Mesenchymal Stem Cell-Derived Exosome-Containing Linc00632 Regulates GATA Binding Protein-3 Expression in Allergic Rhinitis by Interacting with Enhancer of Zeste Homolog 2 to Inhibit T Helper Cell 2 Differentiation

<b><i>Background:</i></b> Allergic rhinitis (AR) is regarded as one of the most common allergic disease of nasal mucosa affecting many people worldwide. Long noncoding RNAs are critical modulators affecting AR progression, whereas the pathogenesis of Linc00632 in the development of AR remains unclear. <b><i>Methods:</i></b> T helper cell 2 (Th2) differentiation of CD4⁺ T cells was measured by flow cytometry. Real-time quantitative PCR assay and Western blot were applied to determine the levels of RNA and proteins, respectively. The interleukin (IL)-4 and IL-13 levels were quantitatively assessed through ELISA. Subcellular fractionation was conducted to detect the cellular localization of Linc00632. RNA immunoprecipitation experiment was employed to validate the interaction relationship between Linc00632 and enhancer of zeste homolog 2 (EZH2). Chromatin immunoprecipitation assay was used for determination of protein-DNA interactions. <b><i>Results:</i></b> The expression of Linc00632 was significantly decreased by 4 times in nasal mucosa of AR patients. Human umbilical cord mesenchymal stem cell-derived exosome dramatically inhibited Th2 differentiation, decreased GATA binding protein-3 (GATA-3) protein expressions and IL-4 levels by about 2 times in CD4⁺ T cells. Knockdown Linc00632 partially reversed the effects of exosomes on Th2 differentiation, IL-4 and IL-13 levels, and GATA-3 expression. Linc00632 overexpression could suppress Th2 differentiation of CD4⁺ T cells, reduced IL-4 and IL-13 levels, and GATA-3 expressions roughly 2 times. Linc00632 repressed the expression of GATA-3 by interacting with EZH2. GATA-3 overexpression partially reversed the effect of Linc00632 on Th2 differentiation of CD4⁺ T cells. <b><i>Conclusion:</i></b> Linc00632 acted as a suppression factor in Th2 differentiation by inhibiting the expression of GATA-3 via interacting with EZH2, which might provide a new insight for understanding the action mechanism of Linc00632 in AR.

Effect of Coptischinensis, Glycyrrhiza uralensis, and Fermented Glycine max Extract as Proactive Therapy for Atopic Dermatitis

The aim of this study was to investigate the effect of the Coptis chinensis, Glycyrrhiza uralensis, and fermented Glycine max (3Hb) extract on lipid barrier recovery and the alleviation of atopic dermatitis (AD). The 3Hb extract was administered to lipid barrier-eliminated mice (3HbT) for 5 days. Subsequently, the effect of the 3Hb extract on general skin features and the regulation of filaggrin, inflammatory response, Th2 differentiation, and the skin micro-environment for defense, was evaluated. In the 3HbT, filaggrin was effectively recovered. The clinical skin score was significantly lower in the 3HbT compared with control groups. In addition, significant decreases in pH and TEWL as well as in the levels of kallikrein 7, PAR-2, TSLP, IL-4, Fc ε receptor, and phosphate-NF-κB p65 were observed in the 3HbT, compared with the other control groups. Further, compared with control groups, the 3HbT showed a significant increase in those of claudin, cathelicidin, TLR, and NHE-1. Our results indicated that the 3Hb extract effectively recovered filaggrin. Through the recovery of filaggrin, inflammation and the Th2 differentiation process can be regulated, and microenvironments for defense can be recovered. Therefore, we confirmed the potential of the 3Hb extract for use in the proactive therapy of AD.

CD11b+ lung dendritic cells at different stages of maturation induce Th17 or Th2 differentiation

Dendritic cells (DC) in the lung that induce Th17 differentiation remain incompletely understood, in part because conventional CD11b+ DCs (cDC2) are heterogeneous. Here, we report a population of cDCs that rapidly accumulates in lungs of mice following house dust extract inhalation. These cells are Ly-6C+, are developmentally and phenotypically similar to cDC2, and strongly promote Th17 differentiation ex vivo. Single cell RNA-sequencing (scRNA-Seq) of lung cDC2 indicates 5 distinct clusters. Pseudotime analysis of scRNA-Seq data and adoptive transfer experiments with purified cDC2 subpopulations suggest stepwise developmental progression of immature Ly-6C+Ly-6A/E+ cDC2 to mature Ly-6C–CD301b+ lung resident cDC2 lacking Ccr7 expression, which then further mature into CD200+ migratory cDC2 expressing Ccr7. Partially mature Ly-6C+Ly-6A/E–CD301b– cDC2, which express Il1b, promote Th17 differentiation. By contrast, CD200+ mature cDC2 strongly induce Th2, but not Th17, differentiation. Thus, Th17 and Th2 differentiation are promoted by lung cDC2 at distinct stages of maturation.

Tissue-based IL-10 signalling in helminth infection limits IFNγ expression and promotes the intestinal Th2 response

Type 2 immunity is activated in response to both allergens and helminth infection. It can be detrimental or beneficial, and there is a pressing need to better understand its regulation. The immunosuppressive cytokine IL-10 is known as a T helper 2 (Th2) effector molecule, but it is currently unclear whether IL-10 dampens or promotes Th2 differentiation during infection. Here we show that helminth infection in mice elicits IL-10 expression in both the intestinal lamina propria and the draining mesenteric lymph node, with higher expression in the infected tissue. In vitro, exogenous IL-10 enhanced Th2 differentiation in isolated CD4+ T cells, increasing expression of GATA3 and production of IL-5 and IL-13. The ability of IL-10 to amplify the Th2 response coincided with its suppression of IFNγ expression and, in vivo, we found that, in intestinal helminth infection, IL-10 receptor expression was higher on Th1 cells in the small intestine than on Th2 cells in the same tissue, or on any Th cell in the draining lymph node. In vivo blockade of IL-10 signalling during helminth infection resulted in an expansion of IFNγ+ and Tbet+ Th1 cells in the small intestine and caused a coincident decrease in IL-13, IL-5 and GATA3 expression by intestinal T cells. Together our data indicate that IL-10 signalling promotes Th2 differentiation during helminth infection at least in part by regulating competing Th1 cells in the infected tissue.

Effect of Thymoquinone on Th1 and Th2 Balance in Rats Infected with Mycobacterium tuberculosis

Thymoquinone is an active compound in Nigella sativa which has potential immunomodulatory effect. Mycobacterium tuberculosis could alter the Th1 and Th2 balance by stimulating phagocyte IL-1β production, and subsequent Th2 differentiation. We aim to evaluate the effect of thymoquinone (TQ) in restore the Th1 and Th2 balance in Mycobacterium tuberculosis infection. Four groups of rats were infected with virulent Mycobacterium tuberculosis strain H37Rv. Thymoquinone at different doses was given to three groups, and one group left without treatment. Additional one group was either infected or treated with TQ. We measure IL-1β, IL-4 and IFN-γ levels using ELISA 14 days after TQ treatment. We found there were increased IL-1β, IL-4 and IFN-γ level after Mycobacterium tuberculosis infection, but we observe no significant effect of TQ treatment to Th1 and Th2 balance. We conclude that TQ could not restore Th1 and Th2 balance in rats infected with Mycobacterium tuberculosis.

The CDK8 inhibitor DCA promotes a tolerogenic chemical immunophenotype in CD4 + T cells via a novel CDK8-GATA3-FOXP3 pathway.

Immune health requires innate and adaptive immune cells to engage precisely balanced pro- and anti-inflammatory forces. We employ the concept of chemical immunophenotypes to classify small molecules functionally or mechanistically according to their patterns of effects on primary innate and adaptive immune cells. The high-specificity, low-toxicity cyclin dependent kinase 8 (CDK8) inhibitor DCA exerts a distinct tolerogenic profile in both innate and adaptive immune cells. DCA promotes T reg and Th2 differentiation, while inhibiting Th1 and Th17 differentiation, in both murine and human cells. This unique chemical immunophenotype led to mechanistic studies showing that DCA promotes T reg differentiation in part by regulating a previously undescribed CDK8-GATA3-FOXP3 pathway that regulates early pathways of Foxp3 expression. These results highlight previously unappreciated links between T reg and Th2 differentiation and extend our understanding of the transcription factors that regulate T reg differentiation and their temporal sequencing. These findings have significant implications for future mechanistic and translational studies of CDK8 and CDK8 inhibitors.

Decidual NR2F2-Expressing CD4+ T Cells Promote TH2 Transcriptional Program During Early Pregnancy

A unique immunotolerant microenvironment with Th2 bias in the decidua provides an essential security for successful pregnancy. The disorganized maternal-fetal immune tolerance contributes to more than 50% of unexplained recurrent spontaneous abortion (RSA). How the Th2 bias is developed at the maternal-fetal interface remains undefined. NR2F2, a member of steroid/thyroid nuclear receptor superfamily, is endowed with diverse importance in cell-fate specification, organogenesis, angiogenesis, and metabolism. Here, we showed that NR2F2 was absolutely highly expressed in decidual CD4+T(dCD4+T) cells, but not in peripheral circulating CD4+T cells during early pregnancy. Decidual NR2F2-expressing CD4+T cells dominantly produced Th2 cytokines. In unexplained RSA patients, NR2F2 expression in dCD4+T cells was significantly decreased, accompanied with disordered phenotype of dCD4+T cells. Furthermore, overexpression of NR2F2 promoted the Th2 differentiation of naive CD4+T cells. Immunoprecipitation experiment confirmed the binding relationship between GATA-3 and NR2F2, which implied GATA-3 may be an important interactive element involved in the immunoregulatory process of NR2F2. This study is the first to reveal a previously unappreciated role for NR2F2-mediated dCD4+T cells in maternal-fetal immune tolerance and maintenance of normal pregnancy, in the hope of providing a potential biomarker for prediction and prevention of clinical unexplained RSA.