Development of recombinant S- typhimurium

Suppl
I - 1998
Development of recombinant S- typhimurium
as a model for S. typhi-based vaccine vectors
Vaccine and Molecular BiologY
t87
VMB.6
S.J. Dunstant, C.P. Simmonsl'2, R.A. Strugnelll'2
Abstrak
membuka kesempatan untuk
Keberhasilan uji serta peredaran vaksin baru berasaL dari mutan S. typhi yang dilemahkan telah
perlu
memperhatikan analisis
ini
semacam
vaksin
yang
Pengembangan
baru.
mengembangkan vaksin-vaksin bivalen atau multivalen
bahwa rekombinan s'
menjamin
prometer
dsb)
untuk
gen,
(closis
kekuatan
yang
gen
dipakm
ekspresi
yang teliti terhadap sistem optimal
model
ini
mengembangkan
Penelitian
typhi menimbulkan respons maksimal dibandingknn antigen heterolog yang diekpresi bersamanya.
(TT)
tetanus
c
toksin
digunakan
ltang
gai
antigen
model
fragmen
,àkri, biuol", S. typhimurium yang dicoba pada mencit tifuid. seb
Pada
yang
dilemahkan'
typhimvitm
protein
dari
s.
rekombinan
dengan
diekspresikan
pa'da
biitt
mencit
ternyata sangat imunogenik
jumlah plasmid dan stabilitastrya, kekuatnn
penelitian ni aitetiti berbagai parameter yang penting seperti mutasi yang mungkin ada,
SL 1344 akan
prometer dan efek translasi. Mutan gen yang isogenik seperrr purA, aroA, aroA"/aroD, htrA dan ompR darl S. typhimurium
pula
dibantlingknn
dan
penetrasi
linrfuid
ke
organ
dart
pertumbuhan
it
vivo
hal
dalam'
clibandingkan tlengan mutan cyalcrp dari SR lI
kecuali mutan
Salmonella
rekombinan
pada
seluruh
tlitemuknn
TT
spesifik
antibodi
protektif
dari
Efek
C.
clengan imurtisasi fragmen
tidak
purA. Reaksl immtogenisitas terbukti tidak ada hubungannya dengan kemampuan kuman melnkukan penetrasi ke limpa dan
maksimaL
plak
Imunogenitas
Peyer
kuman
daLam
koLonisasi
tlengan
berkorelasi
berhubungan pukt lengan respons spesifi.k sel T; namun
jumlah )tang rendah,
dari konstruksi plasmid yang'mengandung fragmen C berkorelasi dengan stabilitas plasmid; dan replikon dengan
oleh col El plasmid
yang
diekspresiknn
c
namun stabil (mis. pRSF NIT) akàn sangat efektif sebagai wahana pembawavaksin. Fragmen
plttsmid pIC21H
yang
clerivat
sedangkan
tinggi,
dalam
level
TT
spesifik
pBR 322 clm derivatnya pAT 153 dapat menginduksi antibodi
juga
Gen ,pagC,
in
vivo.
stabil
tidak
TT,
dan
imun
spesifik
justru
respon
memacu
tidak
ternyata
jumlah
tiuggi
plnsmid/sel sangat
yang
yorrg diregulasi in vivo dibandingkan dengan promoter constitutive ,trc tlalam hal ekspresi
pio*ot"r
yang
*"rr1roko,,
clan
,kaLG
,nirB
ln vivo antigen reporter firefly luciferase, dan dalam hal imunogenitas konstruksifragmen C tnvivo. Promoter in vivo yang terkuat
digu(ptrc) tentyàa kurang efektif pada imunisasi dibandingkan dengan promoter yatxg diregulasi in vivo (opagC)' Informasi ini akan
tujuan akhir
nakan untuk secara optimàl mengekspresikan antigen heterolog lain pada mtûan rekombinan S. typhimurium, dengan
mengembangknn vaksin bivalen S. typhi yang baru.
Abstract
oPportutlity
The successfuIl testing and cLevelopment of a new live, attenuated mutant (s) o/S. typhi as a typhoid vaccine creates an
of
optimal exanalysis
require
careful
wiIL
vaccines
of
such
Development
for eleveloping novel bivalent or multivalent human vaccines.
the co-exagainst
response
maximal
typhi
elicit
a
S.
the
recombinant
that
to
ensLtre
promoter
strength,...)
(gene
closage,
pression system
antigen. We have ntodelled the construction of bivalent vaccines ln S. typhimurium uslng the murine model of
(TT), is highly immunogenic in mice when exfever TIte moclel àntigen userl in these studies, the C fragment of tetaruts toxin
including
background mutation, plasmid copy num.
Parameters
typhimurium
S.
attenuated
pinrr"d o, a recombinant protein from live,
purA, aroA, aroA/aroD, htrA
bers and stability, promoter strength/regulation and translation effects were examined in this study. Isogenic
growth and penetration into
in
vivo
of
SR-ll
for
and ompR mutants o/S. typhimurfum 5L1344 were compared with the cyalcrp mutant
were
elicited by all recombinant
antibody
TT-s1tecific
levels
of
Protective
against
C
centraliymphoid organs, and immmisation
fragment.
ceII reSalmonell.ae except the purA mutant. Immunogenicity did not correLate with penetration into the spleen or specific splenic T
with
correlated
plasmid
of
C
constructs
fragment
spotxses, but did correlate with Peyer's patch colonisation. Maximal immunogenicity of
pressecL
'typhoid heterologous
plasmid stability, anl low copy, but stable replicons (eg. pRSF 1010) were effective in vaccine delivery. C fragment expressed from Co 1 E I
plasmù;s pBRj22 and its eleriyatit,e pAT153, ùtdr.tcedttery high let,els of Tt-speëific antibody whereas tlæ high copy derivative ptC2qH
rlkl not elicit TT- specific immune ,"|rponr" ancl was highly unsmble in t,ivo. In vivo-regr'Llated promoters (pPagC, onitB and ,katG) were
compared with the constitutiye promoter ttrc for in vivo expression of the reporter antigen firefly luciJ'erase, ancl in vivo immunogenicity
of C fragment consÛucts. The strongest in vivo promoter (otrc) was Less effective at immunisalion th(]n the in vivo regulatetl promoter
express other heterologous antigens from recombinanr S. typhimurium
,pugô. Wr^ationfrom tlrcse studies wiII be used to optimally
s.
typhi
vaccines.
mutants, with the ultimate aim of deveLoping novel bivaLent
I
Department of Microbiology antl Immunology,
University of M elbourne
ZCRC
1o, Vaccine Technology, University of Melbourne,
Parkville, Vic. 3052, Australia.
Reproduced with permission of the copyright owner. Further reproduction prohibited without
permission.
Minerva Access is the Institutional Repository of The University of Melbourne
Author/s:
Dunstan, SJ; Simmons, CP; Strugnell, RA
Title:
Development of recombinant S. Typhimurium, as a model for S. Typhi-based vaccine vectors
Date:
1998-01-01
Citation:
Dunstan, SJ; Simmons, CP; Strugnell, RA, Development of recombinant S. Typhimurium, as
a model for S. Typhi-based vaccine vectors, Medical Journal of Indonesia, 1998, 7 pp. 187 ?
Persistent Link:
http://hdl.handle.net/11343/191232
File Description:
Published version