Tumor necrosis factor alpha small interfering RNA decreases herpes simplex virus-induced inflammation in a mouse model.

BACKGROUND: Anti-TNFalpha antibodies have been used for treating inflammation in patients. But, more effective and safer drugs need to be developed for improved future therapeutic use. OBJECTIVES: To inhibit the expression of TNFalpha, we used small interfering RNAs (siRNAs) to reduce over expression of TNFalpha in vitro in cell cultures and in an in vivo Behcet's disease-like (BD) mouse model for amelioration of chronic inflammation. METHODS: TNFalpha siRNA was injected intraperitoneally twice with a 1-week interval. To compare the efficacy of TNFalpha siRNA versus an anti-TNFalpha antibody, Infliximab and Etanercept were administered to symptomatic mice with inflamed tissue. RESULTS: Intraperitoneal delivery of TNFalpha siRNA effectively decreased BD symptoms in 18 of 32 cases (56.3%). Scrambled siRNA treatment decreased BD symptoms in 2 of 19 cases (10.5%). Infliximab was effective in 11 of 27 cases (40.7%) and Etanercept was also effective in 9 of 25 cases (36.0%) at the end of the second week after treatment. TNFalpha siRNA reduced serum levels of TNFalpha (1.57 +/- 0.43pg/ml), compared to levels in mice not injected (84.02 +/- 24.59pg/ml) (p<0.01) or scramble injected (118.89 +/- 20.08pg/ml) (p<0.01). After single injection of TNFalpha siRNA, improvement of BD symptoms showed at 9 +/- 7th day on an average, contrary, in Infliximab injected group, improvement was apparent at 15 +/- 4th day after injection (p<0.05). CONCLUSION: We show that siRNAs can be employed to inhibit cytokine gene expression in an in vivo disease mouse model. This inhibition may, therefore, be attributed to the improvement of inflammatory symptoms.

Rebamipide affects the efficiency of colchicine for the herpes simplex virus-induced inflammation in a Behcet's disease mouse model.

Rebamipide inhibits free radicals derived from activated neutrophils and decreases the inhibiting inflammatory cytokine. Behcet's disease (BD) is a chronic, multi-systemic inflammatory disorder with arthritic, gastrointestinal, mucocutaneous, ocular, vascular, and central nervous system involvement. This disease has a chronic course with periodic exacerbations and progressive deterioration. To study the effect of rebamipide treatment to BD-like mice, combination treatment with rebamipide and colchicine was compared to colchicine treatment. Colchicine is one of the most frequently prescribed medicine to the patients with BD. For each BD mouse, 200 microl gastric fluid or 2 microg colchicine or 150 microg rebamipide or 2 microg colchicine plus 150 microg rebamipide was treated orally once per day. Treatment was done for 5 consecutive days. Two hour or 20 days after last administration, spleens were isolated for RT-PCR and real time PCR, and serum was collected for ELISA. In the combination treated group, TNF alpha, MIP-1 alpha, p22 phox, p47 phox, and gp91 phox mRNA expressions were lower than rebamipide treated or colchicine treated groups by reverse transcriptase PCR. NADPH oxidase subunits mRNA were markedly downregulated compared to the colchicine treated group by real time PCR. At 20 days after administration, combination treatment decreased 23.5% of the severity score compared to before administration. In contrast, colchicine treatment decreased 14.3% of the severity score compared to before administration. Rebamipide helped the function of colchicine to improve the HSV induced BD-like symptoms by inhibiting the expression of NADPH oxidase in vivo mouse model.

The Behcet's Disease Quality of Life: reliability and validity of the Korean version.

PURPOSE: The Behcet's Disease Quality of Life (BD-QoL) is a BD-specific measure developed in the UK. The aim of this study was to adapt the BD-QoL for use in Korea. PATIENTS AND METHODS: The translation was based on the guidelines for cross-cultural adaptation. A total of 201 Korean patients with BD participated in this study. To evaluate the psychometric properties, internal consistency and test-retest reliability were used. Factor analysis was performed to examine the construct validity. To provide further evidence for validity, the correlation of BD-QoL with the Clinical Activity Form for Korean Patients with BD (BDCAF-K) and the Center for Epidemiologic Studies-Depression (CES-D) scales was assessed. RESULTS: The Korean version had high internal consistency (Cronbach's alpha, 0.93) and test-retest reliability (r = 0.835). Factor analysis of the questionnaire revealed one interpretable factor as a general health-related quality of life factor. The Korean version significantly correlated with scores of CES-D (r = 0.749, p = 0.000), self-rating scale of well-being over the past 28 days (r = 0.446, p = 0.000), and BDCAF-K score (r = 0.502, p = 0.000). CONCLUSION: Adaptation of the BD-QoL for use in Korea was successful. Together with the BDCAF-K, it may be a valuable tool for assessing the influence of interventions in BD patients and outcome in clinical trials.

Behçet's disease combined with various types of fistula.

Behçet's disease (BD) is a chronic, relapsing, multisystem disorder, characterized by recurrent oral ulcer, genital ulcers, eye lesion, and skin lesion. The underlying pathology is nonspecific vasculitis of all vessel sizes, and severe vasculitis can result in fistula formation of neighboring tissues due to a necrotic process. Herein, eleven cases of BD combined with fistula are presented. In the present study, various types of fistula were associated; enterocutaneous fistula in six patients, and rectovaginal fistula in two. The other three patients showed aortoduodenal fistula, urethrovaginal fistula and urethrocutaneous fistula. They were treated with a corrective operation, but the prognoses were poor due to frequent relapses.

Activation of extracellular signal regulated kinase 1/2 in human dermal microvascular endothelial cells stimulated by anti-endothelial cell antibodies in sera of patients with Behçet's disease.

Anti-endothelial cell antibodies (AECA) have been detected in the sera of patients with Behçet's disease (BD). The isotype of AECA from BD is IgM recognizing 44 kDa antigen (IgM-AECA) of human dermal microvascular endothelial cells (HDMEC). After stimulation of HDMEC with AECA-positive sera from BD patients, the expression of intercellular cell adhesion molecule-1 (ICAM-1) on HDMEC increases significantly. Mitogen-activated protein kinase (MAPK) cascade is one of protein kinase families activated by a wide spectrum of extracellular stimuli. There are several subtypes, including extracellular signal regulated kinase (ERK)1/2, c-Jun NH(2) terminal kinase (JNK), and p38 cascades, and they regulate various cellular processes such as cell growth, differentiation, and inflammation. We examined the involvement of MAPK as a signal transduction pathway in the IgM-AECA-induced ICAM-1 expression. We used enzyme-linked immunosorbent assay (ELISA) and fluorescence-activated cell sorting (FACS) for detecting the induction of ICAM-1 on HDMEC. We also examined the production of tumor necrosis factor alpha (TNFalpha) or interleukin-1alpha (IL-1alpha) by HDMEC after stimulation with IgM-AECA, and checked the involvement of MAPK by Western blot assay. IgM-AECA cocktail from 8 patients with BD induced expression of the ICAM-1 on HDMEC. Neither TNFalpha nor IL-1alpha was detected by ELISA, FACS or reverse transcriptase-polymerase chain reaction in activated HDMEC cultures. IgM-AECA cocktail activated ERK1/2 and showed peak activities at 5 min after the stimulation. Specific MAPK/ERK kinase inhibitor PD98059 inhibited IgM-AECA-induced ERK1/2 activities and ICAM-1 expression on HDMEC at a concentration of 60 microM. IgM-AECA can play a pathogenic role in induction of vasculitis and inflammatory lesions of BD by directly activating endothelial cells, not by production of TNFalpha or IL-1alpha from HDMEC. ERK1/2 are involved in expression of ICAM-1 on HDMEC stimulated with IgM-AECA.

Combined treatment with colchicine and Herba Taraxaci (Tarazacum mongolicum Hand.-Mazz.) attenuates Behcet's disease-like symptoms in mice and influences the expressions of cytokines.

Herbal medicine, Herba Taraxaci (Tarazacum mongolicum Hand.-Mazz.), was administered to mice with Behcet's disease (BD)-like symptoms induced by herpes simplex virus (HSV). BD is a chronic recurrent inflammatory disease. Herba Taraxaci (6 mg) was administered alone or in combination with 2 microg of colchicine to BD-like mice. Colchicine is a drug that is widely used as a medication for BD patients. The water extracts of Herba Taraxaci were administered orally once per day for 20 days. Eighty percent (8 of 10) of mice treated with Herba Taraxaci combined with colchicine showed improvement in mucocutaneous symptoms compared to 0% (0 of 10) of the nontreated group and 30% (3 of 10) treated with colchicine alone. Cytokine expression in spleen tissue collected from treated mice was analyzed by RT-PCR and FACS. Treatment with Herba Taraxaci induced IL-4 mRNA, and spleen from mice receiving the combined treatment (Herba Taraxaci and colchicines) showed an increased number of splenocytes staining with anti-IL-10 (46.8+/-6.80) compared to Herba Taraxaci (35.4+/-2.17) (p<0.05) or colchicine alone (26.2+/-4.47) (p<0.001). These results suggest that the Herba Taraxaci may be an effective complementary agent in the treatment of BD.

Thalidomide upregulates macrophage inflammatory protein-1alpha in a herpes simplex virus-induced Behçet's disease-like animal model.

The mechanism of action of thalidomide in the treatment of patients with Behçet's disease (BD) is poorly understood. There is some evidence to suggest that certain immunological abnormalities are associated with the pathogenesis of BD. A BD-like mouse model induced by herpes simplex virus (HSV) inoculation shows similar immunological abnormalities. In this study, thalidomide was administered in order to understand the mechanism for the improvement in symptoms in BD-like mice. Eight out of ten thalidomide-treated mice showed improvement but none of ten placebo-treated mice (P < 0.005). The improvements were seen in mucocutaneous symptoms. The mice were sacrificed on the 6th day, and the spleens subjected to RT-PCR, FACS, Western blot and immunohistochemical analysis. IL-2, IL-4, IL-6, IL-10, IFN-gamma, TNFalpha, TGFbeta, MCP-1, RANTES, perforin, IP-10, FasL, FasR and MIP-lalpha were determined. Among these, TNFalpha, MIP-1alpha, perforin and Fas were influenced by thalidomide treatment. These results suggest that thalidomide can attenuate HSV-induced BD-like symptoms in mice through the downregulation of TNFalpha (P < 0.005) and the upregulation of MIP-1alpha (P < 0.005), perforin (P < 0.05) and FasR (P < 0.1).

Psychosocial Aspects of Acne Vulgaris: A Community-based Study with Korean Adolescents.

BACKGROUND: Acne vulgaris is a distressing condition that affects the majority of adolescents, but the impact of acne vulgaris on the psychological aspects in this age group is poorly understood. OBJECTIVE: The purpose of this study was to determine the prevalence of acne, and the level of emotional, social, and functional impairments among Korean adolescents with acne. METHODS: Five hundred four middle school students (13~16 years) participated. The severity of acne was graded by visual examination using the Korean Acne Grading System. Self-reported questionnaires, including subjective acne severity rating, the Self Image Questionnaire, the Rosenberg Self-Esteem Questionnaire, the Index of Peer Relations, and the Beck Depression Inventory were used to assess psychologic status. RESULTS: There was a prevalence of acne in 78.9% of the study samples, with 10.2% of students having moderate-to-severe acne. Acne was more prevalent and severe in boys than girls. Participants with severe acne and girls had higher levels of emotional and social impairments. The longer the acne persisted, the more stress the students felt. The degree of stress and extent of self-image impairment were related to subjective severity more than objective grading. CONCLUSION: Acne is a common disorder among Korean adolescents and appears to have a considerable impact on mental health. Dermatologists should be aware of the importance of basic psychosomatic treatment in conjunction with early medical, educational intervention in the management of acne.

Using pCIN-mIL-4 DNA vector to express mRNA and protein and to improve herpes simplex virus-induced Behcet's disease symptoms in mice.

Behcet's disease (BD) is a chronic, recurrent, inflammatory, multisystemic disorder characterized primarily by vasculitis. The etiopathogenesis of BD involves immunogenetics, infectious organisms (streptococcus, herpes simplex virus), immunoregulation and vascular dysfunctions. We previously found that immunoregulation associated with viral infection was important to the development of BD-like symptoms. Recently, we demonstrated that Th2 cytokines up-regulated by Th2 adjuvant were efficient in attenuating or improving these BD-like symptoms. In order to directly augment IL-4 expression, a DNA vector (pCIN-mIL-4) was administered to BD-like mice using the Helios gene gun system. Two injections of the pCIN-mIL-4 vector, spread over 2 weeks, attenuated or improved the mucocutaneous symptoms of 10 out of 12 BD-like mice in our study. The improved mucocutaneous symptoms were crust in face, ulcer in mouth, scruff, back, genital and erythema. This improvement also correlated with induction of IL-4 mRNA in lymph nodes, protein in serum and intracellular IL-4 staining in splenocytes. Normal control mice (n = 10) injected with the pCIN-mIL-4 vector expressed IL-4 mRNA and showed more splenocytes stained with anti-IL-4 antibody (5.77 +/- 0.92%) than did mice injected with the pCIN control vector (3.34 +/- 0.25%; p = 0.02). These findings indicate that an IL-4 DNA vector could be used to express mRNA and protein in vivo and further suggest that such an IL-4 DNA vector could be used as a therapeutic treatment in recurrent inflammation shifted to T helper type 1 cytokine production.

Therapeutic effects of gemcitabine on cutaneous manifestations in an Adamantiades-Behçet's disease-like mouse model.

The aim of this work was to study the effects and side effects of gemcitabine (2',2'-difluorodeoxycytidine, dFdC), a pyrimidine synthesis inhibitor, on skin lesions of a herpes simplex virus (HSV)-induced Adamantiades-Behçet's disease (ABD)-like mouse model. For the dose-escalation study, ICR mice were treated intraperitoneally with dFdC over 5 days. For the efficacy study, ICR mice were inoculated with HSV and classified as having ABD according to a revised Japanese classification, and then 18 ABD mice were randomly assigned to placebo, 0.06 or 0.12 microg of dFdC/day over 5 days. Serum levels of interleukin-4 (IL-4), IL-6, IL-10, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) were determined using enzyme-linked immunosorbent assay. After application of 3 microg of dFdC over 5 days, alanine aminotransferase increased (P = 0.032), but all other kidney and liver parameters were unchanged. In ABD mice, 5 days of dFdC treatment with 0.06 or 0.12 microg of dFdC/day resulted in a dose-dependent improvement of cutaneous manifestations by more than 60% (P = 0.017). There was no significant change in cytokine levels, and none of the cytokine levels correlated with response to treatment. Moreover, dFdC shows promising effects to improve cutaneous lesions in the HSV-induced ABD-like mouse model. In this animal model, effects of dFdC on the cytokine profile remained inconclusive.

Clinical features of patients with Behçet's disease and epididymitis.

PURPOSE: Epididymitis is a rare manifestation of Behçet's disease but its clinical significance is still not fully understood. We evaluated the clinical significance of epididymitis in patients with Behçet's disease. MATERIALS AND METHODS: Of 780 male patients attending our clinics between 1985 and 2002 who were diagnosed with Behçet's disease by international criteria or who had the complete or incomplete type of Behçet's disease by Japanese criteria 36 were identified with epididymitis. Clinical data on these patients were reviewed. RESULTS: The 36 patients were categorized into the complete (13 or 36.1%) and incomplete (23 or 63.9%) types of Behçet's disease with an average age at onset of 27.4 years. The frequency of individual symptoms were oral ulcers and cutaneous involvement in all 36 cases (100%), genital ulcers in 32 (88.9%), arthritis in 18 (50.0%), ocular involvement in 17 (47.2%), central nervous system involvement in 2 (5.6%), gastrointestinal ulcer in 1 (2.8%) and a positive pathergy test in 4 (11.1%). A significantly higher number of patients with epididymitis had genital ulcers (p <0.05), cutaneous involvement (p <0.001), arthritis (p <0.05), central nervous system involvement (p <0.05) and a positive pathergy test (p <0.05) compared with the other 744 with Behçet's disease without epididymitis who served as controls. CONCLUSIONS: To our knowledge there has been no controlled study of the clinical significance of epididymitis in Behçet's disease. Our results suggest a tendency toward severe Behçet's disease manifestations in patients with epididymitis, prompting physicians to evaluate closely and meticulously treat such patients.

Human alpha-enolase from endothelial cells as a target antigen of anti-endothelial cell antibody in Behçet's disease.

OBJECTIVE: To identify and recombine a protein of the human dermal microvascular endothelial cell (HDMEC) that specifically reacts with anti-endothelial cell antibody (AECA) in the serum of patients with Behçet's disease (BD), and to evaluate the usefulness of this protein in BD. METHODS: The proteomics technique, with 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry, was used to identify and recombine HDMEC antigen. Western blotting and enzyme-linked immunosorbent assay (ELISA) of recombinant protein isolated by gene cloning were performed on serum from healthy controls, patients with BD, and patients with other rheumatic diseases (rheumatoid arthritis, systemic lupus erythematosus, and Wegener's granulomatosis). RESULTS: Eighteen of 40 BD patients had serum IgM antibody to HDMEC antigen. The purified protein that reacted with AECA in BD patient sera was found to be alpha-enolase by 2-dimensional gel electrophoresis followed by immunoblotting and MALDI-TOF mass spectrometry. Recombinant alpha-enolase protein was isolated and refined by gene cloning. On Western blots, AECA-positive IgM from the sera of patients with active BD reacted strongly with recombinant human alpha-enolase. BD patient sera positive for anti-alpha-enolase did not react with human gamma-enolase. On dot-blotting, reactivity to human alpha-enolase was detected only in the IgM-positive group. Fifteen of the 18 AECA-positive sera that were positive for the HDMEC antigen showed reactivity to recombinant alpha-enolase IgM antibody by ELISA. CONCLUSION: The alpha-enolase protein is the target protein of serum AECA in BD patients. This is the first report of the presence of IgM antibodies to alpha-enolase in endothelial cells from the serum of BD patients. Although further studies relating this protein to the pathogenesis of BD will be necessary, alpha-enolase and its antibody may prove useful in the development of new diagnostic and treatment modalities in BD.