Immunological characteristics of a synthetic peptide associated with a catalytic domain of mutans streptococcal glucosyltransferase.
Immunological characteristics of a synthetic peptide associated with a catalytic domain of mutans streptococcal glucosyltransferase.
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与变形链球菌葡萄糖基转移酶催化结构域相关的合成肽的免疫学特征。
DOI：
10.1128/iai.62.12.5470-5476.1994
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发表时间：
1994
影响因子：
3.1
通讯作者：
Eastcott,J
中科院分区：
文献类型：
--
作者：
Smith,DJ;Taubman,MA;King,WF;Eida,S;Powell,JR;Eastcott,J
测量由四个拷贝的合成21-mer肽DANFDSIRVDAVDNVDADLLQ组成的多抗原肽构建体的免疫原性。该肽的组成来源于含有参与催化的天冬氨酸的变异链球菌葡糖基转移酶（GTF）的N-末端区域中的序列。将该肽（CAT）构建体合成为赖氨酸骨架上的四聚体，并皮下注射到Sprague-Dawley大鼠中用于多克隆抗体形成或腹腔注射到BALB/c小鼠中，然后将脾细胞与Sp2/0Ag 14小鼠骨髓瘤细胞融合用于单克隆抗体形成。所得大鼠抗血清和小鼠单克隆抗体与CAT和来自远缘链球菌和变形链球菌的天然GTF同工酶反应（在酶联免疫吸附试验和Western印迹[免疫印迹]分析中）。功能性抑制S.用免疫球蛋白M抗CAT单克隆抗体（> 80%抑制）和大鼠血清（约17%抑制）证明了远缘牛GTF-I。该单克隆抗体制剂也适度地抑制了S.变形杆菌GTF混合物。这些结果表明，CAT肽含有B细胞表位，这些表位类似于完整的变形链球菌GTF，并有可能引发抗体，可以抑制GTF功能。因此，该肽构建体内的序列可能具有包含在合成龋齿疫苗中的价值。
The immunogenicity of a multiple antigenic peptide construct consisting of four copies of the synthetic 21-mer peptide DANFDSIRVDAVDNVDADLLQ was measured. The composition of this peptide was derived from a sequence in the N-terminal region of mutans streptococcal glucosyltransferases (GTFs) containing an aspartic acid implicated in catalysis. The peptide (CAT) construct was synthesized as a tetramer on a lysine backbone and subcutaneously injected into Sprague-Dawley rats for polyclonal antibody formation or intraperitoneally injected into BALB/c mice, and then spleen cell fused with Sp2/0Ag14 murine myeloma cells for monoclonal antibody formation. The resulting rat antisera and mouse monoclonal antibodies reacted with CAT and with native GTF isozymes from Streptococcus sobrinus and Streptococcus mutans (in enzyme-linked immunosorbent assay and Western blot [immunoblot] analyses). Functional inhibition of the water-insoluble glucan synthetic activity of S. sobrinus GTF-I was demonstrated with an immunoglobulin M anti-CAT monoclonal antibody (> 80% inhibited) and with rat sera (approximately 17% inhibited). The monoclonal antibody preparation also modestly inhibited the water-soluble glucan synthetic activity of an S. mutans GTF mixture. These results suggest that the CAT peptide contains B-cell epitopes that are similar to those of intact mutans streptococcal GTFs and has the potential to elicit antibody that can inhibit GTF function. Thus, sequences within this peptide construct may have value for inclusion in a synthetic dental caries vaccine.