Antitumor effects on mouse melanoma elicited by local secretion of interleukin-12 and their enhancement by treatment with interleukin-18

ESDR I JSID I SID Abstracts

1072

1069 ANTITUMOR

EFFECTS

SECRETION

OF INTERLEUKIN-12

TREATMENT

MELANOMA AND THEIR

WITH INTERLEUKIN-16.

&X&&a&u

Ichihab$,

Kobe University To investigate

the mechanism

IL-12.

(BIMLIZ)

analysis

Only at the periphery

marked retardation

the antitumor

of not

effect of locally scnctcd

on day6 showed

that NK cells and of B16/IL12

tumors

CD4+T cells and CDX+T cells accumulated

of bath transfectcd (IL18)

of tumor growth

mice, dcplction

at the center and periphery

whereas

(IL-

markedly

and untransfected

tumors.

Systemic

inhibited the growth of B16jIL12,

it did not influence the tumor growth of parental 816 cells in VIVO. these

nevertheless

IeSUkS Suggest

mediated

Urology?,

cells. IL-12 gcnc-

mice. In syngencic

of specimens

more densely

than that of parental 816 tumors, SpZScly

HOrikawaL&

and Depaament

Blh melanoma

showed

in syngcneic

lmmunohistochemical

txeatment with interleukin-18

BY

Tatsuva

of antitumor effect of locally sccrctcd interleukin-12

subcutaneously

accumulated

BY LOCAL

ENHANCEMENT Nag&.

of Dermatology’

only NK cells but also CDS+T cells diminished

maaophages

ELlClTED

Kobe, Japan.

the IL-12 gene into mow

816 melanoma

when implanted

fliroshi

Depahnent

School of Medicine,

I?), wc introduced transfcztcd

ON MOUSE

thatlocal IL-12

primarily

scaetion

can retard the gowth

by NK cells and presumably

effect is augmented

by systemic

administration

of BI6 melanoma

by C!DS+T cells, and ulat its antitumor of a novel cytokinc,

IL-18.

1073

1070 PLATELET-ACTIVATING MELANOMA

F4CTOR

RECEPTOR

IS IzXPRESSED

CELLS AND PLATELET-ACTIVATING

ON HUMAN

FACTOR ENHANCED

MELANOMA CELL GROWTH Takenori Takahashil. Katsunon Moo’. Iwao Ando’, ~tsusbt Kulotal, Koxhxo Nakmnum~. Savun Sat$. Kazuhlko Kumc3. Takao Stum& pnd Kun,h,ko

Tamah*,

Teikyo Umversity, of Bwhemlstry,

‘Department

Kanagawa.

The University

Platelet-activating

of Dermatology,

The Mumnokuchl

Japan, and %epartment

of Dermatology

Hospital 01

and 3Depxtment

of Tokyo, Tokyo. Japan

factor (PAF) receptor transgemc

nuce spootancously

bore melanocyuc

tumors (Ishli et al., EMBO J. 1997). Thus, we first studled the PAF receptor (PAFR) Next, we enprrssmn by RT-PCR. All aght melanoma cell hues showed DAFR expressmn dctcrmmcd wtbout

the effect of PAF on melanoma cell

Melanoma

PAF (C16) and tested for then pmhferauon

cells were mcubated ~5th or

or apoptosls by [JHI-thymidme

uptake assay. MTT [3-(4.5-d~methyl-2-thiazolyl~-2,5-d~phenyl tetrazolium brouudel The PAF-putwed colorunetnc bmassay, or apoptoais detection based on FACS analysts melanoma cells showed the higher probfrrauon activity compared with the control melanoma

cells, whereas no significant

normal kemtmcytes, Induced an mcreased phase

melaoacytes propatmn

Both the proliferation

mhiblled

by a P.4F antagomst

trypsm,zed PAF:P/%ER

1071

A SHIFT IN CUTANEOUS CYTOKINE MILIEU TOWARD A TH2 RESPONSE WITH CHRONICITY IS A UNIVERSALLY APPLIED PARADIGM INDEPENDENT OF GENETIC BACKGROUNDS. mhiohara. Hid&i Kitaeaki and Fur&&i Nakavama, Department of Dermatology, Kyorin University School of Medicine, Japan. Different mwine genetic backgrounds arc known to profoundly influence the diion of T helper (Th) phenotype development: BALBlc mice arc more Th 2 inclined and Our previous studies indicated that the frequencies of antigen susceptible to Leishmania. administration can also alter the directionof Th phenotypes: rcpc&delicitation of contact hypersensitivity (CH) in BALB/c mice leads to a shift in cutaneous cytoklne milieu from a Th 1 to a Th 2.dominated rcsponsc which is associated with the development of cxlyHowever, it was unknown whether the shift is due to the intrinsic type responses. tendency to develop toward a Th 2 response in BALB/c mice. In this study, we have asked whether a similar shift in the type of immune responses and the patterns of cytokine production could be observed in C57BW6 (B6) mice known to be lowRepeated application of OX in B6 responders in terms of Th Z-dependent parameters. mice also resulted in a site-restricted shift in the time course from DTH to an early-type To investigate responses, except for the absence of immediate-type wheal responses. the strain differences between BALB/c and B6 mice in the kinetics and magnitude of cytokine production, the temporal sequence of cytokine gene expression after each Surprisingly, no elicitation of CH was assessed in the acute vs chronic lesions. fundamental differences in their relative balance of Th 1 and Th 2 cytokines were found between BALBlc and B6 mice in either the acute or chronic lesions, although the magnitude of each cytokine production in BALB/c was much greater than that in B6 mice. These results indicate that the relative balance between Th 1 and Th 2 cytokines rather than simply their absolute levels would detetine the type of the immune responses and that a shift toward a Th 2 response by repeated elicitation of CH is a now universally applied paradigm.

melanoma

enhancement was observed m prohferatlon ot By cell cycle analysis. PAF stlmulatmn

or flbroblasls

of S phase and concomitant enhancement (‘I CV309)

decrease

01 GOiGl

and the cell cycle regulation Furthermore.

cells wils delayed by PAF treatmeet

apoptosis

were

of non-adherent

These rcsulfs suggest a

system ts workmp on melanoma cells m then prohfcratmn

THE EFFECT OF THE CHEMOKINE ANTAGONIST MET-RANTES ON HUMAN EOSINOPHILS. Jijm Elmer, Holeer Petetine. Renate H6chstettcr Daniela Kimrma. Timothv N. C. Wells*. Alexander Kaou and Aman& E. I. Proudfoot* Depalrment of Dermatology. Hamover Medical School, Hannover, Germany; *Geneva Biomedical Institute, Glaxo Wellcome R. D., Geneva, Switzerland. Eosinophils are predominant effector cells not only in allergic diseases but also in connective tissue diseases. The recruitment of wsinophils to the side of inflammation and release of reactive oxygen species leading to tissue damage and propagation of the inflammatory response arc mediated by chemokines. Thus, agents that would be able to inhibit or antagonize chemokine-Induced eosinophil activation arc interesting as therapeutical agents. The purpose of this study was to investigate the effect of a chemokine receptor antagonist. Met-RANTES, on its effect on human eosinophil effcctor functions in response to RANTES, MCP-3 and eotaxin. MctRANTES had no intrinsic activip on [Ca”], transients in eosinophils and was able to inhibit dose-dependently [Ca ‘I, transients in eosinophils following stimulation wth RANTES, MCP-3 and eotaxin. In addition Met-RANTES dose-dependently inhibited actin polymerization and release of reactive oxygen species in eosinophils following stimulation wth RANTES, MCP-3 and eotaxin. The results of this study concluded that Met-RANTES is an effective and powerful antagonist of effcctor functions of human eosinophils and is therefore of interest for a new therapeutical approach to prevent the invasion and destructive power of eosinophils in diseases that are accompanied by eosmophil infiltration ach as atopic dermatitis and connective tissue diseases.

and survival iu YWO

1074 CHARACTERIZATION OF THE RECEPTORS FOR TEE CXC CHEMOKINE IL8 ON HUMAN EOSMOPRILS Heme. 0 Gbd. R Hochstctter. D Klmrme. R. Smolank,. A Kacc. J Elsner. Department of Dermatology, Haonaver Mcd,cal School Department of Immunology, Unwcrs,ty of Gottrngeh Germany

, FRG, ’

Chemokmes play an uxpmtant role m anmctmg granuloc~ mto the side of mtlammauon Two subfmmhes ofchcmokmcs differ m tbcr bmlogw actwitv to stmwlatc d&rent km& of dfecmr cells chemokmes such as IL-8 a;ld ENAachvate pmdommandy neutrcphds, CC chcmolunos such as RANTES and eotaxln actnate predommantly eosmophds However, Whereas CXC

c~ntr~~ws,al results have been pubbsbed m the past regarding the funchond response of IL-8 m eosmqlhd actlvatmn. parlhxlarly 111zlll.xglC Lseases In this study, we Investigated the functronal

cvldcnce and exprewon of bath IL-8 receptor typer on human easrnophds To mvcsllgate whether neutrophd contimatmn m&t be responsible for Ihe reported IL-8 et&t on eormophlls, highly puntied human eosmophils were codammatcd wth puntied human neutmpixls from the same donor m tierent concentrahons. IL-8 d,d not mduce B rapId and tnnsm,,t release of cytosobc free Ca” (lC$*],) m eosm@ds that were contmnmated wth 0% of ncutrophils Lnterestmgly, as Me as 5% aCneutropi,d ccmtxmnatlcm WBSenough to mduce a small increase of [Ca”], m msmaphds fo,,owmg snmulahcm wtb IL-8

Fmihmmme, pm-mcubatmn of eosmophds wth TNFa,

IFNN~and IL-4 had no

effect on [Ca”]> tnmsmnts followmg stimulation with K.-b’ RT-PCR expznments revealed that eosmophds, m contrast to ncuhophils, &d not express mRNA for the IL-8 receptors, IL-SRAltL-8RB

In a&bon, flaw cy?omew expenments wth mAb’s agamrt the two IL-8 receptors dcmcnssated no cxpresrmn of IL-IRA and IL-8RB on lug& puntied and cymkmmxcbvatc., eosm~phds, rcspecuvcly In summary, th,s study revealed that IL-BRA and IL-ORB represent CXC chcmokme receptors that are not cxprcssed on human eosmoph~ls we” atic, stimulauon wtb THI and TH2-i,ke cytokmes The CXC-chemokme IL-8, (herefore, does not play an unportant role for attractmg eosmophds to the s,te of mtlammatm,, and actlvatmn m catam chmmc m&mmatory dxeascs