Immune System Genes Research Articles

Autoimmunity and immunodeficiency associated with monoallelic LIG4 mutations via haploinsufficiency

Biallelic mutations in LIG4 encoding DNA-ligase 4 cause a rare immunodeficiency syndrome manifesting as infant-onset life-threatening and/or opportunistic infections, skeletal malformations, radiosensitivity and neoplasia. LIG4 is pivotal during DNA repair and during V(D)J recombination as it performs the final DNA-break sealing step. This study explored whether monoallelic LIG4 missense mutations may underlie immunodeficiency and autoimmunity with autosomal dominant inheritance. Extensive flow-cytometric immune-phenotyping was performed. Rare variants of immune system genes were analyzed by whole exome sequencing. DNA repair functionality and T-cell-intrinsic DNA damage tolerance was tested with an ensemble of invitro and in silico tools. Antigen-receptor diversity and autoimmune features were characterized by high-throughput sequencing and autoantibody arrays. Reconstitution of wild-type versus mutant LIG4 were performed in LIG4 knockout Jurkat T cells, and DNA damage tolerance was subsequently assessed. A novel heterozygous LIG4 loss-of-function mutation (p.R580Q), associated with a dominantly inherited familial immune-dysregulation consisting of autoimmune cytopenias, and in the index patient with lymphoproliferation, agammaglobulinemia, and adaptive immune cell infiltration into nonlymphoid organs. Immunophenotyping revealed reduced naive CD4+ T cells and low TCR-Vα7.2+ T cells, while T-/B-cell receptor repertoires showed only mild alterations. Cohort screening identified 2 other nonrelated patients with the monoallelic LIG4 mutation p.A842D recapitulating clinical and immune-phenotypic dysregulations observed in the index family and displaying T-cell-intrinsic DNA damage intolerance. Reconstitution experiments and molecular dynamics simulations categorize both missense mutations as loss-of-function and haploinsufficient. This study provides evidence that certain monoallelic LIG4 mutations may cause human immune dysregulation via haploinsufficiency.

Positive and negative emotion are associated with generalized transcriptional activation in immune cells

Alterations in immune system gene expression have been implicated in psychopathology, but it remains unclear whether similar associations occur for intraindividual variations in emotion. The present study examined whether positive emotion and negative emotion were related to expression of pro-inflammatory and antiviral genes in circulating leukocytes from a community sample of 90 adolescents (Mage = 16.3 years, SD = 0.7; 51.1% female). Adolescents reported their positive emotion and negative emotion and provided blood samples twice, five weeks apart. Using a multilevel analytic framework, we found that within-individual increases in positive emotion were associated with reduced expression of both pro-inflammatory and Type I interferon (IFN) response genes, even after adjusting for demographic and biological covariates, and for leukocyte subset abundance. By contrast, increases in negative emotion were related to higher expression of pro-inflammatory and Type I IFN genes. When tested in the same model, only associations with positive emotion emerged as significant, and increases in overall emotional valence were associated with both lower pro-inflammatory and antiviral gene expression. These results are distinct from the previously observed Conserved Transcriptional Response to Adversity (CTRA) gene regulation pattern characterized by reciprocal changes in pro-inflammatory and antiviral gene expression and may reflect alterations in generalized immunologic activation. These findings highlight one biological pathway by which emotion may potentially impact health and physiological function in the context of the immune system, and future studies can investigate whether fostering positive emotion may promote adolescent health through changes in the immune system.

Different effects of pesticides on transcripts of the endocrine regulation and energy metabolism in honeybee foragers from different colonies

Honeybees are important pollinators of many crops and contribute to biological biodiversity. For years, a decline in bee populations has been observed in certain areas. This decline in honeybees is accompanied by a decrease in pollinator services. One factor contributing to the decline of bee colonies is the exposure to pesticides. Pesticide exposure of bees, among other effects, can negatively affect orientation, memory, immune system function and gene expression. Among the altered expressed genes are transcripts of endocrine regulation and oxidative phosphorylation. Endocrine regulation plays an important role in the development of nurse bees into foragers and oxidative phosphorylation is involved in energy metabolism. Most of these transcriptional changes were investigated using mixed aged honeybees derived from the same colony. Experiments using nurse bees or foragers of the same age but from different colonies are rare. In the present study, effects of the two pesticides chlorpyrifos and pyraclostrobin on the expression of transcripts linked to endocrine regulation and oxidative phosphorylation in foragers of the same age from three different colonies are investigated to fill this gap. These two pesticides were selected because negative effects at sublethal concentrations on bees are known and because they are found in pollen and nectar of crops and wild plants. For this purpose, 20–22 days old foragers of three different colonies were exposed to different sublethal concentrations of the selected fungicides for 24 h, followed by analysis of the expression of buffy, vitellogenin, hbg-3, ilp-1, mrjp1, 2 and 3, cox5a, cox5b and cox17. Some significant changes in gene expression of both endocrine regulation transcripts and oxidative phosphorylation were shown. Furthermore, it became clear that forager bees from different colonies react differently. This is especially important in relation to the risk analysis of pesticides. In addition, it could be shown that the expression of hbg-3 in the brain of bees is a robust marker to distinguish nurse bees from foragers at the molecular biological level. In summary, this study clearly shows that pesticides, which are often detected in pollen and nectar, display negative effects at sublethal concentrations on bees and that it is important to use bees from different colonies for risk assessment of pesticides.

The Genetic Variant TNFA (rs361525) Is Associated with Increased Susceptibility to Developing Dengue Symptoms.

Dengue virus (DENV) is the causal agent of dengue fever. The symptoms and signs of dengue vary from febrile illness to hemorrhagic syndrome. IFITM3 and TNFA are genes of the innate immune system. Variants IFITM3 (rs12252 T>C) and TNFA (rs1800629 G > A and rs361525 G>A) might alter gene expression and change the course of the disease. Our first objective was to determine whether these variants were associated with the susceptibility and severity of dengue. The second was to assess the association of these variants with each symptom. We studied 272 cases with suspected dengue infection, of which 102 were confirmed dengue cases (DENV+) and 170 were dengue-like cases without DENV infection (DENV-). Samples of 201 individuals from the general population of Mexico were included as a reference. Genotyping was performed by the polymerase chain reaction-restriction fragment length polymorphism technique. Odds ratios and confidence intervals were calculated using Pearson's chi-square test and later adjusted for age and sex with a binary logistic regression model. Haldane correction is applied when necessary. We found a significantly higher frequency of the A allele of TNFA rs361525 in both the DENV+ and DENV- groups compared with the general population. Focusing on DENV+ and DENV-, the frequency of the A allele of TNFA rs361525 was higher in the DENV+ group. A broad spectrum of symptoms was related to the A allele of both TNFA variants. We conclude that TNFA rs361525 increases the susceptibility to symptomatic dengue but can also be associated with susceptibility to other dengue-like symptoms from unknown causes.

Transcriptome Analysis of Immune-Related Gene in Hybrid Yellow Catfish in Reaction to Peptidoglycan Challenge

Hybrid yellow catfish (<i>Tachysurus vachelli x Tachysurus fulvidraco</i>) is one of the most important freshwater fish in China, intensive culturing has caused increased susceptibility to bacteria, viruses, and parasites. However, little is known about its adaptation mechanisms to pathogen infection. To better understand its immune system and immune related gene under pathogen infection, the transcriptome was analyzed by comparing the data of hybrid yellow catfish spleen stimulated by PGN and PBS. Fish in experimental group were injected with 100 μl baccilus subminis PGN (1mg/ml, 69,554, Sigma) intraperitoneally, and the fish in control group injected with 100 ul phosphate buffered solution (PBS). After 24 h injection, spleens were collected and immediately frozen in liquid nitrogen, then stored in -80°C refrigerator. Total RNA from spleens of hybrid yellow catfish were extracted using Trizol reagent (Sangon, China) according to the manufacturer’s instructions. The libraries were constructed using TruSeq Stranded mRNA LT Sample Prep Kit (Illumina, USA) and sequenced on an Illumina HiSeq X Ten platform. There were 285 differentially expressed genes (DEGs) between PGN treated experimental groups and PBS control groups at 24 h, including 213 up regulated and 72 down regulated DEGs. The expression of qRT-PCR from five up regulated DEGs and five down regulated DEGs were consistent with RNA-Seq. In the analysis of GO enrichment, DEGs in the spleens of PGN treated hybrid yellow catfish were significantly enriched in GO items, including Biological Process, Cellular Component, and Molecular Function items. The identified DEGs and enriched GO terms and KEGG pathways were useful for understanding the physical and biochemical performances, and the transcriptome data greatly enriched the genetic information of hybrid yellow catfish.

Comparative genomic diversity analysis of copy number variations (CNV) in indicine and taurine cattle thriving in Europe and Indian subcontinent

Copy number variations (CNVs) include deletions, duplications, and insertions that are larger than 50 bp in size causing structural variation responsible for diversity, adaptation, and breed development. Indian cattle breeds are highly diverse from the taurine breeds. The pattern of CNVRs in 191 animals belonging to 39 cattle breeds (four Indicine and 35 Taurine) was studied based on Illumina 777K BovineHD chip data. The Indicine breeds revealed 2590 CNVs and 335 copy number variation regions (CNVRs) in autosomes. Out of the identified CNVs, 50 were found to be novel. Structure analysis revealed admixed nature of Siri. Neighbor joining tree from CNVR data showed that hot (Kankrej and Hallikar) and cold (Ladakhi and Siri) adapted cattle breeds clustered separately. CNVR of Indian and European breeds revealed that Balkan and Italian breeds of Podolian group are admixed with Indian cattle breeds corroborating indicine introgression (6.1–13.5%). CNVRs spanning the regions of olfactory receptors and immune system genes were identified. AMOVA revealed 9% variation among populations which is 2% greater than SNP based studies showing higher inclusion of variation by CNVR. Detailed analysis of CNVs/CNVRs in Indian cattle adapted to hot and cold climate, and their diversity among worldwide cattle is presented in this study.

Effects of feeding a Saccharomyces cerevisiae fermentation product and ractopamine hydrochloride to finishing beef steers on growth performance, immune system, and muscle gene expression.

The objective of this study was to determine impacts on immune parameters, anti-oxidant capacity, and growth of finishing steers fed a Saccharomyces cerevisiae fermentation product (SCFP; NaturSafe; Diamond V, Cedar Rapids, IA) and ractopamine hydrochloride (RAC; Optaflexx; Elanco Animal Health, Greenfield, IN). Angus-crossbred steers (N = 288) from two sources were utilized in this 90-d study. Steers were blocked by source, stratified by initial body weight to pens of six steers, and pens randomly assigned to treatments (16 pens per treatment). Three treatments compared feeding no supplemental SCFP (control; CON) and supplemental SCFP for 57 d (SCFP57), and 29 d (SCFP29) before harvest. Supplementation of SCFP was 12 g per steer per d, and all steers were fed RAC at 300 mg per steer per d for 29 d before harvest. Blood samples were collected from3 steers per pen, and muscle samples were collected from 1 steer per pen at 57, 29 (start of RAC), and 13 (midRAC) days before harvest. Blood was analyzed from 2 steers per pen for ferric reducing anti-oxidant power (FRAP). Muscle gene expression of myokines, markers of anti-oxidant and growth signaling were assessed. Individual animal BW were also collected on 57, 29, 13, and 1 d before being harvested at a commercial facility (National Beef, Tama, IA). Data were analyzed using the Mixed procedure of SAS 9.4 (Cary, NC) with pen as the experimental unit. The model included fixed effects of treatment and group. Increased BW compared to CON was observed days -29, -13, and -1 in SCFP57 steers (P ≤ 0.05), with SCFP29 being intermediate days -13 and -1. Overall G:F was improved in SCFP29 and SCFP57 (P = 0.01). On day -29, FRAP was greater in SCFP57 than CON (P = 0.02). The percent of gamma delta T cells and natural killer cells in both SCFP29 and SCFP57 was greater than CON on day -13 (P = 0.02). There were no treatment × day effects for muscle gene expression measured (P ≥ 0.25). Interleukin 6 tended to decrease in SCFP29 and SCFP57 on day -13 (P = 0.10). No other treatment effects were observed for muscle gene expression. Muscle gene expression of interleukin 15 was increased (P = 0.01), and expression of interleukin 8 was decreased (P = 0.03) due to RAC feeding. Increased growth in SCFP-fed cattle may be related to changes in anti-oxidant capacity and the immune system.

The Immune Ghosts of Pandemics Past

Klunk J, Vilgalys TP, Demeure CE, et al. Evolution of immune genes is associated with the Black Death. Nature 2022;611:312–319. Our immune system exists to protect us from the external environment and specifically from the complex threat posed by microbial pathogens. It is well-known that microbes have rapidly evolved to either avoid, adapt, or co-opt host biology to survive and multiply. Conversely, infectious diseases are postulated to provide a strong selective pressure driving human genetic evolution, with exposure to pathogens driving immune system adaptation. However, exactly how and which specific loci and related immune genes have changed in response to a specific pathogen and functionally connecting signatures to protection or susceptibility in human populations has proved to be a challenging problem. The recent study by Klunk et al sets out to identify specific genes that may have directly evolved because of pathogen-induced selective pressure. To achieve this, they looked back at one of the most devastating pandemics in human history—the Black Death caused by the bacteria Yersinia pestis, which killed an estimated 30%–50% of the population in Europe. In a remarkable feat of molecular archeology, the study team identified human remains in two closely related European populations, in London and Denmark, devastated by the Black Death. To identify potential loci significantly selected for by this devastating disease, they obtained DNA samples from people who died before, during, and after the pandemic at various burial sites around London. After targeted DNA sequencing of a curated list of immune system genes and loci along with neutral regions of the genome as reference, they first verified that the pandemic did indeed induce a significant selection pressure on the immune system, with the identification of 245 significantly altered variants. After further filtering based on allele frequency patterns before and after the pandemic, they identified 35 potential target loci. Using their identified Denmark cohort to cross-validate these variants they eventually settled on 4 significant and validated loci related to 6 immune genes that potentially conferred either susceptibility or protection from Y. pestis infection. Although all 4 loci suggested interesting potential functional consequences, the clearest candidate was a putatively protective variant close to the gene ERAP2, an aminopeptidase important for antigen presentation, viral clearance, and cytokine responses and a known risk factor for Crohn’s disease. The study team went on to conduct a series of functional studies in monocyte-derived macrophages comparing the ERAP2 gene haplotype selectively favored by Y. pestis infection against the alternative deleterious prepandemic haplotype. Here, they found that macrophages from individuals possessing the selected ERAP2 allele exhibit a different cytokine response to Y. pestis infection and are better able to limit replication in cells, providing a functional corelate for the evolved host response. This study represents a wonderful journey into the history of the biological “arms race” between our immune system and the microbial environment, identifying some the specific changes brought about by a global disease pandemic. These findings illuminate our understanding of our immune responses to pathogens and highlight the evolution of immune system components such as ERAP2, CTLA4, and ICOS, which are associated with chronic inflammatory diseases including Crohn’s disease and autoimmune enteropathies. The study is also highly prescient in the setting of the global SARS-CoV-2 pandemic, and the current, likely highly accelerated, evolution of our immune system.

Immune system gene polymorphisms associated with severe dengue in Latin America: a systematic review.

One of the main challenges in the clinical management of dengue is the early identification of cases that could progress to severe forms of the disease. A biomarker that may enable this identification is the presence of genetic polymorphisms in genes associated with immune responses. The objective of this study was to perform a systematic review of the Latin American literature on these genes. An electronic literature search was carried out in PubMed, Scopus, Lilacs, and the Virtual Health Library, and reference lists of systematic reviews in the area. Case-control studies conducted in Latin American countries examining at least one form of genetic polymorphism related to immune responses against severe dengue were included. In total, 424 articles were identified and 26 were included in this systematic review. Of the 26 selected articles, 16 reported polymorphisms associated with the risk of developing severe dengue (Risk); Similarly, 16 articles reported polymorphisms associated with a decreased risk of severe dengue (Protective). The final analysis revealed that multiple polymorphisms in immune system genes were early markers of the progression of dengue in Latin Americans and found that polymorphisms of the TNF-alpha gene may have a critical role in dengue pathogenesis.

Association of IFITM1 Promoter Methylation with Severity of SARS-CoV-2 Infection.

During viral infections such as SARS-CoV-2, epigenetic changes within the promoter region of the immune system genes would possibly occur and have an effect on the immune system response as well as disease outcome. We aimed to evaluate and compare the methylation level of the IFITM1 gene promoter in different stages of COVID-19 disease with a healthy control group. In this cross-sectional study, 75 COVID-19 patients (25 mild, 25 severe, and 25 critical in addition to 25 age- and gender-matched healthy volunteers) have been included. DNA was extracted from the peripheral white blood cells using a commercial DNA extraction kit. PCR was performed using two types of primers designed for the methylated and unmethylated forms of the IFITM1 gene promoter. The mean age of the patient and healthy volunteer groups was 52.733 ± 13.780 and 49.120 ± 12.490, respectively. Out of a hundred participants, 52 were male. The results demonstrated that severe (p = 0.03, OR 6.729) and critical (p = 0.001, OR 11.156) patients were much more likely to show methylation of the IFITM1 gene in contrast with mild patients. Moreover, IFITM1 methylation was significantly higher in COVID-19 patients in comparison with the healthy volunteer group (p = 0.004, OR 3.17). Furthermore, IFITM1 methylation in male patients with critical status, (p = 0.01) was significantly higher than in male patients with mild status. In addition, IFITM1 methylation of male (p = 0.03) and female (p = 0.01) critical patients was considerably higher compared to males and females of volunteer group. Increased methylation of the IFITM1 gene in the severe and critical stage of COVID-19 diseases may indicate the role of SARS-CoV-2 infection in increasing methylation of this antiviral gene. This might be involved in suppressing the immune system, promoting SARS-CoV-2 replication and disease outcome.

Spectrum of auto-inflammatory diseases in Morocco: a monocentric experience.

Auto-inflammatory diseases (AIDs) result from mutations in genes of the innate immune system leading to periodic multisystemic inflammation. We aimed to describe the clinical, biological and molecular features (when available) and outcomes of Moroccan patients with AIDs. Patient data were collected retrospectively and analysed over a 13-year period. Among 30 patients, 60% had FMF, 16% mevalonate kinase deficiency (MKD) and 24% other AIDs. The mean age at first consultation was 6.9 years, and the mean diagnostic delay was 3 years. Consanguinity was reported in 16 cases. IgA vasculitis was associated with 33% of FMF patients, in whom the main clinical features were fever (88.8%), abdominal pain (100%), arthralgias (88.8%) and arthritis (50%), and the most frequent mutation was M694V (66%). All FMF patients were treated with colchicine. Most MKD patients were confirmed by elevated urinary mevalonic acid levels, and four of five MKD patients received targeted therapy. Chronic recurrent osteomyelitis patients were confirmed by radiological and histological analysis. Two cases of Marshall syndrome were diagnosed according to validated criteria. A case of familial pustular psoriasis was diagnosed based on histological analysis and a patient with Muckle-Wells syndrome by clinical features. The outcome was favourable in 76%, partial in 13%, and three deaths were reported. FMF and MKD are the most reported diseases. AIDs are probably underestimated because they are unknown to clinicians. The aim of this work is to raise awareness among paediatricians about AIDs and create a network for best practice.

Colony Transport Affects the Expression of Some Genes Related to the Apis mellifera L. Immune System

Migratory beekeeping can harm the bee colonies if not executed properly. Here, colonies of Apis mellifera were transported (for one or two hours) or not, following proper technical standards. To analyze gene expression (defensin-1, abaecin, and HSP70), forager bees were collected immediately, 24, and 72 hours after transportation. Bee mortality and population growth were measured before and after transportation. This study concludes that transporting honey bee colonies for 2 hours promotes immune system gene expression, although there are no significant changes in bee mortality and population growth of the colonies.

Prediction of uncontrolled course of bronchial asthma in children based on polymorphisms of genes of signaling molecules of the immune system and detoxification genes

Aim. To study the possibility of predicting the asthma control at various stages of the development of the disease, possibly on the basis of taking into account the genetic polymorphisms of Toll-like receptors, cytokines and detoxification system genes using the statistical method of learning neural networks.Materials and methods. We ex­amined 167 children with bronchial asthma. The degree of asthma control was determined, the following mutations were detected: TLR2-Arg753Glu, TLR4-Asp299Gly, TLR4-Ghr399Ile, TLR9-T1237C, TLR9-A2848G; IL4-C589T, IL6- C174G, IL10-G1082A, IL10-C592A, IL10-C819T, IL12B-A1188C, TNFa-G308A; GSTM, GSTT, GSTM/GSTT, GSTP1 Ile105Val, GSTP1 Ala114Val, by PCR. The STATISTICA Automated Neural Networks package was used to model neural networks.Results. The model is based on the MLP (15-9-3) multilayer perceptron architecture with a layer of 15 input neurons (by the number of analyzed variables), a hidden intermediate layer of 9 neurons and an output layer of 3 neurons by the number of values of the classified variable (control). The training algorithm was chosen by BFGS as the most adequate to the classification task. The error function is traditionally chosen as the sum of squared deviations. The activation function of output neurons is Softmax. The activation function of the intermediate layer is hyperbolic. The volume of the training sample was 88 sets. The volume of samples for testing and quality control of the model was 36 sets. The resulting model was able to predict 79.01% of the correct values of the target variable (the degree of asthma control).Conclusion. The application of the developed program makes it possible to predict the possibility of uncontrolled or partially controlled asthma at any stage of the disease, including preclinical and pre-nosological for groups with a high risk of asthma. This allows you to individually adjust the measures of secondary and even primary prevention of asthma within the personal­ization of therapeutic approaches.

Progressive multifocal leukoencephalopathy genetic risk variants for pharmacovigilance of immunosuppressant therapies.

Progressive multifocal leukoencephalopathy (PML) is a rare and often lethal brain disorder caused by the common, typically benign polyomavirus 2, also known as JC virus (JCV). In a small percentage of immunosuppressed individuals, JCV is reactivated and infects the brain, causing devastating neurological defects. A wide range of immunosuppressed groups can develop PML, such as patients with: HIV/AIDS, hematological malignancies (e.g., leukemias, lymphomas, and multiple myeloma), autoimmune disorders (e.g., psoriasis, rheumatoid arthritis, and systemic lupus erythematosus), and organ transplants. In some patients, iatrogenic (i.e., drug-induced) PML occurs as a serious adverse event from exposure to immunosuppressant therapies used to treat their disease (e.g., hematological malignancies and multiple sclerosis). While JCV infection and immunosuppression are necessary, they are not sufficient to cause PML. We hypothesized that patients may also have a genetic susceptibility from the presence of rare deleterious genetic variants in immune-relevant genes (e.g., those that cause inborn errors of immunity). In our prior genetic study of 184 PML cases, we discovered 19 candidate PML risk variants. In the current study of another 152 cases, we validated 4 of 19 variants in both population controls (gnomAD 3.1) and matched controls (JCV+ multiple sclerosis patients on a PML-linked drug ≥ 2 years). The four variants, found in immune system genes with strong biological links, are: C8B, 1-57409459-C-A, rs139498867; LY9 (alias SLAMF3), 1-160769595-AG-A, rs763811636; FCN2, 9-137779251-G-A, rs76267164; STXBP2, 19-7712287-G-C, rs35490401. Carriers of any one of these variants are shown to be at high risk of PML when drug-exposed PML cases are compared to drug-exposed matched controls: P value = 3.50E-06, OR = 8.7 [3.7-20.6]. Measures of clinical validity and utility compare favorably to other genetic risk tests, such as BRCA1 and BRCA2 screening for breast cancer risk and HLA-B*15:02 pharmacogenetic screening for pharmacovigilance of carbamazepine to prevent Stevens-Johnson Syndrome and Toxic Epidermal Necrolysis. For the first time, a PML genetic risk test can be implemented for screening patients taking or considering treatment with a PML-linked drug in order to decrease the incidence of PML and enable safer use of highly effective therapies used to treat their underlying disease.

Immunogenetic losses co-occurred with seahorse male pregnancy and mutation in tlx1 accompanied functional asplenia.

In the highly derived syngnathid fishes (pipefishes, seadragons & seahorses), the evolution of sex-role reversed brooding behavior culminated in the seahorse lineage's male pregnancy, whose males feature a specialized brood pouch into which females deposit eggs during mating. Then, eggs are intimately engulfed by a placenta-like tissue that facilitates gas and nutrient exchange. As fathers immunologically tolerate allogenic embryos, it was suggested that male pregnancy co-evolved with specific immunological adaptations. Indeed, here we show that a specific amino-acid replacement in the tlx1 transcription factor is associated with seahorses' asplenia (loss of spleen, an organ central in the immune system), as confirmed by a CRISPR-Cas9 experiment using zebrafish. Comparative genomics across the syngnathid phylogeny revealed that the complexity of the immune system gene repertoire decreases as parental care intensity increases. The synchronous evolution of immunogenetic alterations and male pregnancy supports the notion that male pregnancy co-evolved with the immunological tolerance of the embryo.

Metabolic, antioxidant, and immune response of broodstock males ofPenaeus brasiliensisLatreille, 1817 fed diets supplemented by vitamin C

AbstractControlling oxidative stress in shrimps under culture conditions is of vital importance to ensure that males have an optimal physiological status. The use of diets supplemented with vitamins are widely used for this purpose. We evaluated the effects of vitamin C on the metabolic, antioxidant, and immune responses of Penaeus brasiliensisLatreille, 1817 broodstock males under culture conditions. A formulated diet was used as a control (16 mg kg–1 of total ascorbic acid (vitamin C) content) and three diets were prepared with three vitamin C levels: 322 mg kg–1 (Diet A), 628 mg kg–1 (Diet B), and 934 mg kg–1 (Diet C). Triglycerides, cholesterol, glucose, and prophenoloxidase, total proteins, and quantity and type of hemocytes in the hemolymph were determined. We also quantified the antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (Gpx) and lipid peroxidation in the reproductive tract. Differentially expressed genes of the immune system, namely those of α-2 macroglobulin (α2M), peneidin (Pen), hemocyanin, prophenoloxidase, and the genes encoding the antioxidant enzymes SOD, CAT, and Gpx were quantified in the tract. Sperm quantity and quality were recorded. In males fed Diet C, the lowest cholesterol values and triglycerides obtained in the hemolymph were not significantly different from those fed the control diet (P &amp;lt; 0.05); those fed Diet C showed the highest amount of hyaline hemocytes, the lowest CAT enzyme activity, and the highest Gpx activity. The immune system genes only showed differences between α2M and Pen; sperm quantity and quality were not affected by diet. The results indicated that Diet C (13.9 g kg–1 vitamin C) was effective for the maintenance of P. brasiliensis broodstock males because it controlled cholesterol and triglyceride levels; there was less CAT activity and less activation of the immune system mediated by hemocytes.

Major locus on ECA18 influences effectiveness of GonaCon vaccine in feral horses

Contraceptive vaccines are used to reduce birth rates in wild and feral animal populations. While the immunocontraceptive GonaCon-Equine has proven effective in reducing fertility among female feral horses, there is individual variation in the duration of infertility following treatment. To identify genetic factors influencing the effectiveness of GonaCon-Equine, we conducted a genome-wide association study of 88 mares from a feral population genotyped using the Illumina GGP Equine 70k SNP array. Contraceptive treatment schedules and long-term foaling rates have been recorded for each individual. We used mixed linear models to control for relatedness among mares. We found a significant association (p < 5 ×10-8) with a locus on equine chromosome 18. The most likely candidate genes in this region are STAT1 and STAT4, which are both involved in immune system function. Variation in STAT function could affect the immune response to the vaccine, leading to variation in contraceptive efficacy. Additional SNPs reaching a less stringent threshold of significance (p < 5 ×10-6) were located on other chromosomes near known immune system genes, supporting the hypothesis that variation in immunocontraceptive efficacy can be attributed to genetic variation in immune response rather than fertility genes.

Microplastic-Contaminated Feed Interferes with Antioxidant Enzyme and Lysozyme Gene Expression of Pacific White Shrimp (Litopenaeus vannamei) Leading to Hepatopancreas Damage and Increased Mortality

Microplastic pollution can interfere with aquatic animal health and nonspecific immunity, increasing the potential for pathogen infection in crustaceans. However, the long-term effects of microplastics on crustacean immunity are less understood, especially regarding their toxicity in Pacific white shrimp (Litopenaeus vannamei). Effects of high-density polyethylene microplastics (HDPE-MPs) in feed on the mortality rate, hepatopancreas, and nonspecific immune system gene expression of Pacific white shrimp are presented. The LC50 at day 28 of HDPE-MP exposure was determined as 3.074% HDPE-MP in feed. A significant upregulation of the superoxide dismutase (SOD) and glutathione peroxidase (GPx) genes was observed in shrimp that were fed with 0.1 and 0.5% of HDPE-MP; then, they were downregulated significantly, except for the SOD gene expression of shrimp fed with 0.1% of HDPE-MP. The lysozyme (LYZ) gene was upregulated significantly in shrimp that were fed with 0.5, 1, and 3% HDPE-MP for 7 days and downregulated significantly in HDPE-receiving groups for at least 14 days. Significant histopathological changes in the hepatopancreas were observed in the treatment groups. The histopathological score of each lesion was correlated with the increase in HDPE-MP concentration. This study shows that the ingestion of HDPE microplastics can alter the expression of nonspecific immune system genes and damage the hepatopancreas in Pacific white shrimp.

Immunogenetic Determinants of Susceptibility to Head and Neck Cancer in the Million Veteran Program Cohort.

Several inherited variations in immune system genes are significantly associated with susceptibility to head and neck cancer, which could help improve personalized cancer risk estimates.

Identification of Predictive Models Including Polymorphisms in Cytokines Genes Associated with Post-Transplant Complications after Identical HLA-Allogeneic Stem Cell Transplantation

Introduction Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curative treatment for patients with hematologic malignances. Immune system genes like cytokines play a role in inflammatory process that occur during graft versus host disease (GVHD). Therefore, the analysis of polymorphisms that affect the expression or activity of these genes could be used as biomarkers to predict the development of complications after allo-HSCT. The aim of this study was to identify new polymorphisms in genes related to the immune response, mainly cytokines, that could be related to the development of post-transplant complications. Methods We retrospectively selected 90 patients with hematological malignancies who received an allo-HSCT from an HLA-identical sibling donor from 2000 to 2015. The genotyping was performed using a custom NGS panel (73 interleukins and 59 chemokines genes) in a HiSeq platform (Illumina, USA). The bioinformatics analysis was carried out with the GeneSystems software (Sistemas Genómicos, Spain). We analyzed variants located in coding regions, splicing sites, UTR, and 5’ upstream and 3'downstream regions (+/-200pb). The polymorphisms selected corresponding to non-synonymous variants, with depth ≥30X in the canonical isoform with an allele frequency ≥0.4, with a minor allele frequency ≥1% and represented in at least 5% in our cohort. Bayesian logistic regression models (BLR) were used to select the most relevant variables for each post-transplant complication. The models with the highest AUC value, sensitivity and specificity value and the lowest number of genetic variants used were selected. Finally, the score obtained from the chosen predictive model for each complication was used to classify patients as low or high risk according to the selected cutoff point. Results The cumulative incidence (CI) rates for aGVHD of grades II-IV and III-IV at 100 days after transplantation were 56.7% and 23.3%, respectively. The CI for cGVHD and moderate-severe cGVHD was 43.3% and 27.8%, respectively, at 3 years after allo-HSCT. The CI of TRM at 3 years was 24.4% and OS at 5 years was 30%. Using filters defined previously, we detected 737 polymorphisms (644 in non-coding regions and 93 in coding regions). We developed BLR models for II-IV and III-IV aGVHD, cGVHD, moderate-severe cGVHD, TRM, relapse and OS (Table 1). Stratification of the cohort of patients according to the risk of develop each complication is showed in Figure 1. Most polymorphisms found are located in regulatory regions, which until have been little studied, but these regions are involved in the expression genes, therefore could be affecting the function of these genes. Conclusions We have identified new genetic polymorphisms correlate with the risk of developing complications after allo-HSCT from an HLA-identical sibling donor. Based on these data, we have developed a genetic score that encompasses polymorphisms of greater relevance. In this way, patients at greatest risk for developing post-transplant complication who could benefit from personalized management through immunosuppression and other drugs. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal