Alopecia areata serum inhibits the growth of normal dermal papilla cells

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Poster Presentations

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A NOVEL MODEL OF SPONTANEOUS MIXED CONNECTIVE TISSUE DISEASE (MCTD)-LIKE SKIN LESIONS IN AGED MRL MICE. Fukuml Furukawa. Masahlro Takleawa. Takao Tachibana. Yuii Horieuchl. and Sadao Imamura. Department of Dermatology, Hamamatsu Univerwy School of Medicine, Hamamatsu. and Faculty of MedIcme. Kyoto Umversity. Kyoto Japan Epidermal nuclear stammgs (ENS) by direct immunofluorescent (IF) methods are observed m the skin of mixed connective tissue disease (MCTD) and also systemic lupus erythematosus. The mechanism of ENS IS soli controversial. Recemly, we found that seed MRL/Mo-+/+(MRL/n) mice showed the hieh incidence of ENS Usme this newly eslabbshed model mouse. we examioded the possibility that this mouse wll be a model for MCTD. MRLln mu were maintanned under convermonal conditions. Skm biopsy specimens were oblamed from the back skm of 15 aged (IS-20 mo) male. 18 young ( 1~9 mo) male. 15 aged (IO- I4 ma) female, and I3 young ( 1-9 mo) female mace. Direct IF slodres revealed rhar 8 out of 15 aged male and 6 our of 15 aged female mice had ENS, but young male or female mice did not. ENS positive MRL/n mice showed also nuclear deposits of Ig m the kidney. Anti-RNP antibodies as well as anti-DNA anctbodles were demonstrated by Improved ELISA methods. but no1 by lmmunodlffwon methods. In wuo binding assay of antibodies to nucleus of PAM212 cells in culture showed the low incidence (4/30) binding to nucleus but there was no as~oaatmn teween the hindmg and ENS posllivity. Aged MRL/n mu wdl give us a new clue for undersmndmg the mechanism of ENS in skin specimens of collagen disease includmg MCTD.

BEHCET‘S DISEASE: TREATMENT WITH INTERFERON GAMMA Eerherd Fierlbeck. Ottmar Bowhiitr. J&a Wehner-Caroli. Gernot uiversitv of Tuebintren. Behqet’s disease is a chronic systemic inflammatory disease, its cause is unknown. lmmunosuppressanfs may be effective treatments. We describe the therapeutic potentials of recombinant IFN-gamma. We admitted a total of 16 patients with Behcet’s disease in en open Study with IFN-Qemme. In the first two weeks we administered daily 50 100 pg recombinant IFN-gamma subcutaneously, thereafter injections were QiVen three times a week. The study period was fixed et 6 months. After 6 months 13 of the 16 patients were evaluable. In one patient the therapy had to be discontinued after three months because of deterioration of the ocular symptoms. Three patients withdrew because of family affairs. The therapy was well tolerated, only mild influenza-like symptoms occured. Laboratory chemical analysis showed substance caused side effects such es reversible increase of monocytes. serum trislyceride concentration end lactat dehydrogenase activity. Under the long-term treatment the mucocutaneous symptoms became less pronounced or cleared completely. The ocular symptoms did not ameliorate. Some weeks after discontinuation of the treatment exacerbation of the disease appeared. The results suggest that IFN-gamma is a promising remedy for patients with arthritis and mucocutaneous lesions. The mode of action of IFN in Behcet‘s disease remains unknown.

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BORRELIA BURGDORFERI DNA IN MORPHOEA AND LICHEN SCLEROSIS ET ATROPHICUS: DETECTION BY THE POLYMERASE CHAIN REACTION. NJ Levell. KJ Cannl. TA Leslie. ML ~1~1~0 M Dowd. Dermatology Department, Umversity College London Medical School. ‘Medical Microbiology Department. Charing Cross and Wcsrminster Medical School. London. UK. Early studies have indicated serological evidence of Borrelia burgdorfen (BB) infecrion in certain patients wirh morphoea and lichen sclerosis et arrophicus (LS et A). This has been refuted in later sludies which found negative serology. Detection of Borrelia flagella DNA usmg the polymerasc chain reaction (PCR) on skin biopsied from a patient with morphoea following a tick bite has been reported and two reccm studies of patients with LS et A have given conflicting repons of the presence of Borrelia DNA. Four patients (3 male, 1 female; age 35 64) with morPhma and 1 female aged 54 with morphoea and vulva1 LS et A were investigated for BB infection using an enzyme linked immunosorbant assay (ELISA) and western immunoblotting on semm and a nested PCR technique on mine with primers specific for the BB outer surface protein A antigen. The serological tests and PCR were negative in the 4 patients with morphoea. The patiem with morphoa and LS et A bad a positive PCR result and a weakly positive ELISA. Subsequent PCR of biopsied lesional skin from this patienr was also positive. There was no history of tick bites. There was clinical improvement, but not clearance. of the vulval LS et A after 8 weeks treatment wth oral doxycycline but little change in the cutaneous lesions of morphoea. The detection of BB DNA from this pntient indicates thar BB infection may be involved in rhe pathogenesis of a subgroup of pntients wth morphoea and LS et A.

ALOPECIA AREATA SERUM INHIBITS THE GROWTH OF NORMAL DERMAL PAPILLA CELLS N.S. Calver. S M Parkm, 1s Macdonald Hull. 1W J Cunliffe and V.A Randall. Departments of Biomedical Saences. Umvers~tv of Bradford, Bradford, BD7 1DP. UK and ‘Denatolow. The General Infwmarv. Leeds. LSl 3EX. UK The aetiology of Alopecia Areeta (AA) is unknown. although autoantibodies to hair follicle components have been found Since the dermal papilla is believed to regulate the hair folhcle, d would be a prime site for any fectors inhibdmg hair growth. Therefore, we have investigated the effect of serum from AA patients on cultured normal dermal papilla cells to detenme whether it contams any cytotoxlc factors Primary denal paplIla lines (n=ti) were dewed from normal scalp and their ~rotih invesbgeted ln the presence of pooled normal end individual samples of AA serum. Cell growth was measured after 4 days m 96 v&l mlcmtitre plates by the S(4,6D~methylthiszol-2-yl>2,5diphenyltetrezokum bromide (Mm) conversion assay and “,e resutts checked by haemocytometry There was a SiQniflCent increase I” Q& of dermel papilla cells r” the prese”a, of foetal bovine serum or normal human ~81~rn validetmg the essey Pooled AA serum mhibiied growth significentty by 67%; alopecia tot&is and alopeaa universalls ser”m, when compared to “ormel ser”m, had no siQ”,fica”t effect These results suggest Met AA patient serum contams factor(s) which are cytotoxic to dermal pap,lla cells. These need further investiQet,o” to identity the spemic components tilch may well be mvolved in me pethology of AA

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HERPES SIMPLEX “,R”S DETECTION BY FOLYMERASE CHAIN REACTION IN INTESTINAL ULCER OF PATIENTS WITH BEHCET’S DISEASE. Eu&%

AUTOANTIBODIES IN ALOPECIA AREATA. VA Rendall, N.S. Celver. Is_ Macdanald Hull. S.M. Paddn end lW.J. Cunhffe Departments of Biomedical Sciences. Unwersity of Bradford. Bradford, UK and ‘Dermatology, The General Infirmary. Leeds, UK. The aetiolow of Alooecia Areete (AA) is unknown, although it is generally regarded es i; euto immune disease. Thor study was designed to determine whether patients produce autoanbbodles to components of the elOpeCiC scalp or normal heir follicle Longitudinal fmzen scalp sections from active edQes of alopecic IeSiOnS and occlpito-parietel region of normal controls were incubated with serum and/or anti-Human immunoglobulin fluomchmme conjugate. No fluorescence. except that of the vaswlature. wes identified in normal scalp sections wth normal serum. Alooecic scelo sections from IQ oatients showed increeoed fluorescence from th; outer mbi ð in 76.9% df ceses; rubQmuPa showed fluorescence to other rwlp keretinocyte structures. Fluorescence we6 also detected from mcubation with me fluorochrome conjugete clone in 67.6%. Normal scalp SeCtiOns incubated with alopecic serum showed similar immunofluorescance patterns. to patient sections. These results demonstrate the presence of autoantibodies to normal heir follicular components, particulerly the outer root sheath. in AA patients. They imply that AA autoentibodies react with normal structures and not abnormalities in alopecic follides. The fluorescence obtained with the WnjuQete alone indicates that sutoantibodies ere deposited wimin the lesion prior to biopsy. Although these autoantibodies may be involved in the aetiolo~y of AA, they could be produced es the result of damage to effected tissues.

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University, Suwon, and Yonsei University College of Medicine, Seoul, Korea Behcet’s disease has been described as a chronic, recurrent. multisystemic inflammatory disease in which gastrointestinal involvement is occasionally seen. ‘Ihe etiopathogenesis of int&inal manifestation is not clear, but we have previously reported a Possible etiopathogenic role of herpes simplex virus(HSV) DNA in the formation of oral ulceration in Behqet’s disease. However, the clinical and morphologic characteristics of intestinal-BehGet’s disease suggest a correlation with inflammatory bowel disease, making differential diagnosis very difficult. To demonstrate the possible role of HSV DNA m the pathogen=& of intestin&Beh~et’s disease as well as Crohn’s disease, we applied polymerase chain reaction (F’CR) to detect HSV DNA from the lesions of both disease. We obtained seven paraffin-embedded tissue blocks from seven patients with intestinal ulcer who were diagnosed as Behqet’s disease and thirteen blocks from patients with Crohn’s disease. All specimens from patients with Behqet’s disease showed a positive reaction to PCR, in contrast to only two positive reactions out of thirteen specimens from patients with Crohn’s disease It can be deduced that HSV could be considered one of the etiopathogenic factors in gastrointestinal lesions in Behcet’s disease, and although differential diagnosis between Behqet’s disease and Crohn’s dlsease is difficult, detection of HSV DNA could contribute to the solution of the problem.