Anti-inflammatory Trained Immunity Mediated by Helminth Products Attenuates the Induction of T Cell-Mediated Autoimmune Disease

Recent studies have suggested that the innate immune system can display characteristics of immunological memory and this has been called “innate immune memory” or “trained immunity.” Certain fungal products have been shown to induce epigenetic imprinting on monocytes/macrophages that results in heightened inflammatory responses to subsequent stimuli. Here we report that innate immune cells can be trained to be more anti-inflammatory following exposure to products of a helminth pathogen. Macrophages trained in vitro with Fasciola hepatica total extract (FHTE) had enhanced IL-10 and IL-1RA, but reduced TNF production upon re-stimulation with FHTE or TLR ligands and this was reversed by inhibitors of DNA methylation. In contrast, macrophages trained with β-glucan or Bacillus Calmette–Guérin had enhanced TNF production upon re-stimulation with Pam3cys or LPS. Furthermore, FHTE-trained macrophages had enhanced expression of markers of alternative activated macrophages (AAM). Macrophages from mice treated with FHTE expressed markers of AAM and had heightened IL-10 and IL-1RA production in response to FHTE or TLR ligands and had suppressed TNF and IL-12p40 production. Macrophages from mice treated with FHTE had reduced APC function and inhibited IL-17 production and the encephalitogenic activity of T cells in the experimental autoimmune encephalomyelitis (EAE) model. In addition, mice pre-treated with FHTE were resistant to induction of EAE and this was associated with a significant reduction in IL-17-producing γδ and CD4 T cells infiltrating the CNS. Our findings reveal that cells of the innate immune system can be trained in vitro or in vivo to be more anti-inflammatory by exposure to helminth products and this protects mice against the induction of a T cell-mediated autoimmune disease

γδ T lymphocytes: An effector cell in autoimmunity and infection

γδ T cells are non-conventional lymphocytes which show several properties of innate immune cells. They present a limited TCR repertoire and circulate as cells with a pre-activated phenotype thus being able to generate rapid immune responses. γδ T cells do not recognize classical peptide antigens, their TCRs are non-MHC restricted and they can respond to pathogen-associated molecular patterns and to cytokines in absence of TCR ligands. They also recognize self-molecules induced by stress, which indicate infection and cellular transformation. All these features let γδ T cells act as a first line of defense in sterile and non-sterile inflammation. γδ T cells represent 1-10% of circulating lymphocytes in the adult human peripheral blood, they are widely localized in non-lymphoid tissues and constitute the majority of immune cells in some epithelial surfaces, where they participate in the maintenance of the epithelial barriers. γδ T cells produce a wide range of cytokines that orchestrate the course of immune responses and also exert high cytotoxic activity against infected and transformed cells. In contrast to their beneficial role during infection, γδ T cells are also implicated in the development and progression of autoimmune diseases. Interestingly, several functions of γδ T cells are susceptible to modulation by interaction with other cells. In this review, we give an overview of the γδ T cell participation in infection and autoimmunity. We also revise the underlying mechanisms that modulate γδ T cell function that might provide tools to control pathological immune responses.Fil: Shiromizu, Carolina Maiumi. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; ArgentinaFil: Jancic, Carolina Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Medicina Experimental. Academia Nacional de Medicina de Buenos Aires. Instituto de Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Microbiología. Cátedra de Microbiología, Parasitología e Inmunología; Argentin

PLoS Genet

All-trans retinoic acid (ATRA) is instrumental to male germ cell differentiation, but its mechanism of action remains elusive. To address this question, we have analyzed the phenotypes of mice lacking, in spermatogonia, all rexinoid receptors (RXRA, RXRB and RXRG) or all ATRA receptors (RARA, RARB and RARG). We demonstrate that the combined ablation of RXRA and RXRB in spermatogonia recapitulates the set of defects observed both upon ablation of RAR in spermatogonia. We also show that ATRA activates RAR and RXR bound to a conserved regulatory region to increase expression of the SALL4A transcription factor in spermatogonia. Our results reveal that this major pluripotency gene is a target of ATRA signaling and that RAR/RXR heterodimers are the functional units driving its expression in spermatogonia. They add to the mechanisms through which ATRA promote expression of the KIT tyrosine kinase receptor to trigger a critical step in spermatogonia differentiation. Importantly, they indicate also that meiosis eventually occurs in the absence of a RAR/RXR pathway within germ cells and suggest that instructing this process is either ATRA-independent or requires an ATRA signal originating from Sertoli cells

Distinct metabolic programmes established in the thymus control effector functions of gd T cell subsets in tumour microenvironments

International audienceMetabolic programming controls immune cell lineages and functions, but little is known about γδ T cell metabolism. Here, we found that γδ T cell subsets making either interferon-γ (IFN-γ) or interleukin (IL)-17 have intrinsically distinct metabolic requirements. Whereas IFN-γ+ γδ T cells were almost exclusively dependent on glycolysis, IL-17+ γδ T cells strongly engaged oxidative metabolism, with increased mitochondrial mass and activity. These distinct metabolic signatures were surprisingly imprinted early during thymic development and were stably maintained in the periphery and within tumors. Moreover, pro-tumoral IL-17+ γδ T cells selectively showed high lipid uptake and intracellular lipid storage and were expanded in obesity and in tumors of obese mice. Conversely, glucose supplementation enhanced the antitumor functions of IFN-γ+ γδ T cells and reduced tumor growth upon adoptive transfer. These findings have important implications for the differentiation of effector γδ T cells and their manipulation in cancer immunotherapy

Functional analysis of the retinoic acid signaling pathway during spermatogenesis

L’acide rétinoïque (AR) est requis pour de nombreuses fonctions physiologiques parmi lesquelles la reproduction. Il est synthétisé par des rétinaldéhyde déshydrogénases (RALDH1 à 3) et il active la transcription de gènes cible en se liant à ses récepteurs nucléaires RAR (α,β,γ). L’objet de mon travail de thèse a été d’étudier les fonctions de l’AR, produit dans les cellules de Sertoli testiculaire, sur la spermatogenèse de la souris. Ainsi, par l’étude de l’inactivation des gènes des RALDH spécifiquement dans les cellules de Sertoli murines grâce à la mutagenèse somatique, j’ai montré que les cellules de Sertoli dirigent la première différenciation des cellules germinales grâce à leur production d’AR qui est ensuite dispensable au bon déroulement de la spermatogenèse. L’induction de cette première spermatogenèse est possible par l’activation sélective de RAR qui est à la base d’une pléiade de voies de signalisation. J’ai également confirmé le rôle crucial de la voie de signalisation de l’AR dans les cellules de Sertoli pour la spermiation, dernière étape du processus de spermatogenèse, et donc pour la fertilité. Enfin, j’ai démontré la nécessité in vivo de la signalisation par l’AR pour la méiose, qui contrôle l’expression de Stra8, un gène essentiel pour la progression méiotique. Cette régulation se fait de manière cellulaire autonome et requiert la fixation d’un RAR sur son élément de réponse localisé dans la région promotrice de Stra8.Retinoic acid (RA) is required for many physiological functions including reproduction. It is synthesized by retinaldehyde dehydrogenases (RALDH1 to 3) and activates transcription of target genes by binding to its nuclear receptors (RAR α,β,γ ). The purpose of my thesis was to study the functions of RA produced in testicular Sertoli cells in spermatogenesis in the mouse. Thus, by studying the effects of RALDH selective inactivation in murine Sertoli cells by somatic mutagenesis, I showed that Sertoli cells direct the first differentiation of germ cells through their production of RA, which is then dispensable for the proper conduct of spermatogenesis. Induction of the first spermatogenesis is possible by selective activation of RAR, which is at the basis of several signaling pathways. I also confirmed the crucial role of the RA pathway in Sertoli cells for spermiation, the final stage of spermatogenesis, and therefore fertility. Finally, I demonstrated the need of an in vivo RA signaling for meiosis, which controls the expression of Stra8, a gene essential for meiotic progression. This regulation is done cell-autonomously and requires the binding of a RAR on its response element located in the promoter region of Stra8

Functional analysis of the retinoic acid signaling pathway during spermatogenesis

L’acide rétinoïque (AR) est requis pour de nombreuses fonctions physiologiques parmi lesquelles la reproduction. Il est synthétisé par des rétinaldéhyde déshydrogénases (RALDH1 à 3) et il active la transcription de gènes cible en se liant à ses récepteurs nucléaires RAR (α,β,γ). L’objet de mon travail de thèse a été d’étudier les fonctions de l’AR, produit dans les cellules de Sertoli testiculaire, sur la spermatogenèse de la souris. Ainsi, par l’étude de l’inactivation des gènes des RALDH spécifiquement dans les cellules de Sertoli murines grâce à la mutagenèse somatique, j’ai montré que les cellules de Sertoli dirigent la première différenciation des cellules germinales grâce à leur production d’AR qui est ensuite dispensable au bon déroulement de la spermatogenèse. L’induction de cette première spermatogenèse est possible par l’activation sélective de RAR qui est à la base d’une pléiade de voies de signalisation. J’ai également confirmé le rôle crucial de la voie de signalisation de l’AR dans les cellules de Sertoli pour la spermiation, dernière étape du processus de spermatogenèse, et donc pour la fertilité. Enfin, j’ai démontré la nécessité in vivo de la signalisation par l’AR pour la méiose, qui contrôle l’expression de Stra8, un gène essentiel pour la progression méiotique. Cette régulation se fait de manière cellulaire autonome et requiert la fixation d’un RAR sur son élément de réponse localisé dans la région promotrice de Stra8.Retinoic acid (RA) is required for many physiological functions including reproduction. It is synthesized by retinaldehyde dehydrogenases (RALDH1 to 3) and activates transcription of target genes by binding to its nuclear receptors (RAR α,β,γ ). The purpose of my thesis was to study the functions of RA produced in testicular Sertoli cells in spermatogenesis in the mouse. Thus, by studying the effects of RALDH selective inactivation in murine Sertoli cells by somatic mutagenesis, I showed that Sertoli cells direct the first differentiation of germ cells through their production of RA, which is then dispensable for the proper conduct of spermatogenesis. Induction of the first spermatogenesis is possible by selective activation of RAR, which is at the basis of several signaling pathways. I also confirmed the crucial role of the RA pathway in Sertoli cells for spermiation, the final stage of spermatogenesis, and therefore fertility. Finally, I demonstrated the need of an in vivo RA signaling for meiosis, which controls the expression of Stra8, a gene essential for meiotic progression. This regulation is done cell-autonomously and requires the binding of a RAR on its response element located in the promoter region of Stra8

Etude fonctionnelle de la voie de signalisation de l'acide rétinoïque au cours de la spermatogenèse

L acide rétinoïque (AR) est requis pour de nombreuses fonctions physiologiques parmi lesquelles la reproduction. Il est synthétisé par des rétinaldéhyde déshydrogénases (RALDH1 à 3) et il active la transcription de gènes cible en se liant à ses récepteurs nucléaires RAR (a,b,g). L objet de mon travail de thèse a été d étudier les fonctions de l AR, produit dans les cellules de Sertoli testiculaire, sur la spermatogenèse de la souris. Ainsi, par l étude de l inactivation des gènes des RALDH spécifiquement dans les cellules de Sertoli murines grâce à la mutagenèse somatique, j ai montré que les cellules de Sertoli dirigent la première différenciation des cellules germinales grâce à leur production d AR qui est ensuite dispensable au bon déroulement de la spermatogenèse. L induction de cette première spermatogenèse est possible par l activation sélective de RAR qui est à la base d une pléiade de voies de signalisation. J ai également confirmé le rôle crucial de la voie de signalisation de l AR dans les cellules de Sertoli pour la spermiation, dernière étape du processus de spermatogenèse, et donc pour la fertilité. Enfin, j ai démontré la nécessité in vivo de la signalisation par l AR pour la méiose, qui contrôle l expression de Stra8, un gène essentiel pour la progression méiotique. Cette régulation se fait de manière cellulaire autonome et requiert la fixation d un RAR sur son élément de réponse localisé dans la région promotrice de Stra8.Retinoic acid (RA) is required for many physiological functions including reproduction. It is synthesized by retinaldehyde dehydrogenases (RALDH1 to 3) and activates transcription of target genes by binding to its nuclear receptors (RAR a,b,g ). The purpose of my thesis was to study the functions of RA produced in testicular Sertoli cells in spermatogenesis in the mouse. Thus, by studying the effects of RALDH selective inactivation in murine Sertoli cells by somatic mutagenesis, I showed that Sertoli cells direct the first differentiation of germ cells through their production of RA, which is then dispensable for the proper conduct of spermatogenesis. Induction of the first spermatogenesis is possible by selective activation of RAR , which is at the basis of several signaling pathways. I also confirmed the crucial role of the RA pathway in Sertoli cells for spermiation, the final stage of spermatogenesis, and therefore fertility. Finally, I demonstrated the need of an in vivo RA signaling for meiosis, which controls the expression of Stra8, a gene essential for meiotic progression. This regulation is done cell-autonomously and requires the binding of a RAR on its response element located in the promoter region of Stra8.STRASBOURG-Bib.electronique 063 (674829902) / SudocSudocFranceF

Retinoic acid expression associates with enhanced IL-22 production by ?? T cells and innate lymphoid cells and attenuation of intestinal inflammation.

Retinoic acid (RA), a vitamin A metabolite, modulates mucosal T helper cell responses. Here we examined the role of RA in regulating IL-22 production by ?? T cells and innate lymphoid cells in intestinal inflammation. RA significantly enhanced IL-22 production by ?? T cells stimulated in vitro with IL-1? or IL-18 and IL-23. In vivo RA attenuated colon inflammation induced by dextran sodium sulfate treatment or Citrobacter rodentium infection. This was associated with a significant increase in IL-22 secretion by ?? T cells and innate lymphoid cells. In addition, RA treatment enhanced production of the IL-22-responsive antimicrobial peptides Reg3? and Reg3? in the colon. The attenuating effects of RA on colitis were reversed by treatment with an anti-IL-22 neutralizing antibody, demonstrating that RA mediates protection by enhancing IL-22 production. To define the molecular events involved, we used chromatin immunoprecipitation assays and found that RA promoted binding of RA receptor to the IL-22 promoter in ?? T cells. Our findings provide novel insights into the molecular events controlling IL-22 transcription and suggest that one key outcome of RA signaling may be to shape early intestinal immune responses by promoting IL-22 synthesis by ?? T cells and innate lymphoid cells

Development of a quantitative fluorescence-based ligand-binding assay

A major goal of biology is to develop a quantitative ligand-binding assay that does not involve the use of radioactivity. Existing fluorescence-based assays have a serious drawback due to fluorescence quenching that accompanies the binding of fluorescently-labeled ligands to their receptors. This limitation of existing fluorescence-based assays prevents the number of cellular receptors under investigation from being accurately measured. We have developed a method where FITC-labeled proteins bound to a cell surface are proteolyzed extensively to eliminate fluorescence quenching and then the fluorescence of the resulting sample is compared to that of a known concentration of the proteolyzed FITC-protein employed. This step enables the number of cellular receptors to be measured quantitatively. We expect that this method will provide researchers with a viable alternative to the use of radioactivity in ligand binding assays