Hironori Matsushima - Academia.edu (original) (raw)

Papers by Hironori Matsushima

Supplemental Figure S1. Effects of VBL on human DCs. Human DC cultures were incubated for 24 h wi... more Supplemental Figure S1. Effects of VBL on human DCs. Human DC cultures were incubated for 24 h with 0.3 μM VBL or vehicle alone and then examined for the surface expression (median fluorescence intensity) of the indicated markers and cell viability within CD1a + populations (PI uptake) (A). Supernatants harvested from the above human DC cultures were examined for the indicated cytokines and chemokines (B). Following 24 h culture with 0.3 μM VBL or vehicle alone, human DCs were incubated for 10 min with FITC-DX at 37 o C. Data shown are the means ± SD (n = 3) of FITC fluorescent signals within the CD11c + populations. Statistically significant differences compared with the vehicle-treated control are indicated with asterisks (*P < 0.05, **P < 0.01, ***P < 0.001). All the results are representative of two (C) or three independent experiments (A, B).

Despite the crucial roles dendritic cells (DC) play in host immunity against cancer, the pharmaco... more Despite the crucial roles dendritic cells (DC) play in host immunity against cancer, the pharmacologic effects of many chemotherapeutic agents have remained mostly unknown. We recently developed a DC biosensor clone by engineering the stable murine DC line XS106 to express the yellow fluorescent protein (YFP) gene under the control of interleukin (IL)-1B promoter. In this study, the resulting XS106 pIL1-YFP DC clone was used to screen 54 anticancer drugs. Each drug was tested at five concentrations (0.1-10 Mmol/L) for its effects on YFP expression, cell viability, and granulocyte macrophage colonystimulating factor-dependent growth. Our unbiased systematic screening unveiled a striking heterogeneity among the tested anticancer drugs in their effects on the three functional variables. Interestingly, 15 drugs induced significant YFP expression at subcytotoxic concentrations and were thus categorized as ''DC-stimulatory'' anticancer drugs. These drugs were subsequently found to induce at least one of the characteristic maturational changes in mouse bone marrowderived DCs. For example, vinblastine, a prototypic drug of this class, induced the production of IL-1B, IL-6, and IL-12, increased surface expression of CD40, CD80, CD86, and MHC class II, and augmented T cell-stimulatory capacity of DCs. Not only do these results illustrate the differential pharmacologic effects of commonly used chemotherapeutic agents on DCs, they may also provide a conceptual framework for rationale-based selection and combination of anticancer drugs for clinical application.

Supplementary Figure 2 from Dual Therapeutic Efficacy of Vinblastine as a Unique Chemotherapeutic... more Supplementary Figure 2 from Dual Therapeutic Efficacy of Vinblastine as a Unique Chemotherapeutic Agent Capable of Inducing Dendritic Cell Maturation

Supplementary Figure Legend from Classification of Chemotherapeutic Agents Based on Their Differe... more Supplementary Figure Legend from Classification of Chemotherapeutic Agents Based on Their Differential In vitro Effects on Dendritic Cells

Our recent unbiased functional screen of 54 chemotherapeutic drugs unveiled striking heterogeneit... more Our recent unbiased functional screen of 54 chemotherapeutic drugs unveiled striking heterogeneity in their effects on dendritic cells (DC). Most notably, vinblastine (VBL) was found to induce phenotypic and functional maturation of DCs in vitro. Here, we sought to determine whether VBL exhibits “dual” therapeutic efficacy in living animals by directly killing tumor cells and by boosting host immunity via DC maturation. Local injection of VBL in a low dose into the skin of C57BL/6 mice induced in situ maturation of epidermal Langerhans cells. When coinjected with a model antigen, ovalbumin (OVA), VBL enhanced OVA-specific cellular and humoral immune responses. When injected directly into the OVA cDNA–transduced E.G7 tumors, VBL augmented clonal expansion of OVA-reactive CD8 T cells and CTL activities. In B16 melanoma model, intratumor VBL injection induced apoptosis of melanoma cells, phenotypic maturation of tumor-infiltrating DCs, and significant CTL activities. Although complete ...

Supplementary Movie 1 from Dual Therapeutic Efficacy of Vinblastine as a Unique Chemotherapeutic ... more Supplementary Movie 1 from Dual Therapeutic Efficacy of Vinblastine as a Unique Chemotherapeutic Agent Capable of Inducing Dendritic Cell Maturation

Journal of Immunology, Jul 1, 2004

Recent studies have revealed that murine bone marrow-derived cultured mast cells (BMMC), which ar... more Recent studies have revealed that murine bone marrow-derived cultured mast cells (BMMC), which are phenotypically immature mast cells, express functional TLR2 and TLR4 that recognize distinct pathogen-associated molecules. However, it remains relatively uncertain whether mast cells express other TLR. We recently established a method to obtain large numbers of murine fetal skin-derived cultured mast cells (FSMC); these cells exhibit important features of connective tissue type mast cells. Working with FSMC and BMMC, the TLR mRNA expression profiles were compared between both cell types. Although TLR2 and TLR4 mRNA were detected in both cells at comparable levels, TLR3, TLR7, and TLR9 mRNA were expressed by FSMC at higher levels than by BMMC, suggesting distinct TLR expression profiles among different mast cell populations. With respect to their functional aspects, FSMC, but not BMMC, dose dependently produced proinflammatory cytokines (TNF-␣ and IL-6) and chemokines (RANTES, MIP-1␣, and MIP-2) in response to poly(I:C), R-848, and CpG oligodeoxynucleotide, which are TLR3, TLR7, and TLR9 activators, respectively. Interestingly, these TLR activators failed to induce degranulation and IL-13 production by both mast cells, although peptidoglycan and LPS (TLR2 and TLR4 activators, respectively) induced IL-13 production by both cells. Mast cells, thus, may have potential to recruit other immune cells to the infected sites by responding to various bacterial and viral components through TLR signaling pathways, presumably being involved in initiating innate immunity and subsequently linking innate and acquired immune responses.

The Journal of Immunology, May 1, 2012

Journal of immunology (Baltimore, Md. : 1950), 2015

Rapid enhancement of phagocyte functionality is a hallmark of neutrophil priming. GeneChip analys... more Rapid enhancement of phagocyte functionality is a hallmark of neutrophil priming. GeneChip analyses unveiled elevated CD54, dectin-2, and IL-1β mRNA expression by neutrophils isolated from inflammatory sites. In fact, CD54 and dectin-2 protein expression was detected on neutrophils recovered from skin, peritoneal, and lung inflammation lesions but not on those in bone marrow or peripheral blood. Neutrophils increased CD54 and dectin-2 mRNA during migration in Boyden chambers and acquired CD54 and dectin-2 surface expression after subsequent exposure to GM-CSF. Neutrophils purified from IL-1β promoter-driven DsRed-transgenic mice acquired DsRed signals during cell migration or exposure to GM-CSF. CD54 and dectin-2 were expressed by DsRed(+) (but not DsRed(-)) neutrophils in GM-CSF-supplemented cultures, and neutrophils recovered from inflammatory sites exhibited strong DsRed signals. The dynamic process of neutrophil priming was studied in chemically induced inflammatory skin lesions...

PLoS Pathogens, 2012

Neutrophil abscess formation is critical in innate immunity against many pathogens. Here, the mec... more Neutrophil abscess formation is critical in innate immunity against many pathogens. Here, the mechanism of neutrophil abscess formation was investigated using a mouse model of Staphylococcus aureus cutaneous infection. Gene expression analysis and in vivo multispectral noninvasive imaging during the S. aureus infection revealed a strong functional and temporal association between neutrophil recruitment and IL-1b/IL-1R activation. Unexpectedly, neutrophils but not monocytes/macrophages or other MHCII-expressing antigen presenting cells were the predominant source of IL-1b at the site of infection. Furthermore, neutrophil-derived IL-1b was essential for host defense since adoptive transfer of IL-1bexpressing neutrophils was sufficient to restore the impaired neutrophil abscess formation in S. aureus-infected IL-1bdeficient mice. S. aureus-induced IL-1b production by neutrophils required TLR2, NOD2, FPR1 and the ASC/NLRP3 inflammasome in an a-toxin-dependent mechanism. Taken together, IL-1b and neutrophil abscess formation during an infection are functionally, temporally and spatially linked as a consequence of direct IL-1b production by neutrophils.

PLoS ONE, 2013

Neutrophils contribute to innate host immunity by functioning as professional phagocytes, whereas... more Neutrophils contribute to innate host immunity by functioning as professional phagocytes, whereas dendritic cells (DCs) are prototypic antigen presenting cells (APCs) responsible for the induction of adaptive immune responses. We have demonstrated recently that neutrophils trans-differentiate into a unique population, termed "neutrophil-DC hybrids," expressing surface markers of both neutrophils and DCs and exhibiting dual functionality of both phagocytes and APCs. Although the hybrid cells emerged in significant numbers in murine bone marrow (BM) culture in the presence of GM-CSF, mechanisms regulating their development remained mostly unknown. In this study, we tested a total of 61 cytokines for their potentials to regulate neutrophil-DC hybrid formation using a newly developed BM micro-culture system combined with semi-automated FACS analysis. Several cytokines including GM-CSF were found to promote the generation of neutrophil-DC hybrids defined by the phenotype of CD11c + /MHC II + /Ly6G +. When tested in the presence of GM-CSF, hybrid cell development was enhanced by IL-4 and suppressed by interferon-γ (IFNγ) in dose-dependent fashions. We next determined in vivo impacts of IL-4 and IFNγ on the development of neutrophil-DC hybrids in thioglycollate-induced peritonitis lesions. Intraperitoneal administrations of IL-4/anti-IL-4 antibody complex (IL-4C) significantly increased the number of hybrids recovered from the lesions. By contract, recovery of hybrids was reduced by recombinant IFNγ. With regard to function, those hybrid cells recovered from IL-4C-treated mice and IFNγ-treated mice showed potent abilities to capture E.coli. These observations imply that emergence of neutrophil-DC hybrids in inflammatory sites is tightly regulated by local cytokine milieus.

Peptides, 1998

Substance P (SP) has been shown to induce phosphatidylinositol (PI) hydrolysis and Ca 2ϩ mobiliza... more Substance P (SP) has been shown to induce phosphatidylinositol (PI) hydrolysis and Ca 2ϩ mobilization in AR42J cells. In this study, we confirmed the expression of NK 1 but not NK 2 or NK 3 receptors in this cell line, and further investigated signaling pathways via NK 1 receptors and their desensitization. The activation of NK 1 receptors by SP affected neither basal cyclic AMP level nor cyclic AMP accumulation induced by secretin and forskolin, although it stimulated PI hydrolysis. Furthermore, SP induced Ca 2ϩ mobilization even in the absence of extracellular Ca 2ϩ , though maximal response was reduced. U73122, a phospholipase C (PLC) inhibitor, nearly abolished Ca 2ϩ response to SP. In addition, SP-induced Ca 2ϩ signaling and PI hydrolysis rapidly desensitized following short exposure to SP, which did not affect the Ca 2ϩ amount in intracellular Ca 2ϩ stores or Ca 2ϩ responses to carbachol and gastrin releasing peptide-10. These findings suggested that NK 1 receptors do not couple to adenylate cyclase, although they induce PI response, and that NK 1 receptors induce both intracellular Ca 2ϩ release and Ca 2ϩ influx through PLC activation. Ca 2ϩ signaling and PI hydrolysis through NK 1 receptors desensitized rapidly after the stimulation, maybe dependent on the modification of NK 1 receptors.

Journal of Investigative Dermatology, 2012

As a skin-resident member of the dendritic cell (DC) family, Langerhans cells (LCs) are generally... more As a skin-resident member of the dendritic cell (DC) family, Langerhans cells (LCs) are generally regarded to function as professional antigen presenting cells. Here we report a simple method to visualize the endocytotic activity of LCs in living animals. BALB/c mice received subcutaneous injection of FITC-conjugated dextran (DX) probes into the ear skin and were then examined under confocal microscopy. Large numbers of FITC + epidermal cells became detectable 12-24 h after injection as background fluorescence signals began to disappear. Most (>90%) of the FITC + epidermal cells expressed Langerin, and >95% of Langerin + epidermal cells exhibited significant FITC signals. To assess intracellular localization, Alexa Fluor 546-conjugated DX probes were locally injected into IAβ-EGFP knock-in mice and Langerin-EGFP-DTR mice-three dimensional rotation images showed close association of most of the internalized DX probes with MHC class II molecules, but not with Langerin molecules. These observations support the current view that LCs constantly sample surrounding materials, including harmful and innocuous antigens, at the environmental interface. Our data also validate the potential utility of the newly developed imaging approach to monitor LC function in wild-type animals. Recent advances in mouse genetic engineering and intravital confocal imaging have enabled direct visualization of LC behaviors in living animals.

Journal of Investigative Dermatology, 2010

IL-1 is a prototypic inflammatory cytokine that has pathogenic roles in various skin disorders. A... more IL-1 is a prototypic inflammatory cytokine that has pathogenic roles in various skin disorders. Although Langerhans cells (LCs) have been reported to express IL-1b mRNA upon application of contact sensitizers, it remains unclear whether other cell types produce IL-1b in skin. Thus, we sought to directly identify IL-1bproducing cells in living animals by construction of transgenic mice expressing DsRed fluorescence protein gene under the control of IL-1b promoter. Little DsRed fluorescence signal was detected in skin under steadystate conditions. Striking increases in DsRed signal were observed after topical application of a contact sensitizer, oxazolone, which also induced markedly elevated IL-1b mRNA and protein expression. DsRed signal was expressed primarily by CD45 þ /CD11b þ myeloid leukocytes in both epidermal and dermal compartments and was detected only in small fractions of epidermal LCs. Interestingly, DsRed þ cells emerged preferentially as clusters around hair follicles. Intravital confocal imaging experiments revealed highly motile potentials of DsRed þ cells-they constantly crawled around hair follicles via amoeba-like movements with a mean velocity of 1.0 ± 0.4 mm min-1 (epidermis) or 2.7 ± 1.4 mm min-1 (dermis). The newly developed in vivo imaging system represents a useful tool for studying spatial regulation of IL-1b production in skin.

Journal of Investigative Dermatology, 2003

We describe a novel culture system for generating large numbers of murine skin-associated mast ce... more We describe a novel culture system for generating large numbers of murine skin-associated mast cells and distinguish their characteristics from bone marrow-derived cultured mast cells. Culture of day 16 fetal skin single cell suspensions in the presence of interleukin-3 and stem cell factor allowed expansion and maturation of mast cells in the presence of stromal cells. The average yield of mast cells after 2 wk was 7.3 million cells per fetus at a purity of 96%. These fetal skin-derived cultured mast cells increased their histamine content in a time-dependent manner to 3.6 pg per cell after 2 wk and 6.7 pg per cell after 4 wk. Phenotypic analyses revealed much greater expression of CD49b and CD81 and lesser expression of CD77 and CD102 on fetal skin-derived cultured mast cells as compared with bone marrow-derived cultured mast cells. These findings suggest a close similarity between fetal skin-derived cultured mast cells and freshly isolated cutaneous mast cells. Connective tissue mast cell characteristics of fetal skin-derived cultured mast cells were evidenced by: (1) their greater histamine content than bone marrow-derived cultured mast cells; (2) the presence of heparin; and (3) their degranulation in response to compound 48/80 and substance P. Importantly, fetal skin-derived cultured mast cells secreted greater amounts of interleukin-13 but much less MIP-1beta and interleukin-6 than bone marrow-derived cultured mast cells in response to ionomycin. Thus fetal skin-derived cultured mast cells have many characteristics distinct from bone marrow-derived cultured mast cells and can be used as a model of cutaneous mast cells to discern their functions.