5 research outputs found
Synthesis and Biological Evaluation of α‑Galactosylceramide Analogues with Heteroaromatic Rings and Varying Positions of a Phenyl Group in the Sphingosine Backbone
We
designed and synthesized seven α-GalCer analogues with
a pyrazole moiety and varying positions of a phenyl group in the sphingosine
backbone to polarize cytokine secretion. On the basis of in vitro
and in vivo biological evaluations, we found that analogue <b>5</b> induced greater polarization toward Th2 and greater secretion of
the immunomodulatory cytokine, IL-4, over secretion of pro-inflammatory
cytokines, IFN-γ and IL-17. Treatment of a single dose of analogue <b>5</b> markedly ameliorated disease pathogenesis in an animal model
of an inflammatory demyelinating disease of the central nervous system,
compared to that of KRN7000 (<b>1</b>). Therefore, this new
α-GalCer analogue <b>5</b> is a novel iNKT ligand that
stimulates the selective secretion of anti-inflammatory cytokines
and regulates autoimmune diseases by reducing Th1 and Th17 responses
Additional file 1: of IL-1β induces IL-6 production and increases invasiveness and estrogen-independent growth in a TG2-dependent manner in human breast cancer cells
Figure S1. Representative photos of TG2-overexpressing MCF7 cells (MCF7_TG2) and control vector-transfected MCF-7 cells (MCF7_Cont). Figure S2. Effect of immunostimulants on TG2-overexpressing MCF7 cell (TG2) and control vector-transfected MCF-7 cell (Cont) IL-6 expression as measured by ELISA. Cells were treated with toll-like receptor agonists; lipopolysaccharide (LPS, 1 μg/ml), Pam3Cys (Pam, 5 μg/ml), peptidoglycan (PGN, 50 μg/ml), and CPG-ODN (CPG, 1 mM), and bleomycin (BLM, 1 μg/ml) for 48 h. Figure S3. IL-1β increased stem-cell-like phenotypes in a TG2-dependent manner. MCF7_Cont and MCF7_TG2 cells were treated with IL-1β for 6 days, and cell surface expression of CD24 and CD44 was analyzed by flow cytometry analysis. Figure S4. MCF7_Cont and MCF7_TG2 cells were treated with IL-1β (10 ng/ml) for the indicated times. IRAK1 and IRAK2 were detected by Western blot. Figure S5. MCF7_TG2 cells were treated with IL-1β (10 ng/ml) for 30 min. Cell lysates were immunoprecipitated with an anti-TG2 antibody, and TG2, IRAK1, TRAF6, and MyD88 were detected by Western blot. (DOC 1319 kb
Treatment of Sepsis Pathogenesis with High Mobility Group Box Protein 1‑Regulating Anti-inflammatory Agents
Sepsis
is one of the major causes of death worldwide when associated
with multiple organ failure. However, there is a critical lack of
adequate sepsis therapies because of its diverse patterns of pathogenesis.
The pro-inflammatory cytokine cascade mediates sepsis pathogenesis,
and high mobility group box proteins (HMGBs) play an important role
as late-stage cytokines. We previously reported the small-molecule
modulator, inflachromene (<b>1d</b>), which inhibits the release
of HMGBs and, thereby, reduces the production of pro-inflammatory
cytokines. In this context, we intraperitoneally administered <b>1d</b> to a cecal ligation and puncture (CLP)-induced mouse model
of sepsis and confirmed that it successfully ameliorated sepsis pathogenesis.
On the basis of a structure–activity relationship study, we
discovered new candidate compounds, <b>2j</b> and <b>2l</b>, with improved therapeutic efficacy in vivo. Therefore, our study
clearly demonstrates that the regulation of HMGB1 release using small
molecules is a promising strategy for the treatment of sepsis
<i>In Vivo</i> Differentiation of Therapeutic Insulin-Producing Cells from Bone Marrow Cells <i>via</i> Extracellular Vesicle-Mimetic Nanovesicles
The current diabetes mellitus pandemic constitutes an important global health problem. Reductions in the mass and function of β-cells contribute to most of the pathophysiology underlying diabetes. Thus, physiological control of blood glucose levels can be adequately restored by replacing functioning β-cell mass. Sources of functional islets for transplantation are limited, resulting in great interest in the development of alternate sources, and recent progress regarding cell fate change <i>via</i> utilization of extracellular vesicles, also known as exosomes and microvesicles, is notable. Thus, this study investigated the therapeutic capacity of extracellular vesicle-mimetic nanovesicles (NVs) derived from a murine pancreatic β-cell line. To differentiate insulin-producing cells effectively, a three-dimensional <i>in vivo</i> microenvironment was constructed in which extracellular vesicle-mimetic NVs were applied to subcutaneous Matrigel platforms containing bone marrow (BM) cells in diabetic immunocompromised mice. Long-term control of glucose levels was achieved over 60 days, and differentiation of donor BM cells into insulin-producing cells in the subcutaneous Matrigel platforms, which were composed of islet-like cell clusters with extensive capillary networks, was confirmed along with the expression of key pancreatic β-cell markers. The resectioning of the subcutaneous Matrigel platforms caused a rebound in blood glucose levels and confirmed the source of functioning β-cells. Thus, efficient differentiation of therapeutic insulin-producing cells was attained <i>in vivo</i> through the use of extracellular vesicle-mimetic NVs, which maintained physiological glucose levels
Functional Manipulation of Dendritic Cells by Photoswitchable Generation of Intracellular Reactive Oxygen Species
Reactive oxygen species (ROS) play
an important role in cellular
signaling as second messengers. However, studying the role of ROS
in physiological redox signaling has been hampered by technical difficulties
in controlling their generation within cells. Here, we utilize two
inert components, a photosensitizer and light, to finely manipulate
the generation of intracellular ROS and examine their specific role
in activating dendritic cells (DCs). Photoswitchable generation of
intracellular ROS rapidly induced cytosolic mobilization of Ca<sup>2+</sup>, differential activation of mitogen-activated protein kinases,
and nuclear translocation of NF-κB. Moreover, a transient intracellular
ROS surge could activate immature DCs to mature and potently enhance
migration <i>in vitro</i> and <i>in vivo</i>.
Finally, we observed that intracellular ROS-stimulated DCs enhanced
antigen specific T-cell responses <i>in vitro</i> and <i>in vivo</i>, which led to delayed tumor growth and prolonged
survival of tumor-bearing mice when immunized with a specific tumor
antigen. Therefore, a transient intracellular ROS surge alone, if
properly manipulated, can cause immature DCs to differentiate into
a motile state and mature forms that are sufficient to initiate adaptive
T cell responses <i>in vivo</i>