Normal Immune System Research Articles

Improvement of the Immunity System Through Sports: Novel Regulatory Mechanisms for Hypertension.

Hypertension and its resulting target organ damage is a complex process associated with a range of physiological and molecular factors, including immune regulation. The profound effects of exercise on normal immune system function and the development and progression of hypertension are well known. This review aims to create new avenues for preventing and treating hypertension and its associated target organ damage. This narrative review emphasizes the role of exercise training in the prevention/treatment of hypertension development through immune response modulation and presents current perspectives on the available scientific evidence. Several studies have shown that exercise regulates hypertension by altering immune cells, which is partly attributable to the anti-inflammatory effects of exercise training. Regular exercise modifies immune modulation and could represent a new mechanism for regulating hypertension. Although the utilization of exercise training and the immune system in conjunction for treating and preventing hypertension is still in its early stages, current scientific literature indicates numerous potential physiological links between exercise training, the immune system, and hypertension.

Mechanism of formation and significance of antimitochondrial autoantibodies in the pathogenesis of primary biliary cholangitis

Primary biliary cholangitis (PBC) is a chronic cholestatic progressive liver disease associated with cholangiopathies. The detection of antimitochondrial autoantibodies (AMAs) plays an important role in the diagnosis of classical PBC. AMAs are formed against the antigenic component associated with the dihydrolipoyl transacetylase of pyruvate dehydrogenase complex (E2 PDC) localized on the inner membrane of mitochondria. The loss of immune tolerance of E2 PDC in PBC is thought to be the cause of the mechanism of AMA formation and immune-mediated destruction of biliary epithelial cells (BECs) of the small- and medium-sized intrahepatic bile ducts. E2 PDC is not only present in BECs, but is also abundant in the mitochondria of all nucleated cells. The question remains as to why E2 PDC of only small BECs is the target of autoimmune attack. There is no evidence that AMAs have a deleterious effect on BECs. New scientific data has emerged that explains the damage to BECs in PBC by the defect of the biliary bicarbonate (HCO3–) “umbrella” that protects BECs from the detergent action of bile acids under physiological conditions. Disruption of HCO3– production by BECs in PBC leads to changes in the pH of hepatic bile, accompanied by accumulation of bile acids in the small BECs. The detergent action of bile acids leads to damage of membrane structures of BECs and their apoptosis, development of ductulopenia, and intrahepatic cholestasis. For the first time, it has been suggested that under the influence of bile acids, the E2 PDC antigen may undergo conformational changes that alter its immunological properties. E2 PDC becomes a neoantigen that is recognized by the normal (“healthy”) immune system as a foreign antigen, leading to the production of AMAs. For the first time, the authors of this review provide an explanation for why only small BECs are damaged in PBC.

Immune-dysregulation harnessing in myeloid neoplasms.

Myeloid malignancies arise in bone marrow microenvironments and shape these microenvironments in favor of malignant development. Immune suppression is one of the most important stages in myeloid leukemia progression. Leukemic clone expansion and immune dysregulation occur simultaneously in bone marrow microenvironments. Complex interactions emerge between normal immune system elements and leukemic clones in the bone marrow. In recent years, researchers have identified several of these pathological interactions. For instance, recent works shows that the secretion of inflammatory cytokines such as tumor necrosis factor-α (TNF-α), from bone marrow stromal cells contributes to immune dysregulation and the selective proliferation of JAK2V617F+ clones in myeloproliferative neoplasms. Moreover, inflammasome activation and sterile inflammation result in inflamed microenvironments and the development of myelodysplastic syndromes. Additional immune dysregulations, such as exhaustion of T and NK cells, an increase in regulatory T cells, and impairments in antigen presentation are common findings in myeloid malignancies. In this review, we discuss the role of altered bone marrow microenvironments in the induction of immune dysregulations that accompany myeloid malignancies. We also consider both current and novel therapeutic strategies to restore normal immune system function in the context of myeloid malignancies.

The kynurenine pathway in patients with rheumatoid arthritis during tumor necrosis factor α inhibitors treatment.

The importance of the kynurenine pathway in normal immune system function has led to an appreciation of its possible contribution to autoimmune disorders such as rheumatoid arthritis (RA). The aim of the study was to evaluate the effect of treatment with tumor necrosis factor α (TNF-α) inhibitors on the activity of the kynurenine pathway in patients with RA. This was an investigator-initiated, prospective, observational study. The study was performed on 30 RA patients (Caucasian, 11 male, 19 female; mean age 45 ±16 years) treated with TNF-α inhibitors. All patients were assessed before and after 6 months of therapy. As a control group, age- and sex-matched, 20 healthy volunteers were recruited. Disease activity was evaluated by the Modified Disease Activity Score with 28-joint count (DAS28). Inflammatory markers were assessed routinely by the hospital central laboratory. Serum concentrations of kynurenine, serotonin and tryptophan were measured with specific immunoassays. To estimate indoleamine 2,3-dioxygenase (IDO) activity, kynurenine-to-tryptophan ratio was calculated. The results of our study showed changes in tryptophan metabolism in RA patients, compared with healthy controls. Surprisingly, RA patients had statistically significant decreased kynurenine-to-tryptophan ratio (p = 0.003), which could indicate diminished IDO activation in RA. Moreover, we found no significant changes in kynurenine-to-tryptophan ratio after treated with TNF-α inhibitors (p = 0.490), despite disease remission. Additionally, tryptophan metabolism activity did not correlate with objective markers of inflammation. The RA patients had altered tryptophan metabolism, compared with healthy controls. The mechanisms affecting tryptophan metabolism in RA may be complex. We believe that continuing elucidation of pathophysiological pathways relevant in RA offer substantial hope for the development of specific pharmacotherapy for treatment of RA - especially for comorbidity of RA and depression.

P-385 Personalized immunotherapy in RIF patients with different endometrial immune profiles

Abstract Study question Does immunotherapy effective in patients with recurrent implantation failure (RIF) according to the type of endometrial immune profile? Summary answer Immunotherapy based on the profile of the endometrial immune system has significant results in increasing the pregnancy rate and live birth rate. What is known already Personalized medicine in the area of infertility is in its early stages, and considering the promising results of personalization of treatment in other disorders, including cancer, it is expected to have acceptable results in infertility as well. Given that one of the key factors that affects the infertility of RIF patients is the imbalance of the endometrial immune system, efforts have been made to correct this disorder to increase the chances of fertility. Studies have classified RIF patients into different groups based on endometrial immune system profiles: Balanced, High, Low, and Mixed endometrial immune system activity. Study design, size, duration The study selected 267 RIF patients who were referred to Valiasr Hospital in Tabriz from Aug 2020-Sep 2023, and divided them into five groups: unclassified RIF patients (n = 89), patients with normal and balanced endometrial immune system profile (n = 63), and patients with high (n = 53), low (n = 38) and mixed (n = 24) profiles of endometrial immune system. Based on type of disorder identified, the appropriate immunotherapy was suggested for each group, including vaginal probiotics, lymphocyte therapy and Tacrolimus. Participants/materials, setting, methods The study classified RIF patients based on the endometrial immune system profiles, by the gene expression level of IL-18, IL-15, TWEAK, Fn-14, and CD56. The study included a group of unclassified RIF patients, who underwent routine IVF/ET cycles without immunotherapy. The Balanced group received no immunotherapy. For the Low endometrial immune group, probiotics and lymphocyte therapy were used. High group received probiotics and Tacrolimus, and for Mixed group, probiotics, Tacrolimus, and lymphocyte therapy were used. Main results and the role of chance The primary and secondary endpoints of this study were clinical pregnancy and delivery of a live born infant, respectively. The results showed that RIF patients without classification based on the profile of the endometrial immune system who went through the routine process of IVF/ET cycles had a 25.8% and 23.5% pregnancy and live birth rates, respectively. The pregnancy and live birth rates of the Balanced group was 28.5% and 25.3% (P=NS) while it was significantly higher in the High (56.6%, 54.7%), Low (47.3%, 44.7%) and Mixed groups (37.5%, 29.1%), respectively. According to the obtained results, it was found that the appropriate immunotherapy treatment based on the endometrial immune activity profiles in RIF patients has significantly increased the effective implantation rate in the IVF/ET cycles (P < 0.05). Limitations, reasons for caution It is necessary to examine a larger number of patients. Further work is needed to explore other parameters that are affected by endometrial immune system disorder and that can help classify RIF patients and recommend the most effective treatment. Wider implications of the findings For RIF patients, accurate identification of the cause of the disorder can be very effective for choosing the best treatment method. Personalized immunotherapy for RIF patients with immunologically abnormalities can lead to very promising and acceptable results while having few complications. Trial registration number Not Applicable

Methylation of T and B Lymphocytes in Autoimmune Rheumatic Diseases.

The role of abnormal epigenetic modifications, particularly DNA methylation, in the pathogenesis of autoimmune rheumatic diseases (ARDs) has garnered increasing attention. Lymphocyte dysfunction is a significant contributor to the pathogenesis of ARDs. Methylation is crucial for maintaining normal immune system function, and aberrant methylation can hinder lymphocyte differentiation, resulting in functional abnormalities that disrupt immune tolerance, leading to the excessive expression of inflammatory cytokines, thereby exacerbating the onset and progression of ARDs. Recent studies suggest that methylation-related factors have the potential to serve as biomarkers for monitoring the activity of ARDs. This review summarizes the current state of research on the impact of DNA and RNA methylation on the development, differentiation, and function of T and B cells and examines the progress of these epigenetic modifications in studies of six specific ARDs: systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, systemic sclerosis, juvenile idiopathic arthritis, and ankylosing spondylitis. Additionally, we propose that exploring the interplay between RNA methylation and DNA methylation may represent a novel direction for understanding the pathogenesis of ARDs and developing novel treatment strategies.

Abstract 4188: Kyinno PBMC humanized mouse platform: Pioneering service in biomedical research

Abstract Tumor immunotherapy is the use of the human immune system to kill tumors. Compared with traditional chemotherapy and radiotherapy, immunotherapy has significant advantages such as significant efficacy, long-lasting effects, and low toxicity. Wild-type mice are widely used as preclinical models for immunotherapy due to their normal immune system, but they have many shortcomings, such as only being able to inoculate mouse tumor cells, differences between mouse and human immune systems, etc., which cannot accurately simulate the interaction between drugs and tumor cells in the human body. Humanized mouse models include PBMCs humanized mouse models, HSC humanized mouse models, and human embryonic bone, liver, and thymus tissue (BLT) transplantation models. Among them, the PBMCs humanized mouse model is widely used due to its simple preparation, short cycle, and high levels of hCD3+ T cells in the peripheral blood of mice. To assist in the development of immunotherapy, KYINNO has established a PBMC humanized mouse platform that can perform in vivo drug efficacy verification of various immunotherapeutic drugs such as CD3 bispecific antibodies or multi-specific antibodies, immune checkpoint inhibitors, etc. We screen many PBMC donors and select PBMC donors with light GvHD and good hCD3+ T cell reconstruction effects. The drug administration window of the PBMC humanized model has been extended to 6 weeks. Platform verification has been performed on subcutaneous and intravenous injection models of various tumors. Overall, we successfully established the PBMC humanized mouse platform, which is a powerful tool for immunotherapy. Citation Format: Tongtong Liu, Guojin Wu, Feng He, Longtan Luo, Zhengcheng Guo, Shuliang Li, Jinying Ning, Feng Hao. Kyinno PBMC humanized mouse platform: Pioneering service in biomedical research [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4188.

Current insights into microbiome-based therapy. Review

Current insights into microbiome-based therapy. Review

Mitochondria-Targeted Nitronyl Nitroxide Radical Nanoparticles for Protection against Radiation-Induced Damage with Antioxidant Effects.

Radiotherapy is a non-invasive method that is widely applied to treat and alleviate cancers. However, radiation-induced effects in the immune system are associated with several side effects via an increase in oxidative stress and the inflammatory response. Therefore, it is imperative to develop effective clinical radiological protection strategies for the radiological protection of the normal organs and immune system in these patients. To explore more effective radioprotective agents with minimal toxicity, a mitochondria-targeted nitronyl nitroxide radical with a triphenylphosphine ion (TPP-NIT) was synthesized and its nanoparticles (NPs-TPP-NIT) were prepared and characterized. The TPP-NIT nanoparticles (NPs-TPP-NIT) were narrow in their size distribution and uniformly distributed; they showed good drug encapsulation efficiency and a low hemolysis rate (<3%). The protective effect of NPs-TPP-NIT against X-ray irradiation-induced oxidative damage was measured in vitro and in vivo. The results show that NPs-TPP-NIT were associated with no obvious cytotoxicity to L-02 cells when the concentration was below 1.5 × 10-2 mmol. NPs-TPP-NIT enhanced the survival rate of L-02 cells significantly under 2, 4, 6, and 8 Gy X-ray radiation exposure; the survival rate of mice was highest after 6 Gy X-ray irradiation. The results also show that NPs-TPP-NIT could increase superoxide dismutase (SOD) activity and decrease malondialdehyde (MDA) levels after the L-02 cells were exposed to 6.0 Gy of X-ray radiation. Moreover, NPs-TPP-NIT could significantly inhibit cell apoptosis. NPs-TPP-NIT significantly increased the mouse survival rate after irradiation. NPs-TPP-NIT displayed a marked ability to reduce the irradiation-induced depletion of red blood cells (RBCs), white blood cells (WBCs), and platelets (PLTs). These results demonstrate the feasibility of using NPs-TPP-NIT to provide protection from radiation-induced damage. In conclusion, this study revealed that NPs-TPP-NIT may be promising radioprotectors and could therefore be applied to protect healthy tissues and organs from radiation during the treatment of cancer with radiotherapy.

Development of a Fully Immunocompetent Adoptive Transfer In Vivo Model of Npm1 cA;Idh2 R140Q;Flt3 ITD AML

AML is a highly heterogeneous disease, both genetically and phenotypically. This includes both intra- and inter-patient heterogeneity that significantly impacts prognosis and treatment responses. Mouse models are a powerful tool to further our understanding of fundamental disease biology and to advance novel therapies. While human cell line (CDX) and primary patient (PDX) xenografts are reproducible in vivo models, these models are by necessity immunodeficient. Transgenic mouse models provide an opportunity to study disease development driven by human relevant mutations within a fully immunocompetent system. However, drug development studies often require large numbers of mice and predictable time of disease development that is infeasible using primary transgenic animals. We therefore sought to develop a transgenic mouse model representative of human disease that was able to consistently be adoptively transferred into fully immunocompetent recipients. Our laboratory has interest in multiple targeted therapeutics and chose to focus on a model that may have sensitivity to multiple translationally relevant targeted drugs. We crossed the previously established Npm1 cA (Vassiliou et al 2011), Flt3 ITD (Lee et al 2007) and Idh2 R140Q (Shih et al 2017) Cre-inducible alleles to generate a triple mutant AML model ( Npm1 cA;Idh2 R140Q;Flt3 ITD). This model develops an aggressive AML with a median survival of 44 days. Complete blood counts at endpoint demonstrate significantly elevated white cells as well as profound anemia and thrombocytopenia. Npm1 cA;Idh2 R140Q;Flt3 ITD mice also have splenomegaly and infiltration of immature myeloid cells into both lymphoid and non-lymphoid tissues. Flow cytometry analysis of the blood, spleen and bone marrow shows a significant expansion of CD11b+cKit+ cells characteristic of an AML blast phenotype. Given the aggressive nature of this model, we sought to determine whether Npm1 cA;Idh2 R140Q;Flt3 ITD AML cells could be adoptively transferred into either immunodeficient or fully immunocompetent recipients without prior preconditioning. NOD- Prkdc em26Cd52Il2rg em26Cd22/NjuCrl (NCG) and B6.SJL-Ptprca Pepcb/BoyJ (PepBoy) recipients were injected with 1x10 6 bulk splenocytes via the tail vein. Recipient mice quickly develop a lethal AML that faithfully recapitulates the disease observed in the primary transgenic donors. Circulating leukemia cells are detectable from approximately 2 weeks post adoptive transfer (post-AT) and median survival of PepBoy recipients ranged from 17 to 60 days depending on donor (Figure 1). Comprehensive immune profiling using a 36-specificity spectral flow panel showed that in PepBoy recipients, the donor derived cells accurately recapitulate the diversity of the AML immunophenotype of the primary transgenic donor. Notably, cells from male donors do not establish detectable disease in immunocompetent female recipients. This is associated with an increased number of T cells in the female recipients of male cells suggesting immune mediated control of AML development. To assess the response to targeted therapies in a fully immune competent model, we adoptively transferred of our Npm1 cA;Idh2 R140Q;Flt3 ITD to into PepBoy recipients. Mice were randomized to treatment groups based on peripheral blood disease burden at 14 days post AT (0.2-1% of CD45+ cells). Mice received daily oral gavage of either vehicle, 100 mg/kg venetoclax (BCL2 inhibitor), 100 mg/kg enasidenib (IDH2 inhibitor) or 30 mg/kg gilteritinib (FLT3 inhibitor). All three drugs resulted in a significant reduction in circulating tumor burden after 14 days of treatment. Enasidenib did not prolong survival compared to vehicle whereas venetoclax treatment resulted in a significant survival advantage (median 47 days vs 40 in the vehicle). This model was most sensitive to gilteritinib with a median survival of 68 days. (Figure 2). Together these findings demonstrated that the Npm1cA;Idh2 R140Q;Flt3 ITD model is an aggressive model of AML. Adoptive transfer of splenocytes into either immunodeficient NCG or fully immunocompetent PepBoy recipients faithfully recapitulates the disease of primary mice in a reproducible manner. This model therefore provides an in vivo model conducive to drug development studies and provides the opportunity to understand the impact of both AML development and therapeutics on the normal immune system.

Effects of Bovine Lactoferrin on the Maintenance of Respiratory and Systemic Physical Conditions in Healthy Adults-A Randomized, Double-Blind, Placebo-Controlled Trial.

We investigated the effects of bovine lactoferrin (LF) on the maintenance of the respiratory and systemic physical conditions. A randomized, double-blind, placebo-controlled trial was conducted. Healthy adults at Kyushu University of Health and Welfare ingested a placebo or bovine LF (200 mg/day) for 12 weeks. The primary endpoints were the total respiratory and systemic symptom scores. The secondary endpoint was the activity of plasmacytoid dendritic cells (pDCs) in peripheral blood. A total of 157 subjects were randomized (placebo, n = 79; LF, n = 78), of whom, 12 dropped out. The remaining 145 participants were included in the full analysis set (placebo group, n = 77; LF group, n = 68). The total scores for respiratory and systemic symptoms during the intervention were significantly lower in the LF group than in the placebo group. The expression of CD86 and HLA-DR on pDCs was significantly higher in the LF group than in the placebo group at week 12. Adverse events were comparable between the groups, and no adverse drug reactions were observed. These results suggest that orally ingested LF supports the normal immune system via maintaining pDC activity, and maintains respiratory and systemic physical conditions in healthy adults.

Tumour induction in BALB/c mice for imaging studies: An improved protocol.

Dealing with nude mice, which lack thymus and therefore are sensitive to unsterile conditions, needs special care and laboratory conditions. For preclinical studies, especially tumour imaging purposes, in which therapeutic properties of drugs or therapeutic compounds are not studied, mice with normal immune system can be a favourable alternative if they carry tumours of interest. In the current study, we introduce an optimized protocol for induction of human tumours in BALB/c mice for preclinical studies. Immune system of BALB/c mice was suppressed by administration of cyclosporine A (CsA), ketoconazole and cyclophosphamide. The tumours of MDA-MB-231, A-431 and U-87-MG human cancer cells were induced by subcutaneous injection of the cells to the immunosuppressed mice. Tumour size was calculated weekly. Histopathological and metastatic analyses were performed using haematoxylin and eosin staining. The combination of the three drugs was found to suppress immune system and decrease the numbers of white blood cells, including lymphocytes. At the eighth week, tumours with a dimension of approximately 1400 mm3 developed. Large atypical nuclei with scant cytoplasm were found to exist using histopathological analysis. No metastasis was observed in the tumour-bearing mice. A combination of CsA, ketoconazole and cyclophosphamide can be used to suppress the immune system in BALB/c mice and induce tumours with significant size.

Intercellular Adhesion Molecule 1: More than a Leukocyte Adhesion Molecule.

Intercellular adhesion molecule 1 (ICAM-1) is a transmembrane protein in the immunoglobulin superfamily expressed on the surface of multiple cell populations and upregulated by inflammatory stimuli. It mediates cellular adhesive interactions by binding to the β2 integrins macrophage antigen 1 and leukocyte function-associated antigen 1, as well as other ligands. It has important roles in the immune system, including in leukocyte adhesion to the endothelium and transendothelial migration, and at the immunological synapse formed between lymphocytes and antigen-presenting cells. ICAM-1 has also been implicated in the pathophysiology of diverse diseases from cardiovascular diseases to autoimmune disorders, certain infections, and cancer. In this review, we summarize the current understanding of the structure and regulation of the ICAM1 gene and the ICAM-1 protein. We discuss the roles of ICAM-1 in the normal immune system and a selection of diseases to highlight the breadth and often double-edged nature of its functions. Finally, we discuss current therapeutics and opportunities for advancements.

83 Assessing Exposure to Fungal Bioaerosols in Transport Environments: Analysing Fungal Composition of Passive Dust Samples Collected in UK Railway Stations

Abstract Bioaerosols consist of a complex mixture of airborne microorganisms including fungi, bacteria, pollen, particulates and by-products of cells. While exposure to diverse microorganisms is essential for normal immune system development, bioaerosol inhalation has been associated with respiratory allergy and inflammation. Bioaerosols are ubiquitous, yet their composition within different environments is not well understood. Such information is essential for assessing exposure and associated health impacts. Here we focus on transport environments, namely railway stations (RSs), which pose a potential source of occupational and community exposure. Over 1200 passive dust samples were taken from 17 RSs across the UK from 03/2014-05/2015. Geographical Information System methods were used to provide information on population characteristics of the immediate surrounding area of the RSs. Together with information on RS layout and passenger numbers, this informed the selection of 250 samples, representing 9 RSs of varied geographical location, layout and footfall for further analysis. High throughput sequencing (HTS) with a metabarcoding approach targeting the ITS2 region was used to analyse the fungal composition of the sample subset. Such HTS techniques provide an opportunity to measure a wider range of microorganisms than traditional culture or microscopy techniques. RS characteristics were varied and included indoor/enclosed and mostly outdoor layouts. Annual footfall varied from approximately 10-148 million. The fungal composition of RS over time and relationships with different RS characteristics will be presented, identifying those driving any differences in composition.

Abstract 21: A novel bladder cancer syngeneic mouse model using gene-edited organoids derived from bladder urothelium

Abstract In recent years, immuno-checkpoint inhibitors have expanded the range of treatment options in advanced bladder cancer, but there are still many issues to be overcome, such as drug resistance, effect of combination therapy and prediction of adverse events. To address these issues, there is an urgent need for useful animal models with normal immune system. About half of the human muscle invasive bladder cancers (MIBCs) show p53 mutations in which over 60% are missense mutations. Additionally, the downregulation of tumor suppressor gene PTEN is known in human MIBC. We also investigated cell of origin of chemical-induced mouse bladder cancer using lineage tracing method and found that Krt5 expressing cells with Trp53 alteration efficiently give a rise of high-grade MIBC with squamous differentiation. Based on these results, we have been working on establishing MIBC mouse model using genetically engineered mouse (GEM) in which Cas9 and target gene mutation express specifically in Krt5 expressing cells by Cre-LoxP system. We have also used CRISPR/Cas9 gene editing technique and 3D organoid culture system. We generated organoids from the GEM bladder urothelium with Krt5CreERT2/+; LSL-Cas9; LSL-Trp53R172H/+. For further gene editing, we infected adeno-associated virus with sgRNA to the organoids. The organoid with Trp53R172H/+ and Pten knock-out showed tumorigenicity in immunodeficient mice. The organoid-derived tumors are histologically similar with the basal-squamous subtype of human bladder cancer. Moreover, these tumors exhibited wild type (WT) Trp53 loss of heterozygosity in genomic level. Similarly, we confirmed tumorigenicity in immunodeficient mice with organoids from Trp53flox/flox; Ptenflox/flox and Trp53R172H/flox; Ptenflox/flox mice, but not from Trp53R172H/flox;Pten+/+ mice or Trp53R172H/+; Ptenflox/flox mice, suggesting that both loss of WT Trp53 and Pten loss are required for tumor formation in this setting. RNA sequencing showed Pten loss is associated with up-regulation of metabolism related gene sets and loss of WT Trp53 is associated with up-regulation of proliferation related gene sets. Furthermore, Trp53R172H/flox; Ptenflox/flox organoids showed higher engraftment rate in immunocompetent mice with less infiltration of CD8+ cells compared with Trp53flox/flox; Ptenflox/flox organoids, suggesting that gain of function of mutant Trp53 may affect immune microenvironment. Our data illustrates the usefulness of clinically relevant syngeneic bladder cancer mouse model from GEM-derived organoids for the research of tumor immunity. Citation Format: Akihiro Hamada, Yuki Kita, Takeshi Sano, Ryosuke Ikeuchi, Hideaki Takada, Kenji Nakamura, Toru Sakatani, Takayuki Goto, Atsuro Sawada, Shusuke Akamatsu, Takashi Kobayashi. A novel bladder cancer syngeneic mouse model using gene-edited organoids derived from bladder urothelium [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 21.

Microbiome Contributions to Health.

The human microbiome is vast and is present in spaces previously thought to be sterile such as the lungs. A healthy microbiome is diverse and functions in an adaptive way to support local as well as organism health and function. Furthermore, a normal microbiome is essential for normal immune system development rendering the array of microbes that live in and on the human body key components of homeostasis. A wide array of clinical conditions and interventions including anesthesia, analgesia, and surgical intervention may derange the human microbiome in a maladaptive fashion with bacterial responses spanning decreased diversity to transformation to a pathogenic phenotype. Herein, we explore the normal microbiome of the skin, gastrointestinal tract, and the lungs as prototype sites to describe the influence of the microbiomes in each of those locations on health, and how care may derange those relations.

Combined Cutaneous &amp; Visceral Forms of Leishmaniasis; Case Report

This is a three years old child from Himreen rural areas the patient had fever, pallor, &amp; hepatosplenomegaly and two ulcers on the face. The patient responded to Pentostam. While we used in Iraq to see leishmaniasis as either pure cutaneous or pure visceral forms of leishmaniasis, this is a case with combined cutaneous &amp; visceral forms of leishmaniasis in a patient with normal immune system. Which may explained by the leishmaniae species that primarily cause cutaneous disease &amp; rarely cause visceral disease or by primarily viscerotropic leishma'nia that primarily cause visceral disease which may also cause skin lesions or by mixed infections by two different species of leishmaniae, one of them is dermatotropic, &amp; the other one is viscerotropic.

At low levels, inorganic mercury interference with antigen signaling is associated with modifications to a panel of novel phosphoserine sites in B cell receptor pathway proteins

Epidemiological studies indicate that human and animal exposure to environmental mercury (Hg) disrupts normal immune system function, but the molecular mechanism responsible for this is still unresolved. We have previously utilized phospho-proteomic mass spectrometry to demonstrate that in the absence of B Cell Receptor (BCR) stimulation, exposure of B cells to Hg induces significant changes to a great many elements of the BCR signaling pathway in a concentration dependent manner. In this report, we have extended those initial findings by utilizing mass spectrometry to evaluate in detail the effect of low-level Hg exposure on BCR induced phospho-proteomic changes. Specifically, murine WEHI-231 B lymphoma cells were exposed to environmentally relevant levels of Hg with or without concomitant BCR stimulation. The cellular phospho-proteomes were then profiled by LC-MS/MS. We found that for low-level exposures, Hg interference with signal transduction across the BCR pathway was predominantly associated with modification of phosphorylation of 12 phosphosites located on seven different proteins. Nine sites were serine, two sites tyrosine and one site threonine. Most of these sites are novel, in the sense that only the two tyrosine and one of the serine sites have previously been reported to be associated with BCR signaling.

AN OVERVIEW OF SECONDARY IMMUNODEFICIENCY STATES IN CHILDREN ADMITTED TO PAEDIATRIC INTENSIVE CARE UNIT IN A TERTIARY CARE HOSPITAL IN GUNTUR DISTRICT OF ANDHRA PRADESH

Objective: To review the different causes of secondary immunodeciencies in children requiring prolonged hospital stay or recurrent hospital admissions and provide clinicians with an updated overview of potential factors that contribute to immunodeciency in a tertiary care hospital over 18 months. Background Immunocompromised status is the common cause of morbidity and mortality in hospital-admitted children. Secondary immunodeciency contributes to most of the immunocompromised states rather than primary, resulting from a variety of extrinsic factors that can affect a host with an intrinsically normal immune system, including undernutrition, infectious agents, drugs, metabolic diseases, and environmental conditions. In routine clinical practice, these arise from a number of conditions, such as treatment with glucocorticoids and immuno-modulatory drugs, surgery and trauma, extreme environmental conditions, and chronic infections, such as those caused by HIV. The most common cause of immunodeciency is malnutrition, affecting many communities around the world with restricted access to food resources [1]. Methods This is a retrospective observational study using hospital records of children aged 1 month to 12 years, admitted to PICU from January 2021 to August 2022 in a tertiary care hospital in the Guntur district of Andhra Pradesh, India, who were underweight, or on some immunosuppressive agents and those who had hospital stay lasting more than 14 days and who needed &gt;14 days of intravenous antibiotic. Results In children less than 1 year, protein energy malnutrition especially SAM is the most common cause of secondary immunodeciency. Among children older than 1 year, children using steroids for nephrotic syndrome constitute the major cause. As secondary immunodeciency Conclusion disorders are far more common than primary immunodeciency disorders. PEM is the most common cause in children less than 1 year, Exclusive breastfeeding should be advised to all children, and attention to nutritional supplementation should be given as early as possible. Even brief exposure to steroids may land children into a state of immunosuppression, so judicious use of steroids is mandatory and regular follow-up is necessary.

Determination of Genotypes in Cytomegalovirus (CMV) Strains Obtained from Pediatric and Adult Immunocompromised Patients

Background and Purpose: Cytomegalovirus causes asymptomatic disease in individuals with normal immune system; and leads to serious complications in immunocompromised individuals and fetus. In CMV, gB is the most studied glycoprotein in terms of genotyping. Up to now, four different gB genotypes (gB 1-4) of CMV have been identified. &#x0D; In this study, it was aimed to determine the genotypes of CMV strains obtained from patients with immune deficiency.&#x0D; Methods: Twenty children and 29 adults, 49 patients who were followed in the Department of Adult Hematology and Pediatric Hematology were included in the study. DNA isolation was performed from samples with CMV DNA level of 1000 IU / ml and above, and 474 bp region from the gB region of the virus was amplified by nested PCR. This region was sequenced by the Sanger (ABI 3500 Prism) sequencing. Next generation sequencing (NGS) method was applied to the samples that CMV genotype could not be determined by Sanger sequencing.&#x0D; Results: Distribution of CMV genotypes of patients determined by Sanger sequencing; while it was determined as 18/49 (36.7%) type 1, 5/49 (10.2%) type 2, 5/49 (10.2%) type 3 and 1/49 (2%) type 4; 14/49 (28.5%) of them were found as mixed genotypes. CMV genotype could not be determined in 6 patients by Sanger sequencing and CMV genotype of these 6 patients were found as mixed genoype by NGS. Mixed genotype was detected in 20 (40.9%) of 49 patients, in total by Sanger sequencing and NGS. &#x0D; Conclusion: It is remarkable that the most common genotype is mixed genotype. It was concluded that it is appropriate to study the next generation sequencing method in patients whose CMV genotype cannot be determined by Sanger sequencing.