Elsaid M M, Vitor R W, Frézard F J, Martins M S
Departamento de Parasitologia, Universidade Federal de Minas Gerais, Belo Horizonte, MG, 31270-901, Brasil.
Mem Inst Oswaldo Cruz. 1999 Jul-Aug;94(4):485-90. doi: 10.1590/s0074-02761999000400010.
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.
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。