Enterovirus spectrum from the active surveillance of hand foot and mouth disease patients under the clinical trial of inactivated Enterovirus A71 vaccine in Jiangsu,China, 2012‐2013

Epidemiological data from active surveillance on human enterovirus, which could cause hand, foot, and mouth disease, were limited. An active surveillance system was used to investigate the enterovirus spectrum and the incidence of different enteroviruses in infants aged 6–35 months in Jiangsu Province from 2012 to 2013. Fifty‐nine infants were randomly selected from 522 non‐EV‐A71/CV‐A16 HFMD patients. We collected 173 throat swabs and 174 rectal swabs from these infants. RT‐PCR was used to amplify 5'‐UTR and VP1 regions of enteroviruses and the serotypes were determined by the sequence comparison using BLAST. Twenty‐one non‐EV‐A71/CA16 enterovirus serotypes were detected in those infants. E16, E18 were firstly reported in HFMD patients. The four top common non‐EV‐A71/CV‐A enteroviruses among infants were CV‐B3, CV‐A10, CV‐A6, and E9 with the HFMD incidence rates at 1.4%, 0.84%, 0.56%, and 0.47%, respectively. Over 20.8% patients were co‐infected with multiple enteroviruses. Neither the course of sickness nor clinical symptoms of the co‐infected patients was more severe than those infected with single enterovirus. Two patients were infected different enterovirus successively within 2 months. Several new enterovirus serotypes and multiple models of infection associated with HFMD were discovered through the active surveillance system. These data provide a better understanding of the viral etiology of HFMD. J. Med. Virol. 87:2009–2017, 2015. © 2015 Wiley Periodicals, Inc.

     

Enteroviruses in the pathogenesis of type 1 diabetes

The question if enteroviruses could cause beta-cell damage and type 1 diabetes has become more and more relevant when recent studies have provided new evidence supporting this scenario. One important observation is the recent discovery of IFIH1 as a risk gene for type 1 diabetes. This gene is an innate immune system receptor for enteroviruses offering one possible mechanism for the diabetogenic effect of enteroviruses. This is further emphasized by the observations suggesting that the innate immune system is activated in the pancreatic islets of type 1 diabetic patients and that the innate immune system is important for the defense against the virus and for the regulation of adaptive immune system. Important progress has also been gained in studies analyzing pancreas tissue for possible presence of enteroviruses. Several studies have found enteroviruses in the pancreatic islets of type 1 diabetic patients using various methods. The virus seems to be located in the islets while exocrine pancreas is mostly uninfected. One recent study found the virus in the intestinal mucosa in the majority of diabetic patients. Enteroviruses can also infect cultured human pancreatic islets causing either rapid cell destruction or a persistent-like noncytolytic infection. Combined with all previous, epidemiological findings indicating the risk effect of enteroviruses in cross-sectional and prospective studies, these observations fit to a scenario where certain diabetogenic enterovirus variants establish persistent infection in gut mucosa and in the pancreatic islets. This in turn could lead to a local inflammation and the breakdown of tolerance in genetically susceptible individuals. This is also supported by mouse experiments showing that enteroviruses can establish prolonged infection in the pancreas and intestine, and some virus strains cause beta-cell damage and diabetes. In conclusion, recent studies have strengthened the hypothesis that enteroviruses play a role in the pathogenesis of type 1 diabetes. These findings open also new opportunities to explore the underlying mechanism and get closer to causal relationship.     

Comparative and correlative assessments of cytokine,complement and antibody patterns in paediatric type 1 diabetes

One of the most widespread and effective environmental factors is the infection with enteroviruses (EVs) which accelerate β cell destruction in type 1 diabetes (T1D). This study represented a comparison between diabetic EV⁺ and EV^– children as well as correlation analysis between autoantibodies, T1D markers, cytokines, complement activation products and anti‐coxsackievirus (CV) immunoglobulin (Ig)G. EV RNA was detected in Egyptian children with T1D (26·2%) and healthy controls (0%). Detection of anti‐CV IgG in T1D‐EV⁺ resulted in 64% positivity. Within T1D‐EV⁺, previously diagnosed (PD) showed 74 versus 56% in newly diagnosed (ND) children. Comparisons between populations showed increased levels of haemoglobin A1c (HbA1c), C‐reactive protein (CRP), nitric oxide (NO), glutamic acid decarboxylase and insulin and islet cell autoantibodies [glutamic acid decarboxylase autoantibodies (GADA), insulin autoantibodies (IAA) and islet cell cytoplasmic autoantibodies (ICA), respectively], interferon (IFN)‐γ, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, IL ?10, IL ?12, IL ?17, C3d and sC5–9 in T1D‐EV⁺ versus T1D‐EV^–. Conversely, both IL‐20 and transforming growth factor (TGF‐β) decreased in T1D‐EV⁺ versus EV^–, while IL‐4, ?6 and ?13 did not show any changes. Correlation analysis showed dependency of accelerated autoimmunity and β cell destruction on increased IFN‐γ, IL‐12 and IL‐17 versus decreased IL‐4, ?6 and ?13. In conclusion, IFN‐γ, IL‐12 and IL‐17 played an essential role in exacerbating EV⁺‐T1D, while C3d, sC5b ?9, IL‐10 and ?20 displayed distinct patterns.     

Elevated systemic glutamic acid level in the non‐obese diabetic mouse is Idd linked and induces beta cell apoptosis

Although type 1 diabetes (T1D) is a T‐cell‐mediated disease in the effector stage, the mechanism behind the initial beta cell assault is less understood. Metabolomic differences, including elevated levels of glutamic acid, have been observed in patients with T1D before disease onset, as well as in pre‐diabetic non‐obese diabetic (NOD) mice. Increased levels of glutamic acid damage both neurons and beta cells, implying that this could contribute to the initial events of T1D pathogenesis. We investigated the underlying genetic factors and consequences of the increased levels of glutamic acid in NOD mice. Serum glutamic acid levels from a (NOD×B6)F₂ cohort (n = 182) were measured. By genome‐wide and Idd region targeted microsatellite mapping, genetic association was detected for six regions including Idd2, Idd4 and Idd22. In silico analysis of potential enzymes and transporters located in and around the mapped regions that are involved in glutamic acid metabolism consisted of alanine aminotransferase, glutamic‐oxaloacetic transaminase, aldehyde dehydrogenase 18 family, alutamyl‐prolyl‐tRNA synthetase, glutamic acid transporters GLAST and EAAC1. Increased EAAC1 protein expression was observed in lysates from livers of NOD mice compared with B6 mice. Functional consequence of the elevated glutamic acid level in NOD mice was tested by culturing NOD. Rag2^−/− Langerhans’ islets with glutamic acid. Induction of apoptosis of the islets was detected upon glutamic acid challenge using TUNEL assay. Our results support the notion that a dysregulated metabolome could contribute to the initiation of T1D. We suggest that targeting of the increased glutamic acid in pre‐diabetic patients could be used as a potential therapy.     

Pathogenic T helper type 17 cells contribute to type 1 diabetes independently of interleukin‐22

We have shown that pathogenic T helper type 17 (Th17) cells differentiated from naive CD4⁺ T cells of BDC2·5 T cell receptor transgenic non‐obese diabetic (NOD) mice by interleukin (IL)‐23 plus IL‐6 produce IL‐17, IL‐22 and induce type 1 diabetes (T1D). Neutralizing interferon (IFN)‐γ during the polarization process leads to a significant increase in IL‐22 production by these Th17 cells. We also isolated IL‐22‐producing Th17 cells from the pancreas of wild‐type diabetic NOD mice. IL‐27 also blocked IL‐22 production from diabetogenic Th17 cells. To determine the functional role of IL‐22 produced by pathogenic Th17 cells in T1D we neutralized IL‐22 in vivo by using anti‐IL‐22 monoclonal antibody. We found that blocking IL‐22 did not alter significantly adoptive transfer of disease by pathogenic Th17 cells. Therefore, IL‐22 is not required for T1D pathogenesis. The IL‐22Rα receptor for IL‐22 however, increased in the pancreas of NOD mice during disease progression and based upon our and other studies we suggest that IL‐22 may have a regenerative and protective role in the pancreatic islets.     

Aqueous extracts of Syzygium brazzavillense can inhibit the infection with coxsackievirus B4 in vitro

Traditional practitioners commonly use plant crude extracts to treat various diseases in patients with symptoms that can be seen during enterovirus infections. In this study, the antienteroviral activity of medicinal plants from the Republic of Congo has been evaluated in vitro. Through an ethnopharmacological approach, seven plants grouped into six families were identified. Aqueous and organic extracts of various organs from these plants were prepared. The organic extracts at subcytotoxic concentrations did not inhibit the cytopathic effect (CPE) induced by coxsackievirus (CV)B1-5, CVA6, poliovirus type 1, and enterovirus 71. The aqueous extract of Syzygium brazzavillense, but not those of other plants, inhibited the CPE induced by CVB3 and CVB4 at 30 µg/mL (CC₅₀; 2800 µg/mL, IC₅₀; 0.8 µg/mL) and by CVB2 and poliovirus type 1 at higher concentrations. When aqueous extract of this plant was mixed with CVB4, the replication of the virus was inhibited. In conclusion, aqueous extracts of Syzygium brazzavillense can inhibit the infection with CVB4 and other enteroviruses in vitro. The present ethnopharmacological investigation helped to identify a plant with potential properties useful to combat enterovirus infections.     

The non‐inherited maternal HLA haplotype affects the risk for type 1 diabetes

The aim was to test the hypothesis that the human leucocyte antigen (HLA) haplotype that is not inherited from the mother, that is, the non‐inherited maternal antigen (NIMA) affects the risk for type 1 diabetes (T1D). A total of 563 children with T1D and 286 non‐diabetic control children from Sweden were genotyped for DRB1, DQA1 and DQB1 alleles. The frequency of positively (DR4‐DQA1*0301‐B1*0302 and DR3‐DQA1*0501‐B1*0201), negatively (DR15‐DQ A1*0102‐B1*0602) or neutrally (all other) T1D associated HLA haplotypes were compared between NIMA and non‐inherited paternal antigen (NIPA). All comparisons were carried out between HLA‐matched patients and controls. The frequency of positively associated NIMA was higher among both DR4/X‐positive healthy individuals compared wit DR4/X‐positive patients (P < 0.00003) and DR3/X‐positive healthy individuals compared with DR3/X‐positive patients (P < 0.009). No such difference was observed for NIPA. High‐risk NIMA was increased compared to NIPA among healthy DR3/X‐ and DR4/X‐positive children (P < 0.05). There was no difference in frequency of positively associated haplotypes between patient NIMA and NIPA. The NIMA but not the NIPA affects the risk for T1D, suggesting that not only the inherited but also non‐inherited maternal HLA haplotypes, perhaps through microchimerism or other mechanisms, may influence the risk for the disease.     

Enterovirus genotypes among patients with severe acute respiratory illness,influenza‐like illness,and asymptomatic individuals in South Africa, 2012‐2014

Enteroviruses can cause outbreaks of severe acute respiratory illness (SARI) and EV‐A, ‐B, ‐C, and ‐D species have different pathogenic profiles and circulation patterns. We aimed to characterize and determine the prevalence of enterovirus genotypes among South African patients with respiratory illness and controls during June 2012 to July 2014. Syndromic SARI and influenza‐like illness (ILI) surveillance was performed at two sentinel sites. At each site nasopharyngeal/oropharyngeal specimens were collected from SARI and ILI patients as well as controls. Specimens were tested for enterovirus by real‐time PCR. Positive specimens were further genotyped by sequencing a region of the VP1 gene. The prevalence of enterovirus was 5.8% (87/1494), 3.4% (103/3079), and 3.4% (46/1367) among SARI, ILI, and controls, respectively (SARI/controls, P = 0.002 and ILI/control, P = 0.973). Among the 101/236 (42.8%) enterovirus‐positive specimens that could be genotyped, we observed a high diversity of circulating enterovirus genotypes (a total of 33 genotypes) from all four human enterovirus species with high prevalence of Enterovirus‐B (60.4%; 61/101) and Enterovirus‐A (21.8%; 22/101) compared to Enterovirus‐C (10.9%; 11/101) and Enterovirus‐D (6.9%; 7/101) (P = 0.477). Of the enterovirus genotypes identified, Echovirus 30 (9.9%, 10/101), Coxsackie virus B5 (7.9%, 8/101) and Enterovirus‐D68 (6.9%, 7/101) were most prevalent. There was no difference in disease severity (SARI or ILI compared to controls) between the different enterovirus species (P = 0.167). We observed a high number of enterovirus genotypes in patients with respiratory illness and in controls from South Africa with no disease association of EV species with disease severity.

     

The type 1 diabetes resistance locus B10 Idd9.3 mediates impaired B‐cell lymphopoiesis and implicates microRNA‐34a in diabetes protection

NOD.B10 Idd9.3 mice are congenic for the insulin‐dependent diabetes (Idd) Idd9.3 locus, which confers significant type 1 diabetes (T1D) protection and encodes 19 genes, including microRNA (miR)‐34a, from T1D‐resistant C57BL/10 mice. B cells have been shown to play a critical role in the priming of autoantigen‐specific CD4⁺ T cells in T1D pathogenesis in non‐obese diabetic (NOD) mice. We show that early B‐cell development is impaired in NOD.B10 Idd9.3 mice, resulting in the profound reduction of transitional and mature splenic B cells as compared with NOD mice. Molecular analysis revealed that miR‐34a expression was significantly higher in B‐cell progenitors and marginal zone B cells from NOD.B10 Idd9.3 mice than in NOD mice. Furthermore, miR‐34a expression in these cell populations inversely correlated with levels of Foxp1, an essential regulator of B‐cell lymphopoiesis, which is directly repressed by miR‐34a. In addition, we show that islet‐specific CD4⁺ T cells proliferated inefficiently when primed by NOD.B10 Idd9.3 B cells in vitro or in response to endogenous autoantigen in NOD.B10 Idd9.3 mice. Thus, Idd9.3‐encoded miR‐34a is a likely candidate in negatively regulating B‐cell lymphopoiesis, which may contribute to inefficient expansion of islet‐specific CD4⁺ T cells and to T1D protection in NOD.B10 Idd9.3 mice.     

Detection of enterovirus in the thyroid tissue of patients with graves' disease

The etiology and pathogenesis of Graves' disease (GD) are still unknown, although it is thought that both genetic and environmental factors are important. Some indirect evidence implies that a viral infection may be a possible etiologic factor in autoimmunity. The main objective of this study was to examine direct evidence of the presence of enteroviruses (EVs) in the thyroid tissue of patients with GD. Thyroid tissue from 22 patients with newly diagnosed GD was obtained by core needle biopsy, while tissue from 24 patients with chronic GD and 24 control subjects without any autoimmune thyroid diseases was collected during neck surgery. Formalin‐fixed, paraffin‐embedded thyroid tissue samples were examined for the presence of enterovirus capsid protein using immunohistochemistry and for enterovirus RNA using in situ hybridization. Enterovirus capsid protein was detected in 17 (37%) patients and in 4 (17%) control subjects (P = 0.103). Enterovirus RNA was identified in thyroid tissue from nine (20%) patients, but in none of the control subjects (P = 0.016). Eight (90%) of the nine virus RNA positive patients were also positive for enterovirus protein. This is the first study to analyze thyroid tissue for EVs, including patients with untreated, newly diagnosed GD. The results suggest that EVs are more frequently present in thyroid tissue of patients than controls. Further studies are indicated to explore this association to find out if a low‐grade chronic enteroviral infection might be involved in the pathogenesis of GD and if this could offer new therapeutic and preventive opportunities. J. Med. Virol. 85:512–518, 2013. © 2012 Wiley Periodicals, Inc.     

Role of Viruses and Other Microbes in the Pathogenesis of Type 1 Diabetes

Type 1 diabetes is caused by an immune-mediated destruction of insulin producing beta-cells in the pancreas. The risk of the disease is determined by interactions between more than 40 different susceptibility genes and yet unidentified environmental factors. The rapidly increasing incidence indicates that these environmental agents have a significant role in the pathogenesis. Microbes have associated with both increased and decreased risk reflecting their possible role as risk or protective factors. Two main hypotheses have been proposed to explain these effects: the hygiene hypothesis suggests that microbial exposures in early childhood stimulate immunoregulatory mechanisms which control autoimmune reactions (analogy with allergy), while the triggering hypothesis suggests that specific microbes damage insulin producing cells. Certain viruses, particularly enteroviruses, are currently the main candidates for such risk microbes. Enteroviruses cause diabetes in animals and have associated with increased risk of type 1 diabetes in epidemiological studies. They have also been detected in the pancreas of diabetic patients. Possible protective effect of microbes has been studied in animal models and in epidemiological studies, where certain enteral microbes (e.g. hepatitis A virus and Helicobacter pylori) and patterns of gut microbiome have associated with low risk of type 1 diabetes. In conclusion, these microbial effects offer attractive possibilities for the development of preventive interventions for type 1 diabetes based on the elimination of triggering agents (e.g. enterovirus vaccines) or use of protective microbes as probiotics.     

Role of enterovirus infections in IgE sensitization

Among other infectious agents, enteroviruses have been associated with protection against allergic diseases. The aim of the present study was to confirm these findings using a highly sensitive and specific neutralization antibody assay and to investigate whether the protective effect is related to certain enterovirus serotypes. Antibodies against 12 enterovirus serotypes were measured in 60 children who were positive for allergen‐specific IgE and in 190 control children. Echoviruses seemed to be more protective than coxsackie‐B‐viruses and echovirus 11 had the strongest independent protective effect (P = 0.001; OR = 0.35, 95% CI: 0.18–0.67). The results support previous observations suggesting that infections by certain enterovirus types are associated with protection against IgE sensitization. J. Med. Virol. 84:268–271, 2012. © 2011 Wiley Periodicals, Inc.     

Immune responses against enterovirus A71 infection: Implications for vaccine success

Enterovirus A71 (EV‐A71) from the Picornaviridae family is an important emerging pathogen causing hand, foot, and mouth disease (HFMD) outbreaks worldwide. EV‐A71 also caused fatal neurological complications in young children especially in Asia. On the basis of seroepidemiological studies from many Asian countries, EV‐A71 infection is very common. Children of very young age are particularly vulnerable. Large‐scale epidemics that occur every 3 to 4 years are associated with accumulation of an immunologically naive younger population. Capsid proteins especially VP1 with the presence of major B‐ and T‐cell epitopes are the most antigenic proteins. The nonstructural proteins mainly contribute to T‐cell epitopes that induce cross‐reactive immune responses against other enteroviruses. Dominant epitopes and their neutralization magnitudes differ in mice, rabbits, and humans. Neutralizing antibody is sufficient for immune protection, but poorer cellular immunity may lead to severe neurological complications and deaths. Some chemokines/cytokines are consistently found in severely ill patients, for example, IL‐6, IL‐10, IL‐17A, MCP‐1, IL‐8, MIG, IP‐10, IFN‐γ, and G‐CSF. An increase in white cell counts is a risk factor for severe HFMD. Recent clinical trials on EV‐A71 inactivated vaccine showed >90% efficacy and a robust neutralization response that was protective, indicating neutralizing antibody correlates for protection. No protection against other enteroviruses was observed. A comprehensive understanding of the immune responses to EV‐A71 infection will benefit the development of diagnostic tools, potential therapeutics, and subunit vaccine candidates. Future development of a multivalent enterovirus vaccine will require knowledge of correlates of protection, understanding of cross‐protection and memory T‐cell responses among enteroviruses.     

Administration of Sulfatide to Ameliorate Type I Diabetes in Non‐Obese Diabetic Mice

The endogenous glycosphingolipid sulfatide is a ligand for CD1d‐restricted type II natural killer T (NKT) lymphocytes. Through the action of these cells, sulfatide treatment has been shown to modulate the immune response in mouse models for autoimmune diseases, infections and tumour immunity. Sulfatide exists naturally in different organs including the pancreas, where sulfatide co‐localizes with insulin within the Langerhans islet β‐cells, targets for the immune destruction in type 1 diabetes (T1D). Human T1D patients, but not patients with type 2 diabetes nor healthy individuals, have autoantibodies against sulfatide in serum, suggesting that sulfatide induces an immune response in the natural course of T1D in humans. Here, we investigate sulfatide as an autoantigen and a modulator of autoimmune disease in the murine model for T1D, the non‐obese diabetic (NOD) mice. We demonstrate that aged NOD mice displayed serum autoantibody reactivity to sulfatide; however, this reactivity did not correlate with onset of T1D. Repeated administration of sulfatide did not result in an increase in serum reactivity to sulfatide. Moreover, a multidose sulfatide treatment of female NOD mice initiated at an early (5 weeks of age), intermediate (8 weeks of age) or late (12 weeks of age) phase of T1D progression did not influence the incidence of disease. Thus, we demonstrate that a fraction of NOD mice develop autoantibody reactivity to sulfatide; however, we fail to demonstrate that sulfatide treatment reduces the incidence of T1D in this mouse strain.     

Gut immune deficits in LEW.1AR1‐iddm rats partially overcome by feeding a diabetes‐protective diet

The gut immune system and its modification by diet have been implicated in the pathogenesis of type 1 diabetes (T1D). Therefore, we investigated gut immune status in non‐diabetes‐prone LEW.1AR1 and diabetes‐prone LEW.1AR1‐iddm rats and evaluated the effect of a low antigen, hydrolysed casein (HC)‐based diet on gut immunity and T1D. Rats were weaned onto a cereal‐based or HC‐based diet and monitored for T1D. Strain and dietary effects on immune homeostasis were assessed in non‐diabetic rats (50–60 days old) and rats with recent‐onset diabetes using flow cytometry and immunohistochemistry. Immune gene expression was analysed in mesenteric lymph nodes (MLN) and jejunum using quantitative RT‐PCR and PCR arrays. T1D was prevented in LEW.1AR1‐iddm rats by feeding an HC diet. Diabetic LEW.1AR1‐iddm rats had fewer lymphoid tissue T cells compared with LEW.1AR1 rats. The percentage of CD4⁺ Foxp3⁺ regulatory T (Treg) cells was decreased in pancreatic lymph nodes (PLN) of diabetic rats. The jejunum of 50‐day LEW.1AR1‐iddm rats contained fewer CD3⁺ T cells, CD163⁺ M2 macrophages and Foxp3⁺ Treg cells. Ifng expression was increased in MLN and Foxp3 expression was decreased in the jejunum of LEW.1AR1‐iddm rats; Ifng/Il4 was decreased in jejunum of LEW.1AR1‐iddm rats fed HC. PCR arrays revealed decreased expression of M2‐associated macrophage factors in 50‐day LEW.1AR1‐iddm rats. Wheat peptides stimulated T‐cell proliferation and activation in MLN and PLN cells from diabetic LEW.1AR1‐iddm rats. LEW.1AR1‐iddm rats displayed gut immune cell deficits and decreased immunoregulatory capacity, which were partially corrected in animals fed a low antigen, protective HC diet consistent with other models of T1D.     

CD40‐mediated signalling influences trafficking,T‐cell receptor expression,and T‐cell pathogenesis,in the NOD model of type 1 diabetes

CD40 plays a critical role in the pathogenesis of type 1 diabetes (T1D). The mechanism of action, however, is undetermined, probably because CD40 expression has been grossly underestimated. CD40 is expressed on numerous cell types that now include T cells and pancreatic β cells. CD40⁺ CD4⁺ cells [T helper type 40 (TH40)] prove highly pathogenic in NOD mice and in translational human T1D studies. We generated BDC2.5.CD40^−/− and re‐derived NOD.CD154^−/− mice to better understand the CD40 mechanism of action. Fully functional CD40 expression is required not only for T1D development but also for insulitis. In NOD mice, TH40 cell expansion in pancreatic lymph nodes occurs before insulitis and demonstrates an activated phenotype compared with conventional CD4⁺ cells, apparently regardless of antigen specificity. TH40 T‐cell receptor (TCR) usage demonstrates increases in several Vα and Vβ species, particularly Vα3.2⁺ that arise early and are sustained throughout disease development. TH40 cells isolated from diabetic pancreas demonstrate a relatively broad TCR repertoire rather than restricted clonal expansions. The expansion of the Vα/Vβ species associated with diabetes depends upon CD40 signalling; NOD.CD154^−/− mice do not expand the same TCR species. Finally, CD40‐mediated signals significantly increase pro‐inflammatory Th1‐ and Th17‐associated cytokines whereas CD28 co‐stimulus alternatively promotes regulatory cytokines.     

A survey of known immune epitopes in the enteroviruses strains associated with acute flaccid myelitis

Enteroviruses are potentially linked to the emergence of Acute Flaccid Myelitis (AFM), a rare but very serious condition that affects the nervous system. AFM has been associated with coxsackievirus A16, enterovirus A71 (EVA71) and enterovirus D68 (EVD68). Little is known about host-pathogen interactions for these viruses, and whether immune responses may have a protective or immunopathological role in disease presentations. Towards addressing this issue, we used the Immune Epitope Database to assess the known inventory of B and T cell epitopes from enteroviruses, focusing on data related to human hosts. The extent of conservation in areas that are targets of B and T cell immune responses were examined. This analysis sheds light on regions of the enterovirus polypeptide that can be probed to induce a specific or cross-reactive B or T cell the immune response to enteroviruses, with a particular focus on coxsackievirus A16, EVA71 and EVD68. In addition, these analyses reveal the current gap-of-knowledge in the T and B cell immune responses that future studies should aim to address.     

Hyperglycemia Affects the Expression of Inflammatory Genes in Peripheral Blood Mononuclear Cells of Patients with Type 2 Diabetes

ABSTRACT

Objective: Innate immune system dysregulation and chronic low-grade inflammation are associated with the pathogenesis of type 2 diabetes (T2D). The aim of this study was to investigate the effect of hyperglycemia on mRNA expression of four inflammatory genes in peripheral blood mononuclear cells (PBMCs) of pre-diabetic and diabetic individuals.

Methods: In a case-control study, quantitative real-time PCR was used to analyze changes in IL-1β, IL1R1, IL-6, and IL6ST gene expression in PBMCs of 30 T2D patients with high blood glucose levels, 24 diabetic and nondiabetic individuals with moderately high blood glucose levels and 30 controls with normal blood glucose levels.

Results: In T2D patients with high blood glucose, IL-1β expression showed a 2.69-fold increase (p = 0.0380), while IL-6 expression levels were 3.45 fold lower (p = 0.0045) versus control subjects. Moreover, compared with control group the expression of IL1R1 and IL-6 genes both were downregulated in individuals with moderately high blood glucose levels by 2.38 (p = 0.0365) and 4.34 fold (p = 0.0027), respectively. In addition, hemoglobin A1C (A1C) levels were positively correlated with IL-1β expression and fasting plasma glucose (FPG) levels showed a positive correlation with IL-1β and a negative correlation with IL-6 expression.

Conclusion: The observed changes in both IL-1β and IL-6 mRNA levels in PBMCs may contribute to the development of inflammatory processes involved in the pathogenesis of T2D. Downregulation of IL1R1 in individuals with mild hyperglycemia may indicate an attempt to reduce the pro-inflammatory effects of IL-1β via auto-stimulation.