Mycobacterium tuberculosis produces D-serine under hypoxia to limit CD8+ T cell-dependent immunity in mice.

Adaptation to hypoxia is a major challenge for the survival of Mycobacterium tuberculosis (Mtb) in vivo. Interferon (IFN)-γ-producing CD8+ T cells contribute to control of Mtb infection, in part by promoting antimicrobial activities of macrophages. Whether Mtb counters these responses, particularly during hypoxic conditions, remains unknown. Using metabolomic, proteomic and genetic approaches, here we show that Mtb induced Rv0884c (SerC), an Mtb phosphoserine aminotransferase, to produce D-serine. This activity increased Mtb pathogenesis in mice but did not directly affect intramacrophage Mtb survival. Instead, D-serine inhibited IFN-γ production by CD8+ T cells, which indirectly reduced the ability of macrophages to restrict Mtb upon co-culture. Mechanistically, D-serine interacted with WDR24 and inhibited mTORC1 activation in CD8+ T cells. This decreased T-bet expression and reduced IFN-γ production by CD8+ T cells. Our findings suggest an Mtb evasion mechanism where pathogen metabolic adaptation to hypoxia leads to amino acid-dependent suppression of adaptive anti-TB immunity.

A hybrid hydrogel constructed using drug loaded mesoporous silica and multiple response copolymer as an intelligent dressing for wound healing of diabetic foot ulcers.

Traditional wound dressings have poor mechanical properties and a single function, which cannot achieve rapid healing of diabetic wounds in a unique physiological microenvironment. In order to develop multifunctional hydrogel dressings with appropriate biological activity to accelerate wound healing and obtain better clinical therapeutic effects, herein we report a hybrid system based on drug loaded mesoporous silica and injectable polymer hydrogels mixed with hypoglycemic drug metformin (Met) as a dressing for diabetic wounds. Firstly, a copolymer with the phenylboronic acid group in the side group, poly(acrylamide-co-dimethylaminopropylacrylamide-co-methacrylamidophenylboronic acid) (abbreviated as PB), was prepared. PB was mixed with polyvinyl alcohol (PVA) to obtain an injectable hydrogel (named PP) with pH/glucose dual responsiveness, which was formed through the combination of the phenylborate group of PB and o-diol of PVA. In another reaction, polydopamine-modified mesoporous silica nanoparticles (MSN@PDA) were prepared and used to adsorb antibiotic tetracycline hydrochloride (TH) to obtain drug-loaded MSN@PDA-TH nanoparticles. Subsequently, the hybrid hydrogel dressing (abbreviated as PP/MSN@PDA-TH/Met) was obtained by mixing PB, PVA, Met and MSN@PDA-TH. The self-healing, rheological and adhesive properties of the hybrid hydrogel were characterized. The results show that the hydrogel dressing has good physical properties. Met and TH were released in vitro in different pH media and glucose environments. The results show that the hydrogel dressing has dual responsiveness towards pH and glucose, and can continuously release metformin and tetracycline, which is conducive to accelerating wound healing. The antimicrobial activity, ROS clearance ability and biocompatibility of the hydrogel dressing were evaluated. The results indicate that the hydrogel dressing was multifunctional. Finally, a full-thickness wound repair model of diabetic mice induced by streptozotocin (STZ) was established. The hybrid hydrogel dressing was applied to the wound surface of mice. The wound healing testing on diabetic mice confirmed that the wound covered with the hybrid hydrogel dressing was completely healed with the formation of the new skin and hair within 9 days to 12 days. Histological analysis indicates that, compared to the PBS control, the hydrogel dressing did not cause significant inflammation in the wound, and a large number of blood vessels, glands and hair follicles appeared. This study provides a good strategy for multi-drug synergistic treatment of diabetic foot ulcers.

Rapid, sensitive and cost-effective determination of immune checkpoint inhibitor activity using a magnetic bead-based binding assay.

Immune checkpoint Inhibitors (ICIs) are effective immunno-therapeutic agents for cancer. Rapid and sensitive determination of the blocking activity of ICIs is important for ICIs development and immunological research. Among various immune checkpoint (IC) binding assays, cell-based binding assays are widely regarded, and the functional ELISA is a convenient alternative. However, these methodologies are limited by time-consuming preparation of cell lines stably expressing IC molecules, or long turnaround time with high cost. In this study, two magnetic bead based binding assays were developed to evaluate activity of ICIs, which was determined by a soluble ligand/bead immobilized receptor based binding assay (sL/bR binding assay) that assessed efficacy to block binding of one soluble IC ligand on its cognate receptor immobilized beads, or by a soluble receptor/bead immobilized ligand based binding assay (sR/bL binding assay) that assessed efficacy to block binding of soluble IC receptor on its cognate ligand immobilized beads. Half maximal inhibitory concentration (IC50) values of ICIs were calculated to determine ICIs activity. The sL/bR binding assay accurately determined the activity of two TIGIT blocking antibodies, since the relative blocking activity of two TIGIT antibodies determined by the sL/bR binding assay established in this study and that by the cell based binding assay were almost identical. In contrast, the sR/bL binding assay showed significantly improved sensitivity to determine activity of two PD-1 blocking antibodies than the sL/bR binding assay that was tested in this study and previous reports. Moreover, both amount of the used recombinant protein of ICI receptor/ligand and turnaround time of the two binding assays were more than 10 times less than those of the functional ELISA. These data indicate that the two magnetic bead based binding assays are sensitive, rapid and cost-effective methods to determine blocking activity of ICIs.