Activated T-helper Cells Research Articles

In silico designing of multi-epitope vaccine against canine parvovirus using reverse vaccinology.

Canine parvovirus (CPV-2) is a highly contagious virus affecting dogs worldwide, posing a significant threat. The VP2 protein stands out as the predominant and highly immunogenic structural component of CPV-2. Soon after its emergence, CPV-2 was replaced by variants known as CPV-2a, 2b and 2c, marked by changes in amino acid residue 426 of VP2. Additional amino acid alterations have been identified within VP2, with certain modifications serving as signatures of emerging variants. In Brazil, CPV-2 outbreaks persist with diverse VP2 profiles. Vaccination is the main preventive measure against the virus. However, the emergence of substitutions presents challenges to conventional vaccine methods. Commercial vaccines are formulated with strains that usually do not match those currently circulating in the field. To address this, the study aimed to investigate CPV-2 variants in Brazil, predict epitopes, and design an in silico vaccine tailored to local variants employing reverse vaccinology. The methodology involved data collection, genetic sequence analysis, and amino acid comparison between field strains and vaccines, followed by the prediction of B and T cell epitope regions. The predicted epitopes were evaluated for antigenicity, allergenicity and toxicity. The final vaccine construct consisted of selected epitopes linked to an adjuvant and optimized for expression in Escherichia coli. Structural predictions confirmed the stability and antigenicity of the vaccine, while molecular docking demonstrated interaction with the canine toll-like receptor 4. Molecular dynamics simulations indicated a stable complex formation. In silico immune simulations demonstrated a progressive immune response post-vaccination, including increased antibody production and T-helper cell activity. The multi-epitope vaccine design targeted prevalent CPV-2 variants in Brazil and potentially other regions globally. However, experimental validation is essential to confirm our in silico findings.

Mathematical modeling of interactions between colon cancer and immune system with a deep learning algorithm

Colon cancer is a complex disease with genetically unstable cell lines. In order to better understand the complexity of colon cancer cells and their metastatic mechanisms, we develop a mathematical model in this study. The model is based on a system of fractional-order differential equations and Fractional-Cancer-Informed Neural Networks (FCINN). The model captures a dynamic network of interactions between dendritic cells (DCs), cytotoxic T-cells (CD8+\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$8^+$$\\end{document}), helper T-cells (CD4+\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$4^+$$\\end{document}), and colon cancer cells through fractional differential equations. By varying the fractional order between 0 and 1, we can classify patients into different groups based on their immune patterns. The goal of this paper is to identify different immune patterns and cancer cell behaviors, as well as the parameters that play an important role in metastasis, control, or elimination of cancer cells in the model. However, several parameters in the model are difficult to estimate in a patient-specific manner. To address this challenge, we use FCINN as an effective deep-learning tool for parameter estimation and numerical simulation of the model. Our findings suggest that the most effective factors in controlling the progression and preventing metastasis of colon cancer are the initial number of cancer cells, the inhibiting rates of tumor cells by DCs, the source of DCs, and the activation of helper T-cells by DCs. These findings suggest that DCs can be used as an immunotherapy tool for the control and treatment of colon cancer.

Single-Nucleotide Polymorphisms of the PAR2 and IL-17A Genes Are Significantly Associated with Chronic Pain.

Patients with chronic pain are affected psychologically and socially. There are also individual differences in treatment efficacy. Insufficient research has been conducted on genetic polymorphisms that are related to individual differences in the susceptibility to chronic pain. Autoimmune disorders can lead to inflammation and chronic pain; therefore, we focused on the autoimmune-related protease-activated receptor 2 (PAR2/F2RL1) and interleukin 17A (IL-17A/IL17A) genes. PAR2 and IL-17A are associated with autoimmune diseases that lead to chronic pain, and PAR2 regulates T-helper (Th) cell activation and differentiation. We hypothesized that the PAR2 and IL-17A genes are associated with chronic pain. The present study used a case-control design to statistically examine associations between genetic polymorphisms and the vulnerability to chronic pain. The rs2243057 polymorphism of the PAR2 gene and rs3819025 polymorphism of the IL-17A gene were previously reported to be associated with pain- or autoimmune-related phenotypes. Thus, these polymorphisms were investigated in the present study. We found that both rs2243057 and rs3819025 were significantly associated with a susceptibility to chronic pain. The present findings revealed autoimmune-related genetic factors that are involved in individual differences in chronic pain, further aiding understanding of the pathomechanism that underlies chronic pain and possibly contributing to future personalized medicine.

155 Impact of Ambient Temperature on Bovine Viral Diarrhea Virus and Infectious Bovine Rhinotracheitis Vaccination Response

Abstract Vaccines are created to help the immune system fight pathogens faster and more effectively, but little is known on how season of the year impacts the immune system response to vaccination. This research was conducted to evaluate how ambient temperature (summer vs fall) impacted antibody production and immune cell type changes in Brahman and Brahman-influenced cows. Cows (n = 18) were immunized (2 mL I.M.) with a combination Modified Live Vaccine for Bovine Viral Diarrhea Virus (BVDV) and Infectious Bovine Rhinotracheitis Virus (IBR) on d 0 in either July 2022 (Summer; n = 12) or November 2022 (Fall; n = 6). Blood samples (30 mL/cow) were collected on d -3, 0, 2, 4, 6, 8, 10, 14 post vaccination and transferred to 50 mL conical tubes containing EDTA to separate plasma and peripheral blood mononuclear cells (PBMC). Density gradient centrifugation was used to isolate PBMC. Serum samples were also collected (10 mL/cow) on d -3, 0, 14. Cells were incubated with propidium iodide and antibody surface cell markers for CD4, CD8, CD25, CD14, CD86, and CD335. An Amnis FlowSight flow cytometer was used to determine the percentage of each cell type in the PBMC. Concentrations of antibodies for BVDV and IBR were determined in serum samples collected pre-vaccination (d -3 and 0) and on d 14. Differences in antibody titers and leukocyte populations were analyzed as repeated measures using the MIXED model procedure in SAS. There were no differences in IBR (P = 0.2) or BVDV (P = 0.47) antibody concentrations between seasons. Percentage of antigen presenting cells (CD14+) was greater in the Summer compared with the Fall (P = 0.01; Summer=11.7 ± 1%; Fall = 7.3 ± 1.1%), but natural killer cells (CD335+) were greater in the Fall compared with the Summer (P = 0.002; Summer = 0.6 ± 0.6%; Fall = 4.4±0.7%). There was a significant treatment by time interaction for antigen presenting cells (P = 0.04), T-helper cells (CD4+; P = 0.01), activated T-helper cells (CD4+CD25+; P ≤ 0.0001), activated cytotoxic T-cells (CD8+CD25+; P = 0.03), activated T-cells (CD25+; P ≤ 0.0001), and natural killer cells (P ≤ 0.0001), but there was no difference between seasons in activated antigen presenting cells (CD14+CD86+; P = 0.19). From pre-vaccination to d 14, natural killer cells were elevated during the Fall compared with the Summer on all days (P = 0.04). Antigen presenting cells were greater in the Summer compared with the Fall on d 4, 10 and 14 (P < 0.02). T-helper cells, activated T-helper cells, activated cytotoxic T-cells, and activated T-cells increased post-vaccination in the Fall, but did not change during the Summer. Thus, ambient temperature (summer vs fall) did not directly impact production of antibodies for IBR or BVDV but did induce changes in specific immune cells following vaccination.

Bifidobacteria shape antimicrobial T-helper cell responses during infancy and adulthood

Microbial infections early in life are challenging for the unexperienced immune system. The SARS-CoV-2 pandemic again has highlighted that neonatal, infant, child, and adult T-helper(Th)-cells respond differently to infections, and requires further understanding. This study investigates anti-bacterial T-cell responses against Staphylococcus aureus aureus, Staphylococcus epidermidis and Bifidobacterium longum infantis in early stages of life and adults and shows age and pathogen-dependent mechanisms. Beside activation-induced clustering, T-cells stimulated with Staphylococci become Th1-type cells; however, this differentiation is mitigated in Bifidobacterium-stimulated T-cells. Strikingly, prestimulation of T-cells with Bifidobacterium suppresses the activation of Staphylococcus-specific T-helper cells in a cell-cell dependent manner by inducing FoxP3+CD4+ T-cells, increasing IL-10 and galectin-1 secretion and showing a CTLA-4-dependent inhibitory capacity. Furthermore Bifidobacterium dampens Th responses of severely ill COVID-19 patients likely contributing to resolution of harmful overreactions of the immune system. Targeted, age-specific interventions may enhance infection defence, and specific immune features may have potential cross-age utilization.

Cell-type-specific role of P2Y2 receptor in HDM-driven model of allergic airway inflammation.

Allergic airway inflammation (AAI) is a chronic respiratory disease that is considered a severe restriction in daily life and is accompanied by a constant risk of acute aggravation. It is characterized by IgE-dependent activation of mast cells, infiltration of eosinophils, and activated T-helper cell type 2 (Th2) lymphocytes into airway mucosa. Purinergic receptor signaling is known to play a crucial role in inducing and maintaining allergic airway inflammation. Previous studies in an ovalbumin (OVA)-alum mouse model demonstrated a contribution of the P2Y2 purinergic receptor subtype (P2RY2) in allergic airway inflammation. However, conflicting data concerning the mechanism by which P2RY2 triggers AAI has been reported. Thus, we aimed at elucidating the cell-type-specific role of P2RY2 signaling in house dust mite (HDM)-driven model of allergic airway inflammation. Thereupon, HDM-driven AAI was induced in conditional knockout mice, deficient or intact for P2ry2 in either alveolar epithelial cells, hematopoietic cells, myeloid cells, helper T cells, or dendritic cells. To analyze the functional role of P2RY2 in these mice models, flow cytometry of bronchoalveolar lavage fluid (BALF), cytokine measurement of BALF, invasive lung function measurement, HDM re-stimulation of mediastinal lymph node (MLN) cells, and lung histology were performed. Mice that were subjected to an HDM-based model of allergic airway inflammation resulted in reduced signs of acute airway inflammation including eosinophilia in BALF, peribronchial inflammation, Th2 cytokine production, and bronchial hyperresponsiveness in mice deficient for P2ry2 in alveolar epithelial cells, hematopoietic cells, myeloid cells, or dendritic cells. Furthermore, the migration of bone-marrow-derived dendritic cells and bone-marrow-derived monocytes, both deficient in P2ry2, towards ATP was impaired. Additionally, we found reduced levels of MCP-1/CCL2 and IL-8 homologues in the BALF of mice deficient in P2ry2 in myeloid cells and lower concentrations of IL-33 in the lung tissue of mice deficient in P2ry2 in alveolar epithelial cells. In summary, our results show that P2RY2 contributes to HDM-induced airway inflammation by mediating proinflammatory cytokine production in airway epithelial cells, monocytes, and dendritic cells and drives the recruitment of lung dendritic cells and monocytes.

Mining Autoimmune-Disorder-Linked Molecular-Mimicry Candidates in Clostridioides difficile and Prospects of Mimic-Based Vaccine Design: An In Silico Approach.

Molecular mimicry, a phenomenon in which microbial or environmental antigens resemble host antigens, has been proposed as a potential trigger for autoimmune responses. In this study, we employed a bioinformatics approach to investigate the role of molecular mimicry in Clostridioides difficile-caused infections and the induction of autoimmune disorders due to this phenomenon. Comparing proteomes of host and pathogen, we identified 23 proteins that exhibited significant sequence homology and were linked to autoimmune disorders. The disorders included rheumatoid arthritis, psoriasis, Alzheimer's disease, etc., while infections included viral and bacterial infections like HIV, HCV, and tuberculosis. The structure of the homologous proteins was superposed, and RMSD was calculated to find the maximum deviation, while accounting for rigid and flexible regions. Two sequence mimics (antigenic, non-allergenic, and immunogenic) of ≥10 amino acids from these proteins were used to design a vaccine construct to explore the possibility of eliciting an immune response. Docking analysis of the top vaccine construct C2 showed favorable interactions with HLA and TLR-4 receptor, indicating potential efficacy. The B-cell and T-helper cell activity was also simulated, showing promising results for effective immunization against C. difficile infections. This study highlights the potential of C. difficile to trigger autoimmunity through molecular mimicry and vaccine design based on sequence mimics that trigger a defensive response.

Characteristics of T- and B-lymphocytes of blood at different stages of pregnancy and peculiarities of immunity in women with preeclampsia

The study of immune mechanisms during pregnancy is an important area of research aimed at understanding the immunogenesis of pregnancy and its complications. One such complication is nephropathy, which requires accurate prediction of its course and appropriate therapy.
 The present study aimed to assess the blood levels of T-lymphocytes (T-helper, T-suppressor/cytotoxic), B-lymphocytes, and markers of their activation in pregnant women, with a particular focus on patients with preeclampsia (PE).
 Methods. Using a cellular cytofluorimeter and the corresponding test systems, we determined the relative levels of immunocompetent blood cells such as CD3+, CD4+, CD8+, CD19+-l with the expression of HLA-DR, CD25, CD5 activation markers in 436 non-pregnant women and 514 pregnant women. We analyzed the characteristics of these indicators at different stages of pregnancy, specifically before (1-4 groups) and after (5 group) 20 weeks. In the second stage, we performed an analysis of the indicators in 107 women with PE, 14 women after PE, and 54 pregnant women in the reference group without complications at 20+ weeks. This analysis included T- and B-cell studies of these patient groups under in vitro conditions with autoserum and inactivated serum.
 Results. The first weeks of pregnancy were characterized by a high relative level (%) in the blood of T-l and subpopulations of T-h and Ts/c, as well as their activation according to the expression data of DR, CD25 (for CD4+-l), and CD5 (for CD19+-l). These indicators significantly decrease from 20 weeks of pregnancy and become comparable to the levels in non-pregnant women. The percentages of CD3+4+25+-l and CD19+5+-l are below the normal range, indicating a decrease in T-helper cell and B-cell activation during this period of pregnancy. In patients with PE after 20 weeks of pregnancy, the relative levels of T-l, B-l, T-h, and Ts/c did not differ from the reference group. Incubation of T- and B-l of women with PE with autoserum in vitro almost tripled the number of T- and B-l, and the blocking effect was greater than in women without PE, with a significant difference between them; the average ratio T-l/T-las exceeded the reference in the subgroup of active manifestations of PE.
 Conclusion. Studying the relative levels of T-l, T-h, T-s/c, and B-l in the blood, as well as markers of their activation, at different stages of pregnancy with in vitro detection of features of the humoral immune response as predictors of NP allows determining individual components of immunogenetic and prognostic markers for personalized therapy during pregnancy.

Factor VIII trafficking to CD4+ T cells shapes its immunogenicity and requires several types of antigen-presenting cells

AbstractDespite >80 years of clinical experience with coagulation factor VIII (FVIII) inhibitors, surprisingly little is known about the in vivo mechanism of this most serious complication of replacement therapy for hemophilia A. These neutralizing antidrug alloantibodies arise in ∼30% of patients. Inhibitor formation is T-cell dependent, but events leading up to helper T-cell activation have been elusive because of, in part, the complex anatomy and cellular makeup of the spleen. Here, we show that FVIII antigen presentation to CD4+ T cells critically depends on a select set of several anatomically distinct antigen-presenting cells, whereby marginal zone B cells and marginal zone and marginal metallophilic macrophages but not red pulp macrophages (RPMFs) participate in shuttling FVIII to the white pulp in which conventional dendritic cells (DCs) prime helper T cells, which then differentiate into follicular helper T (Tfh) cells. Toll-like receptor 9 stimulation accelerated Tfh cell responses and germinal center and inhibitor formation, whereas systemic administration of FVIII alone in hemophilia A mice increased frequencies of monocyte-derived and plasmacytoid DCs. Moreover, FVIII enhanced T-cell proliferation to another protein antigen (ovalbumin), and inflammatory signaling–deficient mice were less likely to develop inhibitors, indicating that FVIII may have intrinsic immunostimulatory properties. Ovalbumin, which, unlike FVIII, is absorbed into the RPMF compartment, fails to elicit T-cell proliferative and antibody responses when administered at the same dose as FVIII. Altogether, we propose that an antigen trafficking pattern that results in efficient in vivo delivery to DCs and inflammatory signaling, shape the immunogenicity of FVIII.

Astaxanthin, a Carotenoid without Vitamin A Activity, Augments Antibody Responses in Cultures Including T-helper Cell Clones and Suboptimal Doses of Antigen

Astaxanthin, a carotenoid without vitamin A activity, enhances T-dependent antigen (Ag)-specific humoral immune responses. We examined carotenoid actions on T-helper (Th) cell activity in a direct manner with reconstitution experiments; spleen Th cells were replaced with Ag-specific Type 1 and Type 2 (Th1 and Th2) Th cell clones. The Ag for the Th1 and Th2 clones were pigeon cytochrome C and rabbit γ-globulin, respectively. Astaxanthin and β-carotene augmented the number of IgM antibody (Ab)-secreting cells when unprimed B cells were incubated with Th clones and stimulated with suboptimal doses of Ag specific for each Th clone. The number of IgG Ab-secreting cells were greater with use of in vivo primed B cells than with unprimed B cells in both Th clones. Astaxanthin but not β-carotene augmented the number of IgG Ab-secreting cells when primed B cells and Th cell clones were stimulated with suboptimal doses of Ag specific for each Th clone. In the presence of optimal doses of Ag for each Th clone, neither carotenoid augmented the number of Ab-secreting cells. Astaxanthin and β-carotene may enhance the actions of both Th1 and Th2 cells for humoral immune responses with suboptimal Ag challenges; certain carotenoids may help maintain Ag-mediated immune responses at optimal levels.

Critical Role of the Transcription Factor AKNA in T-Cell Activation: An Integrative Bioinformatics Approach.

The human akna gene encodes an AT-hook transcription factor, the expression of which is involved in various cellular processes. The goal of this study was to identify potential AKNA binding sites in genes that participate in T-cell activation and validate selected genes. Here we analyzed ChIP-seq and microarray assays to determine AKNA-binding motifs and the cellular process altered by AKNA in T-cell lymphocytes. In addition, we performed a validation analysis by RT-qPCR to assess AKNA's role in promoting IL-2 and CD80 expression. We found five AT-rich motifs that are potential candidates as AKNA response elements. We identified these AT-rich motifs in promoter regions of more than a thousand genes in activated T-cells, and demonstrated that AKNA induces the expression of genes involved in helper T-cell activation, such as IL-2. The genomic enrichment and prediction of AT-rich motif analyses demonstrated that AKNA is a transcription factor that can potentially modulate gene expression by recognizing AT-rich motifs in a plethora of genes that are involved in different molecular pathways and processes. Among the cellular processes activated by AT-rich genes, we found inflammatory pathways potentially regulated by AKNA, suggesting AKNA is acting as a master regulator during T-cell activation.

Probiotic Bacillus Strains Enhance T Cell Responses in Chicken

Banning antibiotic growth promotors and other antimicrobials in poultry production due to the increasing antimicrobial resistance leads to increased feeding of potential alternatives such as probiotics. However, the modes of action of those feed additives are not entirely understood. They could act even with a direct effect on the immune system. A previously established animal-related in vitro system using primary cultured peripheral blood mononuclear cells (PBMCs) was applied to investigate the effects of immune-modulating feed additives. Here, the immunomodulation of different preparations of two probiotic Bacillus strains, B. subtilis DSM 32315 (BS), and B. amyloliquefaciens CECT 5940 (BA) was evaluated. The count of T-helper cells and activated T-helper cells increased after treatment in a ratio of 1:3 (PBMCs: Bacillus) with vital BS (CD4+: p < 0.05; CD4+CD25+: p < 0.01). Furthermore, vital BS enhanced the proliferation and activation of cytotoxic T cells (CD8+: p < 0.05; CD8+CD25+: p < 0.05). Cell-free probiotic culture supernatants of BS increased the count of activated T-helper cells (CD4+CD25+: p < 0.1). UV-inactivated BS increased the proportion of cytotoxic T cells significantly (CD8+: p < 0.01). Our results point towards a possible involvement of secreted factors of BS in T-helper cell activation and proliferation, whereas it stimulates cytotoxic T cells presumably through surface contact. We could not observe any effect on B cells after treatment with different preparations of BS. After treatment with vital BA in a ratio of 1:3 (PBMCs:Bacillus), the count of T-helper cells and activated T-helper cells increased (CD4+: p < 0.01; CD4+CD25+: p < 0.05). Cell-free probiotic culture supernatants of BA as well as UV-inactivated BA had no effect on T cell proliferation and activation. Furthermore, we found no effect of BA preparations on B cells. Overall, we demonstrate that the two different Bacillus strains enhanced T cell activation and proliferation, which points towards an immune-modulating effect of both strains on chicken immune cells in vitro. Therefore, we suggest that administering these probiotics can improve the cellular adaptive immune defense in chickens, thereby enabling the prevention and reduction of antimicrobials in chicken farming.

S3241 A Case of Autoimmune Hepatitis in the Setting of the Janssen COVID-19 Vaccine

Introduction: Autoimmune hepatitis (AIH) is an inflammatory autoimmune condition that can result in chronic liver disease, it can be caused by the interaction of hereditary, epigenetic, immune, and environmental triggers. Vaccine related AIH is an uncommon phenomenon that has been reported in the presence of other vaccinations.. We present a unique case of AIH in the setting of the Janssen SARS-CoV-2 vaccine (JJ). Case Description/Methods: A 45-year-old female with a history ofHTN, DM, GERD, and obesity presented from her primary care physician’s office with abnormal liver chemistries in a hepatocellular pattern. Her hepatic profile showed AST 1119, ALT 988, AP 130, and total bilirubin of 1.5 (R-factor 22.8). Given the magnitude and rapidity of the elevated chemistries a broad serologic workup was initiated. She was negative for hepatitis A, B, and C, as well as HSV 1/2, EBV, CMV, HIV, VZV, and adenovirus. Her ferritin and iron studies did not suggest iron overload. Her ceruloplasmin and alpha 1 antitrypsin levels were within acceptable ranges. Her ANA was negative; her smooth muscle antibody was positive (1:320), IgG was elevated to 3,451 mg/dL. Her urine toxin screen was negative. She had chronically been on HCTZ and took occasionally seamoss supplements. She denied using tylenol or any other over the counter or herbal supplements. In the past month she had received a single dose of the Janssen SARS CoV-2 vaccination (Johnson and Johnson). Her liver chemistries prior were AST 17, ALT 14, AP 52, and total bilirubin of 0.3. She subsequently underwent a liver biopsy which showed moderate (grade 3) lobular inflammatory activity, mild (grade 2) periportal activity, and mild (stage 2) fibrosis with an abundance of plasma cells consistent with autoimmune hepatitis. Treatment was initiated with oral prednisone with a resulting improvement in her liver chemistries (Figure). Discussion: Autoimmune hepatitis in the setting of recent administration COVID 19 vaccine administration has been rarely reported. We present a unique case of COVID vaccine induced autoimmune hepatitis from a non-mRNA based vaccine. It has been suggested that possible mechanisms may include molecular mimicry or bystander activation of dormant autoreactive T-helper cells for both tissue-specific and non-tissue-specific reactions. We aim to shed light to this interesting paradigm and to spark the scientific discourse to further delineate potential mechanisms leading to this uncommon complication.Figure 1.: Liver biopsy which showed moderate (grade 3) lobular inflammatory activity, mild (grade 2) periportal activity, and mild (stage 2) fibrosis with an abundance of plasma cells consistent with autoimmune hepatitis.

Assessment of CD4&lt;sup&gt;+&lt;/sup&gt; cells subpopulations with the expressing CD39 and CD73 ectonucleotidases in children with psoriasis

Purinergic signaling modulates systemic and local inflammatory responses in immune-mediated and autoimmune diseases, including psoriasis. Extracellular ATP is an important factor of purinergic regulation, and its levels are regulated by catalytic effects of CD39 and CD73 ectonucleotidases. The aim of the present study was to estimate the number of regulatory T cells (Tregs), activated T-helper cells (Thact), T-helper type 17 (Th17) expressing CD39 and CD73 ectonucleotidases in children with psoriasis vulgaris, depending on age, disease duration and severity of the pathological process. We have examined a total of 114 children with psoriasis vulgaris (70 girls and 44 boys) and 41 healthy children serving as a comparison group (25 girls and 16 boys). The age of children with psoriasis was 12.5 (10.1-15.8) years, and 12.4 (7.4-16.1) years for the comparison group. The severity of psoriasis was assessed by the PASI and BSA indices. The number of cells with CD39 and CD73 expression on Tregs, Thact and Th17 was estimated by flow cytofluorimetry. The highest number of CD39-expressing cells was found in the Tregs and CD73-expressing cells in Thact, both in children with psoriasis and in the comparison group. The number of CD39+Th17 was lower in children with psoriasis, but CD39+CD73+Thact and CD39+CD73+Th17 were higher than in comparison group (p &lt; 0.05). There was a decreased number of CD73+Tregs, CD39+Thact, CD39+Th17, CD39+CD73+Thact and CD39+CD73+Th17 with age in healthy children (p &lt; 0.05). In patients with psoriasis, the number of CD73+Th17 increased with age. A decrease in CD73+Th17, and an increase in CD39+CD73+ Tregs with higher PASI and BSA indices were detected. An increased PASI (&gt; 10) showed patients with both high and low CD39+Tregs, with CD39+Tregs being reduced in 48% of cases, increased in 35% and normal values in only 17% of cases. Monitoring the numbers of Tregs, Thact and Th17 cells expressing CD39 and CD73 in children with psoriasis may be used to evaluate chronic inflammation, given the role of CD39 and CD73 ectonucleotidases in shaping the immune response in immune-mediated diseases,

Dinamika Model Matematika Reaksi T-Helper

T cells are a major component of the human immune system. These T cells have a number that varies depending on the body's immune response when fighting bacteria or viruses. However, the condition of excess immune cells in the body can also be dangerous. Theoretical studies on the dynamics of T-Helper cells in the body are needed to get the right simulation in treating patients without conducting medical tests on every patient on a daily basis. This study discusses the dynamics of the mathematical model of the T-Helper reaction with the influence of antigen and IL-2. From this study, two equilibrium points were obtained, namely disease-free equilibrium and endemic equilibrium. The use of parameter values from the experimental results shows that the disease-free equilibrium point is locally unstable, while the endemic equilibrium point is locally stable. The numerical simulation showed that the antigen increased from 1st day to the highest value at 0.926 on the 11th day until on the 20th day it started to be constant towards at the value which is the antigen could be activate the resting T-Helper. The process of activating T-Helper, create IL-2 which can stimulating the proliferation and activity of T-Helper cells, so they can divide the activated cell of T-Helper into two memory cells.

Clinical significance of soluble interleukin-2 receptor measurement in patients with idiopathic retroperitoneal fibrosis.

Idiopathic retroperitoneal fibrosis (iRPF) is a rare chronic fibro-inflammatory disorder of unknown etiology. Activated T-helper cells, which shed soluble interleukin-2 receptor (sIL-2R) into the circulation, may play a pathogenetic role. Hence, measuring sIL-2R may be of value in monitoring disease activity and treatment response in iRPF patients. We performed a prospective inception cohort study of 82 patients with untreated (re)active iRPF stratified by elevated (> 623U/mL) or normal sIL-2R level at baseline and compared disease characteristics among these groups. Baseline and changes in sIL-2R levels following treatment with tamoxifen (TMX) or prednisone (PDN) were analyzed and related to treatment response. Median sIL-2R level was 668 U/mL (IQR 502.8-827.5); 48 patients (59%) had elevated baseline sIL-2R levels. Patients with elevated sIL-2R presented with higher CRP (P = 0.049) and serum creatinine (sCr) levels (P < 0.001) and more often had hydroureteronephrosis (P = 0.01). There was an age and sex adjusted linear association between baseline sIL-2R and both CRP (P = 0.02) and sCr (P < 0.001). Baseline and serial levels of sIL-2R were predictive and concordant, respectively, with clinical response in patients treated with PDN. ROC curve analyses of sIL-2R on a continuous scale and PDN treatment success showed an AUC of 0.84. A serum sIL-2R cut-off value for PDN treatment success of ≤ 703U/mL was found with a sensitivity of 100% and specificity of 72%. Serial measurement of sIL-2R may be of value in monitoring disease activity and PDN treatment response in iRPF patients.

Multi-scale Pan-cancer Integrative Analyses Identify the STAT3-VSIR Axis as a Key Immunosuppressive Mechanism in Head and Neck Cancer.

VSIR is a novel immune checkpoint protein whose expression on tumor cells across cancers remains largely uncharacterized. Here we purposed to decode the pan-cancer biologic and clinical significance of VSIR overexpression in the tumor compartment. We performed multi-omics integrative analyses of 9,735 tumor samples to identify cancers with non-leukocytic expression of VSIR (VSIR High), followed by association with overall survival and immune cell infiltration levels. Orthogonal assessments of VSIR protein expression and lymphocytic infiltration were performed using quantitative immunofluorescence (QIF). Integrative modeling identified a subset of cancer types as being enriched for VSIR High tumors. VSIR High tumors were associated with significantly poorer overall survival in immunogenic ovarian serous adenocarcinoma (SA) and oral cavity squamous cell carcinoma (SCC). QIF assessments in an independent validation cohort confirmed overexpression of VSIR as being associated with poorer overall survival within immunogenic oral cavity SCC. VSIR overexpression was associated with lower CD4 helper T-cell infiltration in both ovarian SA and oral cavity SCC, but did not impact CD8 T-cell infiltration. VSIR overexpressing tumors in both cancer types exhibited significantly higher STAT3 signaling activity. Pharmacologic inhibition of STAT3 signaling resulted in dose-dependent reduction of VSIR expression in ovarian SA and oral cavity SCC cells. The STAT3-VSIR axis is a potentially significant immunomodulatory mechanism in oral cavity and ovarian cancers, whose activation is associated with poorer survival and an immune microenvironment marked by decreased CD4 helper T-cell activity. The role of VSIR as a tumor-intrinsic modulator of resistance to immunotherapy warrants further exploration.

B-cell receptor dependent phagocytosis and presentation of particulate antigen by chronic lymphocytic leukemia cells.

Aim:T-helper cells could play an important role in the pathogenesis of chronic lymphocytic leukemia (CLL), a common B-cell neoplasm. Although CLL cells can present soluble antigens targeted from the B-cell receptor to T-helper cells via major histocompatibility complex (MHC) class II, antigens recognized by some CLL cells may be encountered in a particulate form. Here the ability of CLL cells to internalize and present anti-immunoglobulin M (IgM) beads as a model for the interaction of CLL cells with particulate antigens was investigated.Methods:The effect of anti-IgM beads on antigen presentation pathways was analyzed using RNA-seq and internalization of anti-IgM beads by primary CLL cells was investigated using confocal microscopy and flow cytometry. Antigen presentation was investigated by analyzing activation of a T-cell line expressing a T-cell receptor specific for a peptide derived from mouse κ light chains after incubating CLL cells with a mouse κ light chain-containing anti-IgM monoclonal antibody. Kinase inhibitors were used to characterize the pathways mediating internalization and antigen presentation.Results:Stimulation of surface IgM of CLL cells increased expression of the antigen presentation machinery and CLL cells were able to phagocytose anti-IgM beads. Internalization of anti-IgM beads was associated with MHC class II-restricted activation of cognate T-helper cells. Antigen presentation by CLL cells was dependent on activity of spleen tyrosine kinase (SYK) and phosphatidylinositol 3-kinase delta (PI3Kδ) but was unaffected by inhibitors of Bruton’s tyrosine kinase (BTK).Conclusions:CLL cells can internalize and present antigen from anti-IgM beads. This capacity of CLL cells may be particularly important for recruitment of T-cell help in vivo in response to particulate antigens.

S1965 B Adaptive: A Case of Rituximab-Associated Colitis and SARS-CoV-2 Vaccination

Introduction: Rituximab (RTX) is a monoclonal antibody against the CD20 surface antigen on B-Cell lymphocytes and is used in various hematologic and autoimmune conditions. Our case describes a patient on RTX therapy with pancolitis that coincided with SARS-CoV-2 vaccination (Moderna-mRNA1273). We attempt to explore a potential correlation of mRNA vaccination platforms, RTX therapy and new onset colitis. Case description/methods: A 49 year old male presented with 6 weeks of watery, non-bloody diarrhea, right upper quadrant pain, >10 pound weight loss and fever. History was significant for stage IVa follicular lymphoma in remission on RTX maintenance therapy, splenectomy and SARS-CoV-2 vaccination (Moderna mRNA-1273) 2 days prior to onset of diarrhea. Infectious work up was negative and CT imaging demonstrated diffuse colonic wall thickening with new interval adjacent pancolonic fat stranding. Initial colonoscopy demonstrated diffuse scattered clean based ulcers between 1-8 mm in size throughout the colon and terminal ileum. Biopsies revealed active chronic ileitis with eosinophils, occasional crypt abscess and crypt destruction negative for HSV, CMV or signs of recurrent lymphoma. He was then treated with oral budesonide, bentyl and loperamide but returned 6 weeks later without any relief in abdominal pain or diarrhea. Repeat colonoscopy demonstrated circumferential hyperemia/edema with apthous ulcers throughout the colon. We plan to treat as a Crohn's patient with corticosteroid induction and trial Vedolizumab for maintenance of remission. Discussion: The novel use of mRNA vaccines has been noted to be game changing in its production time as well as its effectiveness at preventing COVID-19 illness. The mechanism of immunity is proposed to be a combination of generation of neutralizing antibodies and T-helper cell activation. To our knowledge, mRNA-1273 has not directly caused colitis. Literature also describes occurrences of fulminant colitis in patients on RTX therapy and is believed to be secondary to dysregulation caused by depletion of CD20+ B cells and upregulation of autoreactive T-cells. While RTX induced colitis may arise de novo, there may be a correlation between the immunogenicity of mRNA vaccines like mRNA-1273 and the induction of colitis in patients while on immunosuppressants. This case demonstrates the increased need for further evaluation of the pathogenesis of RTX induced colitis as well as the immunogenic response seen in COVID-19 vaccination platforms.

Dietary Intake of n-3 PUFA-Enriched Hen Eggs Changes Inflammatory Markers' Concentration and Treg/Th17 Cells Distribution in Blood of Young Healthy Adults-A Randomised Study.

In the present study, we aimed to determine the effects of n-3 polyunsaturated acid (PUFA) supplementation (~1053 mg/per day), i.e., α-linolenic (~230 mg), eicosapentaenoic (~15 mg), and docosahexaenoic acid (~105 mg), through hen eggs, on pro- and anti-inflammatory parameters in healthy individuals (23.8 ± 2.57 years old). Here, we demonstrate differential effects of regular hen eggs (N = 21; W/M = 10/11) and n-3 PUFA-enriched hen eggs (N = 19; W/M = 10/9) consumption on the serum levels of lipid mediators, representation of peripheral T helper cell subsets (recently activated T-helper cells, nTreg, Th17 and non-Th17-IL-17A secreting T-helper lymphocytes) and their functional capacity for cytokine secretion. Both diets significantly altered systemic levels of pro-inflammatory and inflammation resolving lipid mediators; however, only the n-3 PUFAs group showed a significant shift towards anti-inflammatory prostanoids and increased levels of pro-resolving oxylipins. Both study groups showed reduced frequencies of peripheral nTreg lymphocytes and decreased rates of peripheral Th17 cells. Their functional capacity for cytokine secretion was significantly altered only in the n-3 PUFAs group in terms of increased transforming growth factor β-1 and reduced interleukin 6 secretion. Diet supplemented with n-3 PUFAs alters immune response towards inflammation resolving conditions through effects on lipid mediators and cytokine secretion by T lymphocytes in human model without underlying comorbidities.