IMMUNE MODIFICATION OF CYTOMEGALOVIRUS INFECTION IN BONE MARROW

骨髓中巨细胞病毒感染的免疫修饰

• 批准号: 6236630
• 负责人: John D Shanley
• 金额: $ 21.28万
• 依托单位: CITY OF HOPE NATIONAL MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 1998-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6236630
• 关键词: CHO cells; Herpesviridae vaccine; SDS polyacrylamide gel electrophoresis; active immunization; antibody formation; bone marrow transplantation; cell sorting; cellular immunity; cytomegalovirus; disease /disorder model; enzyme linked immunosorbent assay; gene expression; homologous transplantation; host organism interaction; humoral immunity; immunocytochemistry; laboratory mouse; microorganism immunology; nonhuman therapy evaluation; plasmids; transfection /expression vector; virus DNA; virus genetics; virus protein; western blottings

Despite recent progress in prophylaxis and treatment, CMV continues to be a major obstacle to success in allogeneic bone marrow transplantation (BMT). Studies have shown that the outcome of CMV infection in this setting is improved by both early reconstitution of recipient immunity to CMV and reduction of CMV replication. Furthermore, previous studies in both humans and animals have demonstrated that several facets of donor immunity to CMV can be transferred to BMT recipients and that such transfer improves outcome. This proposal will explore the ability to modify donor immunity to CMV by several immunization strategies. We will further test whether immunization immunity an be transferred to BMT recipients and alter the outcome of CMV infection. We hypothesize that immunity to CMV infection can be altered by immunization with either DNA of specific viral genes or with H2-compatible cellular vectors expressing specific gene products. This immunity will be manifested by humoral and cell mediated responses and by alterations in the course of infection. We further hypothesize that this immunization induced alteration in immunity to CMV can be transferred from a bone marrow donor tot he recipient of an allogeneic transplant. There are three specific aims of this project; Specific Aim 1: To develop optimal expression vectors for obtaining maximal expression of HCMV (pp65, pp150, pp28, pp71) and MCMV (gB and gH) proteins in vitro and in vivo. once, an optimal vector is developed, we will insert the herpes simplex-1 thymidine kinase (HSV 1-tk) gene into the expression vector to provide a suicidal constituent. We will also develop new murine gene vectors for study. Specific Aim 2: We will evaluate the expression plasmids containing specific CMV genes (HCMV pp65, pp150, pp28, pp71; MCMV gB, and gH) for their ability to alter host immunity in vivo, using DNA immunization and using H2-compatible cell vectors expression specific viral gene products. Specific Aim 3: Using an established murine model of allogeneic bone marrow transplantation, we will determine the efficacy of transferring immunization-induced immunity from a donor to the transplant recipient. These studies will determine the feasibility of altering donor immunity to CMV using immunization with either DNA of specific viral genes or with H2- compatible cellular vectors expressing specific gene products and the subsequent transfer of immunization induced immunity to recipients of allogenic marrow recipients.

尽管最近在预防和治疗方面取得了进展，但CMV仍然是 异基因骨髓移植成功的主要障碍 （BMT）。 研究表明，在这种情况下，CMV感染的结果 通过受体免疫力的早期重建， CMV和减少CMV复制。 此外，以往的研究 人类和动物都证明了捐赠者的几个方面 对CMV的免疫可以转移到BMT接受者， 改善结果。 该提案将探讨修改捐助者的能力， 通过几种免疫策略获得CMV免疫力。 我们将进一步测试 免疫接种是否可以转移到BMT接受者， CMV感染的后果。 我们假设，对CMV感染的免疫力可以被改变， 用特定病毒基因的DNA或用H2相容的 表达特定基因产物的细胞载体。 这份豁免权将是 表现为体液和细胞介导的反应， 感染的过程。 我们进一步假设，这种免疫诱导 对CMV免疫力的改变可以从骨髓供体转移 他接受了同种异体移植 该项目有三个具体目标：具体目标1： 用于获得HCMV最大表达的最佳表达载体（pp 65， pp 150、pp 28、pp 71）和MCMV（gB和gH）蛋白。 一旦开发出最佳载体，我们将插入单纯疱疹病毒-1 胸苷激酶（HSV 1-tk）基因导入表达载体，以提供一种 自杀成分 我们还将开发新的鼠基因载体， study. 具体目标2：我们将评估表达质粒 含有特异性CMV基因（HCMV pp 65、pp 150、pp 28、pp 71; MCMV gB，和 gH），因为它们能够利用DNA改变体内宿主免疫力 免疫接种和使用H2相容细胞载体表达特异性病毒 基因产物 具体目标3：使用已建立的小鼠模型， 异基因骨髓移植，我们将确定疗效 将免疫诱导的免疫从供体转移到移植物 收件人。 这些研究将确定改变供体免疫力的可行性， CMV使用特定病毒基因的DNA或H2- 表达特异性基因产物的相容性细胞载体， 随后的免疫接种转移诱导了免疫力， 同种异体骨髓受体。