masson’s Trichrome (MT) stained smears were prepared for light microscopy. Specimens for immunofluorescence microscopy were stained using Fluorescein Isothiocyanate (FITC) conjugated polyclonal rabbit anti-sera against human IgG, IgM, IgA, C3, C1q, kappa, lambda. Correlations were evaluated using Spearman’s rank correlation. A p<0.05 was considered as statistically significant. GRAPHPAD PRISM 5 was used for statistical analysis. Results: From a total of 47 cases, 41 cases in immunoglobulin associated group, 4 cases in C3 nephropathy group and 2 cases in pauci-immune group were found. Autoimmune diseases had the strongest association followed by infections and dysproteinemia. Severity and chronicity parameters were more prevalent in the immunoglobulin positive group. The pauci-immune group showed maximum vascular involvement, indicating it to arise from a vascular pathology. Conclusion: Membranoproliferative Glomerulonephritis is not a single disease entity instead this pattern of injury could be seen in a variety of diseased condition.

Recent advances in understanding and treating nephrotic syndrome

Idiopathic nephrotic syndrome (INS) is one of the most common glomerular diseases in children and adults, and the central event is podocyte injury. INS is a heterogeneous disease, and treatment is largely empirical and in many cases unsuccessful, and steroids are the initial mainstay of therapy. Close to 70% of children with INS have some response to steroids and are labelled as steroid-‘sensitive’, and the rest as steroid-‘resistant’ (also termed focal segmental glomerulosclerosis), and single-gene mutations underlie a large proportion of the latter group. The burden of morbidity is enormous, both to patients with lifelong chronic disease and to health services, particularly in managing dialysis and transplantation. The target cell of nephrotic syndrome is the glomerular podocyte, and podocyte biology research has exploded over the last 15 years. Major advances in genetic and biological understanding now put clinicians and researchers at the threshold of a major reclassification of the disease and testing of targeted therapies both identified and novel. That potential is based on complete genetic analysis, deep clinical phenotyping, and the introduction of mechanism-derived biomarkers into clinical practice. INS can now be split off into those with a single-gene defect, of which currently at least 53 genes are known to be causative, and the others. Of the others, the majority are likely to be immune-mediated and caused by the presence of a still-unknown circulating factor or factors, and whether there is a third (or more) mechanistic group or groups remains to be discovered. Treatment is therefore now being refined towards separating out the monogenic cases to minimise immunosuppression and further understanding how best to stratify and appropriately direct immunosuppressive treatments within the immune group. Therapies directed specifically towards the target cell, the podocyte, are in their infancy but hold considerable promise for the near future.

Evaluation of pertussis immunity status in schoolchildren immunized with whole-cell vaccine

SUMMARYIt has recently been reported that the worldwide increase in the number of pertussis cases is a result of the waning of whole-cell vaccine-induced immunity. Thus, in this study, we aimed to investigate the pertussis immunity status of primary and secondary school students in a district of Ankara, Turkey. A total of 997 healthy students, aged 9–17 years, who had been immunized with four doses of whole-cell pertussis vaccine were included in the study. The subjects were divided into two age groups: 9–14 and 15–17 years. To determine the immune status, serum levels of IgG anti-pertussis toxin (aPT) antibody were tested by in-house ELISA and arbitrarily evaluated as non-immune [<10 ELISA units (EU)/ml], immune (10–100 EU/ml), and recent infection (>100 EU/ml). Serum samples of 997 students (559 females, 438 males) aged between 9 and 17 years (mean 13·02±2·25, median 13 years) were tested. Non-immune, immune and recent infection levels of aPT were found in 27·3%, 59·3% and 13·4% of individuals, respectively. The immune group did not have statistically significant differences between males and females (P=0·68). In the 9–14 and 15–17 years age groups, serum aPT antibody levels ⩾10 EU/ml were 73·1% and 72·2%, respectively, which did not represent any statistical difference (P=0·81). Students aged 15–17 years had a higher immunity rate than the 9–14 years group, and the percentage of students with recent infection in the 9–14 years group was higher than the 15–17 years group (P<0·001). The peak age of non-immunized subjects was 9 years (47·0%), and decreased to a minimum at age 12–13 years, and began to increase again from age 13–14 years. In contrast, the ratio of recent infection was least at age 9–10 years, began to increase, and reached a peak at 12 years, and then decreased. On the other hand, it was observed that household size and monthly income were not associated with the immunity status (P=0·65,P=0·37, respectively). The results of the present study show that levels of antibody against pertussis decreased in the younger age groups and, as a result, there is an increase in the number of pertussis cases. Thus, in order to decrease the incidence of pertussis and protect infants, we recommend the application of booster doses at regular intervals.

Sudden presentation of immune-mediated inner ear disease: characterization and acceptance of a cochleovestibular dysfunction

Since the McCabe report, growing indirect evidence has accumulated to indicate the implication of immune mechanisms in the pathogenesis of immune-mediated inner-ear disease (IMIED). A clinical study of a group of patients affected by this condition was performed in order to characterize the immune group, based on a recently reported profile, and compared with the vascular, viral and idiopathic aetiologies of sudden deafness. Patients affected by immune-mediated inner-ear disease had the best and the earliest recovery rate of hearing (p = 0.0028 and p = 0.017, respectively). However, this group of patients also had the higher rate of recurrence (p = 0.034), supporting the typical clinical course of the autoimmune disorders. On the basis of the results the criteria used in the diagnosis of the sudden presentation of the immune-mediated inner ear disease could be accepted leading to the characterization of this condition. Likewise, the role of the supporting cells in the pathogenesis of the IMIED is discussed.