Tcf1 and Lef1 transcription factors establish CD8(+) T cell identity through intrinsic HDAC activity.

The CD4(+) and CD8(+) T cell dichotomy is essential for effective cellular immunity. How individual T cell identity is established remains poorly understood. Here we show that the high-mobility group (HMG) transcription factors Tcf1 and Lef1 are essential for repressing CD4(+) lineage-associated genes including Cd4, Foxp3 and Rorc in CD8(+) T cells. Tcf1- and Lef1-deficient CD8(+) T cells exhibit histone hyperacetylation, which can be ascribed to intrinsic histone deacetylase (HDAC) activity in Tcf1 and Lef1. Mutation of five conserved amino acids in the Tcf1 HDAC domain diminishes HDAC activity and the ability to suppress CD4(+) lineage genes in CD8(+) T cells. These findings reveal that sequence-specific transcription factors can utilize intrinsic HDAC activity to guard cell identity by repressing lineage-inappropriate genes.

Monoglyceride reduces viability of porcine epidemic diarrhoea virus in feed and prevents disease transmission to post-weaned piglets.

Outbreaks of African swine fever virus (ASFv) and porcine epidemic diarrhoea virus (PEDv) have revealed the susceptibility of livestock to disease transmitted through feed. Several viruses, including PEDv, survive in feed and may introduce disease that causes significant morbidity and mortality. In 2013, PEDv, which causes severe diarrhoea and vomiting, reached North America after spreading for decades across Eurasia. The global exchange of ingredients has created demand for products that prevent disease transmission from feed. Formaldehyde-based products are highly effective at inactivating enveloped viruses when applied at 3.25 kg/t. Alternative products to formaldehyde, including carboxylic acids, essential oils and medium chain fatty acids (MCFAs), have exhibited mixed efficacy against PEDv and require application rates higher than formaldehyde. Amphiphilic molecules like MCFAs disrupt the bilayer-lipid membranes that protect viral nucleic acids through the formation of micelles. Monoglycerides form micelles at lower concentrations than MCFAs, which suggests they may be more potent against enveloped viruses. The potential efficacy of monoglycerides against enveloped viruses in feed led to the development and examination of an experimental monoglyceride blend. The proprietary monoglyceride blend significantly (p < .0001) reduced PEDv viability in vitro after application to feed at 1.5, 2.5 and 3.5 kg/t. The monoglyceride was tested in a natural feeding behaviour challenge model in piglets. The feed was contaminated with ice-blocks containing viable PEDv, and the piglets were exposed to PEDv through the feed bin for 20 days. At the end of the 20-day challenge period, all pigs were rectally swabbed and tested for PEDv by qPCR. In the untreated control group 54.8% of the piglets tested positive for PEDv, whereas none of the MCFA-treated feed (10 kg/t inclusion) transmitted PEDv. Strikingly, the monoglyceride-treated groups (1.5, 2.5 and 3.5 kg/t) all exhibited 100% protection from PEDv. These data support the use of this proprietary monoglyceride blend in mitigation and prevention of viral disease transmission to piglets from contaminated feed.

Particulate and solubilized ß-glucan and non-ß-glucan fractions of Euglena gracilis induce pro-and anti-inflammatory innate immune cell responses and exhibit antioxidant properties.

PURPOSE: The purpose of this work was to determine the pro-and anti-inflammatory properties of the single-cell organism Euglena gracilis (EG) and various fractions of its whole biomass. METHODS: Heterotrophically grown EG was tested, along with its aqueous fraction (E-AQ), the intact linear ß-glucan paramylon granules (PAR), and alkaline-solubilized paramylon. Peripheral blood mononuclear cell cultures were treated with the test products and analyzed for a variety of cellular responses. Immune cell activation was evaluated by flow cytometry detection of CD69 levels on CD3-CD56+ NK cells, CD3+CD56+ NKT cells, and monocytes, and cytokines were analyzed from the cell culture supernatants. Antioxidant capacity was measured by Folin-Ciocalteu assay and cellular antioxidant protection and MTT assays. RESULTS: EG and E-AQ were the most effective in driving immune cell responses as measured by CD69 upregulation on NK and NKT cells and proinflammatory (tumor necrosis factor, IL-6, IL-1ß) cytokine production. None of the test products effectively stimulated monocyte. EG and PAR inhibited reactive oxygen species under conditions of oxidative stress. E-AQ contained antioxidants capable of providing cellular antioxidant protection from oxidative damage and protection of mitochondrial function under inflammatory conditions. CONCLUSION: The effects of EG on immune function are only partially attributable to the content of the ß-glucan, paramylon. The regulation of additional cellular responses, such a reactive oxygen species production and resistance to oxidative stress, is likely mediated by currently unknown molecules found in the EG cell.

Microbiota and caspase-1/caspase-8 regulate IL-1ß-mediated bone disease.

A leucine-to-proline missense mutation at residue 98 in the proline-serine-threonine phosphatase interacting protein 2 (Pstpip2) gene leads to autoinflammatory disease that is characterized by splenomegaly, necrosis, and spontaneous development of osteomyelitis in mice (Pstpip2cmo). Disease progression in these mice resembles that of chronic recurrent multifocal osteomyelitis in humans. Our group and others have shown that disease progression in Pstpip2cmo mice is mediated by the cytokine IL-1ß, independently of inflammasomes or IL-1α. Our recent publication highlighted herein establishes that diet-induced changes in intestinal microbiota provide protection against the development of osteomyelitis in Pstpip2cmo mice. Moreover, the proteases caspase-1 and caspase-8 have redundant roles in cleaving IL-1ß and promoting disease. This addendum reviews the current literature on the Pstpip2cmo murine disease model and the clinical significance of the role of PSTPIP2 in regulating autoinflammatory osteomyelitis, which is mediated by innate components of immune cells.