Lundquist Rasmus, Nielsen Leif Kofoed, Jafarshad Ali, Soesoe Daw, Christensen Lars Harder, Druilhe Pierre, Dziegiel Morten Hanefeld
H:S Blodbanken KI2033, Copenhagen University Hospital, DK-2100 Copenhagen, Denmark.
Infect Immun. 2006 Jun;74(6):3222-31. doi: 10.1128/IAI.00928-05.
Immunoglobulins from individuals with immunity to malaria have a strong antiparasitic effect when transferred to Plasmodium falciparum malaria infected patients. One prominent target of antiparasitic antibodies is the merozoite surface antigen 3 (MSP-3). We have investigated the antibody response against MSP-3 residues 194 to 257 (MSP-3(194-257)) on the molecular level. mRNA from peripheral blood leukocytes from clinically immune individuals was used as a source of Fab (fragment antibody) genes. A Fab-phage display library was made, and three distinct antibodies designated RAM1, RAM2, and RAM3 were isolated by panning. Immunoglobulin G1 (IgG1) and IgG3 full-length antibodies have been produced in CHO cells. Reactivity with the native parasite protein was demonstrated by immunofluorescence microscopy, flow cytometry, and immunoblotting. Furthermore, the antiparasitic effect of RAM1 has been tested in vitro in an antibody-dependent cellular inhibition (ADCI) assay. Both the IgG1 and the IgG3 versions of the antibody show an inhibitory effect on parasite growth.
对疟疾具有免疫力的个体的免疫球蛋白在转移至感染恶性疟原虫的患者体内时具有强大的抗寄生虫作用。抗寄生虫抗体的一个主要靶点是裂殖子表面抗原3(MSP-3)。我们在分子水平上研究了针对MSP-3第194至257位氨基酸残基(MSP-3(194-257))的抗体反应。来自临床免疫个体外周血白细胞的mRNA用作Fab(片段抗体)基因的来源。构建了一个Fab噬菌体展示文库,并通过淘选分离出三种不同的抗体,分别命名为RAM1、RAM2和RAM3。已在CHO细胞中产生免疫球蛋白G1(IgG1)和IgG3全长抗体。通过免疫荧光显微镜、流式细胞术和免疫印迹证明了与天然寄生虫蛋白的反应性。此外,已在体外抗体依赖性细胞抑制(ADCI)试验中测试了RAM1的抗寄生虫作用。该抗体的IgG1和IgG3版本均对寄生虫生长显示出抑制作用。