Levels Of Th2 Cytokines Research Articles

P1190 Generation of blood-derived reference samples with endogenous cytokines

Abstract Background Biomarkers have become an integral part of drug development and decision-making. The quantification of cytokines, commonly used to gain insights into pathological processes in indications such as Inflammatory Bowel Diseases (IBD), and for monitoring therapeutic intervention, should rely on robust and performant methods. Relevant reference samples are crucial for evaluating the performance of biomarker assays implemented in the context of clinical studies, but oHow to evaluate the parallelism of a method for biomarkers that are non-detectable in healthy donors (HD)? oDoes the selectivity of the recombinant protein in the biological matrix reflect the endogenous biomarker? oHow does the stability of the recombinant protein compare with endogenous biomarker? oCan you use incurred samples (informed consent, sufficient volume) for evaluating parallelism and stability? Methods To address these questions, we generated samples with endogenous biomarkers, named MAQC, by spiking supernatants of stimulated PBMC in the serum or plasma from HD. Three to four levels of MAQC per cytokine were prepared and used to assess parallelism, selectivity and long-term stability (LTS) at -80°C of pro- and anti-inflammatory cytokines and chemokines by use of V-plex, S-plex kits from MSD and ELLA kits. For all these parameters, accuracy was determined, with a target acceptance of 70-130% of the nominal value for selectivity and LTS or upstream dilution for parallelism. Results We were able to generate serum and plasma reference samples with low- to high-range levels of Th1 (IFN-g, IL-12p70, IL-2, TNF-α, CCL-2), Th2 (IL-4, IL-13, IL-10), Th17 (IL-17A) and Proinflammatory (IL-6, IL-8, IL-1β) cytokines. Selectivity was acceptable for most cytokines, except for IL-13, IL-4, IL-17A and IL-6. For IL-6, a major interference was observed in some samples, likely due to the presence of soluble IL-6 receptor in the original serum sample. For the other 3 cytokines, interference, leading to a bias close to -30%, was observed that could not be lifted by diluting samples. For all cytokines but IL-13, endogenous diluted similarly as recombinant protein from calibrator. Twelve months LTS was demonstrated for all cytokines in MAQC and comparable to recombinant proteins spiked in human serum. Conclusion To conclude, reference samples with endogenous cytokines/chemokines can be produced from primary immune cells. These can be used to assess parallelism and long-term stability when sufficient volume of matrices with high enough concentrations are not available. These would be even more valuable if they could be commercially available for use as quality controls in validation and clinical studies.

Antagonism of CD28 blocks allergic responses in the ovalbumin-induced asthmatic model mice.

Allergen-reactive T helper (Th) 2 cells play a pivotal role in initiating asthma pathogenesis. The absence or interruption of CD28 signaling causes significant consequences for T-cell activation, leading to reduced cell proliferation and interleukin (IL)-2 production. A novel compound, Cyn-1324, exhibits a higher binding affinity to CD28 than CD80. Thus, targeting the CD28-CD80 interaction emerges as a promising therapeutic approach for allergic asthma. However, the impact of CD28 antagonists on allergen-induced asthma remains unreported. In this study, we explored the effects of intranasally administered Cyn-1324 on airway inflammation in the ovalbumin (OVA)-induced murine allergic model. The results revealed a significant reduction in airway hyper-responsiveness (AHR), eosinophil recruitment, and cell infiltration in lung tissues, as well as decreased OVA-specific IgE in serum and Th2 cytokine levels in OVA-stimulated lymphocyte cultures. Additionally, we demonstrated the immunosuppressive effects of Cyn-1324 in vitro, including decreased T-cell proliferation and IL-2 secretion, together with increased p27kip1 expression via inhibiting the PI3K signaling pathway. Notably, Cyn-1324 not only inhibited the NF-κB pathway, but also appeared to suppress p38 activation, which is downstream of CD3 signaling, and reduced calcium-induced NFAT protein expression. These findings suggest that Cyn-1324 alleviates allergic responses by inhibiting the CD28-CD80 interaction and holds promise as an immunosuppressive agent for allergic patients.

CLINICAL CHARACTERISTICS AND SERUM CONCENTRATIONS OF SOME T-HELPER 2-DERIVED CYTOKINES IN PATIENTS WITH GENERALIZED ERYTHEMA MULTIFORME

Objective: To describe the clinical characteristics of patients with erythema multiforme (EM) presenting with generalized skin lesions and to investigate the serum concentrations of certain T-helper 2 (Th2) cytokines in these patients, exploring their correlation with specific clinical features. Subjects and Methods: This cross-sectional descriptive study included 33 patients diagnosed with EM featuring generalized skin lesions at the National Hospital of Dermatology and Venereology from April 2017 to August 2019. Additionally, 32 healthy controls (HCs) participated as the control group. Clinical examinations, medical history interviews, and serum sample collections were conducted for all subjects. The fluorescence covalent microbead immunosorbent assay technique was used to detect multiple cytokines simultaneously: IL-4, IL-5, and IL-13. The Mann-Whitney U test compared the serum cytokine levels between the two groups, with statistical significance set as p<0.05. Pearson's test was applied to evaluate the correlation between two quantitative variables. Results: The mean age of EM patients with generalized lesions was 42.2 years; 30.3% were male, and 69.7% were female. Twenty-two patients (66.7%) had an illness duration of less than one week—a history of medication use before illness onset was reported by 60.6% of patients. Only 15.2% of patients had mucosal lesions. In the EM group, serum levels of IL-4 and IL-5 were 1.55±4.35 pg/ml and 7.12±18.91 pg/ml, respectively, which were lower than those in the HCs group (9.15±10.17 pg/ml and 21.38±20.85 pg/ml, respectively), p<0.001. Serum IL-13 levels were identical in both groups (1.48 pg/ml). No correlation was found between serum IL-4 levels and age (r=0.08; p>0.05), nor between serum IL-5 levels and age (r=-0.055; p>0.05). Conclusion: Th2 cytokine levels in the EM group with generalized skin lesions were lower compared to the HCs group, with no correlation between these cytokine levels and age. Th2 is likely not an important factor in the pathogenesis of generalized EM. Received: 16 May 2023Revised: 24 June 2023Accepted: 22 July 2023

Maternal aspartame exposure alters lung Th1/Th2 cytokine balance in offspring through nuclear factor-κB activation.

Epidemiological evidence suggests that maternal intake of nonnutritive sweeteners is positively associated with early childhood asthma incidence. We investigated the effects of maternal aspartame exposure during pregnancy and lactation on lung Th1/Th2 cytokine balance and intestinal microbiota in offspring and explored the mechanisms that mediate these effects. Pregnant BALB/c mice were randomly divided on gestational day 7 into two dietary intervention groups: control (drinking water only) and aspartame (drinking water+0.25g/L aspartame) groups. The dams nursed their offspring for 3weeks. On postnatal day 21, heart blood samples were collected, and immunoglobulin E levels were measured. Microorganisms from the lower gastrointestinal tract were sampled using a culture-independent approach. Lung tissues were harvested for biochemical analyses. Maternal aspartame exposure increased the body weight of the dams from gestational day 7 to postnatal day 21 and the body weight of the offspring from birth to postnatal day 21. Maternal aspartame exposure significantly increased the levels of Th2 cytokines (interleukin [IL]-4, IL-5, and IL-13) and IL-17 and immunoglobulin E but reduced that of a Th1 cytokine (interferon-γ) in the offspring's lung tissues. The altered Th1/Th2 balance was accompanied by increased lung nuclear factor-κB activation. The bacterial composition and alpha-diversity of the gut microbiota of the offspring did not differ significantly between the control and aspartame groups. Our findings suggest maternal aspartame exposure influences lung Th1/Th2 cytokine balance in offspring through nuclear factor-κB activation.

SPA14 liposomes combining saponin with fully synthetic TLR4 agonist provide adjuvanticity to hCMV vaccine candidate

In the aim of designing and developing a novel saponin-based adjuvant system, we combined the QS21 saponin with low microgram amounts of the fully synthetic TLR4 agonist, E6020, in cholesterol-containing liposomes. The resulting adjuvant system, termed SPA14, appeared as a long-term stable and homogeneous suspension of mostly unilamellar and a few multilamellar vesicles, with an average hydrodynamic diameter of 93 nm, when formulated in citrate buffer at pH 6.0-6.5. When compared in an in vitro human innate immunity construct to AS01B, the QS21/MPL® liposomal adjuvant system of GSK, and with QS21-Liposomes used as benchmarks, SPA14 displayed the strongest immunostimulatory potential based on antigen-presenting cell (APC) activation and cytokine secretion, which was essentially driven by the highly active E6020 agonist in this assay. When tested as an adjuvant in vivo with human cytomegalovirus glycoprotein B (gB) and pentamer complex (PC) as test antigens, SPA14 was generally well tolerated and as active as AS01B for the induction of long-lasting CMV-neutralizing antibodies in mice and non-human primates (NHPs). Both adjuvants promoted the induction of Th-1 responses based on IgG2c production in mice and IFN-γ production in mice and NHPs, but in mice, a higher level of Th-2 cytokines (IL-5) and higher IgG1 over IgG2c secreting cells ratios were obtained with SPA14 indicating that the adjuvant profile of SPA14 could be less Th-1 biased than that of AS01B. From a developability standpoint, SPA14 could be manufactured by a simple and scalable ethanol injection method, owing to the high solubility in ethanol of all its lipidic components, including E6020. Furthermore, E6020 is a single molecule, well-characterized fully synthetic TLR4 agonist constructed in eight synthetic steps from entirely crystalline starting materials and intermediates via an optimized high-yield synthetic route. Overall, our data suggest that SPA14 is a viable, easy-to-manufacture, potent novel adjuvant system that could be broadly applicable as a ready-to-mix adjuvant in the form of a long-term stable liquid formulation.

Sex-specific cytokine, chemokine, and growth factor signatures in T1D patients and progressors.

Numerous studies have reported altered cytokine levels in type 1 diabetes (T1D) patients, yet findings remain inconsistent. In this pilot study, we tested the hypothesis that circulating immune markers exhibit sex-based differences in T1D, both prior to and after disease onset. We analyzed 47-48 cytokine, chemokine, and growth factor levels in two cohorts. To assess post-disease differences, we analyzed serum samples from 25 controls and 25 T1D patients. To examine pre-disease progression, we utilized samples from 21 control children and 16 T1D progressors, collected at age 5 years before disease onset. Across all T1D patients and controls, only macrophage colony-stimulating factor and interleukin (IL)-6 showed significant differences. However, we identified notable alterations when comparing sex-age-matched controls and T1D samples. Female T1D patients exhibited lower levels of inflammatory cytokines (tumor necrosis factor-α, IL-6, IL-1a), Th2 cytokines (IL-4, IL-13), and chemokines (macrophage inflammatory protein (MIP)-1α, regulated upon activation, normal T cell expressed and secreted, MIP-3) compared to female controls, differences that were not observed in males. Notably, IL-22 was lower in female T1D patients compared to female controls, whereas it was higher in male T1D patients compared to male controls. Male T1D patients showed elevated levels of growth factors (epidermal growth factor, platelet-derived growth factor-AB/BB) compared to male controls. In T1D progressors, growth-regulated alpha was lower compared to controls in both sexes. Multiple regression analysis further revealed associations between cytokine levels and factors such as age, BMI, and breastfeeding duration. Overall, our findings serve as a proof of concept, highlighting the importance of sex-specific differences in T1D pathogenesis. However, follow-up studies with larger sample sizes are needed to validate and generalize these results.

Association of systemic inflammation and long-term dysfunction in COVID-19 patients: A prospective cohort.

COVID-19 has significant long-term impacts, including a chronic syndrome known as long-COVID, characterized by persistent symptoms post-recovery. The inflammatory response during acute infection is hypothesized to influence long-term outcomes. This study aimed to identify inflammatory biomarkers predictive of functional outcomes one year after hospital discharge. A prospective cohort study was conducted with 213 COVID-19 patients admitted to ICUs in Southern Brazil between June and November 2020. After exclusions and follow-ups, 109 patients were evaluated for one-year post-discharge. Plasma levels of Th1 (TNF-α, INF-γ, IL-12), Th2 (IL-4, IL-5, IL-6, IL-10, IL-13), and Th17 (IL-17, IL-22) cytokines were measured. Functional outcomes in psychiatric, cognitive, general health, and health perception domains were assessed. Statistical analyses included multivariate regression, regularized partial correlation network analysis, and K-means clustering. We demonstrate that plasma levels of various cytokines, along with demographic and clinical characteristics, can predict four distinct domains of functional outcomes one year following hospital discharge due to COVID-19 and that an hyperinflammatory phenotype was associated with the occurrence of a worse in psychiatric, general health, and health perception domains. The network analysis highlighted complex interconnections among immune markers and clinical variables, elucidating their roles in long-term health. These findings support using biomarkers for patient stratification and indicate potential targets for therapeutic interventions.

Age-specific dynamics of neutralizing antibodies, cytokines, and chemokines in response to La Crosse virus infection in mice.

La Crosse virus (LACV) is a primary cause of pediatric arboviral encephalitis in the United States, particularly affecting children aged 16 years or younger. This age-related susceptibility extends to murine models, where weanling mice (3 weeks old) succumb to LACV infection, while adults (≥6 weeks old) demonstrate resistance. Despite its clinical relevance, the host immune response to LACV is not fully understood. In this study, we investigated the roles of neutralizing antibodies (nAbs), cytokines, and chemokines in weanling and adult mice following infection with 5 × 105 plaque-forming units (PFU) of LACV. Weanling mice demonstrated early disease onset with elevated peripheral viremia, but passive transfer of adult serum, confirmed to have nAbs, to naïve weanlings prior to infection completely rescued them from death. Moreover, adult mice had increased Th1 cytokines, Th9/Th17/Th22/Treg cytokines, and many chemokines. In contrast, weanlings had higher Th2 cytokines, correlating with symptoms onset. Flow cytometry and intracellular cytokine staining further demonstrated that weanling mice produced higher levels of IL-4 by CD4+ and CD8+ T cells compared to adults, regardless of infection status. Conversely, LACV-infected adult mice had increased IFN-γ production by CD8+ T cells compared to uninfected controls. Finally, the adoptive transfer of splenocytes from immune adult mice to naïve weanlings delayed neurological symptoms and improved survival. In conclusion, this study links nAbs and cytokine and chemokine responses to protective immunity in adult mice, contrasting with the pathogenesis seen in weanlings. These findings underscore the importance of further research into innate and adaptive immune mechanisms during LACV infection.IMPORTANCELa Crosse virus (LACV) is a primary cause of pediatric encephalitis in the United States, with an impact on children aged 16 years or younger. This age-related susceptibility is recapitulated in mouse models, where young mice succumb to LACV-induced disease, while adults demonstrate resistance. Our understanding of host responses to LACV remains underexplored. This study sheds light on the dynamics of neutralizing antibodies (nAbs), cytokines, and chemokines following LACV infection in both adult and weanling mice. Our study reveals age-specific variations in viremia, neutralizing antibody titers, survivability, and levels of cytokines and chemokines. Adult mice exhibit significantly elevated levels of Th1 cytokines, contrasting with elevated levels of Th2 cytokines observed in weanling mice, often coinciding with the onset of symptoms. These data reveal age-specific dynamics in cytokines and chemokines associated with protective versus pathogenic immunity, emphasizing the need for further studies on innate and adaptive immunity.

Interconnection of the Gut-Skin Axis in NC/Nga Mouse with Atopic Dermatitis: Effects of the Three Types of Bifidobacterium bifidum CBT-BF3 (Probiotics, Postbiotics, and Cytosine-Phosphate-Guanine Oligodeoxynucleotide) on T Cell Differentiation and Gut Microbiota.

The gut microbiota is an immune system regulator in the gut-skin axis. Dysfunctional interactions between the gut microbiota and the gut immune system can lead to the development of skin diseases such as atopic dermatitis (AD). Probiotics and postbiotics positively affect the balance of the gut microbiota, immune regulation, protection against pathogens, and barrier integrity. This study investigated the effects of probiotic Bifidobacterium bifidum, postbiotic B. bifidum (heat-killed), and cytosine-phosphate-guanine oligodeoxynucleotide (CpG ODN) on the gut microbiota and T cell differentiation in NC/Nga mice induced with AD. 2,4-Dinitrochlorobenzene-induced AD mice had an increased SCORing atopic dermatitis-index and increased mRNA expression levels of Th2 and Th17 cell transcription factors and cytokines, and thymic stromal lymphopoietin (TSLP) cytokine in their mesenteric lymph nodes (mLNs; p<0.05). However, oral administration of the three types of B. bifidum (probiotics, postbiotics, CpG ODN) to AD mice decreased the mRNA expression levels of Th2 and Th17 cell transcription factors and cytokines as well as TSLP cytokine. They increased the mRNA expression levels of regulatory T (Treg) cell transcription factor and cytokine, galectin-9, and filaggrin genes (p<0.05). These effects were more noticeable in the mLNs than in the spleen. In addition, AD mice showed a decrease in Faecalibacterium prausnitzii, Roseburia spp., Leuconostoc citreum, Weissella cibaria, and Weissella koreensis (p<0.05). However, oral administration of the three types of B. bifidum increased Bacteroides spp., Bifidobacterium spp., F. prausnitzii, and Roseburia spp. (p<0.05).

Th17 and T Regulatory Cytokines in Serum, Lesional Skin, and Stimulated Peripheral Blood Mononuclear Cell Culture Supernatants from Type 1 Leprosy Reaction Patients.

There are conflicting reports regarding the roles of T helper-17 (Th17) and T regulatory (Treg) cells in type 1 leprosy reactions (T1Rs). Also, literature on the correlation of immunological parameters with a validated scoring system and the effect of treatment on cytokines is lacking. Adult patients with untreated T1R and nonreactional spectrum-matched controls were included in the study for comparison of levels of Th17 and Treg pathway cytokines in serum, skin lesions (reactional), and peripheral blood mononuclear cells (PBMCs) culture supernatants. Venous blood samples were collected at baseline and after resolution of reaction (post treatment with nonsteroidal anti-inflammatory drugs [NSAIDs] or steroids) for serum cytokine estimation and PBMC stimulation assays, and lesional (reactional) skin biopsy for cytokine messenger RNA (mRNA) estimation. Thirty-two cases of T1R were recruited (23 patients completed follow-up). Serum levels of cytokines were not significantly different between cases and controls or between pre- and post-treatment samples. Tissue mRNA and Mycobacterium leprae (M. leprae) antigen-stimulated PBMC culture supernatant levels of Interleukin (IL)-17A, IL-17F, IL-6, and IL-23 were significantly higher in T1R than in controls. Levels of IL-10 and Transforming Growth Factor-beta (TGF-β) were comparable among the two groups. The levels of all cytokines were significantly reduced after treatment. There was no significant difference in magnitude of the fall between those treated with steroids versus NSAIDs. This study suggests heightened Th17 response in T1R, with a prominent inability of the regulatory cytokines IL-10 and TGF-β to control the associated inflammation. The dynamics of change after resolution of T1R were comparable between NSAID and oral steroid treatment groups.

Dynamics of Peripheral Lymphocyte Subsets from Birth until Old Age

The immune system is inexperienced before birth and tends to be tolerogenic, rather than immunogenic. After birth, the adaptive immune system develops while facing microbial challenges, but it can become impaired as old age progresses and persistent inflammation can lead to chronic morbidity, disability and frailty. To investigate the potential contributions of lymphocyte subsets to immunity from birth until old age, we enumerated circulating innate and conventional lymphocytes and measured serum cytokine levels in 10 cord blood samples and in peripheral blood from 10 healthy term neonates, 23 healthy school-age children, 25 young adults and 11 older subjects. Flow cytometric analysis revealed that B cell frequencies increase during childhood and gradually decrease into adulthood, whereas natural killer cell frequencies increase throughout life. T cell frequencies remained relatively constant throughout life, as did their expression of CD4 and CD8. However, all four innate T cell populations studied—invariant natural killer T cells, mucosa-associated invariant T cells and the Vδ1 and the Vδ2 subsets of γδ T cells—were extremely rare in cord blood and in peripheral blood of neonates, but they expanded after birth reaching highest levels in adulthood. Analysis of serum cytokine levels revealed that proinflammatory and T helper type 1 (Th1) cytokine levels increase in adulthood, whereas Th2 and Th17 cytokine levels remain relatively constant. These changes in lymphocyte numbers and cytokine levels across the lifetime are likely to affect immunocompetence, leaving newborn and elderly people susceptible to infection, cancer and immune-mediated disease.

Unveiling Potent Anti-Asthmatic Effect of Curcumin in Combination with Salmeterol in Swiss Albino Mice

Background: Asthma is a long-term inflammatory respiratory condition marked by alterations in the airways and an increase in inflammatory cell infiltration. It has been observed that Curcumin possesses immune-modulating, anti-inflammatory, and relaxing properties for smooth muscle in the airways. Salmeterol is believed to ease the smooth muscles of the airways. Objective: Swiss Albino mice were used in the research to examine the combination anti-asthmatic effects of Curcumin and Salmeterol in asthma produced by ova albumin and milk induced eosinophilia and leucocytosis. Methods: The mice received pre-treatment with Curcumin (10 mg/kg, 20 mg/kg intraperitoneally) as well as Salmeterol (5 mg/kg) after being stimulated with an Ovalbumin (OVA) challenge and milk. After the induction period, various hematological, biochemical, molecular (ELISA), and histological analyses were performed. Results: The findings demonstrated that the combined treatment decreased the animal’s overall leukocyte and eosinophil numbers in a manner that was dose-dependent. Additionally, the therapy reduced albumin and overall protein amount in serum, BALF and lung tissues, facilitated changes in haematological parameters, and reduced the rise of Th2 cytokines (IL-4, TNF-α, IL-13) levels that is induced by OVA in lungs and BALF, total IgE level in serum. The combined action of Curcumin and Salmeterol reduced OVA-induced inflammatory influx and ultrastructural abnormalities, according to histopathological evaluation. Conclusion: The findings of this investigation demonstrate that curcumin and salmeterol together possess anti-asthmatic effects through suppressing Th2 triggered immune response and possessing an anti-inflammatory effect and anti-allergic effect. Thus combination of treatments might be a novel technique for managing asthma.

The expression and clinical significance of cytokines Th1, Th2, and Th17 in ovarian cancer

BackgroundThis study was to analyze the levels of Th1, Th2 and Th17 cytokines in peripheral blood samples from ovarian cancer (OC) patients. MethodsNinty-five OC patients including 45 OC and 50 benign ovarian disease (BOD) were selected at Zhejiang Cancer Hospital from October 2021 to March 2022; 46 healthy participants were simultaneously selected at Taizhou Municipal Hospital as healthy controls (HC). The expressions of Th1, Th2 and Th17 were compared in all participants. Marker levels were analyzed with age, histological type, tumor size, ovarian number and clinical stage of OC. ResultsThe IL6 and IL8 levels were significantly higher in OC compared to BOD and HC (p < 0.00). The IL-4 expression was significantly higher in OC compared to HC (p < 0.00). The expressions of IL2, IL6 and IL10 were significantly higher in pathological stage III-IV OC compared with pathological stage I-II OC (p < 0.05). Meanwhile, the levels of IL-2 and IL-10 were significantly higher in OC with bilateral ovaries than in OC with single ovary (p < 0.05). AUCs of different markers were to diagnose OC. The findings also implied that the expressions of IL-6, IL-8 and IL-10 were significantly different between OC and control groups (p < 0.05), while the levels of IL-2, IL-4, TNF-α, IFN-γ, IL-12p70, IL-1β and IL-5 between the two groups were not different (p > 0.05). ConclusionsIn peripheral blood from OC patients, the immune system was more dysregulated and immune cells produced more cytokines with contrasting actions. These data showed significant clinical implications for the diagnosis of OC.

Allergo-immunopathology mechanism of thymol-inhibiting airway remodeling in asthmatic mice by regulating TGF-β/Smad3 pathway.

Allergic asthma is an important public health problem and is a complicated respiratory sickness that is characterized by bronchial inflammation, bronchoconstriction, and breathlessness. Asthma is orchestrated by type 2 immune response and remodeling is one of the important outputted problem in chronic asthma. Thymol is a naturally occurring monocyclic phenolic, it has a series of biological properties, and its immunomodulatory and anti-remodeling effects on allergic asthma were evaluated. The OVA-LPS-induced asthmatic mice were treated with thymol. Methacholine challenge test, eosinophil count, and levels of IL-4, IL-5, IL-13, and IL-33 in bronchoalveolar lavage fluid, total and OVA-specific IgE levels in serum, remodeling factors, gene expression of TGF-β, Smad2, Smad3, and lung histopathology were done. Treatment with thymol could control AHR, eosinophil percentage levels of Th2 cytokines and Igs, remodeling factors, expression of TGF-β, Smad2 and Smad3 genes, inflammation, goblet cell hyperplasia, and mucus production in asthmatic mice. Thymol can control asthma pathogens and related remodeling and fibrosis bio-factors and can be a potential treatment of asthma.

Comparison of serum cytokines and chemokines levels and clinical significance in patients with pemphigus vulgaris-A retrospective study.

In this study, we aimed to examine the relationship between the serum cytokine levels of patients with pemphigus vulgaris (PV) and the Pemphigus Disease Area Index (PDAI), along with the presence of anti-desmoglein (Dsg) 1 antibody, anti-Dsg3 antibody and co-infection among patients with pemphigus vulgaris. This retrospective study included 62 PV patients and 59 healthy individuals who attended the Second Affiliated Hospital of Kunming Medical University from November 2014 to November 2022. The serum concentrations of cytokines and chemokines were assessed using the Luminex 200 System (a high-throughput cytokine detection method). Additionally, anti-Dsg1 and anti-Dsg3 antibodies were determined through enzyme-linked immunosorbent assay, while disease severity was evaluated using the PDAI scoring system. The PV group exhibited elevated levels of Th1 cytokines (such as interleukin (IL)-1RA, IL-1β, IL-2, IL-12p70, GM-CSF, TNF-α, IL-18, IFN-γ), Th2 cytokines (IL-5, IL-10, IL-13) and Th17/Th22-related cytokines (IL-17A, IL-22) compared to the healthy control group (p < 0.05). Conversely, the levels of chemokines (macrophage inflammatory protein-1 alpha (MIP-1α), stromal cell-derived factor-1 alpha (SDF-1α), interferon-inducible protein-10 (IP-10), Regulated on Activation in Normal T-Cell Expressed And Secreted (RANTES), growth-regulated on-gene-alpha (GRO-α), MIP-1β) and Th2 (IL-31) were lower in the PV group compared to the healthy control group (p < 0.05). No significant differences were observed in other cytokines and chemokines (p > 0.05). Additionally, IL-7, IFN-γ, IL-18 and GRO-α showed positive correlations with PDAI, IL-6 correlated positively with anti-Dsg3 antibody levels, and IL-12p70, IL-18, and IFN-γ correlated positively with anti-Dsg1 antibody levels. Furthermore, IL-15 exhibited a positive association with skin infections. PV patients have elevated levels of various cytokines and chemokines, and there are different degrees of elevation in cytokines and chemokines associated with the activation of various T cell subsets. PDAI and the Dsg1 antibody levels are mainly related to the Th1-related cytokines.

Fas/FasL-Mediated Apoptosis and Inflammation Contribute to Recovery from HSV-2-Mediated Spinal Cord Infection.

Herpes simplex virus type 2 (HSV-2) is a sexually transmitted pathogen that causes a persistent infection in sensory ganglia. The infection manifests itself as genital herpes but in rare cases it can cause meningitis. In this study, we used a murine model of HSV-2 meningitis to show that Fas and FasL are induced within the CNS upon HSV-2 infection, both on resident microglia and astrocytes and on infiltrating monocytes and lymphocytes. Mice lacking Fas or FasL had a more severe disease development with significantly higher morbidity, mortality, and an overall higher CNS viral load. In parallel, these Fas/FasL-deficient mice showed a severely impaired infection-induced CNS inflammatory response with lower levels of infiltrating CD4+ T-cells, lower levels of Th1 cytokines and chemokines, and a shift in the balance between M1 and M2 microglia/monocytes. In vitro, we confirmed that Fas and FasL is required for the induction of leucocyte apoptosis, but also show that the Fas/FasL pathway is required for adequate cytokine and chemokine production by glial cells. In summary, our data show that the Fas/FasL cell death receptor pathway is an important defense mechanism in the spinal cord as it down-regulates HSV-2-induced inflammation while at the same time promoting adequate anti-viral immune responses against infection.

Variants in the Late Cornified Envelope Gene Locus Are Associated With Elevated T-helper 17 Responses in Patients With Postinfectious Lyme Arthritis.

Postinfectious Lyme arthritis (LA) is associated with dysregulated immunity and autoreactive T- and B-cell responses in joints. Here we explored the role of host genetic variation in this outcome. The frequency of 253 702 single-nucleotide polymorphisms (SNPs) was determined in 147 patients with LA (87 with postinfectious LA and 60 with antibiotic-responsive LA), and for comparison in 90 patients with erythema migrans or the general population (n = 2504). Functional outcome of candidate SNPs was assessed by evaluating their impact on clinical outcome and on immune responses in blood and synovial fluid in patients with LA. Six SNPs associated with late cornified envelope (LCE3) genes were present at greater frequency in patients with postinfectious LA compared to those with antibiotic-responsive LA (70% vs 30%; odds ratio, 2; P < .01). These SNPs were associated with heightened levels of inflammatory Th17 cytokines in serum but lower levels of interleukin 27, a regulatory cytokine, implying that they may contribute to dysregulated Th17 immunity in blood. Moreover, in patients with postinfectious LA, the levels of these Th17 mediators correlated directly with autoantibody responses in synovial fluid, providing a possible link between LCE3 SNPs, maladaptive systemic Th17 immunity, and autoreactive responses in joints. Variation in the LCE3 locus, a known genetic risk factor in psoriasis and psoriatic arthritis, is associated with dysregulated systemic Th17 immunity and heightened autoantibody responses in joints. These findings underscore the importance of host genetic predisposition and systemic Th17 immunity in the pathogenesis of postinfectious (antibiotic-refractory) Lyme arthritis.

In mouse model of mixed granulocytic asthma with corticosteroid refractoriness, Bronchom mitigates airway hyperresponsiveness, inflammation and airway remodeling

BackgroundAsthma is a heterogeneous, inflammatory disease with several phenotypes and endotypes. Severe asthmatics often exhibit mixed granulocytosis with reduced corticosteroid sensitivity. Bronchom is a newly developed Ayurvedic prescription medicine, indicated for the treatment of obstructive airway disorders. The purpose of the present study was to evaluate the in-vivo efficacy of Bronchom in mouse model of mixed granulocytic asthma with steroidal recalcitrance.MethodsHigh-performance thin layer chromatography (HPTLC) and Ultra-high performance liquid chromatography (UHPLC) were employed to identify and quantitate the phytometabolites present in Bronchom. The preclinical effectiveness of Bronchom was assessed in house dust mite (HDM) and Complete Freund’s adjuvant (CFA)-induced mixed granulocytic asthma model in mice. High dose of dexamethasone was tested parallelly. Specific-pathogen-free C57BL/6 mice were immunized with HDM and CFA and nineteen days later, they were intranasally challenged with HDM for four consecutive days. Then the mice were challenged with nebulized methacholine to evaluate airway hyperresponsiveness (AHR). Inflammatory cell influx was enumerated in the bronchoalveolar lavage fluid (BALF) followed by lung histology. Additionally, the concentrations of Th2 and pro-inflammatory cytokines was assessed in the BALF by multiplexed immune assay. The mRNA expression of pro-inflammatory cytokines and Mucin 5AC (MUC5AC) was also evaluated in the lung.ResultsHPTLC fingerprinting and UHPLC quantification of Bronchom revealed the presence of bioactive phytometabolites, namely, rosmarinic acid, gallic acid, methyl gallate, piperine, eugenol and glycyrrhizin. Bronchom effectively reduced AHR driven by HDM-CFA and the influx of total leukocytes, eosinophils and neutrophils in the BALF. In addition, Bronchom inhibited the infiltration of inflammatory cells in the lung as well as goblet cell metaplasia. Further, it also suppressed the elevated levels of Th2 cytokines and pro-inflammatory cytokines in the BALF. Similarly, Bronchom also regulated the mRNA expression of pro-inflammatory cytokines as well as MUC5AC in mice lungs. Reduced effectiveness of a high dose of the steroid, dexamethasone was observed in the model.ConclusionsWe have demonstrated for the first time the robust pharmacological effects of an herbo-mineral medicine in an animal model of mixed granulocytic asthma induced by HDM and CFA. The outcomes suggest the potential utility of Bronchom in severe asthmatics with a mixed granulocytic phenotype.

Recombinant Subunit Vaccine Candidate against the Bovine Viral Diarrhea Virus.

Multivalent live-attenuated or inactivated vaccines are often used to control the bovine viral diarrhea disease (BVD). Still, they retain inherent disadvantages and do not provide the expected protection. This study developed a new vaccine prototype, including the external segment of the E2 viral protein from five different subgenotypes selected after a massive screening. The E2 proteins of every subgenotype (1aE2, 1bE2, 1cE2, 1dE2, and 1eE2) were produced in mammalian cells and purified by IMAC. An equimolar mixture of E2 proteins formulated in an oil-in-water adjuvant made up the vaccine candidate, inducing a high humoral response at 50, 100, and 150 µg doses in sheep. A similar immune response was observed in bovines at 50 µg. The cellular response showed a significant increase in the transcript levels of relevant Th1 cytokines, while those corresponding to the Th2 cytokine IL-4 and the negative control were similar. High levels of neutralizing antibodies against the subgenotype BVDV1a demonstrated the effectiveness of our vaccine candidate, similar to that observed in the sera of animals vaccinated with the commercial vaccine. These results suggest that our vaccine prototype could become an effective recombinant vaccine against the BVD.

Transcriptome and pathophysiological analysis during Bacillus cereus group infection in Pelodiscus sinensis uncovered the importance of iron and toll like receptor pathway

The Chinese softshell turtle (CST; Pelodiscus sinensis) is an important freshwater aquaculture species with high economic value. CST is farmed throughout Asia, especially in China and Taiwan. However, the presence of diseases, particularly those caused by the Bacillus cereus group, poses significant threats to commercial CST farming. A recent study has unveiled new genospecies within the B. cereus group, Bacillus shihchuchen. Within this group, strain QF108–045 stands out for its high virulence, notably leading to severe septicemia in CST. This study aimed to explore the transcriptome and pathophysiological changes in CSTs infected with QF108–045. Upon infection with QF108–045, turtles exhibited upregulation of Toll-like receptor (TLR) 2, 4, and 5, among which, TLR5 was significantly upregulated. Activation of TLR signaling leading to elevated levels of IL-1β, TNFα, and Th17 cytokines, which subsequently activate nuclear factor kappa B (NF-κB). CSTs challenged with QF108–045 exhibited a substantial decrease packed cell volume, indicating severe hemolysis. The notable upregulation of transferrin, ZIP8/14, and HO-1 indicates an active transfer of iron or heme into cells, and melanomacrophages in the spleen showed a positive signal in Perl's Prussian blue staining, implying their importance as essential iron storage cells. However, despite the hemolysis, ferroptosis was inhibited by the upregulation of system Xc− related genes. Additionally, our study revealed a notable increase in TLR5 expression following immunization with recombinant flagellin from QF108–045, coupled with analysis of protein tertiary docking modules. These findings advance our understanding of defense mechanisms in CSTs during QF108–045 infection, providing valuable insights for future vaccine development efforts.