The application of chimeric antigen receptor (CAR)-T cell therapy in patients with advanced solid tumors remains a significant challenge. Simultaneously targeting antigen and the solid tumor microenvironment are two major factors that greatly impact CART cell therapy outcomes. In this study, we engineered CART cells to specifically target B7-H3, a protein commonly found in solid human tumors, using a single-chain variable fragment (scFv) derived from an anti-B7-H3 monoclonal antibody. We tested the antitumor activity of B7-H3 CART cells in mouse models with solid human tumors and determined that B7-H3 CART cells exhibited potent antitumor activity against B7-H3 + tumor cells in vitro and in vivo. In addition, PD-1 decoy receptors were engineered to include extracellular PD-1 fused to the intracellular stimulatory domain of either CD28 or IL-7 receptor, respectively, which were then introduced into B7-H3 CART cells. As a result, these newly modified, superior CART cells exhibited more persistent antitumor activity in B7-H3 + /B7-H1 + tumors in vivo. Our findings indicate that B7-H3 specific CART cells have the potential to treat multiple types of advanced solid tumors.

The role of B7-DC in T-cell responses remains controversial because both coinhibitory and costimulatory functions have been reported in various experimental systems in vitro and in vivo. In addition to interacting with the coinhibitory receptor PD-1, B7-DC has also been shown to bind repulsive guidance molecule b (RGMb). The functional consequences of the B7-DC/RGMb interaction, however, remain unclear. More than a decade ago, we reported that replacement of a murine B7-DC mutant lysine with serine (K113S) at positive 113 resulted in a loss of binding capacity to PD-1. Nevertheless, K113S remained costimulatory for T cells in vitro, implicating a dual functionality for B7-DC in T-cell responses. Here we show that recombinant K113S protein interacts with RGMb with a similar affinity to wild-type B7-DC. More importantly, K113S costimulates CD4 T-cell responses via RGMb and promotes Th1 polarization. RGMb is expressed on the surface of naive mouse T cells, macrophages, neutrophils and dendritic cells. Finally, K113S/RGMb costimulation suppresses Th2-mediated asthma and ameliorates small airway inflammation and lung pathology in an experimental mouse model. Our findings indicate that RGMb is a costimulatory receptor for B7-DC. These findings from the K113S variant provide not only a possible explanation for the B7-DC-triggered contradictory effects on T-cell responses, but also a novel approach to investigate the B7-DC/PD-1/RGMb axis. Recombinant K113S or its derivatives could potentially be developed as an agonist for RGMb to costimulate the Th1 response without triggering PD-1-mediated T-cell inhibition.Cellular & Molecular Immunology advance online publication, 8 May 2017; doi:10.1038/cmi.2017.17.

Yogurt is a source of bioactive compounds and probiotic microorganisms that modify immunity and metabolism to benefit human health beyond nutrition. In this study, we examined the capacity of yogurt to prevent epithelial barrier disruption in vitro. Different preparations of yogurt were added apically to Caco-2 monolayers before IL-1β exposure. Dulbecco's modified Eagle medium containing 25% (vol/vol) low-fat yogurt prevented cytokine-induced transepithelial resistance reduction and increases to paracellular permeability measured with fluorescein isothiocyanate-dextran (4 kDa), whereas nonfat yogurt was unable to decrease paracellular permeability to fluorescein isothiocyanatedextran. Moreover, the concentration of IL-8 in low-fatyogurt-treated inflamed cells was decreased to 252.40 ± 27.24 pg/mL, which was lower than that of untreated, inflamed cells (407.20 ± 50.05 pg/mL), further indicating the anti-inflammatory roles of low-fat yogurt. The low-fat yogurt was able to downregulate the transcription of myosin light-chain kinase (MLCK) gene, but upregulate the expression of tight junction protein ZO-1 (TJP1). These findings indicate that low-fat yogurt can maintain intestinal barrier integrity better than nonfat yogurt after pro-inflammatory cytokine exposure.

T cell quiescence is essential for maintaining a broad repertoire against a large pool of diverse antigens from microbes and tumors, but the underlying molecular mechanisms remain largely unknown. We show here that CD8α is critical for the maintenance of CD8 + T cells in a physiologically quiescent state in peripheral lymphoid organs. Upon inducible deletion of CD8α, both naïve and memory CD8 + T cells spontaneously acquired activation phenotypes and subsequently died without exposure to specific antigens. PILRα was identified as a ligand for CD8α in both mice and humans, and disruption of this interaction was able to break CD8 + T cell quiescence. Thus, peripheral T cell pool size is actively maintained by the CD8α–PILRα interaction in the absence of antigen exposure.