Carlsson J, Udomsangpetch R, Wåhlin B, Ahlborg N, Berzins K, Perlmann P
Department of Immunology, University of Stockholm, Sweden.
Exp Parasitol. 1990 Oct;71(3):314-25. doi: 10.1016/0014-4894(90)90036-c.
For selection of immunogens capable of inducing high levels of antibodies reactive with the Plasmodium falciparum antigen Pf155/RESA, rabbits were immunized with synthetic peptides corresponding to sequences based on the repeat subunits EENVEHDA and (EENV)2 from the C-terminus of this antigen. The antibodies obtained were analyzed with regard to binding to synthetic peptides in ELISA and to reactivity with parasite antigens by immunofluorescence or immunoblotting. All antisera reacted with both the peptides EENVEHDA and (EENV)2 as well as with Pf155/RESA. Antibody fractions specific for each of the two peptides were prepared by affinity chromatography on insolubilized peptides. Strong reactivity with antigens in the membrane of erythrocytes infected with early stages of the parasite as well as reactivity with Pf155/RESA in immunoblotting correlated with reactivity of antibody with (EENV)2. Antibody preparations reactive with EENVEHDA and depleted of (EENV)2 reactivity showed only a weak reactivity with Pf155/RESA but reacted also with P. falciparum polypeptides of 250, 210, and 88 kDa. In immunofluorescence, these antibodies stained mainly the intraerythrocytic parasite. Both EENVEHDA- and (EENV)2-specific antibodies inhibited merozoite reinvasion in P. falciparum in vitro cultures, the latter antibodies being the most efficient. This study defines the specificity and cross-reactivity with other P. falciparum antigens of antibodies to the C-terminal repeats of Pf155/RESA.
为了筛选能够诱导产生与恶性疟原虫抗原Pf155/RESA发生高反应性抗体的免疫原,用基于该抗原C末端重复亚基EENVEHDA和(EENV)2序列的合成肽对兔子进行免疫。通过酶联免疫吸附测定(ELISA)分析所获得的抗体与合成肽的结合情况,并通过免疫荧光或免疫印迹分析其与寄生虫抗原的反应性。所有抗血清均与肽EENVEHDA和(EENV)2以及Pf155/RESA发生反应。通过在固定化肽上进行亲和层析制备了对两种肽各自具有特异性的抗体组分。与感染寄生虫早期阶段的红细胞膜中的抗原具有强反应性以及在免疫印迹中与Pf155/RESA具有反应性,这与抗体与(EENV)2的反应性相关。与EENVEHDA反应且缺乏(EENV)2反应性的抗体制剂与Pf155/RESA仅表现出弱反应性,但也与250、210和88 kDa的恶性疟原虫多肽发生反应。在免疫荧光中,这些抗体主要对红细胞内的寄生虫进行染色。EENVEHDA特异性抗体和(EENV)2特异性抗体均能抑制恶性疟原虫体外培养中的裂殖子再入侵,后者的抗体最为有效。本研究确定了针对Pf155/RESA C末端重复序列的抗体与其他恶性疟原虫抗原的特异性和交叉反应性。