de Velasco E A, Merkus D, Anderton S, Verheul A F, Lizzio E F, Van der Zee R, Van Eden W, Hoffman T, Verhoef J, Snippe H
Eijkman-Winkler Institute for Medical and Clinical Microbiology, Utrecht University, The Netherlands.
Infect Immun. 1995 Mar;63(3):961-8. doi: 10.1128/iai.63.3.961-968.1995.
Improvement of antibody responses to polysaccharides through their linkage to proteins is thought to be mediated by protein-specific T helper (Th) cells. To investigate whether the carrier protein of a conjugate could be substituted by a Th epitope, Streptococcus pneumoniae type 17F polysaccharide (PS) was bromoacetylated and coupled to different peptides via their carboxy-terminal cysteines. Two peptides, one from the mycobacterial 65-kDa heat shock protein (hsp65) and the other from influenza virus hemagglutinin, are well-known Th epitopes. Two other peptides were selected from the pneumolysin sequence by Th epitope prediction methods; one of them was synthesized with cysteine either at the carboxy or the amino terminus. Three conjugates consistently elicited in mice anti-PS immunoglobulin M (IgM) and IgG responses that were not observed upon immunization with derivatized PS without peptide. The same conjugates induced no anti-PS antibody responses in athymic (nu/nu) mice, whereas clear responses were elicited in euthymic (nu/+) controls, demonstrating the thymus-dependent character of these conjugates. Only the three conjugates inducing anti-PS responses were capable of eliciting antipeptide antibodies. One of the immunogenic conjugates was studied in more detail. It induced significant protection and an anti-PS IgG response comprising all subclasses. On the basis of these results and proliferation studies with peptide and conjugate-primed cells, it is concluded that linkage of Th epitopes to PS in the right orientation enhances its immunogenicity in a thymus-dependent manner. Future possibilities for using peptides as carriers for inducing antibody responses to poorly immunogenic saccharide antigens are discussed.
通过将多糖与蛋白质连接来改善抗体反应被认为是由蛋白质特异性辅助性T(Th)细胞介导的。为了研究结合物的载体蛋白是否可以被一个Th表位替代,17F型肺炎链球菌多糖(PS)被溴乙酰化,并通过其羧基末端的半胱氨酸与不同的肽偶联。两种肽,一种来自分枝杆菌65 kDa热休克蛋白(hsp65),另一种来自流感病毒血凝素,是众所周知的Th表位。另外两种肽是通过Th表位预测方法从肺炎溶血素序列中选择的;其中一种在羧基末端或氨基末端合成了半胱氨酸。三种结合物在小鼠中一致地引发了抗PS免疫球蛋白M(IgM)和IgG反应,而用未偶联肽的衍生化PS免疫时未观察到这种反应。相同的结合物在无胸腺(nu/nu)小鼠中未诱导出抗PS抗体反应,而在有胸腺(nu/+)对照中引发了明显的反应,证明了这些结合物的胸腺依赖性特征。只有三种诱导抗PS反应的结合物能够引发抗肽抗体。对其中一种免疫原性结合物进行了更详细的研究。它诱导了显著的保护作用和包含所有亚类的抗PS IgG反应。基于这些结果以及对肽和结合物致敏细胞的增殖研究,可以得出结论,Th表位以正确的方向与PS连接以胸腺依赖的方式增强了其免疫原性。还讨论了使用肽作为载体来诱导对免疫原性差的糖类抗原产生抗体反应的未来可能性。