TLR-5 influences T cell activity: implications for development of Type 1 Diabetes

TLR-5 influences T cell activity: implications for development of Type 1 Diabetes | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article TLR-5 influences T cell activity: implications for development of Type 1 Diabetes Karsten Buschard, Lars Krogvold, Ivan Gerling, Knut Dahl-Jørgensen, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5782494/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract In mammalian and human life, one of the most important problems is to choose the best defence against microorganisms. Innate cells are good against bacteria, and T cells against virus mainly because of antibody production via T helper and B lymphocytes. Toll-like receptor 5 (TLR-5) is a regulator of this choice; when it is highly expressed, T cells are inhibited, and innate cells are favored. In activated pancreatic beta cells, TLR-5 has been found highly expressed and may therefore be protected from T cell destruction e.g., during pregnancy. We investigated mRNA from islets of Langerhans from newly diagnosed T1D patients for TLR-5. Also, we examined for polymorphisms between TLR-5, metabolic parameters, and Type 1 Diabetes (T1D). mRNA for TLR-5 was downregulated by one-third in the newly diagnosed T1D patients compared to controls. Regarding polymorphisms, two associations were found between TLR-5 and monocytes which are cells from the innate immune system. Also, a significant polymorphism was seen concerning TLR-5 and T1D. T cells are important for autoimmune diseases including T1D. In the present study we find low values of mRNA of TLR-5 which enhance T cells. We hope that the findings mentioned in this article may be influential for the understanding of how T1D develops. Health sciences/Endocrinology/Endocrine system and metabolic diseases/Diabetes/Type 1 diabetes mellitus Biological sciences/Immunology/Autoimmunity Biological sciences/Cell biology/Mechanisms of disease TLR-5 Type 1 Diabetes Islets of Langerhans Polymorphisms Autoimmunity T cells. Figures Figure 1 Figure 2 INTRODUCTION Mammalian organisms possess both the innate and the adaptive immune system. Therefore, in each situation with an infection, it is a choice whether it should react with granulocytes or lymphocytes, or both. Bacteria may be fought best with the innate immune cells, virus best with antibodies via T cell stimulation, and cancer by T cells, which on the other hand give risk of autoimmunity. Toll-like receptor 5 (TLR-5) is a crucial component of the innate immune system, primarily stimulated by bacterial flagellin 1 . In 2011, we found that TLR-5 is the only TLR molecule which is upregulated on beta cells following glucose stimulation 2 . Initially, the purpose of this upregulation was unclear, but it was hypothesized to be a defence mechanism for beta cells, which are more vulnerable when being active 3 , 4 . It has now been shown that high TLR-5 boosts the innate immune system and granulocyte activity 5 . In contrast, low TLR-5 expression stimulates T cells and promotes autoimmunity; in mice, TLR-5 deficiency has been linked to increase risk of developing metabolic diabetes 6 . Also, TLR-5 shortage, exacerbates the autoimmune lupus-like disease 7 . In the present study, we aimed to examine TLR-5 expression in the islets of Langerhans in newly diagnosed Type 1 Diabetes (T1D) patients. Additionally, possible polymorphisms related to the TLR-5 system were investigated. RESULTS In newly diagnosed T1D patients, the mRNA level of TLR-5 in the islets of Langerhans was significantly reduced by one-third compared to control individuals (34.1 ± 17.8 vs 50.5 ± 11.2, p = 0.019). Notably, one outlier patient, the oldest in the group, showed a TLR-5 expression of 64.0. Excluding this patient, the statistics revealed an even more pronounced decrease in TLR-5 expression for the newly diagnosed T1D patients, showing a 48% reduction compared to controls (26.6 ± 6.2 vs 50.5 ± 11.2, p = 0.0012). As it can be seen from Fig. 1 , the prediabetic antibody-positive persons do have the same level of TLR-5 mRNA as the controls and tended to be higher than the DiViD patients. Hence the reduction in TLR-5 occurs when the insulitis become malignant and clinical diabetes arrives. Interestingly, the T1D patients with disease duration of 5 years still tend to have lower TLR-5 values than the controls (p = 0.09). There was found no correlation between TLR-5 and C-peptide values for the antibody-positive persons nor for the DiViD patients. Four of the antibody-positive persons had two autoantibodies, which were either GAD, IAA, IA2 or Zink transporter antibodies, and their TLR-5 values were 52.0 ± 12.1. The other eight antibody-positive persons had GAD antibodies only and their TLR-5 values were 47.4 ± 11.1. Hence, no difference between these subgroups. The insulitis score for the DiViD patients has earlier been reported 8 . In Fig. 2 , the TLR-5 values are plotted against the numbers of CD3 T cells per islets adjusted for size. Interestingly, there was a tendency (p = 0.08) for a negative correlation showing the higher TLR-5, the lower T cell numbers. The analysis of polymorphisms associated with the TLR-5 system is presented in Table 1 ; one single nucleotide polymorphism (SNP) was correlated with T1D. Interestingly, the same SNP (rs113292043) and rs5744168 were highly significantly associated with monocyte and neutrophil counts, which are key cells of the innate immune system. DISCUSSION In the present study, we have demonstrated an association between TLR-5 expression in the islets of Langerhans and development of T1D. Additionally, we identified a polymorphism related to T1D. Gram-negative bacteria with flagellin stimulate TLR-5, which enhances granulocyte activity to eliminate these bacteria 9 , 10 . In contrast, this is not an obvious job for T cells which are present to react against tumor cells and virus, and to produce antibodies via T helper and B lymphocytes. The identified polymorphisms between TLR-5 and monocyte count are interesting since monocytes are crucial components of the innate immune system. Monocytes’ primary role in the innate immune system is to phagocytose and destroy infectious agents 11 . New virus infections lead to T cell activation, but interestingly, repeated viral infections increase TLR-5 expression 12 . This might be because memory B lymphocytes are already primed, reducing the need for new T cell activation. Thus, for T1D repeated viral infections might offer protection and there might be no pressure of lower TLR-5 expression; therefore, early-life exposure to viruses can have a beneficial impact on the immune system. Interestingly, a study found a reduced incidence of T1D among children attending pre-school and daycare, likely due to increased exposure to repeated virus infections 13 . The development of T1D requires islet destruction by T cells; this is supported by many studies, e.g., virus-induced diabetes in mice is not occurring in nude mice that lack the thymus-dependent immune system, showing that virus is nontoxic and acts through T cells 14 . Also, treatment with CD3 antibodies are beneficial by inducing regulatory T cells and approved by FDA for delaying progression from stage 2 to stage 3 (overt) T1D 15 . Activation of TLR molecules other than TLR-5 does not seem to inhibit autoimmunity 16 – 19 . During early pregnancy, TLR-5 expression increases 20 . This may take place in order not to destroy paternal antigens including the fetus. The higher expression of TLR-5 during high activity of beta cells can be explained to avoid development of T1D in pregnant women since during (late) pregnancy beta cells produce 40% more insulin. However, there is a 3.8-times increased risk of developing T1D during the last trimester of pregnancy, but not more despite the pressure on the beta cells 21 . To have activated T cells has its price: the higher risk of autoimmunity, but maybe also lower longevity. This is seen in mice in which Flagellin has been used in a study showing higher longevity in mice after intake of this compound 22 . Like TLR-5 the sphingolipid, sulfatide, inhibits T cells 23 , 24 . In a human epidemiological study, we have measured sulfatide in peripheral blood and compared this to longevity 25 . The individuals with the highest content of sulfatide have the lowest mortality. This was significant (0.86, range 0.74–0.99, p = 0.036) dependent on the degree of correction. Thus, also in humans, there is a trend to better longevity when T cells are inhibited. Experience with flagellin due to low hygiene standard seems natural in Russian Karelia, in poor countries, and in the 1950’s including east Germany 26 . These places also have low incidence of T1D 27 . Interestingly, the hygiene theory is all about exposure to activated bacteria which then according to the above is the mechanism for less autoimmunity 28 . Polymorphisms of the TLR5 gene have been associated with innate immune responses in other inflammatory diseases, including SLE 29 . Notably, the rs5744168 SNP of TLR5 generates a stop codon that could affect TLR-5 function 30 , whereas rs113292043 is an intergenic variant with no know functional consequences. The polymorphisms correlate with reduced number of monocytes and neutrophils, and increased risk of T1D. A consequence of the present study is that T1D can develop due to an unfavourable choice of immunological defence by T cells which dispose to an autoimmune reaction. A parallel to serious or deadly COVID-19 infection may be in play. It starts classically as a virus infection with T cells and antibodies, but dramatic autoimmunity can be initiated in the lungs, and steroid or other anti-autoimmune treatment may be instituted (too late) 31 , 32 . Regarding T1D, a treatment strategy could involve flagellin to increase TLR-5 expression, and thereby naturally inhibiting T cells and potentially avoiding autoimmunity. METHODS RNA analysis Human pancreatic tissue utilized in this study was obtained from the DiViD study 33 from newly onset (disease duration 35 days) T1D patients aged 24–35 years (n = 5), and from the nPOD study 34 with a healthy control group (n = 18). Direct to [email protected] for requests to the datasets. The nPOD study consists of pancreases from organ donors following accidental death, as opposed to the living patients in the DiViD study. The nPOD study samples consists of 18 control persons (9F/9M) aged 36.2 ± 15.6 (mean ± SD) years old; 12 antibody-positive persons (5F/7M) aged 20.4 ± 8.4 years old; 19 persons with intermedium diagnosed T1D (10F/9M) aged 33.2 ± 17 years old; and finally, 8 persons with T2D (5F/3M) aged 39.8 ± 13.4 years. Frozen sections of 25 randomly selected pancreatic islets were subjected to laser capture dissection 35 and islets from two to five sections were pooled, and RNA extraction was done using the Arcturus Pico Pure RNA Isolation Kit (Applied Biosystems, Grand Island, NY, USA). RNA quantification was performed using a Bioanalyzer 2100 instrument (Agilent Technologies, Santa Clara, CA, USA). Gene expression analysis was conducted using Affymetrix expression arrays (Gene Chip Human Gene 2.0 ST, Thermo Fisher) and normalized using global scaling 36 . All tissue handling procedures was conducted in the same laboratory, to ensure consistency in the comparison. Furthermore, the analysis was performed using same equipment, and the RNA quality was certified by RIN value measurements (> 3.5). Genetic association analysis GWAS data were retrieved from 37 – 44 , and associations analysed for TLR-5 with innate immune cell numbers and T1D defined by ADA criteria 45 . Blood glucose levels were evaluated in individuals free of diabetes defined by physician diagnosis, medication use, or fasting glucose < 7 mmol/L 46 . Statistical analysis The statistical analysis for RNA expression was conducted using GraphPad Prism 8.0.2 (GraphPad, La Jolla, Ca, USA). For comparison between groups, a one-way ANOVA was employed followed by Dunnett’s multiple comparison test and a 95% CI. Statistical significance was considered when p < 0.05. Ethics statement The DiViD and nPOD studies were approved by The Norwegian government’s regional ethics committee (reference 2009/1907) and by the University of Tennessee Health Science Center’s local institutional review board (reference 10-00848-XM). All methods were carried out in accordance with relevant guidelines and regulations. We confirm that all experimental protocols were approved by The Norwegian government’s regional ethics committee (reference 2009/1907) and by the University of Tennessee Health Science Center’s local institutional review board (reference 10-00848-XM). Finally, we confirm that informed consent was obtained from all subjects and/or their legal guardians. Declarations COMPETING INTERESTS The authors declare no competing interests. Author Contribution Karsten Buschard conceptualized the project and wrote the original manuscript draft. Lars Krogvold, Knut Dahl-Jørgensen, and Ivan Gerling provided the analysed material and performed the RNA expression analysis. Flemming Pociot performed the SNP analysis. All authors edited, reviewed, and approved the final manuscript. Data Availability Data is being deposit with datadryad.org. The protocols used can be obtained upon request to the corresponding author. References Feuillet, V. et al. 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Trans-ethnic and Ancestry-Specific Blood-Cell Genetics in 746,667 Individuals from 5 Global Populations. Cell 182 , 1198-1213.e14 (2020). Table 1 Table 1. Shows polymorphism related to TLR-5. Lead Variant (CHR:POS:REF:ALT) dbSNP Phenotype P-Value 1:223285200:G:A rs5744168 Monocyte percentage Monocyte count Neutrophil percentage Neutrophil count 1.77e-16 3.21e-12 1.40e-4 6.83e-3 40,47 1:223231657:C:T rs113292043 Monocyte count Monocyte percentage Type 1 diabetes 1.21e-14 1.24e-14 1.3e-2 41,47,48 42–44 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5782494","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":404483034,"identity":"5024f774-1b0e-4b7a-abad-2265e9a79c71","order_by":0,"name":"Karsten Buschard","email":"data:image/png;base64,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","orcid":"","institution":"Rigshospitalet","correspondingAuthor":true,"prefix":"","firstName":"Karsten","middleName":"","lastName":"Buschard","suffix":""},{"id":404483035,"identity":"19a5d0bf-7ae6-49a9-bbf2-00c1380b303a","order_by":1,"name":"Lars Krogvold","email":"","orcid":"","institution":"Oslo University Hospital","correspondingAuthor":false,"prefix":"","firstName":"Lars","middleName":"","lastName":"Krogvold","suffix":""},{"id":404483036,"identity":"772ce6e1-118e-487e-994a-969901f37e39","order_by":2,"name":"Ivan Gerling","email":"","orcid":"","institution":"University of Tennessee","correspondingAuthor":false,"prefix":"","firstName":"Ivan","middleName":"","lastName":"Gerling","suffix":""},{"id":404483037,"identity":"b1985918-f2ef-4cf9-b4d3-21dfbb596c7e","order_by":3,"name":"Knut Dahl-Jørgensen","email":"","orcid":"","institution":"Oslo University Hospital","correspondingAuthor":false,"prefix":"","firstName":"Knut","middleName":"","lastName":"Dahl-Jørgensen","suffix":""},{"id":404483038,"identity":"c9bf41be-bd89-4092-aa54-1f619564b25c","order_by":4,"name":"Flemming Pociot","email":"","orcid":"","institution":"Steno Diabetes Center","correspondingAuthor":false,"prefix":"","firstName":"Flemming","middleName":"","lastName":"Pociot","suffix":""},{"id":404483039,"identity":"7b4af3f0-1be9-493f-becb-f3999e88ca73","order_by":5,"name":"Camilla Hartmann Friis Hansen","email":"","orcid":"","institution":"University of Copenhagen","correspondingAuthor":false,"prefix":"","firstName":"Camilla","middleName":"Hartmann Friis","lastName":"Hansen","suffix":""}],"badges":[],"createdAt":"2025-01-07 15:08:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5782494/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5782494/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":74436068,"identity":"225c4d99-2a1b-48e8-a840-f898934f8fa7","added_by":"auto","created_at":"2025-01-22 09:24:18","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":29932,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eToll-like receptor 5 \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e(\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eTLR-5) gene expression\u003c/strong\u003e. CTR: non-diabetic controls (n = 18); AB+: non-diabetic autoimmune antibody-positive donors, nPOD (n = 12); T1D (median disease duration, 35 days): donors with newly diagnosed type 1 diabetes, DiViD (n = 5); T1D (median 5 years): donors with intermedium diagnosed type 1 diabetes, nPOD (n = 20); T2D (median 2 years): donors with type 2 diabetes, nPOD (n = 8); T1D Boxes indicate 25% and 75% quartiles and whiskers 1.5 × interquartile ranges. TLR-5 expression significantly reduced in newly diagnosed T1D patients by one-third compared to control individuals (p=0.019), and 48% reduction without outlier (p=0.0012).\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5782494/v1/49634593ed3ec9d7f83b1ad2.jpg"},{"id":74436081,"identity":"fc532f11-8a3d-4e30-a10b-37709b66e409","added_by":"auto","created_at":"2025-01-22 09:24:18","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":24589,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAssociation between islet TLR5 expression and T lymphocyte numbers. \u003c/strong\u003eNonlinear regression analysis (Spearman) of islet TLR5 gene expression in donors with newly diagnosed Type 1 Diabetes, DiViD (n = 5), and intra-islet CD3+ cells. Each islet is normalized to the area of an IEQ, defined as an islet with a diameter of 150 μm or an average cross-sectional area of 0.010936 mm2.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5782494/v1/12989850a1fc1645c6026152.jpg"},{"id":77737043,"identity":"66e74bef-5e4d-4b29-a980-10b09ab1f732","added_by":"auto","created_at":"2025-03-05 03:23:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":616827,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5782494/v1/f90ee38c-c32f-4251-86d6-3fe0634056ae.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"TLR-5 influences T cell activity: implications for development of Type 1 Diabetes","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eMammalian organisms possess both the innate and the adaptive immune system. Therefore, in each situation with an infection, it is a choice whether it should react with granulocytes or lymphocytes, or both. Bacteria may be fought best with the innate immune cells, virus best with antibodies via T cell stimulation, and cancer by T cells, which on the other hand give risk of autoimmunity.\u003c/p\u003e \u003cp\u003eToll-like receptor 5 (TLR-5) is a crucial component of the innate immune system, primarily stimulated by bacterial flagellin\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. In 2011, we found that TLR-5 is the only TLR molecule which is upregulated on beta cells following glucose stimulation\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Initially, the purpose of this upregulation was unclear, but it was hypothesized to be a defence mechanism for beta cells, which are more vulnerable when being active\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e,\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. It has now been shown that high TLR-5 boosts the innate immune system and granulocyte activity\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. In contrast, low TLR-5 expression stimulates T cells and promotes autoimmunity; in mice, TLR-5 deficiency has been linked to increase risk of developing metabolic diabetes\u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. Also, TLR-5 shortage, exacerbates the autoimmune lupus-like disease\u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eIn the present study, we aimed to examine TLR-5 expression in the islets of Langerhans in newly diagnosed Type 1 Diabetes (T1D) patients. Additionally, possible polymorphisms related to the TLR-5 system were investigated.\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003eIn newly diagnosed T1D patients, the mRNA level of TLR-5 in the islets of Langerhans was significantly reduced by one-third compared to control individuals (34.1\u0026thinsp;\u0026plusmn;\u0026thinsp;17.8 vs 50.5\u0026thinsp;\u0026plusmn;\u0026thinsp;11.2, p\u0026thinsp;=\u0026thinsp;0.019). Notably, one outlier patient, the oldest in the group, showed a TLR-5 expression of 64.0. Excluding this patient, the statistics revealed an even more pronounced decrease in TLR-5 expression for the newly diagnosed T1D patients, showing a 48% reduction compared to controls (26.6\u0026thinsp;\u0026plusmn;\u0026thinsp;6.2 vs 50.5\u0026thinsp;\u0026plusmn;\u0026thinsp;11.2, p\u0026thinsp;=\u0026thinsp;0.0012). As it can be seen from Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the prediabetic antibody-positive persons do have the same level of TLR-5 mRNA as the controls and tended to be higher than the DiViD patients. Hence the reduction in TLR-5 occurs when the insulitis become malignant and clinical diabetes arrives. Interestingly, the T1D patients with disease duration of 5 years still tend to have lower TLR-5 values than the controls (p\u0026thinsp;=\u0026thinsp;0.09). There was found no correlation between TLR-5 and C-peptide values for the antibody-positive persons nor for the DiViD patients. Four of the antibody-positive persons had two autoantibodies, which were either GAD, IAA, IA2 or Zink transporter antibodies, and their TLR-5 values were 52.0\u0026thinsp;\u0026plusmn;\u0026thinsp;12.1. The other eight antibody-positive persons had GAD antibodies only and their TLR-5 values were 47.4\u0026thinsp;\u0026plusmn;\u0026thinsp;11.1. Hence, no difference between these subgroups.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe insulitis score for the DiViD patients has earlier been reported\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. In Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, the TLR-5 values are plotted against the numbers of CD3 T cells per islets adjusted for size. Interestingly, there was a tendency (p\u0026thinsp;=\u0026thinsp;0.08) for a negative correlation showing the higher TLR-5, the lower T cell numbers.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe analysis of polymorphisms associated with the TLR-5 system is presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e; one single nucleotide polymorphism (SNP) was correlated with T1D. Interestingly, the same SNP (rs113292043) and rs5744168 were highly significantly associated with monocyte and neutrophil counts, which are key cells of the innate immune system.\u003c/p\u003e "},{"header":"DISCUSSION","content":"\u003cp\u003eIn the present study, we have demonstrated an association between TLR-5 expression in the islets of Langerhans and development of T1D. Additionally, we identified a polymorphism related to T1D. Gram-negative bacteria with flagellin stimulate TLR-5, which enhances granulocyte activity to eliminate these bacteria\u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e,\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. In contrast, this is not an obvious job for T cells which are present to react against tumor cells and virus, and to produce antibodies via T helper and B lymphocytes.\u003c/p\u003e \u003cp\u003eThe identified polymorphisms between TLR-5 and monocyte count are interesting since monocytes are crucial components of the innate immune system. Monocytes\u0026rsquo; primary role in the innate immune system is to phagocytose and destroy infectious agents\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eNew virus infections lead to T cell activation, but interestingly, repeated viral infections increase TLR-5 expression\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. This might be because memory B lymphocytes are already primed, reducing the need for new T cell activation. Thus, for T1D repeated viral infections might offer protection and there might be no pressure of lower TLR-5 expression; therefore, early-life exposure to viruses can have a beneficial impact on the immune system. Interestingly, a study found a reduced incidence of T1D among children attending pre-school and daycare, likely due to increased exposure to repeated virus infections\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe development of T1D requires islet destruction by T cells; this is supported by many studies, e.g., virus-induced diabetes in mice is not occurring in nude mice that lack the thymus-dependent immune system, showing that virus is nontoxic and acts through T cells\u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Also, treatment with CD3 antibodies are beneficial by inducing regulatory T cells and approved by FDA for delaying progression from stage 2 to stage 3 (overt) T1D\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eActivation of TLR molecules other than TLR-5 does not seem to inhibit autoimmunity\u003csup\u003e\u003cspan additionalcitationids=\"CR17 CR18\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. During early pregnancy, TLR-5 expression increases\u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. This may take place in order not to destroy paternal antigens including the fetus. The higher expression of TLR-5 during high activity of beta cells can be explained to avoid development of T1D in pregnant women since during (late) pregnancy beta cells produce 40% more insulin. However, there is a 3.8-times increased risk of developing T1D during the last trimester of pregnancy, but not more despite the pressure on the beta cells\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eTo have activated T cells has its price: the higher risk of autoimmunity, but maybe also lower longevity. This is seen in mice in which Flagellin has been used in a study showing higher longevity in mice after intake of this compound\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. Like TLR-5 the sphingolipid, sulfatide, inhibits T cells\u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e,\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e. In a human epidemiological study, we have measured sulfatide in peripheral blood and compared this to longevity\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. The individuals with the highest content of sulfatide have the lowest mortality. This was significant (0.86, range 0.74\u0026ndash;0.99, p\u0026thinsp;=\u0026thinsp;0.036) dependent on the degree of correction. Thus, also in humans, there is a trend to better longevity when T cells are inhibited.\u003c/p\u003e \u003cp\u003eExperience with flagellin due to low hygiene standard seems natural in Russian Karelia, in poor countries, and in the 1950\u0026rsquo;s including east Germany\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. These places also have low incidence of T1D\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. Interestingly, the hygiene theory is all about exposure to activated bacteria which then according to the above is the mechanism for less autoimmunity\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003ePolymorphisms of the \u003cem\u003eTLR5\u003c/em\u003e gene have been associated with innate immune responses in other inflammatory diseases, including SLE\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. Notably, the rs5744168 SNP of \u003cem\u003eTLR5\u003c/em\u003e generates a stop codon that could affect TLR-5 function\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e, whereas rs113292043 is an intergenic variant with no know functional consequences. The polymorphisms correlate with reduced number of monocytes and neutrophils, and increased risk of T1D.\u003c/p\u003e \u003cp\u003eA consequence of the present study is that T1D can develop due to an unfavourable choice of immunological defence by T cells which dispose to an autoimmune reaction. A parallel to serious or deadly COVID-19 infection may be in play. It starts classically as a virus infection with T cells and antibodies, but dramatic autoimmunity can be initiated in the lungs, and steroid or other anti-autoimmune treatment may be instituted (too late)\u003csup\u003e\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e,\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eRegarding T1D, a treatment strategy could involve flagellin to increase TLR-5 expression, and thereby naturally inhibiting T cells and potentially avoiding autoimmunity.\u003c/p\u003e"},{"header":"METHODS","content":"\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eRNA analysis\u003c/h2\u003e \u003cp\u003eHuman pancreatic tissue utilized in this study was obtained from the DiViD study\u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e from newly onset (disease duration 35 days) T1D patients aged 24\u0026ndash;35 years (n\u0026thinsp;=\u0026thinsp;5), and from the nPOD study\u003csup\u003e\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e with a healthy control group (n\u0026thinsp;=\u0026thinsp;18). Direct to [email protected] for requests to the datasets. The nPOD study consists of pancreases from organ donors following accidental death, as opposed to the living patients in the DiViD study. The nPOD study samples consists of 18 control persons (9F/9M) aged 36.2\u0026thinsp;\u0026plusmn;\u0026thinsp;15.6 (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) years old; 12 antibody-positive persons (5F/7M) aged 20.4\u0026thinsp;\u0026plusmn;\u0026thinsp;8.4 years old; 19 persons with intermedium diagnosed T1D (10F/9M) aged 33.2\u0026thinsp;\u0026plusmn;\u0026thinsp;17 years old; and finally, 8 persons with T2D (5F/3M) aged 39.8\u0026thinsp;\u0026plusmn;\u0026thinsp;13.4 years. Frozen sections of 25 randomly selected pancreatic islets were subjected to laser capture dissection\u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e and islets from two to five sections were pooled, and RNA extraction was done using the Arcturus Pico Pure RNA Isolation Kit (Applied Biosystems, Grand Island, NY, USA). RNA quantification was performed using a Bioanalyzer 2100 instrument (Agilent Technologies, Santa Clara, CA, USA). Gene expression analysis was conducted using Affymetrix expression arrays (Gene Chip Human Gene 2.0 ST, Thermo Fisher) and normalized using global scaling\u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e. All tissue handling procedures was conducted in the same laboratory, to ensure consistency in the comparison. Furthermore, the analysis was performed using same equipment, and the RNA quality was certified by RIN value measurements (\u0026gt;\u0026thinsp;3.5).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eGenetic association analysis\u003c/h3\u003e\n\u003cp\u003eGWAS data were retrieved from\u003csup\u003e\u003cspan additionalcitationids=\"CR38 CR39 CR40 CR41 CR42 CR43\" citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/sup\u003e, and associations analysed for \u003cem\u003eTLR-5\u003c/em\u003e with innate immune cell numbers and T1D defined by ADA criteria\u003csup\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e. Blood glucose levels were evaluated in individuals free of diabetes defined by physician diagnosis, medication use, or fasting glucose\u0026thinsp;\u0026lt;\u0026thinsp;7 mmol/L\u003csup\u003e46\u003c/sup\u003e.\u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eThe statistical analysis for RNA expression was conducted using GraphPad Prism 8.0.2 (GraphPad, La Jolla, Ca, USA). For comparison between groups, a one-way ANOVA was employed followed by Dunnett\u0026rsquo;s multiple comparison test and a 95% CI. Statistical significance was considered when p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eEthics statement\u003c/h2\u003e \u003cp\u003e The DiViD and nPOD studies were approved by The Norwegian government\u0026rsquo;s regional ethics committee (reference 2009/1907) and by the University of Tennessee Health Science Center\u0026rsquo;s local institutional review board (reference 10-00848-XM). All methods were carried out in accordance with relevant guidelines and regulations. We confirm that all experimental protocols were approved by The Norwegian government\u0026rsquo;s regional ethics committee (reference 2009/1907) and by the University of Tennessee Health Science Center\u0026rsquo;s local institutional review board (reference 10-00848-XM). Finally, we confirm that informed consent was obtained from all subjects and/or their legal guardians.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eCOMPETING INTERESTS\u003c/h2\u003e \u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eKarsten Buschard conceptualized the project and wrote the original manuscript draft. Lars Krogvold, Knut Dahl-J\u0026oslash;rgensen, and Ivan Gerling provided the analysed material and performed the RNA expression analysis. Flemming Pociot performed the SNP analysis. All authors edited, reviewed, and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eData is being deposit with datadryad.org. The protocols used can be obtained upon request to the corresponding author.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eFeuillet, V. \u003cem\u003eet al.\u003c/em\u003e Involvement of Toll-like receptor 5 in the recognition of flagellated bacteria. \u003cem\u003eProc Natl Acad Sci U S A\u003c/em\u003e \u003cstrong\u003e103\u003c/strong\u003e, 12487\u0026ndash;12492 (2006).\u003c/li\u003e\n\u003cli\u003eWeile, C., Josefsen, K. \u0026amp; Buschard, K. Glucose activation of islets of Langerhans up-regulates Toll-like receptor 5: possible mechanism of protection. \u003cem\u003eClin Exp Immunol\u003c/em\u003e \u003cstrong\u003e166\u003c/strong\u003e, 251\u0026ndash;257 (2011).\u003c/li\u003e\n\u003cli\u003eBuschard, K. 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TLR5-deficiency controls dendritic cell subset development in an autoimmune diabetes-susceptible model. \u003cem\u003eFront Immunol\u003c/em\u003e \u003cstrong\u003e15\u003c/strong\u003e, 1333967 (2024).\u003c/li\u003e\n\u003cli\u003eAlajoleen, R. M. \u003cem\u003eet al.\u003c/em\u003e Tlr5 deficiency exacerbates lupus-like disease in the MRL/ lpr mouse model. \u003cem\u003eFront Immunol\u003c/em\u003e \u003cstrong\u003e15\u003c/strong\u003e, (2024).\u003c/li\u003e\n\u003cli\u003eKrogvold, L. \u003cem\u003eet al.\u003c/em\u003e Insulitis and characterisation of infiltrating T cells in surgical pancreatic tail resections from patients at onset of type 1 diabetes. \u003cem\u003eDiabetologia\u003c/em\u003e \u003cstrong\u003e59\u003c/strong\u003e, 492\u0026ndash;501 (2016).\u003c/li\u003e\n\u003cli\u003eMcheik, S., Al-Akl, N. S. \u0026amp; Abdelnoor, A. M. 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K. \u003cem\u003eet al.\u003c/em\u003e Covid-19 a triggering factor of autoimmune and multi-inflammatory diseases. \u003cem\u003eLife Sci\u003c/em\u003e \u003cstrong\u003e319\u003c/strong\u003e, 121531 (2023).\u003c/li\u003e\n\u003cli\u003eKrogvold, L. \u003cem\u003eet al.\u003c/em\u003e Pancreatic biopsy by minimal tail resection in live adult patients at the onset of type 1 diabetes: experiences from the DiViD study. \u003cem\u003eDiabetologia\u003c/em\u003e \u003cstrong\u003e57\u003c/strong\u003e, 841\u0026ndash;843 (2014).\u003c/li\u003e\n\u003cli\u003eCampbell-Thompson, M. \u003cem\u003eet al.\u003c/em\u003e Network for Pancreatic Organ Donors with Diabetes (nPOD): developing a tissue biobank for type 1 diabetes. \u003cem\u003eDiabetes Metab Res Rev\u003c/em\u003e \u003cstrong\u003e28\u003c/strong\u003e, 608\u0026ndash;617 (2012).\u003c/li\u003e\n\u003cli\u003eRichardson, S. J. \u003cem\u003eet al.\u003c/em\u003e Islet cell hyperexpression of HLA class I antigens: a defining feature in type 1 diabetes. \u003cem\u003eDiabetologia\u003c/em\u003e \u003cstrong\u003e59\u003c/strong\u003e, 2448\u0026ndash;2458 (2016).\u003c/li\u003e\n\u003cli\u003eWu, J. \u003cem\u003eet al.\u003c/em\u003e Molecular phenotyping of immune cells from young NOD mice reveals abnormal metabolic pathways in the early induction phase of autoimmune diabetes. \u003cem\u003ePLoS One\u003c/em\u003e \u003cstrong\u003e7\u003c/strong\u003e, (2012).\u003c/li\u003e\n\u003cli\u003eOnengut-Gumuscu, S. \u003cem\u003eet al.\u003c/em\u003e Fine mapping of type 1 diabetes susceptibility loci and evidence for colocalization of causal variants with lymphoid gene enhancers. \u003cem\u003eNat Genet\u003c/em\u003e \u003cstrong\u003e47\u003c/strong\u003e, 381\u0026ndash;386 (2015).\u003c/li\u003e\n\u003cli\u003eRobertson, C. C. \u003cem\u003eet al.\u003c/em\u003e Fine-mapping, trans-ancestral and genomic analyses identify causal variants, cells, genes and drug targets for type 1 diabetes. \u003cem\u003eNat Genet\u003c/em\u003e \u003cstrong\u003e53\u003c/strong\u003e, 962\u0026ndash;971 (2021).\u003c/li\u003e\n\u003cli\u003eSinnott-Armstrong, N. \u003cem\u003eet al.\u003c/em\u003e Genetics of 35 blood and urine biomarkers in the UK Biobank. \u003cem\u003eNat Genet\u003c/em\u003e \u003cstrong\u003e53\u003c/strong\u003e, 185\u0026ndash;194 (2021).\u003c/li\u003e\n\u003cli\u003eWeeks, E. M. \u003cem\u003eet al.\u003c/em\u003e Leveraging polygenic enrichments of gene features to predict genes underlying complex traits and diseases. \u003cem\u003eNat Genet\u003c/em\u003e \u003cstrong\u003e55\u003c/strong\u003e, 1267\u0026ndash;1276 (2023).\u003c/li\u003e\n\u003cli\u003eJurgens, S. J. \u003cem\u003eet al.\u003c/em\u003e Adjusting for common variant polygenic scores improves yield in rare variant association analyses. \u003cem\u003eNat Genet\u003c/em\u003e \u003cstrong\u003e55\u003c/strong\u003e, 544\u0026ndash;548 (2023).\u003c/li\u003e\n\u003cli\u003eChiou, J. \u003cem\u003eet al.\u003c/em\u003e Interpreting type 1 diabetes risk with genetics and single-cell epigenomics. \u003cem\u003eNature\u003c/em\u003e \u003cstrong\u003e594\u003c/strong\u003e, 398\u0026ndash;402 (2021).\u003c/li\u003e\n\u003cli\u003eKurki, M. I. \u003cem\u003eet al.\u003c/em\u003e FinnGen provides genetic insights from a well-phenotyped isolated population. \u003cem\u003eNature\u003c/em\u003e \u003cstrong\u003e613\u003c/strong\u003e, 508\u0026ndash;518 (2023).\u003c/li\u003e\n\u003cli\u003eShirai, Y. \u003cem\u003eet al.\u003c/em\u003e Multi-trait and cross-population genome-wide association studies across autoimmune and allergic diseases identify shared and distinct genetic component. \u003cem\u003eAnn Rheum Dis\u003c/em\u003e \u003cstrong\u003e81\u003c/strong\u003e, 1301\u0026ndash;1312 (2022).\u003c/li\u003e\n\u003cli\u003eElsayed, N. A. \u003cem\u003eet al.\u003c/em\u003e Erratum. 2. Classification and diagnosis of diabetes: Standards of Care in Diabetes-2023. Diabetes Care 2023;46(Suppl. 1):S19-S40. \u003cem\u003eDiabetes Care\u003c/em\u003e \u003cstrong\u003e46\u003c/strong\u003e, 1106 (2023).\u003c/li\u003e\n\u003cli\u003eShroff, R. T. \u003cem\u003eet al.\u003c/em\u003e Immune responses to two and three doses of the BNT162b2 mRNA vaccine in adults with solid tumors. \u003cem\u003eNat Med\u003c/em\u003e \u003cstrong\u003e27\u003c/strong\u003e, 2002\u0026ndash;2011 (2021).\u003c/li\u003e\n\u003cli\u003eVuckovic, D. \u003cem\u003eet al.\u003c/em\u003e The Polygenic and Monogenic Basis of Blood Traits and Diseases. \u003cem\u003eCell\u003c/em\u003e \u003cstrong\u003e182\u003c/strong\u003e, 1214-1231.e11 (2020).\u003c/li\u003e\n\u003cli\u003eChen, M. H. \u003cem\u003eet al.\u003c/em\u003e Trans-ethnic and Ancestry-Specific Blood-Cell Genetics in 746,667 Individuals from 5 Global Populations. \u003cem\u003eCell\u003c/em\u003e \u003cstrong\u003e182\u003c/strong\u003e, 1198-1213.e14 (2020).\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Table 1","content":"\u003cp\u003eTable 1. Shows polymorphism related to TLR-5.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"987\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3492%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLead Variant (CHR:POS:REF:ALT)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.4081%;\"\u003e\n \u003cp\u003e\u003cstrong\u003edbSNP\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.8673%;\"\u003e\n \u003cp\u003e\u003cstrong\u003ePhenotype\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.6993%;\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-Value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.721%;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3492%;\"\u003e\n \u003cp\u003e1:223285200:G:A\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.4081%;\"\u003e\n \u003cp\u003ers5744168\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.8673%;\"\u003e\n \u003cp\u003eMonocyte percentage\u003c/p\u003e\n \u003cp\u003eMonocyte count\u003c/p\u003e\n \u003cp\u003eNeutrophil percentage\u003c/p\u003e\n \u003cp\u003eNeutrophil count\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.6993%;\"\u003e\n \u003cp\u003e1.77e-16\u003c/p\u003e\n \u003cp\u003e3.21e-12\u003c/p\u003e\n \u003cp\u003e1.40e-4\u003c/p\u003e\n \u003cp\u003e6.83e-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.721%;\"\u003e\n \u003cp\u003e\u003csup\u003e40,47\u003c/sup\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 31.3492%;\"\u003e\n \u003cp\u003e1:223231657:C:T\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 13.4081%;\"\u003e\n \u003cp\u003ers113292043\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 25.8673%;\"\u003e\n \u003cp\u003eMonocyte count\u003c/p\u003e\n \u003cp\u003eMonocyte percentage\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eType 1 diabetes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17.6993%;\"\u003e\n \u003cp\u003e1.21e-14\u003c/p\u003e\n \u003cp\u003e1.24e-14\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e1.3e-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 11.721%;\"\u003e\n \u003cp\u003e\u003csup\u003e41,47,48\u003c/sup\u003e\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003cp class=\"MsoNormal\"\u003e\u003csup\u003e\u003cspan lang=\"DA\"\u003e42\u0026ndash;44\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"TLR-5, Type 1 Diabetes, Islets of Langerhans, Polymorphisms, Autoimmunity, T cells.","lastPublishedDoi":"10.21203/rs.3.rs-5782494/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5782494/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn mammalian and human life, one of the most important problems is to choose the best defence against microorganisms. Innate cells are good against bacteria, and T cells against virus mainly because of antibody production via T helper and B lymphocytes. Toll-like receptor 5 (TLR-5) is a regulator of this choice; when it is highly expressed, T cells are inhibited, and innate cells are favored. In activated pancreatic beta cells, TLR-5 has been found highly expressed and may therefore be protected from T cell destruction e.g., during pregnancy. We investigated mRNA from islets of Langerhans from newly diagnosed T1D patients for TLR-5. Also, we examined for polymorphisms between TLR-5, metabolic parameters, and Type 1 Diabetes (T1D). mRNA for TLR-5 was downregulated by one-third in the newly diagnosed T1D patients compared to controls. Regarding polymorphisms, two associations were found between TLR-5 and monocytes which are cells from the innate immune system. Also, a significant polymorphism was seen concerning TLR-5 and T1D. T cells are important for autoimmune diseases including T1D. In the present study we find low values of mRNA of TLR-5 which enhance T cells. We hope that the findings mentioned in this article may be influential for the understanding of how T1D develops.\u003c/p\u003e","manuscriptTitle":"TLR-5 influences T cell activity: implications for development of Type 1 Diabetes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-22 09:23:34","doi":"10.21203/rs.3.rs-5782494/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"cda1f2a9-b673-4954-a5ea-7176ab49907e","owner":[],"postedDate":"January 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":43118907,"name":"Health sciences/Endocrinology/Endocrine system and metabolic diseases/Diabetes/Type 1 diabetes mellitus"},{"id":43118908,"name":"Biological sciences/Immunology/Autoimmunity"},{"id":43118909,"name":"Biological sciences/Cell biology/Mechanisms of disease"}],"tags":[],"updatedAt":"2025-03-05T03:23:15+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-22 09:23:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5782494","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5782494","identity":"rs-5782494","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}