Stevens-Johnson syndrome and toxic epidermal necrolysis (SJS/TEN) are severe, cutaneous adverse drug reactions that are rare but life threatening. Genetic biomarkers for allopurinol-related SJS/TEN ...in Japanese were examined in a genome-wide association study in which Japanese patients (n=14) were compared with ethnically matched healthy controls (n=991). Associations between 890 321 single nucleotide polymorphisms and allopurinol-related SJS/TEN were analyzed by the Fisher's exact test (dominant genotype mode). A total of 21 polymorphisms on chromosome 6 were significantly associated with allopurinol-related SJS/TEN. The strongest association was found at rs2734583 in BAT1, rs3094011 in HCP5 and GA005234 in MICC (P=2.44 × 10(-8); odds ratio=66.8; 95% confidence interval, 19.8-225.0). rs9263726 in PSORS1C1, also significantly associated with allopurinol-related SJS/TEN, is in absolute linkage disequilibrium with human leukocyte antigen-B*5801, which is in strong association with allopurinol-induced SJS/TEN. The ease of typing rs9263726 makes it a useful biomarker for allopurinol-related SJS/TEN in Japanese.
Full text
Available for:
DOBA, EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, IZUM, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UILJ, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Background
Our genomewide association study documented an association between cold medicine‐related Stevens‐Johnson syndrome/toxic epidermal necrolysis (CM‐SJS/TEN) and Ikaros Family Zinc Finger 1 ...(IKZF1). Few studies examined biological and pathological functions of IKZF1 in mucosal immunity. We hypothesized that IKZF1 contributes to the mucocutaneous inflammation.
Methods
Human skin and conjunctival tissues were obtained for immunohistological studies. Primary human conjunctival epithelial cells (PHCjECs) and adult human epidermal keratinocytes (HEKa) also used for gene expression analysis. We also generated K5‐Ikzf1‐EGFP transgenic mice (Ikzf1 Tg) by introducing the Ik1 isoform into cells expressing keratin 5, which is expressed in epithelial tissues such as the epidermis and conjunctiva, and then examined them histologically and investigated gene expression of the epidermis. Moreover, Ikzf1 Tg were induced allergic contact dermatitis.
Results
We found that human epidermis and conjunctival epithelium expressed IKZF1, and in PHCjECs and HEKa, the expression of IKZF1 mRNA was upregulated by stimulation with polyI:C, a TLR3 ligand. In Ikzf1 Tg, we observed dermatitis and mucosal inflammation including the ocular surface. In contact dermatitis model, inflammatory infiltrates in the skin of Ikzf1 Tg were significantly increased compared with wild type. Microarray analysis showed that Lcn2, Adh7, Epgn, Ifi202b, Cdo1, Gpr37, Duoxa1, Tnfrsf4, and Enpp5 genes were significantly upregulated in the epidermis of Ikzf1 Tg compared with wild type.
Conclusion
Our findings support the hypothesis that Ikaros might participate in mucocutaneous inflammation.
Full text
Available for:
BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Prostaglandin (PG) E(2) is produced during inflammatory responses and suppresses the production of cytokines induced by lipopolysaccharide stimulation in macrophages and dendritic cells. In this ...study, we examined the expression of PGE(2) receptors in human conjunctival epithelial cells and investigated whether PGE(2) downregulates polyinosine-polycytidylic acid (polyI:C)-induced cytokine production.
ELISA and quantitative reverse transcription (RT)-PCR were used to examine the effects of PGE(2) on the polyI:C-induced cytokine expressions by primary human conjunctival epithelial cells (PHCjEC). Reverse transcription-PCR was performed to examine the mRNA expression of the PGE(2) receptors EP1, -2, -3 and -4.
PGE(2) significantly attenuated the expressions of chemokine (C-C) motif ligand (CCL) 5, chemokine (C-X-C motif) ligand (CXCL) 10, CXCL11 and interleukin (IL) 6 in PHCjECs. Human conjunctival epithelial cells exhibited expression of EP2, -3 and -4, but not of EP1. EP2 agonist significantly suppressed the polyI:C-induced the expressions of CCL5, CXCL10 and CXCL11 but not of IL-6. EP3 agonist significantly suppressed the expressions of CCL5, CXCL10, CXCL11 and IL-6. On the other hand, EP4 agonist failed to suppress the cytokine production induced by polyI:C stimulation.
Our results show that PGE(2) attenuated the expression of CCL5, CXCL10 and CXCL11 via both EP2 and EP3, and that the expression of IL-6 was attenuated only by EP3.
The authors previously reported that human ocular surface epithelium expressed TLR3 and that its ligand polyI:C stimulated the secretion of IL-6, IL-8 and IFN-β. In this study, to examine ...comprehensive effects of polyI:C stimulation of primary human conjunctival epithelial cells (PHCjECs), the authors performed a gene-expression analysis of the polyI:C-stimulated PHCjECs using oligonucleotide microarrays, GeneChip.
The transcripts upregulated upon polyI:C stimulation in PHCjECs from two individuals were examined using GeneChip. Eleven new upregulated transcripts of interest were confirmed by quantitative real-time PCR (RT-PCR), and seven proteins produced by those transcripts were examined by ELISA or immunoblot analysis in PHCjECs from three other individuals, respectively.
GeneChip analysis showed that 150 transcripts were upregulated more than threefold and that 47 transcripts were upregulated more than 10-fold upon polyI:C stimulation in the PHCjECs. Eleven of the 47 upregulated transcripts (CXCL11, RIG-I, IL28A, CXCL10, CCL5, CCL4, MDA5, IL7R, TSLP, CCL20 and ICAM-1) were significantly upregulated upon polyI:C stimulation by quantitative RT-PCR, and the levels of seven proteins of the transcripts CXCL11, CXCL10, CCL5, CCL20, TSLP, RIG-I and MDA5 were confirmed by ELISA or immunoblot analysis to increase significantly in polyI:C-stimulated PHCjECs.
Our results might show that TLR3 of conjunctival epithelium could not only induce antiviral innate immune responses but also regulate the allergic reactions.
The aim of this study was to use larval, parasitic female and egg antigens from Strongyloides venezuelensis to detect parasite-specific IgG and immune complexes in human serum samples by ...enzyme-linked immunosorbent assay (ELISA). In total, 95 serum samples were analysed, consisting of 30 patients harbouring S. stercoralis larvae, 30 healthy subjects and 35 patients with other parasites. Sensitivity, specificity and diagnostic efficiency were calculated. A significant statistical difference was found in the detection of immune complexes and antibodies in patients harbouring S. stercoralis larvae from larval and eggs antigens, with higher positivity using larval antigen. The larval antigen showed the highest values for sensitivity, specificity and diagnostic efficiency in ELISA from detection of immune complexes. For the first time we used IgG anti-larvae, IgG anti-parasitic females or IgG anti-eggs for immune complex detection. We concluded that the association of antibody and immune complex detection could be used in the diagnosis of human strongyloidiasis.
SUMMARY
The combination of allograft limbal transplantation (ALT) and amniotic membrane transplantation (AMT) has been applied in the treatment of severe ocular surface diseases. The beneficial ...effect of this combination has been thought to result from possible immunosuppressive ability of amniotic membrane (AM). However, the mechanisms of any such ability remain unknown. In this study, we investigated whether human AM has the ability to suppress allo‐reactive T cell responses in vitro. For mixed lymphocyte reaction (MLR), lymphocytes isolated from lymph nodes of C57BL/6 mice (Mls1b, Vβ6+) were cultured with irradiated splenocytes from DBA/2 mice (Mls1a, Vβ6−) with or without human AM. For carboxyfluorescein diacetate succinimidyl ester (CFSE) experiments, responder lymph node cells were labelled with a stable intracellular fluorescent dye and cultured with irradiated stimulator cells. The ratio of responder Vβ6+ T cells was then determined by FACS analysis, and the division profiles of responder Vβ6+ T cells were analysed by CFSE content. Furthermore, Th1 and Th2 cytokine synthesis by allo‐reactive T cells in MLR culture supernatants was determined by enzyme‐linked immunosorbent assay (ELISA). Addition of AM to the MLR culture resulted in the significant inhibition of thymidine incorporation compared with control culture lacking AM. The population of responder CD4+Vβ6+ T cells was significantly reduced in the AM‐treated culture in comparison to control. CFSE analysis revealed less division and lower proliferation of responder CD4+Vβ6+ T cells in cultures with AM than without. In addition, allo‐rective T cell synthesis of both Th1 (IL‐2 and IFNγ) and Th2 (IL‐6 and IL‐10) type cytokine was significantly decreased in the presence of AM. These results indicate that human AM has the ability to suppress allo‐reactive T cells in vitro. This inhibitory effect likely contributes to the success of the ALT‐AMT combination.
Full text
Available for:
BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, SIK, UILJ, UKNU, UL, UM, UPUK
Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are acute severe blistering diseases of the skin and also two of the most devastating ocular surface diseases leading to corneal ...damage and loss of vision. The extreme rarity of cutaneous and ocular surface reactions to drug therapies led us to suspect individual susceptibility. SJS/TEN patients in the acute stage were reported to manifest increased serum levels of Fas Ligand (FasL). Thus, we performed SNP association analysis of the FasL gene.
In 76 Japanese SJS/TEN patients with ocular surface complications and 160 Japanese healthy controls, we examined four SNPs of FasL reported in the Japanese Single Nucleotide Polymorphisms (JSNP) database by sequencing.
The SNP rs.3830150 A/G showed a significant strong inverse association with SJS/TEN. Analysis of the genotype pattern of SNPs rs.3830150 and rs.2639614 (rs.3830150 A/A-rs.2639614 G/G) also manifested a strong inverse association with SJS/TEN.
FasL gene polymorphisms might be associated with SJS/TEN.
In the present study, antigens from parthenogenetic females and eggs of Strongyloides venezuelensis, or anti-parthenogenetic-female and anti-egg antigens were used to detect specific IgG and immune ...complex responses, respectively. Serum samples from experimentally infected immunocompetent and immunosuppressed rats were analysed on days 5, 8, 13 and 21 post-infection (dpi). An enzyme-linked immunosorbent assay (ELISA) was performed using alkaline parasite extract for specific IgG detection, and anti-parthenogenetic-female or anti-egg antigens for immune complex detection. The data were analysed using analysis of variance (ANOVA), followed by a Bonferroni test. When parthenogenetic female or egg extracts were used as antigens, specific IgGs were not detected in either immunocompetent or immunosuppressed rats. When anti-parthenogenetic-female or anti-S. venezuelensis-eggs were used, immune complexes were detected for the duration of the infection in immunosuppressed animals and were only detected between 5 and 13 dpi in immunocompetent animals. The duration of infection was not significantly different between the immunocompetent and immunosuppressed groups when anti-parthenogenetic-female or anti-S. venezuelensis-eggs were used. Parthenogenetic female extracts yielded significant differences between antibody and immune complex responses in immunocompetent rats from 5 to 13 dpi, but only on day 5 dpi in immunosuppressed rats. Exposure to S. venezuelensis egg extract yielded significant differences in both antibody and immune complex detection between immunocompetent and immunosuppressed rats for the duration of the infection. In conclusion, ELISA using alternative antigens may be a successful strategy for identifying immune complexes in serum samples and diagnosing active strongyloidiasis, particularly under conditions of immunosuppression.
Purpose
We reported that PTGER3 SNPs were associated with Stevens‐Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN) with severe ocular complications (SOC). We also reported that about 80% of ...our SJS/TEN patients had taken cold medicines within several days before disease onset and designated them cold medicine related‐SJS/TEN (CM‐SJS/TEN) patients, and that HLA‐A*02:06 was significantly associated with CM‐SJS/TEN in Japanese and Korean populations. Moreover, we documented that HLA‐A*02:06 with TLR3 polymorphisms exerted more than additive effects in SJS/TEN with SOC. In the current study we focused on CM‐SJS/TEN with SOC and analyzed an interactive effect between PTGER3 SNPs and HLA‐A*02:06 in Japanese and Korean populations.
Methods
Samples from 132 Japanese patients with CM‐SJS/TEN with SOC were collected and 221 healthy Japanese volunteers were also recruited as controls. Samples from 30 Korean patients with CM‐SJS/TEN with SOC were collected and 120 healthy Korean volunteers were also recruited as controls. Genotyping of PTGER3 gene SNPs was performed using the TaqMan SNP genotyping assay or the DigiTag2 assay. HLA‐A genotyping was performed using using commercial bead‐based typing kits, WAK Flow.
Results
In Japanese population, we found an interaction with additive effects between HLA‐A*02:06 and the high‐risk genotypes PTGER3 rs1327464 GA or AA (OR = 10.8, p = 2.56 × 10−7). Moreover, we also found an additive effect between HLA‐A*02:06 and the high‐risk genotypes PTGER3 rs1327464 GA or AA (OR = 14.2, p = 5.58 × 10−6).
Conclusions
These finding might show that the combination of these two polymorphisms could give improvements for a genetic testing as compared to using only one susceptibility gene.
Full text
Available for:
BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
PDF
