p66Shc is a negative regulator of FcεRI-dependent signaling in mast cells.

Aggregation of FcεRI on mast cells activates signaling pathways, resulting in degranulation and cytokine release. Release of mast cell-derived inflammatory mediators is tightly regulated by the interplay of positive and negative signals largely orchestrated by adapter proteins. Among these, the Shc family adapter p52Shc, which couples immunoreceptors to Ras activation, positively regulates FcεRI-dependent signaling. Conversely, p66Shc was shown to uncouple the TCR for the Ras-MAPK pathway and prime T cells to undergo apoptotic death. Loss of p66Shc in mice results in breaking of immunologic tolerance and development of lupus-like autoimmune disease, which includes alopecia among its pathological manifestations. The presence of numerous activated mast cells in alopecic skin areas suggests a role for this adapter in mast cells. In this study, we addressed the involvement of p66Shc in FcεRI-dependent mast cell activation. We showed that p66Shc is expressed in mast cells and that mast cells from p66Shc(-/-) mice exhibit enhanced responses following Ag stimulation of FcεRI. Furthermore, using RBL-2H3 cell transfectants, we showed that aggregation of FcεRI resulted in the recruitment of a p66Shc-SHIP1 complex to linker for activation of T cells. Collectively, our data identified p66Shc as a negative regulator of mast cell activation.

Rai acts as a negative regulator of autoimmunity by inhibiting antigen receptor signaling and lymphocyte activation.

Rai (ShcC) belongs to the family of Shc adaptor proteins and is expressed in neuronal cells, where it acts as a survival factor activating the PI3K/Akt survival pathway. In vivo, Rai protects the brain from ischemic damage. In this study, we show that Rai is expressed in T and B lymphocytes. Based on the finding that Rai(-/-) mice consistently develop splenomegaly, the role of Rai in lymphocyte homeostasis and proliferation was addressed. Surprisingly, as opposed to neurons, Rai was found to impair lymphocyte survival. Furthermore, Rai deficiency results in a reduction in the frequency of peripheral T cells with a concomitant increase in the frequency of B cells. Rai(-/-) lymphocytes display enhanced proliferative responses to Ag receptor engagement in vitro, which correlates with enhanced signaling by the TCR and BCR, and more robust responses to allergen sensitization in vivo. A high proportion of Rai(-/-) mice develop a lupus-like autoimmune syndrome characterized by splenomegaly, spontaneous peripheral T and B cell activation, autoantibody production, and deposition of immune complexes in the kidney glomeruli, resulting in autoimmune glomerulonephritis. The data identify Rai as a negative regulator of lymphocyte survival and activation and show that loss of this protein results in breaking of immunological tolerance and development of systemic autoimmunity.

Toxoplasma gondii in women of childbearing age and during pregnancy: seroprevalence study in Central and Southern Italy from 2013 to 2017.

Toxoplasmosis is a worldwide health problem. Infection in pregnant women can result in severe fetal morbidity or in subclinical neonatal infection; most subclinical cases develop ocular and neurological sequelae. The purpose of this serological study was to assess the prevalence of Toxoplasma gondii in two populations of women of childbearing age in Siena (Tuscany, Central Italy) and Bari (Apulia, Southern Italy) between 2013 and 2017 and in a group of pregnant women in Bari in 2016-2017. Serum samples were tested for the presence of specific anti-Toxoplasma gondii IgG antibodies by a commercially available ELISA test. The percentage of seropositive subjects in Bari was significantly higher than in Siena (22.4% vs. 12.4%) and an age-related trend was observed. A low prevalence of T. gondii infection (13.8%) was observed among the pregnant women tested. In addition to showing a significant difference between Central and Southern Italy, this study provides updated data on T. gondii seroprevalence in women during childbearing age and pregnancy. The results confirm a trend toward a decrease, especially in younger people and pregnant women.

TITLE: Toxoplasma gondii chez les femmes en âge de procréer et pendant la grossesse : étude de la séroprévalence dans le centre et le sud de l'Italie de 2013 à 2017. ABSTRACT: La toxoplasmose est un problème de santé mondial. L'infection d'une femme enceinte peut entraîner une morbidité fœtale grave ou une infection néonatale subclinique, et la plupart des cas subcliniques développeront des séquelles oculaires et neurologiques. Le but de cette étude sérologique était d'évaluer la prévalence de Toxoplasma gondii dans deux populations de femmes en âge de procréer à Sienne (Toscane, Italie centrale) et Bari (Pouilles, Italie du Sud) entre 2013 et 2017 et dans un groupe de femmes enceintes à Bari en 2016-2017. Des échantillons de sérum ont été testés pour la présence d'anticorps IgG anti-Toxoplasma gondii spécifiques par un test ELISA commercial. Le pourcentage de sujets séropositifs à Bari était significativement plus élevé qu'à Sienne (22,4 % vs 12,4 %) et une tendance liée à l'âge a été observée. Une faible prévalence de l'infection à T. gondii (13,8 %) a été observée chez les femmes enceintes testées. En plus de montrer une différence significative entre le centre et le sud de l'Italie, cette étude fournit des données actualisées sur la séroprévalence de T. gondii chez les femmes en âge de procréer et pendant la grossesse. Les résultats confirment une tendance à la baisse, notamment chez les jeunes et les femmes enceintes.

Antigenic properties of HCMV peptides displayed by filamentous bacteriophages vs. synthetic peptides.

Several efforts have been invested in the identification of CTL and Th epitopes, as well as in the characterization of their immunodominance and MHC restriction, for the generation of a peptide-based HCMV vaccine. Small synthetic peptides are, however, poor antigens and carrier proteins are important for improving the efficacy of synthetic peptide vaccines. Recombinant bacteriophages appear as promising tools in the design of subunit vaccines. To investigate the antigenicity of peptides carried by recombinant bacteriophages we displayed different HCMV MHCII restricted peptides on the capsid of filamentous bacteriophage (fd) and found that hybrid bacteriophages are processed by human APC and activate HCMV-specific CD4 T-cells. Furthermore we constructed a reporter T-cell hybridoma expressing a chimeric TCR comprising murine alphabeta constant regions and human variable regions specific for the HLA-A2 restricted immunodominant NLV peptide of HCMV. Using the filamentous bacteriophage as an epitope carrier, we detected a more robust and long lasting response of the reporter T-cell hybridoma compared to peptide stimulation. Our results show a general enhancement of T-cell responses when antigenic peptides are carried by phages.

Clinically-relevant cyclosporin and rapamycin concentrations enhance regulatory T cell function to a similar extent but with different mechanisms: an in-vitro study in healthy humans.

Evidence indicates that regulatory T cells (Tregs) are profoundly involved in promoting allograft tolerance after organ transplantation. Since a successful transplantation currently still requires a long-term immunosuppressive treatment, clarifying the specific impact of these drugs on Tregs may be of high clinical relevance. Conflicting results arise from the literature, particularly as concerns cyclosporine (CsA). The specific aim of this work was to evaluate in-vitro the direct effects of clinically-relevant drug concentrations of three widely used immunosuppressive drugs, i.e. CsA, rapamycin (RAPA) and mycophenolic acid (MPA), on Treg activity, number and forkhead/winged helix transcription factor (FoxP3) expression in humans. Tregs (CD4(+)CD25(+)) isolated from healthy donors were cultured in the presence of different concentrations of CsA, RAPA or MPA. The suppressive activity of Tregs was evaluated in mixed lymphocyte reactions with CD4(+)CD25(-) T cells. Phenotype analysis and FoxP3 expression were assessed by flow cytometry. Clinically-relevant CsA and RAPA concentrations significantly enhanced to a similar extent the suppressive activity of Tregs. Although this effect was associated with an increase in Treg number as well as in FoxP3 expression with both drugs, the driving mechanism seemed to be primarily quantitative (i.e. increase of the cell number) for RAPA, whereas mainly qualitative (i.e. increase in FoxP3 levels) for CsA, respectively. Conversely, MPA did not show any effect on Treg function and number. These findings suggest that both RAPA and CsA may be beneficial in promoting Treg-dependent allograft tolerance after organ transplantation.

Simvastatin impairs humoral and cell-mediated immunity in mice by inhibiting lymphocyte homing, T-cell activation and antigen cross-presentation.

Statins block the activity of HMG-CoA reductase, which catalyses the production of mevalonate, an intermediate in cholesterol biosynthesis, which is also a precursor of isoprenoids. In addition to lowering circulating cholesterol, these drugs display anti-inflammatory and immunomodulatory activities in vitro; however, their effects on the development of adaptive immune responses in vivo, as well as the underlying mechanisms, are as yet largely unknown. Here we investigated the outcome of simvastatin treatment on a number of processes, which together orchestrate adaptive immunity to specific antigen. Simvastatin treatment resulted in a marked reduction of T and B cells in spleen, lymph nodes and peripheral blood in mice. This effect could be ascribed principally to an impairment of lymphocyte homing to secondary lymphoid organs. In addition, simvastatin was found to strongly inhibit T-cell responses to the MHCI restricted hen ovalbumin peptide antigen SIINFEKL and to impair ovalbumin uptake and cross-presentation by MHCI. Simvastatin also suppressed antibody responses to immunization with ovalbumin and delayed-type hypersensitivity to allergens. These activities could be largely accounted for by the simvastatin-dependent inhibition of HMG-CoA reductase. The data provide novel mechanistic insight into the activities of simvastatin in the highly complex context of the immune response.