Coronary Endothelial Dysfunction After Kawasaki Disease: Evaluation by Intracoronary Injection of Acetylcholine 11This study was supported in part by Research Grant 6454310 from the Ministry of Education, Tokyo, Japan.

AbstractObjectives. This study sought to assess the endothelial function of long-term coronary artery lesions in patients with Kawasaki disease (KD).Background. The vascular function of the coronary arteries in children with long-term KD remains uncertain. We report our findings of the vascular response of the coronary arteries to intracoronary injection of acetylcholine (ACh) in patients with KD.Methods. A total of 35 patients (25 patients with KD and 10 control subjects) were examined using coronary angiography. Individual arteries were divided into four groups according to the type of the coronary artery lesion: group 1 consisted of 25 sites with regressed aneurysms. These aneurysms had developed in the acute stage but had subsequently regressed and demonstrated normal findings on the follow-up coronary angiogram. Group 2 consisted of 24 sites with persistent aneurysms. Group 3 involved 60 angiographically normal sites in the same patients as those in group 1 or 2. Group 4 consisted of 30 sites in control subjects who had congenital heart disease with normal coronary arteries. During coronary angiography we infused 15 μg of ACh chloride into the coronary artery. The lumen diameters were measured using a cine videodensitometric analyzer to study the distensibility of the coronary artery wall.Results. The mean (±SD) change in diameter was an increase of 11.71 ± 12.34% in group 3 (coronary arteries without lesions in patients with KD) and 12.21 ± 9.71% in the control group, demonstrating marked vasodilation in both groups. In contrast, the changes in the regressed aneurysms of group 1 and in the persistent aneurysms of group 2 were −2.65 ± 12.12% and −0.08 ± 6.51%, respectively, demonstrating no change or mild vasoconstriction. The change in groups 1 and 2 was significantly less than that in group 3 or in the control group. Group 3 showed no significant difference from the control group.Conclusions. These findings suggest that long-term coronary artery lesions, even after aneurysm regression, may have impaired endothelial function. A long-term follow-up study for those patients is essential

Spherical Lactic Acid Bacteria Activate Plasmacytoid Dendritic Cells Immunomodulatory Function via TLR9-Dependent Crosstalk with Myeloid Dendritic Cells

Plasmacytoid dendritic cells (pDC) are a specialized sensor of viral and bacterial nucleic acids and a major producer of IFN-α that promotes host defense by priming both innate and acquired immune responses. Although synthetic Toll-like receptor (TLR) ligands, pathogenic bacteria and viruses activate pDC, there is limited investigation of non-pathogenic microbiota that are in wide industrial dietary use, such as lactic acid bacteria (LAB). In this study, we screened for LAB strains, which induce pDC activation and IFN-α production using murine bone marrow (BM)-derived Flt-3L induced dendritic cell culture. Microbial strains with such activity on pDC were absent in a diversity of bacillary strains, but were observed in certain spherical species (Lactococcus, Leuconostoc, Streptococcus and Pediococcus), which was correlated with their capacity for uptake by pDC. Detailed study of Lactococcus lactis subsp. lactis JCM5805 and JCM20101 revealed that the major type I and type III interferons were induced (IFN-α, -β, and λ). IFN-α induction was TLR9 and MyD88-dependent; a slight impairment was also observed in TLR4-/- cells. While these responses occurred with purified pDC, IFN-α production was synergistic upon co-culture with myeloid dendritic cells (mDC), an interaction that required direct mDC-pDC contact. L. lactis strains also stimulated expression of immunoregulatory receptors on pDC (ICOS-L and PD-L1), and accordingly augmented pDC induction of CD4+CD25+FoxP3+ Treg compared to the Lactobacillus strain. Oral administration of L. lactis JCM5805 induced significant activation of pDC resident in the intestinal draining mesenteric lymph nodes, but not in a remote lymphoid site (spleen). Taken together, certain non-pathogenic spherical LAB in wide dietary use has potent and diverse immunomodulatory effects on pDC potentially relevant to anti-viral immunity and chronic inflammatory disease

Oral administration of Lactococcus lactis subsp. lactis JCM5805 enhances lung immune response resulting in protection from murine parainfluenza virus infection.

When activated by viral infection, plasmacytoid dendritic cells (pDCs) play a primary role in the immune response through secretion of IFN-α. Lactococcus lactis subsp. lactis JCM5805 (JCM5805) is a strain of lactic acid bacteria (LAB) that activates murine and human pDCs to express type I and type III interferons (IFNs). JCM5805 has also been shown to activate pDCs via a Toll-like receptor 9 (TLR9) dependent pathway. In this study, we investigated the anti-viral effects of oral administration of JCM5805 using a mouse model of murine parainfluenza virus (mPIV1) infection. JCM5805-fed mice showed a drastic improvement in survival rate, prevention of weight loss, and reduction in lung histopathology scores compared to control mice. We further examined the mechanism of anti-viral effects elicited by JCM5805 administration using naive mice. Microscopic observations showed that JCM5805 was incorporated into CD11c+ immune cells in Peyer's patches (PP) and PP pDCs were significantly activated and the expression levels of IFNs were significantly increased. Interestingly, nevertheless resident pDCs at lung were not activated and expressions levels of IFNs at whole lung tissue were not influenced, the expressions of anti-viral factors induced by IFNs, such as Isg15, Oasl2, and Viperin, at lung were up-regulated in JCM5805-fed mice compared to control mice. Therefore expressed IFNs from intestine might be delivered to lung and IFN stimulated genes might be induced. Furthermore, elevated expressions of type I IFNs from lung lymphocytes were observed in response to mPIV1 ex vivo stimulation in JCM5805-fed mice compared to control. This might be due to increased ratio of pDCs located in lung were significantly increased in JCM5805 group. Taken together, a specific LAB strain might be able to affect anti-viral immunological profile in lung via activation of intestinal pDC leading to enhanced anti-viral phenotype in vivo

Activation of pDCs in intestine by JCM5805 administration.

<p>Healthy C57BL / 6J mice were divided into control and JCM5805 groups (n = 4 in each group), and mice in the JCM5805 group were orally administered JCM5808 daily for 2 weeks. A. Low density cells prepared from PP of each group were analyzed by FACS. Expression level of cell surface activation marker was evaluated for MHC class II as median fluorescence intensities (M.F.I.) in left panel. Ratio of pDCs to total population was shown in right panel. pDCs was defined as “CD3⁻ Siglec-H⁺ CD11c⁺ in total population”. Short line represents the mean values. *<i>P</i><0.05 (Student’s t test). B, Total mRNA was extracted from PP pDCs from mice in the control (open columns) and JCM5805 groups (dot columns) (n = 8 in each group). <i>Ifnα</i> and <i>Ifnβ</i> gene expressions were measured by qRT-PCR and normalized to <i>Gapdh</i> gene expression. Data are shown as mean ± SD. *<i>P</i><0.05 (Student’s t test). These data are representative of three independent experiments. Each data are mean ± SD.</p

Effects of JCM5805 on mPIV1 infection.

<p>A. Experimental procedure of mPIV1 infection. Mice in the control and JCM5805 groups were fed diet with or without 1 mg / mouse / day of JCM5808 during the study period (day -14 to 15). Mice were intranasally infected with mPIV1 on day 0. On 3 days post-mPIV1 infection, six mice were sacrificed from each group for lung histopathology. Thereafter survival rate, body weight and clinical scores were investigated with remained control mice n = 12, and JCM5805 mice n = 13. B. Survival rate of mice infected with mPIV1. The control (circle) and JCM5805 (square) groups consisted of 12 and 13 mice, respectively. The survival of each animal was monitored daily. <i>P</i><0.001 (Log-Rank test). C. Body weight of mice infected with mPIV1. The control (circle) and JCM5805 (square) groups consisted of 12 and 13 mice, respectively. The body weight of each surviving animal was measured daily. The body weight values are shown as mean ± SD. *<i>P</i><0.05, **<i>P</i><0.01 (Student’s t test). The data shown is representative of two independent experiments.</p

Lung histopathology of mPIV1-infected mice.

<p>A. Representative hematoxylin and eosin (H & E)-stained sections of lung tissues from control and JCM5805 group mice (6 mice per group). Lung tissues were prepared from mice 3 days after infection. Scale bars, 300 μm. B. Histological scoring of lung tissues from mPIV1-infected mice belong to control (open columns) and JCM5805 (dot columns) group. Sections were scored at four levels as follows: 0, no symptoms; 1, low pathogenicity; 2, medium pathogenicity; 3, high pathogenicity. The mean ± SD of the tissues in each group is shown. *<i>P</i><0.05, **<i>P</i><0.01 (Mann-Whitney U test). The data shown is representative of two independent experiments.</p

Effects of Recombination on Hitchhiking Diversity in the Brassica Self-incompatibility Locus Complex

In self-incompatibility, a number of S haplotypes are maintained by frequency-dependent selection, which results in trans-specific S haplotypes. The region of several kilobases (∼40–60 kb) from SP6 to SP2, including self-incompatibility-related genes and some adjacent genes in Brassica rapa, has high nucleotide diversity due to the hitchhiking effect, and therefore we call this region the “S-locus complex.” Recombination in the S-locus complex is considered to be suppressed. We sequenced regions of >50 kb of the S-locus complex of three S haplotypes in B. rapa and found higher nucleotide diversity in intergenic regions than in coding regions. Two highly similar regions of >10 kb were found between BrS-8 and BrS-46. Phylogenetic analysis using trans-specific S haplotypes (called interspecific pairs) of B. rapa and B. oleracea suggested that recombination reduced the nucleotide diversity in these two regions and that the genes not involved in self-incompatibility in the S-locus complex and the kinase domain, but not the S domain, of SRK have also experienced recombination. Recombination may reduce hitchhiking diversity in the S-locus complex, whereas the region from the S domain to SP11 would disfavor recombination

Effect of oral administration of <i>L. lactis</i> JCM5805 to healthy C57BL/6 mice.

<p>Healthy C57BL/6 mice were divided into two groups (n = 8 each). Control group (ctrl) were fed a normal diet, and <i>L. lactis</i> group (JCM5805) were fed a diet containing 1 mg of JCM5805 per day. Two weeks later, low density cells fractions prepared from SPN (A) or MLN (B) derived each group were analyzed for MHCII and CD86 on pDC or mDC. Representative data from three independent experiments are shown.</p