II
中文摘要
弓蟲表面膜蛋白分子SAG1 與SAG2 對蟲體入侵宿主細胞非常
重要，本研究以這兩個蛋白質分子為主角，來發展不同組合的次單位
疫苗，並以BALB/c 小鼠為實驗動物，來評估所發展的各種次單位疫
苗的保護效果。本研究首先根據免疫網路學說製備具有SAG2 內像
（SAG2 internal image）的anti-idiotype IgG（aId-IgG）來作為疫苗，
然後將2 µg、4 µg、8 µg 的aId-IgG 以皮下免疫三組小鼠後，分析小
鼠的免疫反應，發現小鼠淋巴球以弓蟲抗原刺激後可造成明顯的增殖
反應，並分泌大量的IFN-γ，上述的免疫小鼠以1×103 速殖子
（ tachyzoites）作皮下感染攻擊，結果小鼠存活率為75%~88%。另
將弓蟲SAG1 基因與SAG2 基因分別選殖於載體pGEX-6P-1，並以大
腸桿菌BL21 star（DE-3）來表現重組蛋白rSAG1 與rSAG2，然後
將rSAG1 與rSAG2 分別以不同劑量及免疫方式進行動物實驗，發現
小鼠以腹腔免疫重組蛋白10 µg rSAG1 或10 µg rSAG2 所引起的淋巴
球增殖反應較明顯，且可使免疫後的小鼠淋巴球分泌大量IFN-γ及少
量的IL-4，因此反應是偏向Th1 相關的反應；反之以其他不同劑量
的抗原經皮下免疫小鼠後，卻導致IFN-γ分泌量的降低，經免疫反應
分析後，每隻免疫後的小鼠以1×103 速殖子作皮下攻擊感染，結果發
現小鼠腹腔免疫10 µg rSAG1 或10 µg rSAG2 均有64%小鼠可存活。
更進一步將10 µg rSAG1 與10 µg rSAG2 混合後腹腔免疫小鼠，再以
1×103 速殖子進行皮下攻擊感染，小鼠的存活率可達71%，說明混合
重組蛋白rSAG1 與rSAG2 的保護效果比單僅免疫rSAG1 或rSAG2
的結果更好，這結果鼓勵我們製備嵌合蛋白（chimeric protein），即將
SAG1 基因與SAG2 基因先後選殖在同一載體上，使兩段基因接合在
一起，因此所表現的重組蛋白rSAG1/2 將同時具有SAG1 與SAG2
蛋白質的抗原性，以不同劑量的rSAG1/2 進行腹腔免疫小鼠，分析
這些小鼠的免疫反應後，發現以腹腔免疫10 µg rSAG1/2 的小鼠淋巴
III
球在弓蟲抗原TsoAg 的刺激下可造成明顯的增殖反應，而淋巴球也
可以分泌大量的IFN-γ，更驚訝的是在進行皮下感染攻擊之後，竟可
造成80%小鼠的存活，因此rSAG1/2 的保護效果會稍高於混合10 µg
rSAG1 與10 µg rSAG2 的效果。另外，本研究也探討以忌熱性腸毒素
（heat-labile enterotoxin, LT）LTA2B 或LTB 作為腸道黏膜系統的刺激
物來提高黏膜系統抵抗自然感染弓蟲的能力，即將LTA2B 基因與LTB
基因分別選殖後，獲得重組蛋白rLTA2B 與rLTB，然後將rLTA2B 或
rLTB 混合rSAG1/2 進行動物實驗並分析免疫反應，除混合型的抗原
組合外，也討論將LTA2B 基因或LTB 基因和SAG1 基因與SAG2 基因
接合在一起，形成重組蛋白rLTA2B-SAG2/1 或rLTB-SAG1/2 的融合
型抗原，以鼻腔內免疫方式將上述混合型或融合型抗原免疫動物，結
果發現在所有鼻腔內免疫的條件中以4 µg rLTB 混合10 µg rSAG1/2
免疫組以及10 µg rLTB-SAG1/2免疫組的小鼠所展現的淋巴球增殖反
應較好，而這兩組小鼠的腸道IgA 抗體的分泌量也較高，雖然這兩組
小鼠的淋巴球只分泌大量的IL-4，但在以1×103 速殖子皮下感染攻擊
後，在這兩組展現的保護效果（67%）明顯高於其他免疫組，而其餘
的抗原組合方式所表現出來的保護效果均在27%以下，所以利用
rLTB 作為黏膜刺激物的效果優於rLTA2B，且rLTB 不論混合rSAG1/2
或融合rSAG1/2 均可促進黏膜對弓蟲抗原的反應。以不同方式製備
的抗原可因處理方式、抗原組合方式及劑量不同而造成不同的結果，
其中以aId-IgG 的保護效果最好，其次是rSAG1/2，再其次是rSAG1
混合rSAG2，雖然rLTB 混合rSAG1/2 以及rLTB-SAG1/2 所展現的保
護效果不比上述三者高，但卻顯示重大的意義，即自然感染弓蟲的途
徑仍可利用黏膜刺激物來提高抵抗力，所以這些抗原將來均可成為疫
苗發展時的優良候選抗原。

IV
英文摘要
SAG1 and SAG2 are important surface molecules of T. gondii for the
invasion of tachyzoites into host-cells. Previous studies have been
demonstrated they are good candidates for development of vaccines
against toxoplasmosis. Therefore, we used SAG1 and SAG2 to generate
subunit vaccines and evaluated the protective capacity in BALB/c mice.
Anti-idiotype IgG (aId-IgG) with the SAG2 internal image was prepared
from anti-SAG2 monoclonal antibody in accordance with the network
theory. Lymphocytes from mice immunized subcutaneously twice with 2,
4 or 8 µg aId-IgG showed great proliferations and secreted high level of
IFN-γ, which is an important cytokines secreted by Th1 cells. After
challenged subcutaneously with 1×103 tachyzoites, the highest survival
rate reached 88%. Further, SAG1 and SAG2 genes were respectively
cloned and recombinant proteins rSAG1 and rSAG2 were prepared.
Lymphocytes from mice immunized intraperitoneally twice with rSAG1
or rSAG2 displayed apparently Th1-associated responses, while
lymphocytes from mice immunized subcutaneously twice with rSAG1 or
rSAG2 did not. Mice immunized intraperitoneally twice with rSAG1 or
rSAG2 had a survival rate of 64% which was higher than those mice
immunized subcutaneously with rSAG1 or rSAG2. When mice
immunized intraperitoneally twice with rSAG1 mixed with rSAG2,
survival rate was even higher (71%). Therefore, mixed antigens induced
higher protection. Subsequently, SAG1 gene was fused with SAG2 gene
and a recombinant protein rSAG1/2 was expressed from the fused gene.
Th1-associated responses were detected from lymphocytes of mice
immunized intraperitoneally twice with 10 µg rSAG1/2. Interestingly,
80% rSAG1/2-immunized mice survived and it was extremely higher
V
than rSAG1- or rSAG2-immunized mice (64%). In an attempt to
stimulate immune responses against T. gondii infections in the mucosal
system, we chose heat-labile enterotoxin (LT) secreted from toxigenic E.
coli as the stimulator. LTA2B and LTB genes were respectively cloned and
then rLTA2B and rLTB were obtained. Moreover, LTA2B gene or LTB
gene fused with SAG1 and SAG2 genes was performed and then
recombinant proteins rLTA2B-SAG2/1 and rLTB-SAG1/2 were prepared.
Subsequently, mice were immunized intranasally twice with
rLTA2B-SAG2/1, rLTB-SAG1/2, rLTA2B mixed with rSAG1/2, or rLTB
mixed with rSAG1/2. A strong protection (67%) was shown by the group
of mice immunized intranasally with 10 µg rLTB-SAG1/2 or 4 µg rLTB
mixed with 10 µg rSAG1/2. rLTB, rather than rLTA2B, will be a better
candidate for stimulation the mucosal system. In summary, different
survival rates caused by antigens prepared in the study may be attributed
to many factors such as the treatment, the preparation and the dose of
antigen. The highest survival rate is caused by aId-IgG (88%); the second
is shown by rSAG1/2 (80%); the third is resulted from rSAG1mixed
rSAG2 (71%). Although the survival rate raised by rLTB-SAG1/2 or
rLTB mixed rSAG1/2 is slightly low (67%), these data demonstrate
stimulators such as LT could induce anti-Toxoplasma immune response
of the mucosal system. Antigens capable of inducing higher survival rates
in mice should be worthy of further investigation for searching better
vaccine candidates.

總目錄
中文摘要………………………………………………………I
英文摘要…………………………………………………….. III
總目錄………...……………………………………………….V
表目錄………………………………………………………...XI
圖目錄………………………………………………………..XII

第一章 總論…...……………………………………………....1
第1節 弓蟲的發現………………………………………………1
第2節 弓蟲之形態及生活史……………………………………2
第3節 弓蟲之傳播與流行病學…………………………………5
第4節 臨床症狀與病變…………………………………………7
第5節 弓蟲症之治療…………………………………………..10
第6節 弓蟲症之診斷方式……………………………………..12
第7節 弓蟲症之疫學研究……………………………………..16

第二章 研究動機與目的…………………………………..25

第三章 研究內容…………………………….....................26
第1節 Anti-idiotype antibody對小鼠之保護率分析..............27
前言……………….……………………………………………27
材料與方法…………………………………………………….30
1. 單株抗體之製備…………………………………..........30
2. Idiotypic vaccine（anti-idiotype IgG）之製備………...35
3. aId-IgG免疫計劃…………………………………….....36
4. 淋巴球增殖試驗………………………………………..36
5. IFN-

236
考文獻
Achbarou A, Mercereau-Puijalon O, Autheman JM, Fortier B, Camus D,
Dubremetz JF（1991）Characterization of microneme proteins of
Toxoplasma gondii. Mol Biochem Parasitol 47:223-233
Adams JH, Hudson DE, Torii M, Ward GE, Wellems TE, Aikawa M, Miller
LH（1990）The duffy receptor family of Plasmodium knowlesi is
located within the micronemes of invasive malaria merozoites. Cell
Bermudes D, Dubremetz JF, Achbarou A, Joiner KA（1994）Molecular
cloning of a complete cDNA encoding the dense granule protein
GRA3 from Toxoplasma gondii. Mol Biochem Parasitol 68:247-257
Bessiéres MH, Roques C, Berrebi A, Barre V, Cazaux M, Seguela JP
（ 1992 ） IgA antibody response during acquired and congenital
toxoplasmosis. J Clin pathol 45:605-608
Biancifiori F, Rondini C, Grelloni V, Frescura T （ 1986 ） Avian
toxoplasmosis: experimental infection of chicken and pigeon. Comp
Immunol Microbiol Infect Dis 9:337-346
Bonenfant C, Dimier-Poisson I, Velge-Roussel F, Buzoni-Gatel D, Del
Giudice G, Rappuoli R, Bout D（2001）Intranasal immunization with
SAG1 and nontoxic mutant heat-labile enterotoxins protects mice
against Toxoplasma gondii.Infect Immun 69:1605-1612
Bourguin I, Moser M, Buzoni-Gatel D, Tielemans F, Bout D, Urbain J, Leo
O（1998） Murine dendritic cells pulsed in vitro with Toxoplasma
gondii antigens induced protective immunity in vivo. Infect Immun
237
66:4867-4874
Bülow R, Boothroyd JC (1991) Protection of mice from fatal Toxoplasma
gondii infection by immunization with p30 antigen in liposomes. J
Immunol 147:3496-3500
Burg JL, Perelman D, Kasper LH, Ware PL, Boothroyd JC (1988)
Molecular analysis of the gene encoding the major surface antigen of
Toxoplasma gondii. J Immunol 141:3584-3591
Burg JL, Grover CM, Pouletty P, Boothroyd JC（1989）Direct and sensitive
detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase
chain reaction. J Clin Microbiol 27:1787-1792
Burke JM, Roberts CW, Hunter CA, Murray M, Alexander J（1994）
Temporal differences in the expression of mRNA for IL-10 and IFN-γ
in the brains and spleens of C57BL10 mice infected with Toxoplasma
gondii. Parasitol Immunol 16:305-314
Buxton D (1993) Toxoplasmosis: the first commercial vaccine. Parasitol
Today 9:335-337
Canessa A, Del Bono V, Miletich F, Pistoia V (1992) Serum cytokines in
toxoplasmosis: increased levels of interferon-γ in immunocompetent
patients with lymphadenoppathy but not in AIDS patients with
encephalitis. J Infect Dis 165:1168-1170
Cesbron JY, Caron A, Santoro F, Wattre P, Ovlaque G, Pierce RJ,
Delagneau JP, Capron A（1986）Une nouvelle méthode ELISA pour Ie
diagnostic de Ia toxoplasmose: dosage des IgM sériques par
immunocapture avec un anticorps monoclonal anti-Toxoplasma gondii.
Press Méd 15:737-740
238
Cesbron-Delauw MF, Guy B, Torpier G, Pierce RJ, Lenzen G, Cesbron JY,
Charif H, Lepage P, Darcy F, Lecocq JP, Capron A（1989）Molecular
characterization of a 23-kilodalton major antigen secreted by
Toxoplasma gondii. Proc Natl Acad Sci USA 86:7537-7541
Chang GN (1993) Characterization of Babesia gibsoni anti-idiotype
antibody. Chin J Parasitol 6: 13-20
Chang GN (1996) Studies on the immunoprotective potency of Babesia
gibsoni anti-idiotype vaccine in dogs. J Chin Soc Vet Sci 22: 136-144
Channon JY, Suh EI, Seguin RM, Kasper LH（1999）Attachment ligands of
viable Toxoplasma gondii induce soluble immunosuppressive factors
in human monocytes. Infect Immun 67:2547-2551
Chardès T, Bout D (1993) Mucosal immune response in toxoplasmosis. Res
Immunol 144:57-60
Clark IA, Rockett KA, Crowden WB（1991）Proposed link between
cytokines, nitric oxide and human cerebral malaria. Parasitol Today
7:205-207
Clements JD, Yancey RJ, Finkelstein RA（1980）Properties of homogenous
heat-labile enterotoxin from Eschehia colrici. Infect Immun 29:91-97
Cohn JA, McKeeking A, Cohen W, Jacobs J, Holzman RS（1989）
Evaluation of the policy of empiric treatment of suspected Toxoplasma
encephalitisin patients with the acquired immunodeficiency syndrome.
Am J Med 86:521-527
Couvreur G, Sadak A, Fortier B, Dubremetz JF (1988) Surface antigens of
Toxoplasma gondii. Parasitology 97:1-10
Dannemann BR, Vaughan WC, Thulliez P, Remington JS （ 1990 ）
239
Differential agglutination test for diagnosis of recently acquired
infection with Toxoplasma gondii. J Clin Microbiol 28:1928-1933
Dannemann BR, Israelski DM, Leoung GS, McGraw T, Mills J, Remington
JS（1991）Toxoplasma serology, parasitemia and antigenemia in
patients at risk for toxoplasmic encephalitis. AIDS 5:1363-1365
Debard N, Buzoni-Gatel D, Bout D（1996） Intranasal immunization with
SAG1 protein of Toxoplasma gondii in association with cholera toxin
dramatically reduces development of cerebral cysts after oral infection.
Infect Immun 64:2158-2166
Decoster A, Darcy F, Caron A, Capron A（1988） IgA antibodies against
P30 as markers of congenital and acute toxoplasmosis. Lancet
332:1104-1107
Decoster A, Darcy F, Caron A, Vinatier D, Houze de ÍAulonit D, Vittu G,
Niel G, Heyer F, Lecolier B, Delcroix M, Monnier JC, Duhamel M,
Capron A（1992）Anti-P30 IgA antibodies as prenatal markers of
congenital Toxoplasma infection. Clin Exp Immunol 87:310-315
Denkers EY, Caspar P, Sher A（1994）Toxoplasma gondii possesses a
superantigen activity that selectively expands murine T cell receptor
Vβ5-bearing CD8+ lymphocytes. J Exp Med 180:985-994
Derouin F, Thulliez P, Candolfi E, Daffos F, Forestier F（1988）Early
prenatal diagnosis of congenital toxoplasmosis using amniotic fluid
samples and tissue culture. Eur J Clin Microbiol Infect Dis 7:423-425
Desmonts G, Couvreur J（1974）Congenital toxoplasmosis: a prospective
study of 378 pregnancies. N Engl J Med 290:1110-1116
Desmonts G, Naot Y, Remington JS （ 1981 ） An immunoglobulin
240
M-immunosorbent agglutination assay for diagnosis of infectious
diseases: diagnosis of acute congenital and acquired Toxoplasma
infection. J Clin Microbiol 13:859-864
Desmonts G, Forestier F, Thulliez P, Daffos F, Capella-Pavlovsky M,
Chartier M（1985）Prenatal diagnosis of congenital toxoplasmosis.
Lancet 325:500-503
Desolme B, Mévélec MN, Buzoni-Gatel D, Bout D (2000) Induction of
protective immunity against toxoplasmosis in mice by DNA
immunization with a plasmid encoding Toxoplasma gondii GRA4
gene. Vaccine 18:2512-2521
Dubey JP（1986）A review of toxoplasmosis in cattle. Vet Parasitol
22:177-202
Dubey JP, Murrell KD, Fayer R, Schad GA（1986）Distribution of
Toxoplasma gondii tissue cysts in commercial cuts of pork. J Am Vet
Med Assoc 188:1035-1037
Dubey JP, Beattie CP（1988）Toxoplasmosis of animals and man. CRC
Press Florida
Dubey JP（1990）Status of toxoplasmosis in sheep and goats in the United
States. J Am Vet Med Assoc 196:259-262
Dubey JP, Kotula AW, Sharar A, Andrews CD, Lindsay DS （1990）Effect
of high temperature on infectivity of Toxoplasma gondii tissue cysts in
pork. J Parasitol 76:201-204
Dubey JP (1994) Toxoplasmosis. J Am Vet Med Assoc 205:1593-1598
Dubey JP（1996）Strategies to reduce transmission of Toxoplasma gondii to
animals and humans. Vet Parasitol 64:65-67
241
Duermeyer W, Wielaard F, Gruijthuijsen HV, Swinkels J（1980） Enzyme-
linked immunosorbent assay for detection of immunoglobulin M
antibodies against Toxoplasma gondii. J Clin Microbiol 12:805-806
Dupon M, Cazenave J, Pellegrin JL, Ragnaud JM, Cheyron A, Fischer I,
Leng B, Lacut JY（1995）Detection of Toxoplasma gondii by PCR and
tissue culture in cerebrospinal fluid and blood of human
immunodeficiency virus-seropositive patients. J Clin Microbiol
33:2421-2426
Eggers C, Gross U, Klinker H, Schalke B, Stellbrink HJ, Kunze K（1995）
Limited value of cerebrospinal fluid for direct detection of
Toxoplasma gondii in toxoplasmic encephalitis associated with AIDS.
J Neurol 242:644-649
Eichmann K (1978) Expression and function of idiotypes on lymphocytes.
Adv Immunol 26: 195-254
Favre G, Bessiéres MH, Seguela JP（1984）dosage des IgA sériques
spécifiques de Ia toxoplasmose par une méthode ELISA. Application à
120 cas. Bull Sco Fr Parasitol 3:139-142
Frenkel JK, Dubey JP, Miller NL（1970）Toxoplasma gondii in cats: fecal
stages identified as coccidian oocysts. Science 167:893-896
Frenkel JK, Smith DD（1982）Immunization of cats against shedding of
Toxoplasma oocysts. J Parasitol 68:744-748
Frenkel JK (1988) Pathophysiology of toxoplasmosis. Parasitol Today
4:273-278
Finbery RW, Etrl HCJ (1986) Use of T cell-specific anti-idiotypes to
immunise against viral infections. Immunol Rev 90: 129-248
242
Gazzinelli RT, Hakim FT, Hieny S, Shearer GM, Sher A (1991) Synergistic
role of CD4+and CD8+ T lymphocytes in IFN-γ production and
protective immunity induced by an attenuated Toxoplasma gondii
vaccine. J Immunol 146:286-292
Gazzinelli RT, Denkers EY, Sher A (1993a) Host resistance to Toxoplasma
gondii: model for studying the selective induction of cell-mediated
immunity by intracellular parasites. Infect Agents Dis 2:139-149
Gazzinelli RT, Hieny S, Wynn T, Wolf S, Sher A（1993b）IL-12 is required
for the T-cell independent induction of IFN-γ by an intracellular
parasite and induces resistance in T-deficient hosts. Proc Natl Acad Sci
90:6115-6119
Gazzinelli RT, Wysocka M, Hieny S, Kersten T, Carrera L, Cheever A,
Kühn R, Müller W, Trinchieri G, Sher A（1996）In absence of
endogenous IL-10 mice acutely infected with Toxoplasma gondii
succumb to a lethal immune response associated with increased
synthesis of IL-12, IFN-γ and TNF-α. J immunol（in press）
Geary S, Marchlewices BA, Finkelstein RA (1982) Comparisons of
heat-labile enterotoxins from porcine and human strains of
Escherichia coli. Infect Immun 36:215-220
Gill DM（1976）Multiple roles of erythrocytes supernatant in the activation
of adenylate cyclase by Vibrio cholerae toxin in vitro. J Infect Dis
133:S55-S63
Grimwood J, Smith JE（1992）Toxoplasma gondii: the role of a 30 kDa
surface protein in host cell invasion. Exp Parasitol 74:106-111
Grimwood J, Smith JE (1996) Toxoplasma gondii: the role of parasite
243
surface and secreted proteins in host cell invasion. Int J Parasitol
26:169-173
Gross U, Roos T, Appoldt D, Heesemann J（1992）Improved serological
diagnosis of Toxoplasma gondii infection by detection of
immunoglobulin A（IgA）and IgM antibodies against P30 by using the
immunoblot technique. J Clin Microbiol 30:1436-1441
Grzych JM, Capron M, Lambert PH, Dissons C, Torres S, Capron A (1985)
An anti-idiotype vaccine against experimental schistosomiasis. Nature
316: 74-76
Handman E, Goding JW, Remington JS （ 1980 ） Detection and
characterization of membrane antigens of Toxoplasma gondii. J
Immunol 124:2578-2583
Hay J, Hutchison WM （ 1983 ）Toxoplasma gondii-an environmental
contaminant. Ecol Dis 2:33-43
Hedman K, Lappalainen M, Seppada I, Makela O（1989）Recent primary
Toxoplasma infection indicated by a low avidity of specific IgG. J
Infect Dis 159:736-739
Holmgren J (1981) Actions of cholera toxin and the prevention and
treatment of cholera. Nature 292:413-417
Homan WL, Vercammen M, Braekeller JDE, Vershueren H（2000）
Identification of a 200- to 300-fold repetitive 529 bp DNA fragment in
Toxoplasma gondii, and its use for diagnostic and quantitative PCR.
Internal J Parasitol 30:69-75
Hughes HPA（1988）Oxidative killing of intracellular parasites mediated by
macrophages. Parasitol Today 4:340-347
244
Hunter CA, Remington JS（1994）Immunopathogenesis of toxoplasmic
encephalitis. J Infect Dis 170:1057-1067
Huskinson J, Thulliez P, Remington JS（1990）Toxoplasma antigens
recognized by human immunoglobulin A antibodies. J Clin Microbiol
28:2632-2636
Israelski DM, Remington JS（1993）Toxoplasmosis in the non-AIDS
immunocompromised host. Curr Clin Top Infect Dis 13:322-356
Jackson MH, Hutchison WM, Siim JC（1986）Toxoplasmosis in a wild
rodentbpopulation of central Scotland and a possible explanation of
the mode of transmission. J Zool 209:549-557
Jean-Francois MJB, Poskitt DC, MacDonald LM, Turnbull SJ, Yasmeen D
（1993）Production of monoclonal anti-idiotype antibodies which
mimic an M-like protein of Streptococcus equi. Microbiol Immunol 37:
737-742
Jerne NK (1974) Toward a network theory of the immune system. Ann
Immunol 125c: 373-389
Joss AWL（1992）Treatment. In ” Ho-Yen DO, Joss AWL（eds）Human
toxoplasmosis. Oxford University Press, Oxfod, pp:119-143
Kasper LH, Crabb JH, Pfefferkorn ER（1982）Isolation and characterization
of a monoclonal antibody-resistant antigenic mutant of Toxoplasma
gondii. J Immunol 129:1694-1699
Kasper LH, Crabb JH, Pfefferkorn ER（1983）Purification of a major
membrane protein of Toxoplasma gondii by immunoabsorption with a
monoclonal antibody. J Immunol 130:2407-2412
Kasper LH, Currie KM, Bradley MS (1985) An unexpected responseto
245
vaccination with a purified major membrane tachyzoite antigen (P30)
of Toxoplasma gondii. J Immunol 134: 3426-3431
Kasper LH（1989）Identification of stage-specific antigens of Toxoplasma
gondii. Infect Immun 57: 668-672
Kasper LH, Khan IA, Ely KH, Bülow R, Boothroyd JC (1992)
Antigen-specific (p30) mouse CD8+ T cells are cytotoxic against
Toxoplasma gondii-infected peritoneal macrophages. J Immunol
148:1493-1498
Kasper LH, Khan IA (1993) Role of P30 in host immunity and pathogensis
of T. gondii infection. Res Immunol 144:45-48
Khan IA, Eckel ME, Pfefferkorn ER, Kasper LH (1988) Production of γ
interferon by cultured human lymphocytes stimulated with a purified
membrane protein (P30) from Toxoplasma gondii. J Infec Dis
157:979-983
Khan IA, Ely KH, Kasper LH (1991) A purified parasite antigen (P30)
mediates CD8+ T cell immunity against fatal Toxoplasma gondii
infection in mice. J Immunol 147:3501-3506
Kimbita EN, Xuan X, Huang X, Miyazawa, Fukumoto S, Mishima M,
Suzuki H, Sugimoto C, Nagasawa H, Fujisaki K, Suzuki N, Mikami T,
Igarashi I (2001) Serodiagnosis of Toxoplasma gondii infection in cats
by enzyme-linked immunosorbent assay using recombinant SAG1. Vet
Parasitol 102:35-44
Kirisits MJ, Mui E, McLeod R（2000）Measurement of the efficacy of
vaccines and antimicrobial therapy against infection with Toxoplasma
gondii. Internal J Parasitol 30:149-155
246
Kohler H, McNamara M, Ward R（1984）Monoclonal idiotype vaccines. J
Exp Med 159: 655-665
Kuby（2001）Cells and organs of the immune system. In”Immunology”（4th
edn）(ed J Kuby), pp:53-57, W. H. Freeman and Company, New York
Laemmli UK (1970) Cleavage of structural proteins during the assembly of
the head of bacteriophage T4. Nature 227:680-685
Langermans JAM, Van der Hulst MEB, Nibbering PH, Hiemstra PS,
Fransen L, Van Furth R（1992）IFN-γ induced L-arginine-dependent
toxoplasmastatic activity in murine peritoneal macrophages is
mediated by endogenous tumor necrosis factor-α. J Immunol
148:568-574
Lebech M, Lebech AM, Nelsing S, Vuust C, Mathiesen L, Petersen E
（1992）Detection of Toxoplasma gondii-DNA by polymerase chain
reaction in cerebrospinal fluid from AIDS patients with cerebral
toxoplasmosis. J Infect Dis 165:982-983
Lecordier L, Mercier C, Torpier G, Tourvieille B, Darcy F, Liu JL, Maes P,
Tartar A, Capron A, Cesbron-Delauw MF（1993）Molecular structure
of a Toxoplasma gondii dense granule antigen (GRA5) associated with
the parasitophorous vacuole membrane. Mol Biochem Parasitol
59:143-154
Lecordier L, Moleon-Borodowski I, Dubremetz JF, Tourvieille B, Mercier
C, Deslée D, Capron A, Cesbron-Delauw MF（1995）Characterization
of a dense granule antigen of Toxoplasma gondii (GRA6) associated to
the network of the parasitophorous vacuole. Mol Biochem Parasitol
70:85-94
247
LeFichoux Y, Marty P, Chan H, Doucet J（1984）Détection des IgM
antitoxoplasmiques par ISAGA. A propos de 3786 sérologies. Bull
Soc Fr Parasitol 3:15-18
LeFichoux Y, Marty P, Chan H（1987）Les IgA spécifiques dans Ie
diagnostic de Ia toxoplasmose. Ann Pédiatr 34:375-379
Leport C, Raffi F, Matheron S, Katlama C, Regnier B, Saimot AG, Marche
C, Vedrenne C, Vilde JL（1988）Treatment of central nervous system
toxoplasmosis with pyrimethamine/sulfadiazine combination in 35
patients with the acquired immunodeficiency syndrome. Am J Med
84:94-100
Luft BJ, Remington JS（1992）Toxoplasmic encephalitis in AIDS. Clin
Infect Dis 15:211-222
Luft BJ, Hafner R, Korzun AH, Leport C, Antoniskis D, Bosler EM,
Bourland DD, Uttamchandani R, Fuhrer J, Jacobson J（1993）
Toxoplasmic encephalitis in patients with the acquired
immunodeficiency syndrome. N Engl J Med 329:995-1000
Lunden A（1995）Immune responses in sheep after immunization with
Toxoplasma gondii antigens incorporated into iscoms. Vet Parasitol
56:23-35
Lunden A, Parmley SF, Bengtsson KL, Araujo FG (1997). Use of a
recombinant antigen, SAG2, expressed as a glutathione-S-transferase
fusion protein to immune mice against Toxoplasma gondii. Parasitol
Res 83: 6-9
Maggi E, Parronchi P, Manetti R, Simonelli C, Piccinni MP, Rugiu FS, De
Carli M, Ricci M, Romagnani S (1992) Reciprocal regulatory effects
248
of IFN-γ and IL-4 on the in vitro development of human Th1 and Th2
clones. J. Immunol 154:5071-5079
Makioka A, Kobayashi A（1991）Toxoplasmacidal activity of macrophages
activated by recombinant major surface antigen（P30）of Toxoplasma
gondii. Infect Immun 59:2851-2852
Malik MA, Dreesen DW, de la Cruz A（1990）Toxoplasmosis in sheep in
north eastern United States. J Am Vet Med Assoc 196:263-265
Marriott SJ, Roeder DJ, Consigli RA (1987) Anti-idiotypic antibodies to a
polyomavirus monoclonal antibody recognize cell surface components
of mouse kidney cells and prevent polyomavirus infection. J Am Soc
Microbiol 61: 2747-2753
McLeod R, Frenkel JK, Estes RG, Mack DG, Eisenhauer PB, Gibori G
（ 1988 ） Subcutaneous and intestinal vaccination with cysts of
Toxoplasma gondii and acquisition of immunity to peroral and
congenital Toxoplasma challenge. J Immunol 140:1632-1637
McLeod R, Skamene E, Brown CR, Eisenhauer PB, Mack DG（1989）
Genetic regulation of early survival and cyst number after peroral
Toxoplasma gondii infection of A×B/B×A recombinant inbred and
B10 congenic mice. J Immunol 143:3031-3034
McNamara MK, Ward RE, Kohler H (1984) Monoclonal idiotype vaccine
against Streptococcus pneumoniae infection. Science 226: 1325-1326
Mevelec MN, Chardés T, Mercereau-Puijalon O, Bourguin I, Achbarou A,
Dubremetz JF, Bout D（ 1992 ） Molecular cloning of GRA4, a
Toxoplasma gondii dense granule protein, recognized by mucosal IgA
antibodies. Mol Biochem Parasitol 56:227-238
249
Mercier C, Lecordier L, Darcy F, Deslée D, Murray A, Tourvieille B, Maes
P, Capron A, Cesbron-Delauw MF（1993）Molecular characterization
of a dense granule antigen (GRA2) associated with the network of the
parasitophorous vacuole in Toxoplasma gondii. Mol Biochem
Parasitol 58:71-82
Mineo JR, Mcleod R, Mack D, Smith J, Khan IA, Ely KH, Kasper LH
（1993）Antibodies to Toxoplasma gondii major surface protein
(SAG1, P30) inhibit infection of host cells and are produced in murine
intestine after peroral infection. J Immunol 150:3951-3964
Mineo JR, Kasper LH (1994) Attachment of Toxoplasma gondii to host
cells involves major surface protein, SAG1 (P30). Exp Parasitol
79:11-20
Mishima M, Xuan X, Shioda A, Omata Y, Fujisaki K, Nagasawa H,
Mikami T (2001) Modified Protection against Toxoplasma gondii
lethal infection and brain cyst formation by Vaccination with SAG2
and SRS1. J Vet Med Sci 63:433-438
Moss J, Osborne JC, Fishman PH, Nakaya S, Robertson DC（1981）
Escherichia coli heat-labile enterotoxin: ganglioside specificity and
ADP-ribosyltransferase activity. J Biol Chem 256:12861-12865
Murray HW, Rubin BY, Masur H, Roberts RB (1984) Impaired production
of lymphokines and immune (gamma) interferon in the acquired
immunodeficiency syndrome. N Engl J Med 310:883-889
Nicolle C, Manceaux L（1908）Sur une infection àcorps de Leishman (ou
organisms voisins) du gondii. In ”Human Toxoplasmosis” (ed DO
Ho-Yen and AWL Joss), pp:1, Oxford University Press, New York
250
Novati R, Castagna A, Morsica G, Vago L, Tambussi G, Ghezzi S,
Gervasoni C, Bisson C, ďArminoi-Monforte A, Lazzarin A（1994）
Polymerase chain reaction for Toxoplasma gondii DNA in
cerebrospinal fluid of AIDS patients with focal brain lesions. AIDS
8:1691-1694
Parker SJ, Roberts CW, Alexander J (1991) CD8+ T cells are the major
lymphocyte subpopulation involved in the protective immune response
to Toxoplasma gondii in mice. Clin Exp Immunol 84:207-212
Parmley SF, Sgarlato GD, Mark J, Prince JB, Remington JS（1992）
Expression, Characterization, and serological reactivity of
recombinant surface antigen P22 of Toxoplasma gondii. J Clin
Microbiol 27:1127-1133
Parmley SF, Gross U, Sucharczuk A, Windeck T, Sgarlato GD, Remington
JS (1994) Two alleles of the gene encoding surface antigen P22 in 25
strains of Toxoplasma gondii. J Parasitol 80:293-301
Partanen P, Turunen HJ, Paasivuo RTA, Leinikki PO（1984）Immunoblot
analysis of Toxoplasma gondii antigens by human immunoglobulin G,
M and A antibodies at different stages of infection. J Clin Microbiol
20:133-135
Patel B, Young Y, Duffy K, Tanner RP, Johnson J, Holliman RE（1993）
Immunoglobulin-A detection and the investigation of clinical
toxoplasmosis. J Med Microbiol 38:286-292
Petersen E, Nielsen HV, Christiansen L, Spenter J (1998) Immunization
with E. coli produced recombinant Toxoplasma gondii SAG1 with
alum as adjuvant protects mice against lethal infection with
251
Toxoplasma gondii. Vaccine 16:1283-1289
Pfefferkorn ER, Guyre PM（1984）Inhibition of growth of Toxoplasma
gondii in cultured fibroblasts by human recombinant gamma interferon.
Infect Immun 44: 211-216
Pinon JM, Thoannes H, Gruson N（1985）An enzyme-linked immuno-
filtration assay used to compare infant and maternal antibody profiles
in toxoplasmosis. J Immunol Methods 77:15-23
Pinon JM, Thoannes H, Pouletty PH, Poirriez J, Damiens J, Pelletier P
（1986）Detection of IgA specific for toxoplasmosis in serum and
cerebrospinal fluid using a non-enzymatic IgA-capture assay. Diagn
Immunol 4:223-227
Porter SB, Sande MA（1992）Toxoplasmosis of the central nervous system
in the acquired immunodeficiency syndrome. N Engl J Med
327:1643-1648
Prince BJ, Auer KL, Huskinson J, Parmley SF, Araujo FG, Remington JS
(1990) Cloning, expression, and cDNA sequence of surface antigen
P22 from Toxoplasma gondii. Mol Biochem Parasitol 43:97-106
Remington JS, Desmonts G（1990） Toxoplasmosis. In “Infectious diseases
of the foetus and newborn infant” (3rd edn). (ed JS Remington and JO
Klein), pp:89-195,WB Saunders, Philadelphia
Roberts CW, Cruickshank S, Alexander J（1995）Sex determined resistance
to Toxoplasma gondii is associated with temporal differences in
cytokine production. Infect Immun 63:2549-2555
Robertson DC, Kunkel SL, Galligan PH（1980） Structure and function of
E. coli heat-labile enterotoxin:15th Join Conference on Cholera,
252
（DHEW publ no（NIH）80-2003）
Sabin AB, Fedlman HA（1948）Dyes as microchemical indicators of a new
immunity phenomenon affecting a protoznan parasite (Toxoplasma).
Science 108:660-663
Sacks DL, Esser KM, Sher A (1982) Immunization of mice against African
trypanosomiasis using anti-idiotypic antibodies. J Exp Med 155:
1108-1119
Saffer LD, Mercereau-Puijalon O, Dubremetz JF, Schwartzman JD（1992）
Localization of Toxoplasma gondii rhoptry protein by immunoelectron
microscopy during and after host cell penetration. J Protozool
39:526-530
Santoro F, Afchain D, Pierce JR, Cesbron JY, Ovlaque G, Capron A（1985）
Serodiagnosis of Toxoplasma infection using a purified parasite
protein (P30). Clin Exp Immunol 62:262-269
Schedel I, Sutor GC, Hunsmann G, Jurkiewicz E (1999) Phase II study of
anti-CD4 idiotype vaccination in HIV positive volunteers. Vaccine
17:1837-1845
Schoondermark-van de Ven E, Galama J, Kraaijeveld C, van Druten J,
Meuwissen J, Melchers W（1993）Value of polymerase chain reaction
for the detection of Toxoplasma gondii in cerebrospinal fluid from
patients with AIDS. Clin Infec Dis 16:661-666
Sharma SD, Muliena J, Araujo FG, Erlich HA, Remington JS（1983）
Western blot analysis of the antigens of Toxoplasma gondii recognized
by human IgM and IgG antibodies. J Immunol 131:977-983
Sher A, Oswald IP, Hieny S, Gazzinelli RT (1993) Toxoplasma gondii
253
induces a T-independent IFN-γ response in NK cells which requires
both adherent accessory cells and TNF-α. J Immunol 150:3982-3989
Sibley LD, Dobrowolski J, Morisaki JH, Heuser JE（1994）Strategies for
intracellular survival of microbes. Clin Infect Dis 1:245-264
Smith JE (1995) A ubiquitous intracellular parasite: the cellular biology of
Toxoplasma gondii. Int J Parasitol 25: 1301-1309
Splendore A（1908）Uno nuovo protozoa parasite dei conigli:incontrato
nelle lesioni anatomiche d’una malattia che ricorda in molti punti il
Kala-azar dell’uomo. In ”Human Toxoplasmosis” (ed DO Ho-Yen and
AWL Joss), pp:4, Oxford University Press, New York
Stein KE, Soderstrom T (1984) Neonatal administration of idiotype or
anti-idiotype primes for protection against Escherichia coli K13
infection in mice. J Exp Med 160: 1001-1011
Su S, McNamara-Ward M, Apicella MA, Ward RE (1992) A non-toxic
idiotype vaccine against gram-negative bacterial infections. J Immunol
148: 234-238
Subauste CS, Koniaris AH, Remington JS (1991) Murine CD8+ cytotoxic T
lymphocytes lyse Toxoplasma gondii-infected cells. J Immunol
147:3955-3959
Suzuki Y, Orellana MA, Schreiber RD, Remington JS (1988) Interferon-γ:
the major mediator of resistance against Toxoplasma gondii. Science
240:516-518
Suzuki Y, Conley FK, Remington JS（1990）Treatment of toxoplasmic
encephalitis in mice with recombinant gamma interferon. Infect
Immun 58:3050-3055
254
Tanaka M, Okabe T, Sasaki N (1990) Monoclonal internal image
anti-idiotype antibodies of Theileria sergenti merozoite surface
antigen. Jpn. J. Vet. Sci 52: 813-818
Taverne J（1993）Unravelling the cytokine network in malaria. Parasitol
Today 9:38-39
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins
from polyacrylmide gels to nitrocellulose sheets: procedure and some
applications. Proc Natl Acad Sci USA 76:4350-4354
Turunen H, Vuorio KA, Leinikki PO（1983）Determination of IgG, IgM and
IgA antibody responses in human toxoplasmosis by enzyme-linked
immunosorbent assay (ELISA). Scand J Infect Dis 15:307-311
Van Loon AM, Van der Logt JTM, Heessen WFA, Van der Veen J（1983）
Enzyme-linked immunosorbent assay that uses labeled antigens for the
detection of immunoglobulin M and A antibodies in toxoplasmosis:
comparison with indirect immunofluorescence and double-sandwich
enzyme-linked immunosorbent assay. J Clin Microbiol 17:997-1004
Velge-Roussel F, Marcelo P, Lepage AC, Buzoni-Gatel D, Bout DT（2000）
Intranasal immunization with Toxoplasma gondii SAG1 induces
protective cells into both NALT and GALT compartments. Infect
Immun 68:969-972
Waldeland H, Frenkel JK (1986) Live and killed vaccines against
toxoplasmosis in mice. J Parasitol 69:60-65
Walls KW, Bullock SL, Donna KE（1976）Use of the enzyme-linked
immunosorbent assay (ELISA) and its microadaptation for the
serodiagnosis of toxoplasmosis. J Clin Microbiol 5:273-277
Walton BC, Benchoff BM, Brooks WH（1966）Comparison of the indirect
fluorescent antibody test and methylene blue dye test for detection of
antibodies to Toxoplasma gondii . Am J Trop Med Hyg 15:149-152
Wolf A, Cowen D（1937）Granulomatous encephalomyelitis due to an
encephalitozoon (encephalitozoic encephalomyelitis). In ”Human
Toxoplasmosis” (ed DO Ho-Yen and AWL Joss), pp:1-2, Oxford
University Press, New York
Zangerle R, Allerberger F, Pohl P, Fritsch P, Dierich MP (1991) High risk
of developing toxoplasmic encephalitis in AIDS patients seropositive
to Toxoplasma gondii. Med Microbiol Immunol 180:59-66