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用掺入脂质体的不同刚地弓形虫抗原免疫小鼠对弓形虫病的保护作用。

Protection against toxoplasmosis in mice immunized with different antigens of Toxoplasma gondii incorporated into liposomes.

作者信息

Elsaid M M, Vitor R W, Frézard F J, Martins M S

机构信息

Departamento de Parasitologia, Universidade Federal de Minas Gerais, Belo Horizonte, MG, 31270-901, Brasil.

出版信息

Mem Inst Oswaldo Cruz. 1999 Jul-Aug;94(4):485-90. doi: 10.1590/s0074-02761999000400010.

Abstract

Different toxoplasma antigens were entrapped within liposomes and evaluated, in this form, for their ability to protect Swiss mice against toxoplasma infection: soluble tachyzoite antigen (L/TAg), tissue cyst (L/CAg), tachyzoite plus tissue cyst (L/TCAg) or purified antigen of tachyzoite (L/pTAg). The protein used in L/pTAg was purified from tachyzoites using a stage-specific monoclonal antibody which reacted at a molecular weight of 32 kD in SDS PAGE and silver stain using reduced condition. To compare the immuno-adjuvant action of liposomes and of Freund's Complete Adjuvant (FCA), another group of mice was immunized with soluble tachyzoite antigen (STAg) emulsified in FCA (FCA/TAg). Control groups were inoculated with (STAg) alone, phosphate-buffered saline (PBS), FCA with PBS (FCA/PBS) and empty liposomes (L/PBS). Mice were inoculated subcutaneously with these antigens six, four and two weeks before a challenge with 80 tissue cysts of the P strain of Toxoplasma gondii orally. All mice immunized with or without adjuvant showed a humoral response, as measured by Elisa. However, no correlation was found between antibody titer and protection against the challenge. All mice immunized with L/pTAg or L/TCAg survived (100), whereas 80% and 90% of mice from groups which received respectively PBS or FCA/PBS and L/PBS died. All mice immunized with antigens entrapped within liposomes (L/TAg, L/CAg, L/TCAg and L/pTAg) showed low numbers of intracerebral cysts.

摘要

将不同的弓形虫抗原包裹于脂质体中,并以此形式评估其保护瑞士小鼠免受弓形虫感染的能力:可溶性速殖子抗原(L/TAg)、组织包囊(L/CAg)、速殖子加组织包囊(L/TCAg)或纯化的速殖子抗原(L/pTAg)。用于L/pTAg的蛋白质是使用阶段特异性单克隆抗体从速殖子中纯化得到的,该抗体在还原条件下的SDS-PAGE和银染中,在分子量为32 kD处发生反应。为了比较脂质体与弗氏完全佐剂(FCA)的免疫佐剂作用,另一组小鼠用FCA乳化的可溶性速殖子抗原(STAg)进行免疫(FCA/TAg)。对照组分别接种单独的(STAg)、磷酸盐缓冲盐水(PBS)、含PBS的FCA(FCA/PBS)和空脂质体(L/PBS)。在经口用80个刚地弓形虫P株组织包囊攻击前6周、4周和2周,给小鼠皮下接种这些抗原。通过酶联免疫吸附测定法(ELISA)检测,所有接种有或无佐剂的小鼠均显示出体液反应。然而,未发现抗体滴度与抵抗攻击的保护作用之间存在相关性。所有用L/pTAg或L/TCAg免疫的小鼠均存活(100%),而分别接受PBS或FCA/PBS和L/PBS的组中,80%和90%的小鼠死亡。所有用包裹于脂质体中的抗原(L/TAg、L/CAg、L/TCAg和L/pTAg)免疫的小鼠脑内包囊数量均较少。

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