Use of Bacteriophages to Prevent, Diagnose, and Treat Diseases

部分A.目前正在进行三方面的调查。1.使用 噬菌体在治疗人类疾病，因为细菌感染。我们目前正在 处理由克雷伯氏菌引起的感染我们成功地定义了噬菌体 鸡尾酒，照顾噬菌体耐药性，在感染过程中出现。 2.我们已经成功地将克雷伯氏菌改造成具有多个尾部的细菌， 噬菌体的宿主范围能够感染和杀死多个宿主物种。3.我们 我们成功地构建了T7噬菌体衍生物，其中我们克隆了一种肽， 抗生素基因（AMP），当感染给定的宿主时，也会产生大量的抗生素。 抗生素。这使得噬菌体能够通过AMP的作用杀死任何突变的细菌宿主 它对噬菌体有抵抗力然而，AMP的表达一直很差， 杀死噬菌体抗性宿主。但是当AMP的表达与可溶性的 蛋白质，表达合理。目前，我们已经建立了一个噬菌体库， 随机突变的肽抗生素和筛选他们的一个更有效的 抗菌活性。这些发现鼓励我们进一步推进这一进程。B部分。我们 正在使用我们先前开发的噬菌体展示技术， 预防传染病和癌症的疫苗。我们已经建立了 展示CLL白血病、疟疾和Covid 19的潜在抗原肽和蛋白质 病毒我们投入了大量的时间在工程，净化，验证和测试， 多个噬菌体构建体的真实性，这些构建体可能被开发为基于噬菌体的 COVID-19、CLL和疟疾疫苗。目前，我们正在进行多项实验， 分析关于在小鼠模型中测试基于噬菌体的疫苗以产生抗 COVID-19和胶质母细胞瘤我们目前正在制定未来的实验计划， 证明基于噬菌体的COVID-19疫苗的有效性。在这两种情况下，目标是相同的： 制备用于动物免疫原性研究的纯化的工程λ。在第一分支中， 在该项目中，我们用胶质母细胞瘤中存在的EGFRvIII抗原设计了一种噬菌体， 附着在λ噬菌体的D蛋白上。该项目的另一个分支涉及 同样的过程，但抗原是新冠病毒刺突蛋白的片段。这些 这些建筑物是作为先知建造的。然后诱导前噬菌体产生， 将其纯化并浓缩。目前正在测试各自的 动物的免疫原性

Part A. Three lines of investigations are being pursued currently. 1. Use of bacteriophages in treating human diseases because of bacterial infections. We are currently dealing with infections caused by Klebsiella. We have succeeded in defining bacteriophage cocktails that take care of bacteriophage resistance that arises during infection processes. 2. We have succeeded in engineering Klebsiella phages to have multiple tails to increase the host range of a phage to be capable of infecting and killing multiple hosts species. 3. We have succeeded to construct a T7 bacteriophage derivative in which we cloned a peptide antibiotic gene (AMP), which when infects a given host also produces vast amount of the antibiotic. This enables the phage to kill by the action of AMP any mutant bacterial host which becomes resistant to phage. However, the expression of AMP has been poor, which does not kill phage resistant hosts. But when the AMP expression is covalently linked to a soluble protein, the expression was reasonable. Currently, we have made a library of phage carrying random mutations in the peptide anti biotic and screening them for one with more effective anti-bacterial activity. These findings encourage us to pursue the process further. Part B. We are using our previously developed phage display technology using bacteriophage Lambda to make vaccines against infectious diseases as well as against cancer. We already constructed displaying potential antigenic peptides and proteins of CLL leukemia, malaria and Covid 19 virus. We invested a lot of time in engineering, purifying, validating, and testing the authenticity of multiple phage constructs that could potentially be developed as a phage-based vaccine for COVID-19, CLL, and malaria. Currently, we are performing several experiments and analyzing data on testing phage-based vaccines in a mouse model to generate immunity against COVID-19 and glioblastoma. We are currently working in the future experimental plan to demonstrate the efficacy of phage-based COVID-19 vaccine. The goal is the same in both cases: to prepare purified, engineered lambda for animal immunogenecity study. In the first branch of the project, we engineered a phage with the EGFRvIII antigen present in glioblastoma and attached to the D protein of the lambda phage. The other branch of the project involved the same process, but the antigen was a fragment of the covid-19 spike protein. These constructions were made as prophages. The prophages were then induced phage production and phages purified and concentrated. The respective phages are now being tested for immunogenecity in animals.