ECTV Infection Induces Increased Expression of NKG2D Ligands In Vitro and In Vivo

<div><p>(A) MEFs were infected with 0.5 pfu ECTV 189898-p7.5-EGFP for 18 h. Cells were analyzed for staining with the indicated reagents after gating for EGFP⁻ cells (uninfected) and EGFP⁺ cells (infected). Data correspond to one typical experiment from three similar experiments. Shaded area, infected cells stained with isotype-matched control Ig or secondary Ab alone; black line, infected cells stained with the indicated reagent; gray line, non-infected cells stained with the indicated reagent.</p><p>(B) qRT-PCR was performed as described. Data were normalized to the amount of β-actin mRNA. Data are representative of two similar experiments.</p></div

NK Cells Require NKG2D to Control Early Virus Dissemination and for Optimal Cytotoxicity but Not Activation

<div><p>(A) B6 mice were infected with 3,000 pfu ECTV. On day 5 PI, mice were pulsed with BrdU for 3 h and their spleens analyzed by flow cytometry. Plots are gated on CD3ɛ⁻ NK1.1⁺ cells. Data correspond to pools of three mice from three individual experiments.</p><p>(B) Increased viral titers of infected mice with NKG2D blockade in vivo. Intact B6 mice, B6 mice with NKG2D blockade, B6 mice depleted of NK cells, B6 mice depleted of T cells (treated with anti-CD4 + anti-CD8 mAbs), or depleted of T cells and with NKG2D blockade were infected with 3,000 pfu ECTV and the viral titers in spleen were determined on day 3 PI. Data are the average ± SD of three individual mice and are representative of two similar experiments.</p><p>(C) The indicated mice were infected with 3,000 pfu ECTV, and the NK responses in the D-LN were determined at 2 d PI. Upper panel: Dot plot showing the proportion of NK cells (DX5⁺, CD3e⁻) in the D-LN of infected and control uninfected mice. Lower panel: GzB and IFN-γ production by NK cells. Cells correspond to the DX5⁺, CD3ɛ⁻ gate of the upper panels. Data correspond to pools of three mice and are representative of three similar experiments.</p><p>(D) NK cells were purified from spleens (5 d PI) of ECTV-infected intact or NKG2D-blocked mice and stained as indicated. Filled histogram: isotype-matched control Ig; gray line: NKG2D-blocked mice; black line: intact mice.</p><p>(E) NK cells were purified from spleens of intact or NKG2D-blocked mice as indicated and were used as effectors in a 4-h ⁵¹Cr release assay against the indicated targets. Data correspond to pools of three mice and are representative of three experiments.</p><p>(F) As in (D), but the NK cells were purified from untreated mice and a neutralizing anti-NKG2D mAb was added to the in vitro cytotoxicity assays, as indicated.</p></div

NKG2D Uses DAP10 or DAP12 Adapters to Help Resist Mousepox

<div><p>(A) The indicated mice were infected with 3,000 pfu ECTV and the absolute number of lymphocytes in their spleens was determined on day 7 PI by trypan blue exclusion. An uninfected control is also shown. Data are the average ± SD of three pooled mice per group from at least three individual experiments.</p><p>(B) Virus titers in spleen of the indicated mice on day 7 PI. Data are the average ± SD of three pooled mice per group from at least three individual experiments.</p></div

Reduced T Cell Responses in the Absence of NK Cells

<div><p>(A) Intact or NK cell–depleted B6 mice were infected with 3,000 pfu ECTV. T cell proliferation on day 5 PI was determined in the D-LN by using a 3-h BrdU incorporation assay. Upper panel: gated on CD8⁺ T cells; lower panel: gated on CD4⁺ T cells.</p><p>(B) Intact or NK cell–depleted B6 mice were infected with ECTV. Production of IFN-γ and GzB by CD8⁺ T cells in spleen on day 7 PI was determined. Plots are gated on CD8⁺ T cells. Data are representative of three independent experiments.</p></div

Host–Guest Chemistry of Dendrimer–Drug Complexes: 7. Formation of Stable Inclusions between Acetylated Dendrimers and Drugs Bearing Multiple Charges

Drug molecules bearing multiple charges usually form precipitates with cationic dendrimers, which presents a challenge during the preparation of dendrimer inclusions for these drugs. In the present study, fully acetylated polyamidoamine (PAMAM) dendrimers were proposed as stable vehicles for drug molecules bearing two negative charges such as Congo red and indocyanine green. NMR techniques including ¹H NMR and ¹H-¹H NOESY were used to characterize the host–guest chemistry of acetylated dendrimer and these guest molecules. The cationic PAMAM dendrimer was found to form a precipitate with Congo red and indocyanine green, but the acetylated one avoided the formation of cross-linking structures in aqueous solutions. NOESY studies revealed the encapsulation of Congo red and indocyanine green within the interior cavities of PAMAM dendrimers at mild acidic conditions and acetylated dendrimers show much stronger ability to encapsulate the guest molecules than cationic ones. Also, UV–vis–NIR studies suggest that acetylated dendrimers significantly improve the photostability of indocyanine green and prevent the formation of indocyanine green J-aggregates in aqueous solutions. The present study provides a new insight into dendrimer-based host–guest systems, especially for those guest molecules bearing multiple charges

Synthesis and Biological Activities of 5‑Thio-α-GalCers

NKT cells, a unique subset of T cells that recognizes glycolipid antigens presented by CD1d molecules, are believed to produce key cytokines of both Th1 and Th2 T cells and are thus involved in the control of several types of immune response. As an active glycolipid antigen having α-galactosyl ceramide core structure, KRN7000 showed promising immunostimulation activity and was selected as an anticancer drug candidate for further clinical application. In this report, three new KRN7000 structural analogues were designed and synthesized, in which the ring oxygen of the galactopyranose residue is replaced by a sulfur atom along with the variation on the lipid chain. Their abilities for stimulating mouse NKT cells to produce IFN-γ and IL-4 were evaluated both <i>in vivo</i> and <i>in vitro</i>

An Unprecedented M–O Cluster Constructed from Nanosized {[C₅NH₅]₉[H₃₁Mo^V₁₂O₂₄Co^II₁₂(PO₄)₂₃(H₂O)₄]}^2– Anions Exhibiting Interesting Nonlinear-Optical Properties

A novel high-nuclear nanosized cluster modified by conjugated organic ligands (pyridine and imidazole), [C₅NH₅]₈­[C₃H₅N₂]₂­{[C₅NH₅]₉​­[H₃₁Mo₁₂O₂₄Co₁₂​­(PO₄)₂₃­(H₂O)₄]}·12H₂O (<b>1</b>), has been successfully isolated under hydrothermal conditions and structurally characterized. Compound <b>1</b> consists of 12 Co^II and 12 Mo^V ions linked by 23 {PO₄} groups, exhibiting unprecedented nanosized ship-shaped clusters. The magnetic measurements reveal that compound <b>1</b> exhibits dominant antiferromagnetic interactions. Additionally, pyridine and imidazole ligands enhance the delocalized electron effects of clusters, and the third-order nonlinear-optical response of compound <b>1</b> is excellent

Association of the Pro12Ala polymorphism with type 2 diabetes mellitus.

<p>Adjusted for age, sex, BMI, hypertension, and family history of diabetes.</p><p>Abbreviations: CI, confidence interval; OR, odds ratio. The other abbreviations used see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071985#pone-0071985-t001" target="_blank">Table 1</a>.</p

Distribution of allele and genotype frequencies of the Pro12Ala polymorphism in study subjects stratified by BMI.

<p>Adjusted for age, sex, BMI, hypertension, and family history of diabetes.</p><p>The abbreviations used see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071985#pone-0071985-t001" target="_blank">Table 1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071985#pone-0071985-t002" target="_blank">2</a>.</p

Syntheses of new topology BTTB-based metal–organic frameworks in CH₃CN/H₂O mixed solvents

<p>Three complexes, [Zn₂(BTTB)](H₂O)₅ [BTTB = 4,4′,4″,4′″-benzene-1,2,4,5-tetrayltetrabenzoate, also named 1,2,4,5-tetrakis(carboxyphenyl) benzene] (<b>1</b>), [Cd₂(BTTB)(H₂O)₅](H₂O)₃ (<b>2</b>) and [Mn₂(BTTB)(H₂O)₅](H₂O)₂ (<b>3</b>), have been synthesized under hydrothermal conditions in CH₃CN/H₂O. Structural analyses revealed that <b>1</b> has new topologies, i.e. <b><i>nnu1</i></b> (<b><i>nnu</i></b> stands for Nanjing Normal University) or <b><i>nnu2</i></b> depending on the different simplification schemes. Compounds <b>2</b> and <b>3</b> are isostructural and have a <b><i>scu</i></b> (square planar and cubical vertices) topology, which have not been reported for BTTB-based metal–organic frameworks. Thermal stability of <b>1</b>–<b>3</b>, hydrogen adsorption/desorption properties of <b>2</b> and <b>3</b>, and luminescent properties of <b>1</b> and <b>2</b> were investigated. Compounds <b>1</b>–<b>3</b> are stable in air.</p