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Regulation of Ia antigen and costimulatory molecule expression on murine Langerhans cells(LC) in vivo and in vitro
JSID Abstracts/J.
Dermatol. Sci. 8 (1994) 54-88
193
87
1%
CLINICAL EVALUATION
OFIGEANTIBODIES
TOTRICHOPHY-
TONR.,PlTYF7OSPORUMO.ANDCANDlDAA.lN
REGULATION EXPRESSION IN VITRO.
THESERA
FROM PATIENTS WITH ATOPIC DERMATITIS
OF IA ANTIGEN AND COSTIMULATORY ON MURINE LANGERHANS CELLS(LC)
ii OZAWA, S AIBA, S NAKAGAWA and H TAGAMI, Dermatology, Tohoku University School of Medicine,
M.ISHIWA, K.KlTAMURA, C.SUGA,Y.YAMAMOTO, H.KOMATSU. A.SUGIYAM4, S.ONUMA, J.OSAWAand Z.IKEZAWA, Dep Damaml,Yokohama City Uni.Urafune Hospital, Japan
IN
MOLECULE VIVO AND
Department of Sendni, Japan
by a CAP-RAST method in the AD saa(n=216). The muln indicated tlm positive rs~e of anti-C IgE was in 57.6%, anti-P in 53.2% and ant-T in 55.7%. Significant come&ion was observed between anti-T and sn&P(r= 0.546) anti-T and anti-C(r-0.701), and anti-P and anti-C(0.623).
After short term culture, LC up-regalate la anttgen and casthmuktory molecules as well as increase antigen presenting cell function. We have demonstrated in viva that murine LC show the phenotypk smd functional activation after hnpten appliiation similar to that noted in in vitro-activated LC. To elucidate the mechanism of the up-regulation of costimulatory molecules on murine LC in vitro and in viva, we examined the effects of monoclonnl antibodies(mAb1 tu!aiast GM-CSF. TNFa and IL-lp on the up-regulation of ICAM: a&i B? by cultured iC, and also examined the effects of intradermsl injection of GM-CSF, TNFa and IL-lb 00 the costhnuktory mokcuk expression by murine LC in viva When we added anti-GM-CSF or anti-TNFa mAb to the culture of epidcrmsl cells, the up-regulation o( 87 on LC was inhibited, while the increased expression of lo antigen remained unchanged. The addition of snti.IL.la mAb to the culture did not inhibit the upregulation of costimulntcry molecules or la antigen. In contrast, when we injected intrsdermaliy IL-lp or TNFa, LC up-regukted ICAM-I and la antigen, but not 87 significantly. The injection of GM-CSF could not up-regulate ICAM-I, 87 or IP antigen.
194
197
In tropic demmitis(AD),
the impottmce of several fungal allagem
tcichophyton has been repotted as envifomental allagem;
except
high pmalmce
of IgE antibodies to candida a(C) and succesful mponce to oral and topical anti-fungal therapy in some cases of realcitmnt AD In thii paper, IgE mtibodiesa@nst tticbophyton r(T), pityrospotvm o(P) and C WBSmeasured
HLA-B13 IS A MELANOGENESIS-RELATED MOLECULE
The amount of filaggtin in the non-lesional
skin of atopic dermatitis
patients is decreased.
M. MUTO, Y. HAMAMOTO,S. MOGAMI, T. SHIMIZU and C. ASAGAMI, Departmentof Dermatology,Yamaguchi UniversitySchool of Medicine, Ube, Japan It has not been clear whether HLA antigens are associated with melanoaenesis. In order to elucidate involvementof HLA aenes relevant to melanin formation,we performed an HLA populationsurvey, by using peripheralblood lymphocyte obtained from 28 unrelated Jaoanese oatients with melanoma. 10 patients with vitiligo and'120 nohl healthy controls.The results showed that there was a strong association between melanoma and HLA-B13 (25.0% (7/28) jn patients vs 2.5% (3/120) in controls, relative risk = 13.0, p < 0.005). Moreover,the frequencyof HLA-613 antigen was moderately increased in vitiliqo (2/10 = 20.0% vs 3/120 = 2.5%, relative risk = 9.8, p < 0.05). These observations lead to an idea activity
that HLA-B13 controlling
gene is related the melanogenesis
with of
tyrosinase the melanocytes.
As the skin of alopic dermatitis patients is usually dry and scaly, such emollient factors as amino ackts and urocanic acid could be decreased in the stratum comaurn of these patients. The pracursor protein of these emolliinl factors is filaggrin tiich is synthesized in the stratum granulosum. The purpose of this study is to determine the amount of filaggrin synthesized in the non-lesional skin of atopic dermatitis patients. We have determined the amount of filaggrin by two mathods; the one is immunohislochemical method and the another is fmmuno-biochemical method: the biopsiad spedmen with a 3 mm punch was lyophilirsd. extracted. subjwtad lo SDS-PAGE and immuno blotted. The density of a potein band was measured and the net amount of protein from the densitomaterical number was determined according lo the previously-determined standard curve. [Ftesufts] lmmunohi&c&amimlly, the amount of fflaggrin was remarkably decreased in lhs non-tasiinal skin of atopic dermalftis. Its net anwunt in 3 mm non-lesional skin of the atopic dermaliiis patients was around 3.1 ug. but in contrast, around 6.5ug in the normal control. [Conclusion] Fibggrin synthesis was decreased in the non-lesional skin of alopic dsmlat&palisnts.
198
195 INCREASED EXPRESSION ON IN VIVO-ACTIVATED S. AIBA, Il. Dermatology,
Saguchi T, Kusuda S. Takahashi M, Tezuka T Department of Dermatology. Kinki University School of Medicine Osaka, Japan
OZAWA, Toheku
OF CO-STIMULATORY LANGERHANS CELLS
RANT&S
MOLECULES (LC)
S. NAKAGAWA, II. TAGAMI, Department Univ. School of Medicine, Send& Japan.
of
After ha~kn ~~ellutkn. LC Increase Is sotken er~rearlon and enhance -*allge;‘prese.ti~g hmctkln In viva &nikrto tbosc shown is in vitm cuitwed LC. Recently, it km become clear that cortlmuktory wkeuks such as [CAM-l and B7 piay a critical role In nntigea presentation. In this study, we examined the expression of theme eortiatitory mokcrks on In viva-activated LC. We painted various ehemicak 01 the tars of C3H mke. At vartous time periods after cheniai trc&ment, the expression of co-stimuktory mokcuks on LC “” examined bv fior cvtometry. Fresbiv-is&ted LC excrcss [CAM-l, but do not cr&ss B7: Twelv; hrs aft& Lapten pplk&ion, ail the LC up-regnbted ICAM-I and B7. From 411 hn after bapte” nppik~tioa, LC decreased tbelr ICAMand B7 expression. Seventy 1-n hrs afkr hapten qpiiatlon, the expnssi~n of costlauktory aoiceuin become riaihr to that on aan-treated LC. Primary Irrlta&, 10% SLS, tokrqen, 2% DNTB in acetene or snlvent could not up-regnkte Ia antigen or co-stimuktory mokcnks. These data denmmatrate that rpidermai LC np-regukte coltimnktory mokcuks in add&km to la antigen for a ilmlted time period sfler haptea appikation. By the expression of costhnuiatnry mokcuks, epidermal LC “my regukte aiicrgk contact hypcrsensitivlty.
l
EXPRESSED
IN THE
SKIN
OF PATIENTS
WITH
AD
H.YAMADA, R. IZUTANI, J. CHIHARA, T. YUDATE. S. MATUKURA, T. TEZUKA (Dermatology, Surgery, Internal medicine, KINKI Univ. Osaka) RANTES I8 a potent chemcattractanl for various important inflammatory colla much aa ocalncphilr aa well aa mamcry T calls and mcnccytaa, pctontfally recruiting theaa collr from iba circulatlcn to an Inflamed fccua. To oxtent cur undaratanding of the partlclpaticn of ecalncphlla and T calla In ralatlcn. the pcaalbla Invclvamant of RANTES waa axamlned In tha akln of patlanta wlth AD. Wa performed RTPCR aaaay for slmultanecua drtactlcn of mRNAS for RANTES to clarify whether RANTES la praaanca In AD skin. Wa examlnad exoraaalcn of RANTES In the akln. obtained at blcoav. fr6m 10 patlantr with AD and 5 normal akin of cparatldn.‘~ RANTES prlmor was uaad of approprlata doalgn with ampilfy a 243 bp fragmant. Standard prlmara for beta-actln wara uood to aaaaaa the IntaQrlty of mRNA cbtalnad from the aamplaa. RASNTES mRNA waa dotactad In 0 of 10 AD 6kln ti6we aamploa. By contra& normal akln tlrauo aarplo waa noQatlve for RANTES. Our flndlnQa auggaat that RANTES playa a part In T cell and ecalncphlla In AD.
Dermatol. Sci. 8 (1994) 54-88
193
87
1%
CLINICAL EVALUATION
OFIGEANTIBODIES
TOTRICHOPHY-
TONR.,PlTYF7OSPORUMO.ANDCANDlDAA.lN
REGULATION EXPRESSION IN VITRO.
THESERA
FROM PATIENTS WITH ATOPIC DERMATITIS
OF IA ANTIGEN AND COSTIMULATORY ON MURINE LANGERHANS CELLS(LC)
ii OZAWA, S AIBA, S NAKAGAWA and H TAGAMI, Dermatology, Tohoku University School of Medicine,
M.ISHIWA, K.KlTAMURA, C.SUGA,Y.YAMAMOTO, H.KOMATSU. A.SUGIYAM4, S.ONUMA, J.OSAWAand Z.IKEZAWA, Dep Damaml,Yokohama City Uni.Urafune Hospital, Japan
IN
MOLECULE VIVO AND
Department of Sendni, Japan
by a CAP-RAST method in the AD saa(n=216). The muln indicated tlm positive rs~e of anti-C IgE was in 57.6%, anti-P in 53.2% and ant-T in 55.7%. Significant come&ion was observed between anti-T and sn&P(r= 0.546) anti-T and anti-C(r-0.701), and anti-P and anti-C(0.623).
After short term culture, LC up-regalate la anttgen and casthmuktory molecules as well as increase antigen presenting cell function. We have demonstrated in viva that murine LC show the phenotypk smd functional activation after hnpten appliiation similar to that noted in in vitro-activated LC. To elucidate the mechanism of the up-regulation of costimulatory molecules on murine LC in vitro and in viva, we examined the effects of monoclonnl antibodies(mAb1 tu!aiast GM-CSF. TNFa and IL-lp on the up-regulation of ICAM: a&i B? by cultured iC, and also examined the effects of intradermsl injection of GM-CSF, TNFa and IL-lb 00 the costhnuktory mokcuk expression by murine LC in viva When we added anti-GM-CSF or anti-TNFa mAb to the culture of epidcrmsl cells, the up-regulation o( 87 on LC was inhibited, while the increased expression of lo antigen remained unchanged. The addition of snti.IL.la mAb to the culture did not inhibit the upregulation of costimulntcry molecules or la antigen. In contrast, when we injected intrsdermaliy IL-lp or TNFa, LC up-regukted ICAM-I and la antigen, but not 87 significantly. The injection of GM-CSF could not up-regulate ICAM-I, 87 or IP antigen.
194
197
In tropic demmitis(AD),
the impottmce of several fungal allagem
tcichophyton has been repotted as envifomental allagem;
except
high pmalmce
of IgE antibodies to candida a(C) and succesful mponce to oral and topical anti-fungal therapy in some cases of realcitmnt AD In thii paper, IgE mtibodiesa@nst tticbophyton r(T), pityrospotvm o(P) and C WBSmeasured
HLA-B13 IS A MELANOGENESIS-RELATED MOLECULE
The amount of filaggtin in the non-lesional
skin of atopic dermatitis
patients is decreased.
M. MUTO, Y. HAMAMOTO,S. MOGAMI, T. SHIMIZU and C. ASAGAMI, Departmentof Dermatology,Yamaguchi UniversitySchool of Medicine, Ube, Japan It has not been clear whether HLA antigens are associated with melanoaenesis. In order to elucidate involvementof HLA aenes relevant to melanin formation,we performed an HLA populationsurvey, by using peripheralblood lymphocyte obtained from 28 unrelated Jaoanese oatients with melanoma. 10 patients with vitiligo and'120 nohl healthy controls.The results showed that there was a strong association between melanoma and HLA-B13 (25.0% (7/28) jn patients vs 2.5% (3/120) in controls, relative risk = 13.0, p < 0.005). Moreover,the frequencyof HLA-613 antigen was moderately increased in vitiliqo (2/10 = 20.0% vs 3/120 = 2.5%, relative risk = 9.8, p < 0.05). These observations lead to an idea activity
that HLA-B13 controlling
gene is related the melanogenesis
with of
tyrosinase the melanocytes.
As the skin of alopic dermatitis patients is usually dry and scaly, such emollient factors as amino ackts and urocanic acid could be decreased in the stratum comaurn of these patients. The pracursor protein of these emolliinl factors is filaggrin tiich is synthesized in the stratum granulosum. The purpose of this study is to determine the amount of filaggrin synthesized in the non-lesional skin of atopic dermatitis patients. We have determined the amount of filaggrin by two mathods; the one is immunohislochemical method and the another is fmmuno-biochemical method: the biopsiad spedmen with a 3 mm punch was lyophilirsd. extracted. subjwtad lo SDS-PAGE and immuno blotted. The density of a potein band was measured and the net amount of protein from the densitomaterical number was determined according lo the previously-determined standard curve. [Ftesufts] lmmunohi&c&amimlly, the amount of fflaggrin was remarkably decreased in lhs non-tasiinal skin of atopic dermalftis. Its net anwunt in 3 mm non-lesional skin of the atopic dermaliiis patients was around 3.1 ug. but in contrast, around 6.5ug in the normal control. [Conclusion] Fibggrin synthesis was decreased in the non-lesional skin of alopic dsmlat&palisnts.
198
195 INCREASED EXPRESSION ON IN VIVO-ACTIVATED S. AIBA, Il. Dermatology,
Saguchi T, Kusuda S. Takahashi M, Tezuka T Department of Dermatology. Kinki University School of Medicine Osaka, Japan
OZAWA, Toheku
OF CO-STIMULATORY LANGERHANS CELLS
RANT&S
MOLECULES (LC)
S. NAKAGAWA, II. TAGAMI, Department Univ. School of Medicine, Send& Japan.
of
After ha~kn ~~ellutkn. LC Increase Is sotken er~rearlon and enhance -*allge;‘prese.ti~g hmctkln In viva &nikrto tbosc shown is in vitm cuitwed LC. Recently, it km become clear that cortlmuktory wkeuks such as [CAM-l and B7 piay a critical role In nntigea presentation. In this study, we examined the expression of theme eortiatitory mokcrks on In viva-activated LC. We painted various ehemicak 01 the tars of C3H mke. At vartous time periods after cheniai trc&ment, the expression of co-stimuktory mokcuks on LC “” examined bv fior cvtometry. Fresbiv-is&ted LC excrcss [CAM-l, but do not cr&ss B7: Twelv; hrs aft& Lapten pplk&ion, ail the LC up-regnbted ICAM-I and B7. From 411 hn after bapte” nppik~tioa, LC decreased tbelr ICAMand B7 expression. Seventy 1-n hrs afkr hapten qpiiatlon, the expnssi~n of costlauktory aoiceuin become riaihr to that on aan-treated LC. Primary Irrlta&, 10% SLS, tokrqen, 2% DNTB in acetene or snlvent could not up-regnkte Ia antigen or co-stimuktory mokcnks. These data denmmatrate that rpidermai LC np-regukte coltimnktory mokcuks in add&km to la antigen for a ilmlted time period sfler haptea appikation. By the expression of costhnuiatnry mokcuks, epidermal LC “my regukte aiicrgk contact hypcrsensitivlty.
l
EXPRESSED
IN THE
SKIN
OF PATIENTS
WITH
AD
H.YAMADA, R. IZUTANI, J. CHIHARA, T. YUDATE. S. MATUKURA, T. TEZUKA (Dermatology, Surgery, Internal medicine, KINKI Univ. Osaka) RANTES I8 a potent chemcattractanl for various important inflammatory colla much aa ocalncphilr aa well aa mamcry T calls and mcnccytaa, pctontfally recruiting theaa collr from iba circulatlcn to an Inflamed fccua. To oxtent cur undaratanding of the partlclpaticn of ecalncphlla and T calla In ralatlcn. the pcaalbla Invclvamant of RANTES waa axamlned In tha akln of patlanta wlth AD. Wa performed RTPCR aaaay for slmultanecua drtactlcn of mRNAS for RANTES to clarify whether RANTES la praaanca In AD skin. Wa examlnad exoraaalcn of RANTES In the akln. obtained at blcoav. fr6m 10 patlantr with AD and 5 normal akin of cparatldn.‘~ RANTES prlmor was uaad of approprlata doalgn with ampilfy a 243 bp fragmant. Standard prlmara for beta-actln wara uood to aaaaaa the IntaQrlty of mRNA cbtalnad from the aamplaa. RASNTES mRNA waa dotactad In 0 of 10 AD 6kln ti6we aamploa. By contra& normal akln tlrauo aarplo waa noQatlve for RANTES. Our flndlnQa auggaat that RANTES playa a part In T cell and ecalncphlla In AD.