ALLOIMMUNIZATION TO PATERNAL LEUKOCYTES FOR HDN

HDN 父本白细胞同种免疫

• 批准号: 6536330
• 负责人: KENNETH J MOISE
• 金额: $ 7.28万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-09 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6536330
• 关键词: animal breeding; antigen antibody reaction; blood group antigens; blood group incompatibility; blood groups; blood tests; disease /disorder model; erythroblastosis fetalis; erythrocytes; female; flow cytometry; hemoglobin F; histocompatibility; histocompatibility antigens; histocompatibility gene; histocompatibility typing; immunohematology; laboratory rabbit; leukocytes; phagocytosis; prenatal diagnosis; reticuloendothelial system; tissue /cell culture

DESCRIPTION: (Provided by Applicant): Despite the widespread use if prophylactic immune globulin, HDN, secondary to maternal sensitization to the RhD antigen, continues to complicate 6 in 1,000 births in the United States. In severe cases, the intrauterine transfusion of red cells to the fetus can be life-saving. However, despite this technical advancement, 15 to 25% of fetuses still succumb to their anemia. The long-term objective of this investigation is to develop a unique immunologic approach to the treatment of red cell alloimmunization in pregnancy, thereby negating the need for invasive fetal therapy. Such an advance, will allow couples plagued by this disease to more likely achieve a successful outcome to their pregnancy. The first aim of this investigation is to establish an animal model for hemolytic disease of the newborn. Female rabbits will be sensitized to incompatible red cells then bred with a buck with a compatible red cell type producing a control litter. Fetal blood sampling using ultrasound guidance will be used to assess levels of fetal hemoglobin. Neonatal hemoglobins and assessment for hepatosplenomegaly will also be undertaken. The does will be bred a second time with a red cell incompatible buck producing an affected anemic litter. Fetal and neonatal assessments will be repeated. The second aim of the study will be to alloimmunize does to paternal leukocyte antigens. Bucks that are incompatible with specific does at multiple rabbit leukocyte antigen (RLA) loci will be given a series of injections with granulocyte stimulating factor to increase the circulating white blood cell pool. Multiple immunizations of the does will then be undertaken using white cells from these bucks; anti-RLA titers will de assessed using flow cytometry. The third aim of the study will be to prove that immunization to paternal RLA antigens will decrease the severity of the HDN as compared to the second, incompatible breeding. A third breeding will be undertaken with the buck that is incompatible for both red cell and RLA antigens. Fetal and neonatal assessments will be repeated as in the two prior breeding. The fourth aim of the investigation will be to determine if the antibodies to the RLA-A or RLA-D locus are responsible for the protective effect for HDN of paternal anti-RLA antibodies. Specific does will be matched to bucks for either their RLA-A or RLA-D loci before their initial alloimmunization to white blood cells. Incompatible breeding will then be undertaken. The fifth aim of the study will be to investigate the in vitro protective effects of anti-RLA antibodies. A monocyte monolayer assay will be performed using the spleens and sensitized red cells of the pups from the three serial breedings to determine the level of phagocytosis of the fetal reticuloendothelial system.

描述：（由申请人提供）：尽管有广泛使用 预防性免疫球蛋白，HDN，继发于母体敏化的 RHD抗原在美国继续使1,000个分娩中有6个复杂化。 在严重的情况下，红细胞向胎儿输血可能是 救生。 但是，尽管有这种技术进步，但15％至25％ 胎儿仍然屈服于贫血。 这个长期目标 研究是为了开发一种独特的免疫学方法来治疗 红细胞在怀孕中同种免疫，从而消除了侵入性的需求 胎儿疗法。 这样的进步将使夫妻困扰这种疾病 更有可能取得成功的妊娠结局。 第一个目的 这项研究是为了建立一种用于溶血疾病的动物模型 新生儿。 雌性兔子将敏感到不兼容的红细胞 用雄鹿繁殖，带有兼容的红细胞类型，产生控制垃圾。 使用超声指导的胎儿血液采样将用于评估水平 胎儿血红蛋白。 新生儿血红蛋白和评估 还将进行肝肾上腺全瘤。 确实会繁殖一秒钟 与红细胞不兼容的雄鹿产生受影响的贫血垃圾的时间。 将重复胎儿和新生儿评估。 研究的第二个目标 将是对父亲白细胞抗原的同种免疫。 那是雄鹿 在多个兔白细胞抗原（RLA）上与特定的特异性不兼容 将为基因座提供一系列注射粒细胞刺激因子 增加循环的白细胞池。 多次免疫 然后将使用这些雄鹿的白色细胞进行操作。抗RLA 滴度将使用流式细胞仪进行DE评估。 研究的第三个目标将 证明对父亲RLA抗原的免疫将减少 与第二个不兼容的育种相比，HDN的严重程度。 第三 繁殖将与雄鹿一起进行，这两种红色都不兼容 细胞和RLA抗原。 胎儿和新生儿评估将像在 两个先前的繁殖。 调查的第四个目标将是 确定RLA-A或RLA-D基因座的抗体是否负责 父亲抗RLA抗体的HDN的保护作用。 具体 将在他们的RLA-A或RLA-D Loci之前与雄鹿匹配 对白细胞的初始同种免疫。 不兼容的育种将 然后进行。 该研究的第五个目标是调查 抗RLA抗体的体外保护作用。 单核细胞单层测定法 将使用脾脏和幼崽的敏化红细胞进行 确定胎儿吞噬作用水平的三个系列繁殖 网状内皮系统。

项目成果

Paternal leukocyte alloimmunization as a treatment for hemolytic disease of the newborn in a rabbit model.

父本白细胞同种免疫治疗兔模型新生儿溶血病。

• DOI: 10.1067/mob.[phone]
• 发表时间: 2002
• 期刊: American journal of obstetrics and gynecology
• 影响因子: 9.8
• 作者: Whitecar,PaulW;Farb,Rod;Subramanyam,Lalitha;Dorman,Karen;Balu,RukminiB;MoiseJr,KennethJ
• 通讯作者: MoiseJr,KennethJ