Gilardeau Truffinet M, Bossus M, Camus D, Delplace P, Mazingue C, Diesis E, Tartar A, Moreau S, Gras-Masse H, Banic D M
Unite INSERM 42, Villeneuve d'Ascq, France.
Pept Res. 1996 Mar-Apr;9(2):61-70.
The p126 Plasmodium falciparum antigen is processed into two fragments, p50 and p73, the latter one containing the subfragments p47 and p18 when the schizonts rupture. An absence of antibody response against the p126 antigen has been reported recently in H-2b mice and limited to the p73 processed fragment in H-2d mice. Synthetic peptides corresponding to various domains of the molecule have been used to immunize mice in order to overcome the absence of an immune response. Synthetic peptides corresponding to the N-terminus of p50 or p18 as well as to the C-terminus of p47 were unable to induce anti-peptide antibodies when injected carrier-free or coupled to ovalbumin. Synthetic peptides corresponding to the C-terminus of p18 or composed of 6 or 9 serines were able to induce anti-peptide antibodies when injected coupled to a carrier protein. However, none of these antibodies was able to recognize the native p126 molecule. Various synthetic peptides corresponding to the 6-octapeptide [Nt47 (6 x 8)] or the 4-octapeptide [Nt47(4 x 8)] repeat sequence localized at the N-terminus of the p47 have also been used to immunize mice. No antibodies were generated using a carrier-free [Nt47(6 x 8)-Cys]2 or [Nt47 (4 x 8)-Cys]2 peptide, an octameric multiple antigen peptide construct [Nt47(6 x 8)]-MAP or the [Nt47(6 x 8)] coupled to one or two palmitic acids. In contrast, [Nt47(6 x 8)]-Cys coupled to either tetanus toxoid (TT) or ovalbumin (OVA) and [Nt47(4 x 8)]-Cys coupled to OVA induced antibodies against the synthetic peptide and the native p126 molecule in both H-2d and H-2b mice. A multiple antigen peptide construct [Nt47(4 x 8)-MSP-3b]-MAP containing 4 [Nt47(4 x 8)] and 4 [MSP-3b] also induced antibodies against the synthetic peptide [Nt47(4 x 8)-Cys]2 and the native p126 molecule in both H-2d and H-2b mice.
恶性疟原虫的p126抗原在裂殖体破裂时被加工成两个片段,即p50和p73,后者包含亚片段p47和p18。最近报道,H-2b小鼠缺乏针对p126抗原的抗体反应,而H-2d小鼠的抗体反应仅限于p73加工片段。为了克服免疫反应的缺失,已使用与该分子不同结构域相对应的合成肽免疫小鼠。当无载体注射或与卵清蛋白偶联时,与p50或p18的N端以及p47的C端相对应的合成肽无法诱导抗肽抗体。与p18的C端相对应或由6个或9个丝氨酸组成的合成肽在与载体蛋白偶联注射时能够诱导抗肽抗体。然而,这些抗体均无法识别天然p126分子。与位于p47 N端的6个八肽[Nt47(6×8)]或4个八肽[Nt47(4×8)]重复序列相对应的各种合成肽也已用于免疫小鼠。使用无载体的[Nt47(6×8)-Cys]2或[Nt47(4×8)-Cys]2肽、八聚体多抗原肽构建体[Nt47(6×8)]-MAP或与一或两个棕榈酸偶联的[Nt47(6×8)]均未产生抗体。相比之下,与破伤风类毒素(TT)或卵清蛋白(OVA)偶联的[Nt47(6×8)]-Cys以及与OVA偶联的[Nt47(4×8)]-Cys在H-2d和H-2b小鼠中均诱导产生了针对合成肽和天然p126分子的抗体。一种包含4个[Nt47(4×8)]和4个[MSP-3b]的多抗原肽构建体[Nt47(4×8)-MSP-3b]-MAP在H-2d和H-2b小鼠中也诱导产生了针对合成肽[Nt47(4×8)-Cys]2和天然p126分子的抗体。