ENHANCING IMMUNOGENICITY OF EPITOPE INSERTS IN FLAGELLIN

1.已经表明，鞭毛蛋白的高变区， 沙门氏菌菌株（可能用作伤寒疫苗）是一种 用于插入编码表位的DNA的合适基因座， 作为潜在有效的免疫原， 沙门氏菌病 代表IL-1 β的aa 163 -171的九肽具有 被证明具有免疫增强作用。 我们将测试表达式是否 这种九肽增强了对决定因素的免疫应答， 当它与该九肽并置、靠近或在空间上远离时， 插入沙门氏菌疫苗株的高变区。 2.许多多糖抗原被认为是bc T独立的， 感觉到对它们的反应不需要T细胞的帮助;回忆 免疫球蛋白同种型应答不被引发， 限制了这些抗原。 我们建议测试一种方法来转换 这些抗原的寡糖决定簇对T依赖性的影响， 记忆的启发和无限制的类切换。 此外，这一过程将使获得这种抗原成为可能， 表达为肽替代物作为背驮抗原， 鞭毛蛋白的高变区如1中所示。抗沙门氏菌单克隆抗体 Vi将用于筛选随机肽文库中的外源性肽。 - 交叉反应性肽，其将被反向翻译为DNA，以及 插入鞭毛蛋白的高变区表达。 的 基本假设是，在反应中识别的交叉反应性 将通过诱导免疫应答中的交叉反应性来匹配。 特别是这可能会大大提高伤寒的效用 疫苗正在测试，因为它们目前不引起抗Vi。

1. It has been shown that the hypervariable region of flagellin of strains of Salmonella (potentially useful as typhoid vaccines) is a suitable locus for insertion of DNA coding for epitopes which are of interest as potentially effective immunogens for diseases other than salmonellosis. A nonapeptide which represents aa163-171 of Il-1 beta has been shown to be immunoenhancing. We shall test whether the expression of this nonapeptide enhances immune response to determinants which are juxtaposed to, near or spatially removed from this nonapeptide when it is inserted in the hypervariable region of vaccine strains of Salmonella. 2. Many polysaccharide antigens are considered to bc T independent in the sense that response to them does not require T cell help; anamnestic responses are not elicited and the immunoglobulin isotype response is restricted with such antigens. We propose to test a method to convert oligosaccharide determinants of such antigens to T dependence enabling both the elicitation of memory and unrestricted class switching. Furthermore, the process would make it possible to have such antigens expressed as peptide surrogates as piggy-back antigens in the hypervariable region of flagellin as in 1. A monoclonal anti Salmonella Vi will be used to screen a random peptide library for adventitious cross-reactive peptides which will be back-translated to DNA, and inserted for expression in the hypervariable region of flagellin. The essential hypothesis is that cross-reactivity as recognized in reaction will bc matched by cross-reactivity in induction of immune response. Specifically this might greatly enhance the utility of the typhoid vaccines being tested since they do not at present elicit anti-Vi.