- Email: [email protected]
Dynamic analysis of cytokine gene expression and protein production in a zoophilic Schistosomiosis Japonica
Oral Sessions I Parasitology
246
40.
International 47 (Suppl.) (1998) 133-281
Immunology-l (Schistosomosis)
O-0470
DYNAMIC ANALYSIS OF CYTOKINE GENE EXPRESSION AND PROTEIN PRODUCTION IN A ZOOPHILJC SCHISTOSOMIOSIS JAPONICA
o-0472
STLDlEs
ON
KINETlC
A!+TIGENS(CAGS) SCHISTOSOMA EFFICACY
OF
IN
ANALYSIS
THE
JAPONICUM m
OF
RABBITS AND
CIRCULATXNCi
W?ZCTEI
WITI
MoMToRM0
USING
CA09
‘J’BE AS
THE
MARKERS*
Lee -- K*, Lu C*, Lo C*, and Yu C** *Department of Parasitology, National Yang-Ming University and **Section of Allergy, Immunology & Rheumatology, Department of Medicine, Veterans General Hospital, National Yang-Ming University School of Medicine, Taipei, Taiwan In order to investigate the host immune response and to analyze the kinetics of Thl and Th2 cytokine response in Schis~osomajaponi~~~ infection, C57BU6 mice were infected with 50 cercariae of a zoophilic Taiwan strain of S. juponicum and sacrificed 3 to 12 weeks later. The cytokine mRNA expressions were detected with the reverse transcription-polymerase chain reaction and the cytokine measured by the enzyme-linked protein productions were immunosorbent assay. Six cytokines: IFN-y, IL2, R-4, R-5, IL-lo, and TNF- a were evaluated. The results showed that IFN- 7 mRNA level increased significantly at 3-4 wk postinfection, and started to decrease at 5 wk, reaching the preinfection level at 6 wk. The IL-4 and IL-IO mRNA levels, however, began to rise at 7 wk and reached the peak at 8 and 9 wk, then declined at 9 and 12 wk. respectively. The TNF- a response was similar to IL-IO pattern The study on cytokine protein production showed that IFN- 7 is the major immune response before the egg laying. On the other hand, Th2 cytokine IL-4 is the major response after the egg laying when the production of TIN- 7 is suppressed. These findings indicated that the Thl response is involved in the early stage of infection (3-5 wk), while the Th2 response and TNF- a production are the major phenomena associated with the acute stage of infection (7-12 wk) DELETEDTY;E-1 POLARIZATIONIN
O-0471
SCHISTOSOMA
Nobuki Hayash?,
Kiyoshi Matsui’.
Raafat T.Mohamed ***,
Owla***,
and Kazuya Higashino*
lThii
MANSONI
THE LIVER
OF
O-0473
-INFECTEDMICE.
Hiroko Tsutsui**. Somei Kojima”‘.
Yasuko Hycdo*, Kenji Nakanishi**
Yoshio ****
****
Deparmlent of Internal Medicine, **Depiuvnentof
Immunology
Zoology.
and ****Laboratory
Sciences.
Hyogo College of Medicine,Nishinomiya, Japan,
& Medical
of Host Defenses Institute for Advanced Medical
Parasitology, Institute ofMedicalScience,TbeUniversity
“‘Departnwnt
ofTokyo,
of
Tokyo, Japan.
Chronic granulomatous liver Qsease is a lethal change atier infection with Schbrosoma
mansoni
It is well known that Tcells in thespleen of infectedmice
showed chronological changes from ‘Il11 to Tb2 polarizations, which depend on larvalandeggsrages,respectively.
In this study, weinvestigatedtheimmunological
alterations in the liver where the helminth stays. In contrast to immune responses in the spleen
, theliver
lymphocytes were inittally
biased to ‘Ih2 without a Thl
stage and polarized and activatedfurther after oviposition. S.man.wni
-infectedC57BU6
little amount of
Liver lymphocytes from
mice at any stage podwed a huge amount of IL-4 but
IFN-y To clarify the mechanism how the liver prefenrdto ‘I?$ we
tried to identify theTh2driving
cell population including,CD4+
NKI.I+
Tcells.
In the liver of non-treated mice, one fifth of the lymphocytes have CD4 andNKl .I
antigenswith the characteristicsofpredominant II-4 producer.
Unexpectedly. this
cell population gradually reduced after the infection and almost disappeared in the oviposition.
However,Kupffercells from the infectedmice began to increase IL
IO production, but diminished IL-lZ!IL.-18
production. may aidchronic granuloma
formation and Tb2 dominant immune responses in the liver.
STuDltiS UN LINA VACCINE CARRYING Sj32 GENE UF ‘r!!r; tiHlNh’:SE SCHIS’I’OSOMA JAPONICUH hND ITS IMMUNE PRCr’EC’rlurr EFFh’CrS
Shi Y .E..Li Department University,
C.M. of Parasitology, wuhan, China
‘longji
Medical
Aim: To study protective immunity against cercariae challenge in mice immunized with IJNA vaccine of S.japonicum Chinese strain. Method: The plasmids ~BK-LXV containing full length and part cDNA of S.jTZ kDa were constructed. pBKSf32-1 and jnK-Sj’jP-2 were transfoctod into gIH3’1’3 cella. ‘rhe expression of SJ32 in transfected cells was confirmed by IFA. Each BALB/c mouse was vaccinated at weeks 0, 3 and 5 by injection with IOOug of purified DNA into quadriceps muscle. Nice immunized with construct and control were challenged four weeks after final DNA injection, the mice were perfused , and YOI-EQB and eggs in the liver tissue were counted. Results: Perfusion and egg counts showed significant differences either in terms of reduced worm burden 30.91;a-38.56~6, OF reduced egg rate, 55.21~55.713, between the vaccinated and control mice. Conclusion: Sj32 DNA vaccine can result in protection against a subsequent challengs of S.japonicum in mice and may be represented a new approach to develop sununit vaccine.
246
40.
International 47 (Suppl.) (1998) 133-281
Immunology-l (Schistosomosis)
O-0470
DYNAMIC ANALYSIS OF CYTOKINE GENE EXPRESSION AND PROTEIN PRODUCTION IN A ZOOPHILJC SCHISTOSOMIOSIS JAPONICA
o-0472
STLDlEs
ON
KINETlC
A!+TIGENS(CAGS) SCHISTOSOMA EFFICACY
OF
IN
ANALYSIS
THE
JAPONICUM m
OF
RABBITS AND
CIRCULATXNCi
W?ZCTEI
WITI
MoMToRM0
USING
CA09
‘J’BE AS
THE
MARKERS*
Lee -- K*, Lu C*, Lo C*, and Yu C** *Department of Parasitology, National Yang-Ming University and **Section of Allergy, Immunology & Rheumatology, Department of Medicine, Veterans General Hospital, National Yang-Ming University School of Medicine, Taipei, Taiwan In order to investigate the host immune response and to analyze the kinetics of Thl and Th2 cytokine response in Schis~osomajaponi~~~ infection, C57BU6 mice were infected with 50 cercariae of a zoophilic Taiwan strain of S. juponicum and sacrificed 3 to 12 weeks later. The cytokine mRNA expressions were detected with the reverse transcription-polymerase chain reaction and the cytokine measured by the enzyme-linked protein productions were immunosorbent assay. Six cytokines: IFN-y, IL2, R-4, R-5, IL-lo, and TNF- a were evaluated. The results showed that IFN- 7 mRNA level increased significantly at 3-4 wk postinfection, and started to decrease at 5 wk, reaching the preinfection level at 6 wk. The IL-4 and IL-IO mRNA levels, however, began to rise at 7 wk and reached the peak at 8 and 9 wk, then declined at 9 and 12 wk. respectively. The TNF- a response was similar to IL-IO pattern The study on cytokine protein production showed that IFN- 7 is the major immune response before the egg laying. On the other hand, Th2 cytokine IL-4 is the major response after the egg laying when the production of TIN- 7 is suppressed. These findings indicated that the Thl response is involved in the early stage of infection (3-5 wk), while the Th2 response and TNF- a production are the major phenomena associated with the acute stage of infection (7-12 wk) DELETEDTY;E-1 POLARIZATIONIN
O-0471
SCHISTOSOMA
Nobuki Hayash?,
Kiyoshi Matsui’.
Raafat T.Mohamed ***,
Owla***,
and Kazuya Higashino*
lThii
MANSONI
THE LIVER
OF
O-0473
-INFECTEDMICE.
Hiroko Tsutsui**. Somei Kojima”‘.
Yasuko Hycdo*, Kenji Nakanishi**
Yoshio ****
****
Deparmlent of Internal Medicine, **Depiuvnentof
Immunology
Zoology.
and ****Laboratory
Sciences.
Hyogo College of Medicine,Nishinomiya, Japan,
& Medical
of Host Defenses Institute for Advanced Medical
Parasitology, Institute ofMedicalScience,TbeUniversity
“‘Departnwnt
ofTokyo,
of
Tokyo, Japan.
Chronic granulomatous liver Qsease is a lethal change atier infection with Schbrosoma
mansoni
It is well known that Tcells in thespleen of infectedmice
showed chronological changes from ‘Il11 to Tb2 polarizations, which depend on larvalandeggsrages,respectively.
In this study, weinvestigatedtheimmunological
alterations in the liver where the helminth stays. In contrast to immune responses in the spleen
, theliver
lymphocytes were inittally
biased to ‘Ih2 without a Thl
stage and polarized and activatedfurther after oviposition. S.man.wni
-infectedC57BU6
little amount of
Liver lymphocytes from
mice at any stage podwed a huge amount of IL-4 but
IFN-y To clarify the mechanism how the liver prefenrdto ‘I?$ we
tried to identify theTh2driving
cell population including,CD4+
NKI.I+
Tcells.
In the liver of non-treated mice, one fifth of the lymphocytes have CD4 andNKl .I
antigenswith the characteristicsofpredominant II-4 producer.
Unexpectedly. this
cell population gradually reduced after the infection and almost disappeared in the oviposition.
However,Kupffercells from the infectedmice began to increase IL
IO production, but diminished IL-lZ!IL.-18
production. may aidchronic granuloma
formation and Tb2 dominant immune responses in the liver.
STuDltiS UN LINA VACCINE CARRYING Sj32 GENE UF ‘r!!r; tiHlNh’:SE SCHIS’I’OSOMA JAPONICUH hND ITS IMMUNE PRCr’EC’rlurr EFFh’CrS
Shi Y .E..Li Department University,
C.M. of Parasitology, wuhan, China
‘longji
Medical
Aim: To study protective immunity against cercariae challenge in mice immunized with IJNA vaccine of S.japonicum Chinese strain. Method: The plasmids ~BK-LXV containing full length and part cDNA of S.jTZ kDa were constructed. pBKSf32-1 and jnK-Sj’jP-2 were transfoctod into gIH3’1’3 cella. ‘rhe expression of SJ32 in transfected cells was confirmed by IFA. Each BALB/c mouse was vaccinated at weeks 0, 3 and 5 by injection with IOOug of purified DNA into quadriceps muscle. Nice immunized with construct and control were challenged four weeks after final DNA injection, the mice were perfused , and YOI-EQB and eggs in the liver tissue were counted. Results: Perfusion and egg counts showed significant differences either in terms of reduced worm burden 30.91;a-38.56~6, OF reduced egg rate, 55.21~55.713, between the vaccinated and control mice. Conclusion: Sj32 DNA vaccine can result in protection against a subsequent challengs of S.japonicum in mice and may be represented a new approach to develop sununit vaccine.