Reduction In Immune Response Research Articles

Prevalence of Passalurus ambiguus in domestic rabbits (Oryctolagus cuniculus) in Karbala province, Iraq

Commercial rabbit farming is primarily focused on the production of rabbit meat, although the presence of gastrointestinal disorders can impede their productivity. Endoparasites significantly contribute to the development of several diseases in rabbits, leading to higher rates of illness and death. Passalurus ambiguus is a widely distributed nematode that commonly infects the intestines of rabbits and hares. Parasitic disease can have a substantial economic impact on the industry by reducing the commercial value of rabbit meat and skins, impeding growth and development, and leading to mortality. This study aims to examine the frequency of gastrointestinal parasites in domestic rabbits that are bred in Karbala province, Iraq. Throughout the trial, fecal samples were collected from a randomly selected group of 48 live rabbits. The feces are collected directly from the anus of each living animal. The samples were tested and the presence of eggs and nematodes was detected in the infected samples. Analysis of samples was carried out at the Parasitology Laboratory, college of Veterinary Medicine, Kerbala university, using the flotation technique. Feces from 48 rabbits (ages 1-2) years. The feces are taken out of each animal directly from the anus. Upon testing, it was found that 16 out of 48 fecal samples, accounting for a prevalence of 33.33 %, were infected with at least one nematode. Chi-square (χ2) statistics. Age and genders are not statistically significant at P values ≥ 0.05. This study identified the presence of Passalurus ambiguus in rabbits of varying ages residing in Karbala region, Iraq, including both male and female domestic rabbits. Determining the age and gender of rabbits affected by passalurosis would enable the management of the infection level in these animals. Regular use of a consistent and dependable diagnostic method is essential in industrial rabbit farms to detect and monitor gastrointestinal parasite illnesses. Gastrointestinal parasites continue to be significant health issues that lead to the mortality of rabbit litters in production. Stress and a reduction in immune response may be crucial factors driving this phenomenon.

A polyurethane-based hydrophilic elastomer with multi-biological functions for small-diameter vascular grafts

Thrombosis and intimal hyperplasia (IH) are two major problems faced by the small-diameter vascular grafts. Mimicking the native endothelium and physiological elasticity of blood vessels is considered an ideal strategy. Polyurethane (PU) is suitable for vascular grafts in mechanics because of its molecular designability and elasticity; however, it generally lacks the endothelium-like biofunctions and hydrophilicity. To solve this contradiction, a hydrophilic PU elastomer is developed by crosslinking the hydrophobic hard-segment chains containing diselenide with diaminopyrimidine-capped polyethylene glycol (PEG). In this network, the hydrophobic aggregation occurs underwater due to the uninterrupted hard-segment chains, leading to a significant self-enhancement in mechanics, which can be tailored to the elasticity similar to natural vessels by adjusting the crosslinking density. A series of in vitro studies confirm that the hydrophilicity of PEG and biological activities of aminopyrimidine and diselenide give the PU multi-biological functions similar to the native endothelium, including stable catalytic release of nitric oxide (NO) in the physiological level; anti-adhesion and anti-activation of platelets; inhibition of migration, adhesion, and proliferation of smooth muscle cells (SMCs); and antibacterial effect. In vivo studies further prove the good histocompatibility with both significant reduction in immune response and calcium deposition. Statement of significanceConstructing small-diameter vascular grafts similar to the natural vessels is considered an ideal method to solve the restenosis caused by thrombosis and intimal hyperplasia (IH). Because of the long-term stability, bulk modification is more suitable for implanted materials, however, how to achieve the biofunctions, hydrophilicity, and elasticity simultaneously is still a big challenge. In this work, a kind of polyurethane-based elastomer has been designed and prepared by crosslinking the functional long hard-segment chains with PEG soft segments. The underwater elasticity based on hydration-induced stiffening and the multi-biological functions similar to the native endothelium are compatible with natural vessels. Both in vitro and in vivo experiments demonstrate the potential of this PU as small-diameter vascular grafts.

Delivery of Therapeutic Biopolymers Employing Silica-Based Nanosystems

The use of nanoparticles is crucial for the development of a new generation of nanodevices for clinical applications. Silica-based nanoparticles can be tailored with a wide range of functional biopolymers with unique physicochemical properties thus providing several advantages: (1) limitation of interparticle interaction, (2) preservation of cargo and particle integrity, (3) reduction of immune response, (4) additional therapeutic effects and (5) cell targeting. Therefore, the engineering of advanced functional coatings is of utmost importance to enhance the biocompatibility of existing biomaterials. Herein we will focus on the most recent advances reported on the delivery and therapeutic use of silica-based nanoparticles containing biopolymers (proteins, nucleotides, and polysaccharides) with proven biological effects.

MO888: Long-Term Humoral Immune Response Following Full SARS-COV-2 MRNA Vaccination and Booster Dose in Haemodialysis Patients

Abstract BACKGROUND AND AIMS COVID-19 is associated with an increased mortality in maintenance haemodialysis patients. Considering the baseline immunosuppressed status of this population, humoral immune responses to SARS-CoV-2 vaccination remain to be studied. There is no information regarding sustained immune response in this population after a third booster dose following complete m-RNA vaccination. The aim of our study was to describe the humoral immune response in haemodialysis patients following a complete SARS-CoV-2 vaccine schedule and a third booster dose, and explore the factors associated with a sustained immune response. METHOD We conducted a retrospective study in which we included haemodialysis patients with or without (naïve) previous COVID-19 infection. Patients received the following vaccination schedule: two initial doses of m-RNA vaccine with a 4-week interval (complete immunization) followed by a third booster dose at least 4 months after the second initial dose. IgG anti-SARS-CoV-2 spike antibody titles were measured before the first initial vaccine dose and then monthly up to 3 months after the booster dose. We excluded patients without baseline serological samples and those who did not receive the full vaccination schedule. RESULTS The study included 182 haemodialysis patients with a mean age of 68.5 ± 14.5 years, 63.2% were males and 14.3% were under immunosuppression, 67.7% were responders to HBV vaccine and 18.1% had been infected with SARS-CoV-2 prior to vaccination. After the first two vaccine doses, 96.5% developed immediate humoral immune response, and 91.5% remained with positive anti-SARS-CoV-2 spike antibodies after 4 months from complete vaccination. Median antiSARS-CoV-2 spike antibody titles were significantly higher in patients with previous COVID-19 infection and HBV vaccine responders, both immediately (40 000 AU/mL versus 2926 AU/mL, P < 0.001 and 4885 versus 2056 AU/mL, P = 0.003, respectively) and after 4 months from complete vaccination (27 741 versus 663 AU/mL, P < 0.001 and 1356 versus 341 AU/mL, P = 0.001). In the 4 months between complete initial vaccination and the third booster dose, the mean monthly decrease in antiSARS-CoV-2 spike antibody titles was of 31.8% (±18.9) and was significantly lower in patients with prior COVID-19 infection compared with naïve patients (17.2% versus 34.8%, P < 0.001). After the third booster dose, 98.2% of patients showed positive anti-SARS-CoV-2 S antibodies, and this proportion remained stable in the following 3 months. Nevertheless, median anti-SARS-CoV-2 spike antibody titles remained higher in patients with prior COVID-19 both immediately and after 3 months. However, we observed a more sustained humoral response after the booster dose, with a lower mean monthly decrease in antibody titles compared with the initial vaccine schedule (31.8% to 22.6%, P < 0.001). This finding was reciprocated in all groups, regardless of prior serological COVID-19 status, HBV vaccine response, age or sex. Multivariate logistic regression for the risk of a >25% monthly decrease in antibodies showed that prior infection with COVID-19 was a protective factor both after the complete initial vaccination {OR: 0.23, [95% confidence interval (95% CI) 0.06–0.87]} as well as after the third dose (OR: 0.23, 95% CI 0.06–0.81). CONCLUSION Vaccination with m-RNA anti-SARS-CoV-2 is effective in eliciting an immediate humoral response in haemodialysis patients, with a progressive reduction in immune response after 4 months particularly in COVID-19 naive patients. A third booster dose enhances the immune response with significantly higher antibody levels and more sustained humoral immune after 3 months in haemodialysis patients.

Synthesis of Novel MicroRNA-30c Analogs to Reduce Apolipoprotein B Secretion in Human Hepatoma Cells.

Atherosclerosis, a condition characterized by thickening of the arteries due to lipid deposition, is the major contributor to and hallmark of cardiovascular disease. Although great progress has been made in lowering the lipid plaques in patients, the conventional therapies fail to address the needs of those that are intolerant or non-responsive to the treatment. Therefore, additional novel therapeutic approaches are warranted. We have previously shown that increasing the cellular amounts of microRNA-30c (miR-30c) with the aid of viral vectors or liposomes can successfully reduce plasma cholesterol and atherosclerosis in mice. To avoid the use of viruses and liposomes, we have developed new methods to synthesize novel miR-30c analogs with increasing potency and efficacy, including 2'-O-methyl (2'OMe), 2'-fluoro (2'F), pseudouridine (ᴪ), phosphorothioate (PS), and N-acetylgalactosamine (GalNAc). The discovery of these modifications has profoundly impacted the modern RNA therapeutics, as evidenced by their increased nuclease stability and reduction in immune responses. We show that modifications on the passenger strand of miR-30c not only stabilize the duplex but also aid in a more readily uptake by the cells without the aid of viral vectors or lipid emulsions. After uptake, the analogs with PS linkages and GalNAc-modified ribonucleotides significantly reduce the secretion of apolipoprotein B (ApoB) without affecting apolipoprotein A1 (ApoA1) in human hepatoma Huh-7 cells. We envision an enormous potential for these modified miR-30c analogs in therapeutic intervention for treating cardiovascular diseases. This protocol was validated in: J Biol Chem (2021), DOI: 10.1016/j.jbc.2022.101813.

The Cross-Talk between Myeloid and Mesenchymal Stem Cells of Human Bone Marrow Represents a Biomarker of Aging That Regulates Immune Response and Bone Reabsorption

One of the mechanisms that characterizes the aging process of different organs is the accumulation of fat. Different authors have demonstrated that adipose tissue replaces the loss of other cell types, deriving from mesenchymal cells. During aging, there is substitution or trans-differentiation of mesenchymal cells with other cells having the same embryological origin. Newly formed adipocytes were also observed in the trabecular matrix of elderly people’s bones, associated with myeloid cells. In this study, we have investigated the relationship between immature myeloid-derived suppressor cells (I-MDSCs) and mesenchymal stem cells (MSCs) in bone marrow (BM) samples harvested from 57 patients subjected to different orthopedic surgeries. Patients aged from 18 to 92 years were considered in order to compare the cellular composition of bone marrow of young and elderly people, considered a biomarker of immunity, inflammation, and bone preservation. The I-MDSC percentage was stable during aging, but in elderly people, it was possible to observe a strong basal immunosuppression of autologous and heterologous T cells’ proliferation. We hypothesized that this pattern observed in elders depends on the progressive accumulation in the BM of activating stimuli, including cell–cell contact, or the production of different cytokines and proteins that induce the differentiation of bone marrow mesenchymal stem cells in adipocytes. The collected data provided underline the importance of specific biomarkers of aging that promote a reduction in immune response and incremented inflammatory pathways, leading to bone reabsorption in elderly people.

Phospholipase A2 enzymes represent a shared pathogenic pathway in psoriasis and pityriasis rubra pilaris.

Altered epidermal differentiation along with increased keratinocyte proliferation is a characteristic feature of psoriasis and pityriasis rubra pilaris (PRP). However, despite this large degree of overlapping clinical and histologic features, the molecular signatures these skin disorders share are unknown. Using global transcriptomic profiling, we demonstrate that plaque psoriasis and PRP skin lesions have high overlap, with all differentially expressed genes in PRP relative to normal skin having complete overlap with those in psoriasis. The major common pathway shared between psoriasis and PRP involves the phospholipases PLA2G2F, PLA2G4D, and PLA2G4E, which were found to be primarily expressed in the epidermis. Gene silencing each of the 3 PLA2s led to reduction in immune responses and epidermal thickness both in vitro and in vivo in a mouse model of psoriasis, establishing their proinflammatory roles. Lipidomic analyses demonstrated that PLA2s affect mobilization of a phospholipid-eicosanoid pool, which is altered in psoriatic lesions and functions to promote immune responses in keratinocytes. Taken together, our results highlight the important role of PLA2s as regulators of epidermal barrier homeostasis and inflammation, identify PLA2s as a shared pathogenic mechanism between PRP and psoriasis, and as potential therapeutic targets for both diseases.

HMGB1-like dorsal switch protein 1 of the mealworm, Tenebrio molitor, acts as a damage-associated molecular pattern.

High-mobility group box 1 (HMGB1) is a nuclear protein highly conserved in eukaryotes and ubiquitously expressed to regulate transcription and chromatin remodeling. Dorsal switch protein 1 (DSP1) is its insect homolog. A lepidopteran DSP1 acts as a damage-associated molecular pattern (DAMP) in response to immune challenge. The objective of this study was to determine the role of DAMP in the mealworm beetle, Tenebrio molitor, a coleopteran insect. DSP1 of T. molitor (Tm-DSP1) encodes 536 amino acids and shares sequence similarities with Homo sapiens HMGB1 (56.3%) and Spodoptera exigua DSP1 (59.2%). An antisera raised against S. exigua DSP1 was cross-reactive to Tm-DSP1. Like other insect DSPs, Tm-DSP1 has a relatively long N-terminal extension in addition to two conserved HMG box domains. It was expressed in all developmental stages of T. molitor and different larval tissues. Upon immune challenge, its expression level was upregulated. Its RNA interference (RNAi) treatment resulted in a significant reduction in immune responses measured by hemocyte nodule formation against bacterial infection. In addition, the induction of some antimicrobial peptide genes to the immune challenge was suppressed by its RNAi treatment. Interestingly, phospholipase A2 associated with eicosanoid biosynthesis was significantly suppressed in its catalytic activity by the RNAi treatment specific to Tm-DSP1 expression. Without any pathogen infection, injection of a lepidopteran DSP1 induced both cellular and humoral immune responses. These results suggest that Tm-DSP1 in T. molitor can act as a DAMP molecule and mediate immune responses upon immune challenge.

Diagnosis of Myxomatous Mitral Valve Disease (MMVD) in Aged Dogs

Background: Aging is a biological process, which affects many body systems and increases susceptibility to disease that is associated with a reduction in immune response, a decline in cognitive function and a diminished functional reserve of the cardiovascular, pulmonary and renal systems. Diseases of heart or cardiovascular system represents a major component of geriatric medicine among which, myxomatous mitral valve disease is more common among geriatric dogs. The present study is keeping on record about the diagnosis of myxomatous mitral valve disease in geriatric dogs in and around Hyderabad, Telangana state.Methods: A total of1695 geriatric dogs presented to various clinics including university veterinary clinical complex of Hyderabad, Telangana state during 2015 to 2020 were considered for this study. After confirming that they were free from any other systemic disease, medium to large size breed dogs above 8 years with the history of exercise intolerance, dyspnoea and cough were selected. Diagnostic protocol included, auscultation, chest x-ray and 2d-echocardiography to confirm mitral valve degeneration. Result: Myxomatous mitral valve disease was diagnosed in 427/695 geriatric dogs (61.43%) and were showing hyperechoeic thick mitral valve leaflets with kinking and prolapse. Further, echocardiography also revealed dilated left atrium and ventricle, hyperkinesis of interventricular septum, mitral valve degeneration and rupture of chordae tendineae with valuvular regurgitation.

Effective Prediction of Prostate Cancer Recurrence through the IQGAP1 Network.

Simple SummaryProstate cancer (PC) is a leading cause of cancer death in men in the developed countries. PC mortality is related to tumor relapse following curative therapy to primary tumors. It is thus essential to effectively assess the recurrence risk for personalized patient management. However, the current assessment capacity remains insufficient. We examined tumors produced in animals and samples derived from more than 1100 patients. A unique set of gene expression was observed following PC progression and a multigene panel consisting of 27 genes (Sig27gene) was constructed. Sig27gene is novel and robustly predicts PC relapse in two independent patient populations (n = 492 and n = 140) at p < 2 × 10−16. Sig27gene remains an independent risk factor of PC recurrence after adjusting for multiple clinical features. The novel and robust nature of Sig27gene support its translational potential to evaluate the risk of PC relapse in patients with primary PC.IQGAP1 expression was analyzed in: (1) primary prostate cancer, (2) xenografts produced from LNCaP, DU145, and PC3 cells, (3) tumor of PTEN−/− and TRAMP mice, and (4) castration resistant PC (CRPC) produced by LNCaP xenografts and PTEN−/− mice. IQGAP1 downregulations occurred in CRPC and advanced PCs. The downregulations were associated with rapid PC recurrence in the TCGA PanCancer (n = 492, p = 0.01) and MSKCC (n = 140, p = 4 × 10−6) cohorts. Differentially expressed genes (n = 598) relative to IQGAP1 downregulation were identified with enrichment in chemotaxis, cytokine signaling, and others along with reductions in immune responses. A novel 27-gene signature (Sig27gene) was constructed from these DEGs through random division of the TCGA cohort into a Training and Testing population. The panel was validated using an independent MSKCC cohort. Sig27gene robustly predicts PC recurrence at (hazard ratio) HR 2.72 and p < 2 × 10−16 in two independent PC cohorts. The prediction remains significant after adjusting for multiple clinical features. The novel and robust nature of Sig27gene underlie its great translational potential as a prognostic biomarker to predict PC relapse risk in patients with primary PC.

Immunodepressant and oxidant potential of standard leukaemia drug regimen

Chemotherapy improves cancer status but also could carry a risk of harmful side effects. In leukaemia patients, oxidative status and immune system are crucial parameters for patient improvement. The present study is aimed to assess the oxidative stress and the levels of immunoglobulin, including; IgG, IgA, and IgM in acute myelocytic leukaemias (AML) patients. A total of twenty-one patients with AML were enrolled in this study, alongside 24 healthy individuals as a control group. Initially, in both groups, the serum MDA, IgA, IgG, and IgM levels were measured. Then after a month of treatment with standard leukaemia therapy; the measurement of levels of these parameters were repeated for both the patients’ group and the control group for measuring immunoglobulin levels while analysis of MDA was done by the laboratory method. A highly significant rise in the serum MDA level was detected (P&amp;lt;0.001) in patients with AML after treatment with specific cytotoxic drugs was observed in comparison with the control. Moreover, there was a significant modulation in the serum concentration of Ig after starting cytotoxic drugs in comparison with the controls. The study revealed that AML was associated with elevated oxidative stress parameters following treatment with cytotoxic drugs as reflected by the rise in the serum levels of MDA. The decreased serum Ig levels indicate a reduction in immune response in patients with AML due to cytotoxic drugs.

Association of CD46 Cellular Receptor Gene SNP in Measles Vaccine Response

Background: Measles is a highly contagious viral disease. It remains an important cause of death among young children globally, despite the availability of a safe and effective vaccine. Measles transmitted via droplets from the nose, mouth or throat of infected persons. Objective: To evaluate the immune response to measles infection among immunized school aged children, and to detect the cluster of differentiation 46 single nucleotide polymorphism (CD46 SNP) in association with the immune response. Methods: The current study is a cross sectional study including 158 hospitalized patients were previously vaccinated with two doses of measles-mumps-rubella vaccine. The ages of patients were 5-10 years school aged children. All samples were collected from blood collection unit in Al-Imamein Al-Kadhimein Medical city during the period from December 2018 to April 2019. The detection was based on the presence of IgG Antibody to measles. The positive results were considered according to enzyme linked immunosorbant assay and conventional polymerase chain reaction to detect CD46 SNP. Results: Among those 158 subjects' male children count for 62% and female children were 38%. Forty-one (41) immunized children (with mean age 7.98±1.92 years) were low immune response to measles vaccine (MV). On the other hand, only two cases (with mean age 5.50±0.71 years) were negative to measles virus vaccine. Conclusion: Ensuring two doses of MV give protective immune response to MV may declines with aging. CD46 cellular receptor gene SNP (rs7144) may play role in reduction in immune response to MV. Keywords: Measles, MMR, CD46, SNP, Iraq Citation: Al-Gburi SAJ, Kadhim HS, Ghazi HF. Association of CD46 Cellular Receptor Gene SNP in measles vaccine response. Iraqi JMS. 2020; 18(1): 39-46. doi: 10.22578/IJMS.18.1.6

Abstract A31: High aneuploidy levels are linked to a reduced immune-cell abundance in metastatic castrate-resistant prostate cancer

Abstract Single-agent immunotherapeutic approaches are promising for treating castrate-resistant prostate cancer (CRPC) patients; however, to date no significant improvement in survival has been reported. Mixed response to therapy occurs because metastatic lesions are highly heterogeneous. These lesions exhibit distinct genomic changes that are linked to a tumor microenvironment that is often poorly populated by immune cells. In other tumor types, a high percent genome altered (e.g., aneuploidy) associates with an overall reduction in immune response in the primary tumor microenvironment. However, the association between genomic changes and immune cell composition is poorly understood in metastatic prostate cancers. Thus, we quantified the abundance of 22 immune cell types in 330 tumors from two independent public domain CRPC cohorts and 491 primary prostate tumors from TCGA using the Absolute Mode from CIBERSORT. The estimation in CRPC was performed using whole-transcriptome data from West Coast Dream Team (Abida et al., 2019; n=212) and East Coast Dream Team (Robinson et al., 2015; n=117) samples. We did not identify any significant correlations between mutational load and immune cell abundance in the three investigated cohorts. In primary tumors, aneuploidy was weakly negatively correlated with CD8+ and CD4+ T-cell abundance (R=-0.14 and R=-0.09, P&amp;lt;0.05). Overall, metastatic lesions demonstrated a low absolute abundance of the majority of immune cell subtypes, with higher levels of CD8+ T-cells, memory CD4+ T-cells, macrophages, and plasma cells. We found that bone lesions had the lowest abundance of CD8+ T-cells (Kruskal-Wallis, p=0.05) and the highest abundance of macrophages (P&amp;lt;0.0001) compared to lymph node and liver samples. Overall, aneuploidy was negatively correlated with immune cell abundance in both metastatic cohorts (Spearman correlation, P&amp;lt;0.05). Interestingly, lymph node metastases showed the strongest negative correlation between aneuploidy and T-cell abundance in both West and East cohorts (R=-0.36 and R=-0.32, respectively, p&amp;lt;0.01). Our preliminary results show that the extent of chromosomal aberrations in CRPC is significantly associated with underlying immune cell infiltrates. These results have potential implications for future immunotherapy trials in prostate cancer. Citation Format: Thiago Vidotto, Daniela C. Salles, Tamara L. Lotan. High aneuploidy levels are linked to a reduced immune-cell abundance in metastatic castrate-resistant prostate cancer [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2019 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(3 Suppl):Abstract nr A31.

Abstract 2527: Genomic aberrations of ALK in head and neck squamous cell carcinoma

Abstract Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase encoded by the gene ALK, which belongs to a subfamily of the insulin receptor superfamily. ALK signaling has been shown to be involved in cell proliferation, differentiation, and development, etc. Thus far, ALK aberrations are known to be involved in the oncogenesis of non-small cell lung cancer and glioblastoma. Head and neck cancer squamous cell carcinoma (HNSCC) is an aggressive cancer with genomic heterogeneity as revealed by recent whole-exome studies. Though ALK aberrations are found in a notable subset of HNSCC, their biological importance is largely unknown. Comprehensive pan-cancer analysis of 33 cancer types (TCGA, Pan-cancer, Provisional, N=11617, www.cbioportal.org) showed that ALK aberrations (mutations, amplification and gene copy gain) are commonly noted in 27 cancers, with HNSCC ranking 9th having ~22% cases of primary tumors harboring these ALK genomic aberrations. Within the US TCGA HNSCC dataset (Provisional; N=510), 3.5% of HNSCC patient tumors harbor somatic ALK mutations, 0.4% cases with ALK amplification, and 17.4% cases with ALK gain. Further, RNA-seq analysis revealed tumor specific ALK mRNA upregulation in 43 pairs of normal-tumor tissues (~2-fold, P=0.001***), and 457 HNSCC tumors (~1.5-fold, P=0.039*). There are currently 172261 single-nucleotide polymorphism (SNP) on ALK exons in human reported in the dbSNP database. We noted 18.9% of Asian HNSCC patients (N=53) bearing ALK germline variants (N=10) with unknown clinical significance. Interestingly, ALK aberrations (mutations, amplification, gene copy gain) are noted to be significantly associated with TP53 mutations (P&amp;lt;0.0001), HPV-negativity (P=0.0094) and the male gender (P=0.0041; Fisher’s Exact test), but not associated with HNSCC patient survival (P=0.734). Further, patients with ALK mutation and amplification are largely advanced cases (93.33%, Stage III and Stage IV). Gene Set Enrichment Analysis (GSEA) of mRNA expression shows that ALK-altered HNSCC tumors (vs unaltered tumors) appear to have significantly downregulation of immune response gene set [NES (normalized enrichment score)=-6.773, P=0.0026] and inflammatory response gene set (NES=-5.54, P= 0.0022), while nervous system development and cell development signaling gene sets are significantly upregulated (P=0.0015, 0.0016, respectively). Proteomic analysis (TCPA, N=209) showed that ALK-altered HNSCC (vs. -unaltered tumors) have increased PIK3CA and CHEK2 protein expression (P&amp;lt;0.001, 0.0001, respectively), as well as reduced PRAS40(pT246) and VEGFR2 protein expression (P&amp;lt;0.001, 0.001, respectively). In conclusion, ALK aberrations may contribute to HNSCC aggressiveness, potentially via reduction of immune response and activation of PI3K signaling pathway. Future investigations studying the biological effects of ALK aberrations on HNSCC oncogenesis are warranted. Citation Format: Lan Wang, Yuchen Liu, Wenying Piao, Peony Hiu Yan Poon, Chun Kit Yeung, Amy Bik Wan Chan, Chin Wang Lau, Yuxiong Su, Jason Ying Kuen Chan, Vivian Wai Yan Lui. Genomic aberrations of ALK in head and neck squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2527.

Oxidative stress and immunosenescence

Immunosenescence is a process characterized by alterations and progressive deterioration of the structure and function of the immune system.Manifestations of immunosenescence include damage to both innate and adaptive immunity, reduction of immune responses to antigen stimulation, quantitative and functional changes of immune cells and alterations of cytokine activities, all of which may result in attenuated responses to vaccines, enhanced chronic inflammatory reactions, an elevated risk of infections and an increased susceptibility to cancer among old individuals.In recent years, it has been increasingly recognized that oxidative stress plays a vital role in immunosenescence.This review is focused on the influences of oxidative stress on the structure, function and generation of immunecells and the major mechanisms underlying immunosenescence. Key words: Aging; Immunity; Oxidative stress

Immune function and the decision to deploy weapons during fights in the beadlet anemone, Actinia equina.

The ability to mitigate the costs of engaging in a fight will depend on an individual's physiological state. However, the experience of fighting itself may, in turn, affect an individual's state, especially if the fight results in injury. Previous studies have found a correlation between immune state and fighting success, but the causal direction of this relationship remains unclear. Does immune state determine fighting success? Or does fighting itself influence subsequent immune state? Using the beadlet anemone, Actinia equina, we disentangled the cause and effect of this relationship, measuring immune response once pre-fight and twice post-fight. Contrary to previous findings, pre-fight immune response did not predict fighting success, but rather predicted whether an individual used its weapons during the fight. Furthermore, weapon use and contest outcome significantly affected post-fight immune response. Individuals that used their weapons maintained a stable immune response following the fight, whereas those that fought non-injuriously did not. Furthermore, although winners suffered a reduction in immune response similar to that of losers immediately post-fight, winners began to recover pre-fight levels within 24 h. Our findings indicate that immune state can influence strategic fighting decisions and, moreover, that fight outcome and the agonistic behaviours expressed can significantly affect subsequent immunity.

Survival and immune response of the Chagas vector Meccus pallidipennis (Hemiptera: Reduviidae) against two entomopathogenic fungi, Metarhizium anisopliae and Isaria fumosorosea.

BackgroundChagas disease is a key health problem in Latin America and is caused and transmitted by Trypanosoma cruzi and triatomine bugs, respectively. Control of triatomines has largely relied on the use pyrethroids, which has proved to be ineffective in the long term. Alternatively, the use of entomopathogenic fungi has been implemented to control triatomine bugs. These fungi are highly efficient as they induce a reduction in immune response on insects. Meccus pallidipennis is the main triatomine vector of Chagas disease in Mexico. In this work we investigated the effects of two entomopathogenic fungi, Metarhizium anisopliae and Isaria fumosorosea, on M. pallidipennis nymphs in terms of insect survival and immune response.MethodsWe had an infected and a control group for each fungal species and assessed: a) insect survival during 30 days; and, b) phenoloxidase (PO) and prophenoloxidase (proPO; two key traits in insect immune response) at 24, 48, 96 and 144 h. For survival we used Kaplan-Meier survival analysis while for immune response we used factorial, repeated-measures ANOVA for each fungal species.ResultsAnimals treated with M. anisopliae died sooner than animals treated with I. fumosorosea. Infected animals showed lower PO and proPO values than sham individuals, with a clear decrease in these parameters at 24 h with no further changes after this time.ConclusionsOur study widens the possibility of entomopathogenic fungi being used for triatomine control. The negative effect on PO and proPO seems mediated by a down-regulation of the triatomine immune response.

The metabolic cost of mounting an immune response in male brown anoles (Anolis sagrei).

The tradeoff between reproduction and survival is central to life-history theory and is thought to reflect underlying energetic tradeoffs between reproduction and self-maintenance. Immune responses to parasites and pathogens are important components of self-maintenance in many species, but whether these defenses impose significant energetic costs has only been tested in a handful of organisms. We tested for a metabolic cost of mounting an immune response in the male brown anole (Anolis sagrei), a lizard in which we have previously shown that reproduction causes a marked reduction in immune response to the novel antigen phytohaemagglutinin (PHA). We treated captive male anoles with a subcutaneous injection of either PHA, which induces an immune response that manifests as localized swelling, or saline vehicle as a control. Prior to injection and at 24, 48, and 72 hr post-injection, we measured swelling at the site of injection and whole-animal resting metabolic rate (RMR) using stop-flow respirometry. Although we detected a robust swelling response to PHA at 24, 48, and 72 hr post-injection, mean RMR did not differ between treatments at any of these time points. However, within the PHA treatment group, RMR increased with the extent of swelling, suggesting a variable metabolic cost that scales with the magnitude of the induced immune response. Although individual anoles varied considerably in the extent to which they responded to PHA challenge, our results suggest that an immune response can impose a substantial metabolic cost (potentially as much as 63% above baseline RMR) for individuals that do respond maximally. J. Exp. Zool. 323A: 689-695, 2015. © 2015 Wiley Periodicals, Inc.

Single Molecule Förster Resonance Energy Transfer Studies of the Effect of Deglycosylation on the Structure of Immunoglobulin G

The deglycosylation of immunoglobulin G (IgG) antibodies with the bacterial enzyme EndoS has been suggested as a potential treatment for some autoimmune disorders as this process leads to a diminished immune response. The reduction in immune response is thought to arise from weakened binding of effector molecules to the fragment crystallizable (Fc) region of IgG antibodies as a result of a conformational change in the Fc region. The nature of this structural alteration is uncertain due to conflicting results obtained from x-ray crystallography and small-angle x-ray scattering studies. To further examine the impact of deglycosylation on the structure of the Fc region, we have examined both glycosylated and EndoS deglycosylated IgG antibodies using single molecule Forster Resonance Energy Transfer (smFRET). The FRET efficiency histograms obtained from studying freely-diffusing, dye-labeled antibodies suggest that the flexibility of the Fc region increases upon deglycosylation.

Cryopreservation of dental tissue and subsequent isolation of mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are characterized as plate-adherent cells that demonstrate a fibroblast-like growth pattern, express specific cell surface markers (CD13, CD29, CD44, CD73, CD90, CD105, and CD166), and differentiate into mesenchymal lineage cells in vitro (osteocytes, chonrocytes, and adipocytes)1. They were first isolated from bone marrow, but have since been isolated from various tissues, including skin, fat, cartilage, blood, umbilical cord, and dental tissue1,2,3. Recently, dental tissue has emerged as a potential source of autologous MSCs that can be used for tissue regeneration via tissue engineering. Precursor cells in dental tissue were first isolated and characterized from dental pulp, after which cells showing the characteristics of MSCs were isolated from immature wisdom teeth, periodontal ligament, and exfoliated deciduous teeth3,4,5,6. In particular, the dental pulp, follicle, and root apical papilla of immature wisdom teeth have been reported to possess abundant undifferentiated primitive cells2,3,6. Interestingly, stem cells from different dental tissues show similar MSC characteristics, but varying in vitro and in vivo differentiation properties2,3,6. Stem cells from the dental follicle of extracted wisdom teeth exhibit superior osteogenic differentiation potential compared to those from the dental pulp or root apical papilla3,6. In addition, one of the advantages of using dental stem cells from extracted wisdom teeth is that they can be obtained from patients at a relatively late age-from the late teens to mid-twenties. Moreover, recently, a new method for long-term cryopreservation of dental tissue from extracted wisdom teeth has been developed, and its efficacy was confirmed by the >70% cell survival rate of long-term preserved dental tissue7. MSCs have potential clinical applications in the treatment of autoimmune diseases or reduction of immune responses after allotransplantation, as well as in tissue regeneration8. Many researchers have shown that MSCs exert immune suppression and anti-inflammatory activity by dominantly inhibiting T-helper cells while activating regulatory T-cells and suppressing B-lymphocytes8,9. This immunomodulatory effect of MSCs makes them suitable for treatment of severe autoimmune diseases and graft-versus-host disease8. However, most trials for the clinical application of MSCs have used bone marrow-derived MSCs (BMSCs). As mentioned previously, MSCs from dental tissue (DMSCs) exhibit similar characteristics to BMSCs, but have superior osteogenic differentiation potential3,6. Therefore, DMSCs could possibly replace BMSCs in cell therapy and tissue regeneration applications. The new cryopreservation method for dental tissue enables the use of autologous MSCs from preserved dental tissue of extracted wisdom teeth7. These autologous MSCs could reduce unexpected side effects during clinical use, while maintaining similar immunomodulatory efficacy. In conclusion, dental tissue from immature wisdom teeth, including that from the dental follicle, pulp, and root apical papilla, possesses abundant multipotent stem cells that could be useful in both cell therapy and tissue regeneration. Traditionally, these dental tissues have been discarded after extraction. In the future, dental tissue obtained via extraction of impacted teeth should be cryopreserved for use as an autologous stem cell source. The long-term preservation of dental tissue after tooth extraction could thus create a new business model in dentistry, as well as provide greater access to autologous MSCs for patients in need.