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152 UVB-induced disruption of the permeability barrier in murine epidermis dependent upon T-cell mediated epidermal hyperplasia
JSID Abslracts
207
154
151 EFFECT
OF WB-NDUDED
METALLOlliK)NEN
QENE
CMOKHES E)BFtESSK)N.
K. Henada 1 Harhimoto. Department School ot Medicine, HimsakiJapan. Previously prevent
l-l. Nw’umi,
of Dsnatobgy,
Cd** and 1.25-dkhydroxyvitrmin
sunburn
kuratinocyte of induced
lN HAT KEFtAllNOCYlES
cell (SBC) inducttin
by WB
D. Savanura. Hba&d
University
D, wars successfully
in mouse
ski
used to
in viva md promote
survival in vitro after UVB sxposuro. indicating a protective atfact cslktM tnatdbthkmah (MT) with the propetty of ;ntioxidmt
mtivky. The aim of this study is to &firm Cvtokines includthg b-l, l-6. L-l 0, which exposure.
uxpmsabn. tucifemaa trslsfscted
ON
To s-e
MT gene expression by sevarcll se induclbta in the cultured calls
the effect of these
cytokines
on MT gene
wa uasd fetal rat kerrdinocytes (DRL1213) ad performed assay for detemhtiin of fClotuscent intensity in the c&s with kmifar%le reporter v~ctora (pEEM”-lot) conming MT
pmmoter. Although tmetumant with L-B sliihtty increased MT-spacifii fluorosconca, other cytddnes did not show t any signMcant expression. These msulta indicate tha WBinsdiated defenca mechanism against sundamagathrough
L-S n!ay conttibute to the MTexprassion in the skin.
HISTAMINE AUGMENTS UVB-INDUCED
IL-6 PRODUCIION BY HUMKEIQTINOCYTES. S.m ydmdmuw. Department of Dermatology, Hiroshima University School of Medicine, Hiroshima, Japan. Histamine is one of the major mediators implicated in the physiopathology of allergy. In addition to its well known immediate inflammatory properties, it has recently been shown that histamine mediates its effects by participating in a multicellular cytokinr easeade. To further understand the role of histamine in skin inflammation, w investigated whether histamine regulates the production of IL-6 by human kcratinocytes. UVB-irradiated human keratinocytes uere cultured in the absence or presence of histamine (1O’M to lOaM), and IL-6 in the supematants wds measured with an ELISA method, Histamine enhanced UVB-induced synthesis of IL-6 in a dose-dependent manner. and maximum effect was ohserved at IO”M of histamine. Hisramihe alone did not induce synthesis of IL-h. The kinetk? of IL-6 production wds not affected hy histamine. The effecl of histamine \u11sinhibited by Hl and Ha histamine receptor antagonists, and HI and H2 receptor mRNA in human keratinocytes were detected hy RT-PCR technique. These data suggest that histamine interacts with its receptors on keratinocytes and enhances UVB. induced IL-6 production.
152
155
UVB-INDUCED DISRUPTION OF THE FERMEABILITY BARRIER IN MURINE EPIDERMIS DEPENDENT UPON T-CELL MEDIATED EF’IDERMAL HYPERPLASIA. A. Hemtnkei,Y. Uchidsi~, F’M Elies~,and WM. Holler&. ‘Cosmetics Lab.. Kanebo Ltd., Odswnta, Japanand Wept. of Damatdogy, Univ. of Cal% S&d of Medicine & Dermstdogy Service, De@ of VA Medical Ceck~, SW Francisco,CA, USA Since, our previous studtesdemonstmmdboth a proliferative and T-cell mediated response. contribute to UVB-induced permeability barrier disrtptioa. we investigated the relationship betweenthesetwo responsesin this model. We fitst computedthe epidermel prohferntive tesponseto UVB (0.15 J/cm9 in ncrmnl hairless(hrihr) vs. athymic (BALB nude,athy) mice. incorporation of PHI-tbymidine into DNA was increased 420-570% in UVB-treated vs. tmtrentedhr/hr epidermis,an increasswhich ~xecededthe UVB-induced increase in TEWL. In contrast. both the hypeqroliferstive response to WB and subsequentbarrier rdteretion were significantly dimimhed in sthy~compmedwith hr/hr epidermii (i.e., incmasedonly 157% & ltKl%, respecpv,ely, over untreated athy controls), while histology confirmed dtmtntshcd byperplusia in athy vs. hrlhr epidermts. Tmmpiatttstionof FHA-nctivatedimmunecells isdsted from h&r cr normal BAIB mice. into sthy after UVR, mstcsedthe sensitivity of ethy e+ermis to UVB-induced barrier disruption. These data indicate that T cells are a significant factor in acute WE-induced disruption of murine epidermel permeability barrier function, possibly through stimulation of epidermrd bypetpMsia
INIiIBITION OF SUBSTANCE P-INDUCED HISTAMINE RELEASE PROM RAT PERITONEAL MAST CELLS BY g-MBTHOXYPSORALBN PLUS LONG-WAVE ULTRAVIOLET IRRADIATION IN PARALLEL r’ITH&HB SUPPRESSION OF INTRACELLULAR CALCIUM ION wwM. y Ikpsnmml of Dermatology. Gunma University School of Medicine. Maebashi. Japan. Recent advances of science lead us to obtain cultured human mast cells fmm umbilical cord blood cells. However, it is still impossible to develop only mature MCK cells which predominate in the human skin. So. at preseut. MI& cells would correspond closely to the rodent connective tissue mast cells which can be activated by non-speciftc stimuli. In this study, rat peritoneal mast cells
were purified on a Percotlgradientandwe investigatedthe effect of 15 @ml 8-metboxypsoralen(&MOP) plus 0.25-3J/cm*long-waveultraviolet (UVA) irradiation (PUVA) on histaminereleasefrom tbe cells which were challenged with substanceP (SP).SPat a concentrationof 1OsMcauseda significant
histamine release and the amount of histamine nlease reached maximum at 1 mitt after the ehallcnge. g-MOP or UVA alone, witbin the dose range, was not cytotoxic to the cells. respectively.becauseit inducedno histaminerelease from the cells. No remarkable inhibition in SP-induced histamine release was
observedin the cells treatedwith S-MOPor UVA irradiationonly. On dte otherhand,SP-activatedmastcells showeda signiticantlyreducedcapacityfor releasinghistamine following PUVA in a dose-dependent manner of UVA irradiation in prallel with the inhibition of intracellularcalciumion levels, suggestingPUVA inhibits mastcell degranulationthroughthe regulationof
calcium
movement
in the cells.
153
156
UV-INDUCED IL-10 PRODUCTION BY KERATINOCYTES RESULTS FROM DNA DAYAOE. -apt. of Dumatc4ogy. Kyoto Univ. Qr&iat8 school of M8d Kyoto, Japan. IL-10 Is probca by kw8Snocyto8 ln mponre to UV Irradlrtlan 8nd plrys a crucl8l role ln suppres8lon 01 DTH re8pcn88r The ~rpo80 of thl8 8tutty IS to test UN hypothdr
LIGHT TRANSMISSION AND SCATERING CE EF’IDERMAL SHEETS OF HAIRLESS MOUSE. T. Kondoh, Y. Takema. Biological Science Laboratory, Kao Corporation, Tochigi, Japan
thal DNA danu(te stlmulatee krrrUnooyMs to pro&es oytoklnes Utrt mcdlfy uitlcnt stsps tn the Immtmoto5tc pethwuy. me ~toroh etnplcy~ ttwolv88 14 mdcm1~I8888 VW4NSJ,which
rep8lr
pyrlmldln8
dlm8,
enc8p8ul8tod
ln llpo8onw.
Dlmer
r8mov8l by T4N5 w88 conflrmod by NoAb rg8lnrl tymlna dlmorr and by ondonuo*nse setutttvo mttan rrsny. Culture supernrtent
from UFlrr8dlrtW ker8tlnocyt8r orhlbltod 8 50 Y reduction lhelr DTH rr8ponso to rllo8ntlgn. Tr88bnont 01 UV-lrr8Ul8t8tl cells with T4N5 llpo8om~ 8brOg8Wd the wpprnslve l ptlYlty of the aup8rnM8nt, whorrrr control llpo8onw had no effect. The reducllon ol IL-10 ln cultum aupwn8t8nt8 from UV-lrrtilated calls tre8ted with T4N5 Ilposomes but not control Ilposomar was conllrmad by EUSA and thrt Of IL-10 In lBa k8rlthIoCyteS w8S 8tw
8hCWt
bIWtIIr)oh18lCbmdWsy.
w8
Cmdw
m8t
uv
induced DNA d8mapr in kwatlnocylr tr@ers proauction 01 IL10 w 18 8n Immune-modu*tory Dctor of UV-lnduwd Immune 8upprraslon. (ml8 work h88 hem UOM 81 m8 Unhr. Ot Tex88, M.D. Anderson C8nc8r Cmntrr; Chllrman : Dr. M.IKrpke).
We investigated optical properties of epidermis in vitro, which related to skin appearance and light proaction. Epiclermal sheets obtained from midback of albino hairless mice(ICWHR) were mounted on a quarts glass. Their integral transmission were spectrophotometrically scatted at the wave length between 250700nm. A value named straight rate(Rs), which indicates scattering ability, was evaluated from integral transmission and axis angle limmited oansmission(tan B ~0.1). Tbe integral transmission and Rs of epi&rmaJ sheets on saline were 65% and 12% at 50Oum. Shorter wave kngth decreased Rs. The pH or components of buffer influenced Rs. For example, 10% lactic acid incxaased Rs to 2-3
titncs. Wbm free epidermal sheets were moistend on warn, their inkgral transmission and Rs increased as reported previously. However, when wet sheets were dryed on supporter, their values increased, which seemed likely to reflect in viva much more. This altcmarive result might be caused by absence of air surface on the SUppOrter
Side.
207
154
151 EFFECT
OF WB-NDUDED
METALLOlliK)NEN
QENE
CMOKHES E)BFtESSK)N.
K. Henada 1 Harhimoto. Department School ot Medicine, HimsakiJapan. Previously prevent
l-l. Nw’umi,
of Dsnatobgy,
Cd** and 1.25-dkhydroxyvitrmin
sunburn
kuratinocyte of induced
lN HAT KEFtAllNOCYlES
cell (SBC) inducttin
by WB
D. Savanura. Hba&d
University
D, wars successfully
in mouse
ski
used to
in viva md promote
survival in vitro after UVB sxposuro. indicating a protective atfact cslktM tnatdbthkmah (MT) with the propetty of ;ntioxidmt
mtivky. The aim of this study is to &firm Cvtokines includthg b-l, l-6. L-l 0, which exposure.
uxpmsabn. tucifemaa trslsfscted
ON
To s-e
MT gene expression by sevarcll se induclbta in the cultured calls
the effect of these
cytokines
on MT gene
wa uasd fetal rat kerrdinocytes (DRL1213) ad performed assay for detemhtiin of fClotuscent intensity in the c&s with kmifar%le reporter v~ctora (pEEM”-lot) conming MT
pmmoter. Although tmetumant with L-B sliihtty increased MT-spacifii fluorosconca, other cytddnes did not show t any signMcant expression. These msulta indicate tha WBinsdiated defenca mechanism against sundamagathrough
L-S n!ay conttibute to the MTexprassion in the skin.
HISTAMINE AUGMENTS UVB-INDUCED
IL-6 PRODUCIION BY HUMKEIQTINOCYTES. S.m ydmdmuw. Department of Dermatology, Hiroshima University School of Medicine, Hiroshima, Japan. Histamine is one of the major mediators implicated in the physiopathology of allergy. In addition to its well known immediate inflammatory properties, it has recently been shown that histamine mediates its effects by participating in a multicellular cytokinr easeade. To further understand the role of histamine in skin inflammation, w investigated whether histamine regulates the production of IL-6 by human kcratinocytes. UVB-irradiated human keratinocytes uere cultured in the absence or presence of histamine (1O’M to lOaM), and IL-6 in the supematants wds measured with an ELISA method, Histamine enhanced UVB-induced synthesis of IL-6 in a dose-dependent manner. and maximum effect was ohserved at IO”M of histamine. Hisramihe alone did not induce synthesis of IL-h. The kinetk? of IL-6 production wds not affected hy histamine. The effecl of histamine \u11sinhibited by Hl and Ha histamine receptor antagonists, and HI and H2 receptor mRNA in human keratinocytes were detected hy RT-PCR technique. These data suggest that histamine interacts with its receptors on keratinocytes and enhances UVB. induced IL-6 production.
152
155
UVB-INDUCED DISRUPTION OF THE FERMEABILITY BARRIER IN MURINE EPIDERMIS DEPENDENT UPON T-CELL MEDIATED EF’IDERMAL HYPERPLASIA. A. Hemtnkei,Y. Uchidsi~, F’M Elies~,and WM. Holler&. ‘Cosmetics Lab.. Kanebo Ltd., Odswnta, Japanand Wept. of Damatdogy, Univ. of Cal% S&d of Medicine & Dermstdogy Service, De@ of VA Medical Ceck~, SW Francisco,CA, USA Since, our previous studtesdemonstmmdboth a proliferative and T-cell mediated response. contribute to UVB-induced permeability barrier disrtptioa. we investigated the relationship betweenthesetwo responsesin this model. We fitst computedthe epidermel prohferntive tesponseto UVB (0.15 J/cm9 in ncrmnl hairless(hrihr) vs. athymic (BALB nude,athy) mice. incorporation of PHI-tbymidine into DNA was increased 420-570% in UVB-treated vs. tmtrentedhr/hr epidermis,an increasswhich ~xecededthe UVB-induced increase in TEWL. In contrast. both the hypeqroliferstive response to WB and subsequentbarrier rdteretion were significantly dimimhed in sthy~compmedwith hr/hr epidermii (i.e., incmasedonly 157% & ltKl%, respecpv,ely, over untreated athy controls), while histology confirmed dtmtntshcd byperplusia in athy vs. hrlhr epidermts. Tmmpiatttstionof FHA-nctivatedimmunecells isdsted from h&r cr normal BAIB mice. into sthy after UVR, mstcsedthe sensitivity of ethy e+ermis to UVB-induced barrier disruption. These data indicate that T cells are a significant factor in acute WE-induced disruption of murine epidermel permeability barrier function, possibly through stimulation of epidermrd bypetpMsia
INIiIBITION OF SUBSTANCE P-INDUCED HISTAMINE RELEASE PROM RAT PERITONEAL MAST CELLS BY g-MBTHOXYPSORALBN PLUS LONG-WAVE ULTRAVIOLET IRRADIATION IN PARALLEL r’ITH&HB SUPPRESSION OF INTRACELLULAR CALCIUM ION wwM. y Ikpsnmml of Dermatology. Gunma University School of Medicine. Maebashi. Japan. Recent advances of science lead us to obtain cultured human mast cells fmm umbilical cord blood cells. However, it is still impossible to develop only mature MCK cells which predominate in the human skin. So. at preseut. MI& cells would correspond closely to the rodent connective tissue mast cells which can be activated by non-speciftc stimuli. In this study, rat peritoneal mast cells
were purified on a Percotlgradientandwe investigatedthe effect of 15 @ml 8-metboxypsoralen(&MOP) plus 0.25-3J/cm*long-waveultraviolet (UVA) irradiation (PUVA) on histaminereleasefrom tbe cells which were challenged with substanceP (SP).SPat a concentrationof 1OsMcauseda significant
histamine release and the amount of histamine nlease reached maximum at 1 mitt after the ehallcnge. g-MOP or UVA alone, witbin the dose range, was not cytotoxic to the cells. respectively.becauseit inducedno histaminerelease from the cells. No remarkable inhibition in SP-induced histamine release was
observedin the cells treatedwith S-MOPor UVA irradiationonly. On dte otherhand,SP-activatedmastcells showeda signiticantlyreducedcapacityfor releasinghistamine following PUVA in a dose-dependent manner of UVA irradiation in prallel with the inhibition of intracellularcalciumion levels, suggestingPUVA inhibits mastcell degranulationthroughthe regulationof
calcium
movement
in the cells.
153
156
UV-INDUCED IL-10 PRODUCTION BY KERATINOCYTES RESULTS FROM DNA DAYAOE. -apt. of Dumatc4ogy. Kyoto Univ. Qr&iat8 school of M8d Kyoto, Japan. IL-10 Is probca by kw8Snocyto8 ln mponre to UV Irradlrtlan 8nd plrys a crucl8l role ln suppres8lon 01 DTH re8pcn88r The ~rpo80 of thl8 8tutty IS to test UN hypothdr
LIGHT TRANSMISSION AND SCATERING CE EF’IDERMAL SHEETS OF HAIRLESS MOUSE. T. Kondoh, Y. Takema. Biological Science Laboratory, Kao Corporation, Tochigi, Japan
thal DNA danu(te stlmulatee krrrUnooyMs to pro&es oytoklnes Utrt mcdlfy uitlcnt stsps tn the Immtmoto5tc pethwuy. me ~toroh etnplcy~ ttwolv88 14 mdcm1~I8888 VW4NSJ,which
rep8lr
pyrlmldln8
dlm8,
enc8p8ul8tod
ln llpo8onw.
Dlmer
r8mov8l by T4N5 w88 conflrmod by NoAb rg8lnrl tymlna dlmorr and by ondonuo*nse setutttvo mttan rrsny. Culture supernrtent
from UFlrr8dlrtW ker8tlnocyt8r orhlbltod 8 50 Y reduction lhelr DTH rr8ponso to rllo8ntlgn. Tr88bnont 01 UV-lrr8Ul8t8tl cells with T4N5 llpo8om~ 8brOg8Wd the wpprnslve l ptlYlty of the aup8rnM8nt, whorrrr control llpo8onw had no effect. The reducllon ol IL-10 ln cultum aupwn8t8nt8 from UV-lrrtilated calls tre8ted with T4N5 Ilposomes but not control Ilposomar was conllrmad by EUSA and thrt Of IL-10 In lBa k8rlthIoCyteS w8S 8tw
8hCWt
bIWtIIr)oh18lCbmdWsy.
w8
Cmdw
m8t
uv
induced DNA d8mapr in kwatlnocylr tr@ers proauction 01 IL10 w 18 8n Immune-modu*tory Dctor of UV-lnduwd Immune 8upprraslon. (ml8 work h88 hem UOM 81 m8 Unhr. Ot Tex88, M.D. Anderson C8nc8r Cmntrr; Chllrman : Dr. M.IKrpke).
We investigated optical properties of epidermis in vitro, which related to skin appearance and light proaction. Epiclermal sheets obtained from midback of albino hairless mice(ICWHR) were mounted on a quarts glass. Their integral transmission were spectrophotometrically scatted at the wave length between 250700nm. A value named straight rate(Rs), which indicates scattering ability, was evaluated from integral transmission and axis angle limmited oansmission(tan B ~0.1). Tbe integral transmission and Rs of epi&rmaJ sheets on saline were 65% and 12% at 50Oum. Shorter wave kngth decreased Rs. The pH or components of buffer influenced Rs. For example, 10% lactic acid incxaased Rs to 2-3
titncs. Wbm free epidermal sheets were moistend on warn, their inkgral transmission and Rs increased as reported previously. However, when wet sheets were dryed on supporter, their values increased, which seemed likely to reflect in viva much more. This altcmarive result might be caused by absence of air surface on the SUppOrter
Side.