IL-8 production and secretion from normal human epidermal keratinocytes

139

179 PROPERTIES OF PHOSPHORYLATED CYSTATIN a AS A CIMKINENT PFGTEIN OF COFUIFIED EXWEWPE

180 DECREASED COLLAGEN I AND III GENE EXPRESSION ASSOCIATED WITH UNALTERED EXPRESSIOiY OF COLLAGEN "I IN BOTH NORMAL AND FIBROBLASTS CULTURED WITH TUMOR SCLERODEMA NECROSIS FACTOR 0. K.TAKEDA, A.HATAMOCHI, N,ARAt+,;AWA. AND H.UEKI, Departmem of Medical School, 577 Matsushima, Dermatology, Kawasaki

M. TAXAH?!SHIAND T. 'I'!ZWA,Department of K!ematology, Kinki Unlverslty School of Medlcme, 375-2 Ohm-Hiqashi, OsakaSayma, Osaka, Japan %z have Isolated a phosphorylated cyst&m a (P-cyst&in a), which is a cysteine proteinase inhibitor and stained with hematoxylin, frcm newlxxn rat epidemns. In viva location of P-cyst&in c(1s not only keratohyalinules but also the cell mmbrane region of the stratum cornem in the indirect immmofluorescent studies. This finding suggests that P-cyst&in a could 12 a component protein of cornified envelope. Polymrization of P-cyst&in a by a reaction of epiderml transglutaminase (T&se) and the existence of P-cyst&in a in cornified envelope were examined. P-cyst&in u was formed to high mlecular wiqht proten in the presence of 'K&se and Ca2+ ions. Cornified envelo:~?,which was prepared by heat and SDS treamnts of newborn rat stratm corneum, reacted positively with antiP-cystatin u antitody by ELISA and immmohistcchemical techniques. These findings indicate that P-cystatin a is one of ccmpzment protans of cormfied envelope.

scleroderma fibroblasts cultured wit-h TNFa. TNFa inhibited gene expression of collagen I and III, while expression of VI was collagen unaltered in scleroderma fibroblasts. Similar results were obtained in normal fibroblasts cultured with TNFO. results indicate that collagen VI expression is These regulated independently from that of collagen I and III in both normal and scleroderma fibroblasts cultured with TNFa.

181

182

IL-8 PRODUCTION AND SECRETION FROM NORMAL HUMAN EPIDERMAL KERATINOCYTES H. TAKEMATSU’ and H. TAGAMl*, ‘Department of Dermatology, Teikyo University Ichihara Hospital, 3426-3 Anesaki, Ichihara, and *Department of Dermatology, Tohoku University School of Medicine, l-l Seiryo-machi, Sendai Human epidermal keratinocytes constitutively produce leukotactic cytokine IL-B. Immunoreactive IL-R has been found in psoriatic epidermis. suggesting a role of IL-8 as a psoriasis-promoting factor. While normal keratinocytes secrete IL-8 in vitro, we see scant infiltration of neutrophils and mononuclear cells into the normal epidermis in viva. In order to address this issue, we compared the amounts of secreted and cellass&ted IL-S. Normal human keratinocytes were cultured in keratinocyte growth medium. Conditioned medium and cells were collected at appropriate time: and the amounts of IL-8 and neutrophil chemotactic activity in the medun and cells were measured. Time course study showed an increase in IL-8 production which persisted to 11 days. However, the IL8 production per cell decreased with time. The amounts of cell-associated IL-8 remained at the same level. Thus IL-8 production and secretion are intimately related at the growth phase of human epidermal keratinocytes.

183

Okayama 701-01, Japan Kurashiki, studies have demonstrated that tumor necrusis Recent macraphage-derived cytokine, selectively factor a (TNF~), decreases production of collagen I and III, and increases production of collagenase in cultured fibroblasts. We that TNFa inhibited collagen synthesis and reported

a

stimulated collagenase activity in cultured fibroblasts from patients with systemic scleroderma at the meeting last year. I" this study, we measured mRNA levels of not only collagen I and III but also collagen VI in both normal and

RECEPTOR POSITIVE MONOCYTESIN ATOPIC DERRATITIS (AD)

PC c 3.

TAKENAKA,

Y. TANAKA and H. YOSitIDh. Department Dermatology. Nagasaki Unlverslty School of Yedlclne. Sakamoto-macho. Nagasaki, 852, Japan

of i-l

rresence Of IgE. IjC mc:ecuics ad FciR u^x the monocytes isolated from the patients with AD, were assayed by flov cytometry. In patients with AD. each ratlo of FccR and IgE-positive monocytes (16.7%?6.5%. 30.52+9.1:) *as slgnlficantly higher than those in non-atopIc donors. Whereas the proportion of IgG posltlve monocytes from .4D patlent was similar to that in non-atoplc donors. Furthermore. IL-1 release from manacytes were ELI%. As the results, It was demonstrated that

assayed

b;;

the amount of IL-I release from monocytes of the AD patients by stimulation of Dermatophagoldes farlnae-speclflc !gE immune complex was IgE and FccR dependent Our results may suggest the important role of IgE’ and Fc cR+ cells. and may explain the significance that many Fc r.R+ and !p;E+ Langerhans cells were InfIltrated in the dermatltlc lesion of

of the fact macrophages

and AD.

184 THE

EFFECT

DERIVED

OF CYTOKINES

CULTURED

ON PROLIFERATION

MAST

T. TANAKA,

K. NAKAMURA.

Department

of

rmnarn-ku,

Hlroshlma.

Mast

cells

cells

!ncrease

from

lnvestwted cell

prohferatlon

bone

marked IL-3

These IL-3

mast

effects

llgand

results c-kit

suggest

at

Kasurm.

usmg

13Hl

cultured

mast

of

BMMC

by

sites In

dewed

of

epldermal because

observed

m&urn. of cells

lessons

with

know on

in

IL-3

the

the

IntO presence

have

was

c-kit a role

an abilmv

to

we mast

mouse

prol!feratlon wth

the

dermis.

in CBA/J

The

may have

mast

incorporatlon

was

they

the

and

the

to

cytokanes

(BMMC)

co-existence

skin

I”

thvmldlne

ceils

that

order

cell

culture

that

stem

known

accumulation

in dermis, Ilgand.

the

epldermal

m the

been

psor!as~s.

cell

of

vitro.

enhanced

accumulation and

as

proliferation

or c-kit

synerglstlcally cell

the in

l-2-3.

hematopoletlc It has

in dermis

such

marrow-dewed

The of

the

S. YAMAMOTO.

Unlvewty,

multlpotentlal

in number of

and

Hlroshlma

in tissues.

dtsorder

mechanisms

-

Japan

dlfferentiate/prollferate epldermal

E. MORITA.

Dermatology.

dewe

OF BONE MARROW

CELLS.

Ilgand. on

mast

produce