Biology of the Immune Response

该项目的目标是确定和描述某些与癌症发病率增加有关的原发免疫缺陷疾病的遗传基础。虽然这些疾病很少见，但他们的研究在定义人类免疫功能中以前未被怀疑的重要因素方面具有极大的指导意义。免疫生理科进行临床转化研究计划。实验室中对重组免疫刺激分子的基本观察导致了两项临床试验。重组CD40L在CD40L缺乏症患者中的临床试验(00-I-006)和BLySTM在选择性免疫球蛋白A缺乏症患者中的临床试验(OMIM 137100)均已成功完成，并获得了全部患者的收益。目前正在计划对XHIM和选择性IgA缺乏症患者进行进一步的临床试验。对Wiskott-Aldrich综合征(Was)和责任基因(WASP)的研究仍在继续。该假说是，对Wasp蛋白功能的了解(S)将有助于深入了解WAS的分子机制，并为创新的诊断和治疗策略提供重要线索。利用实验室制备的抗WASP的单抗，我们鉴定了几种与WASP蛋白相互作用并在细胞骨架组织和功能中发挥关键作用的蛋白质。我们将这一知识带到了临床上，看看患者可能提供什么见解，从而对WASP蛋白的分子理解产生独特的见解。理解WAS的困难之一是，没有一个中心可以积累足够的患者来定义疾病的全部基因和临床谱系。我们感兴趣的是，患者身上发生的基因突变与观察到的临床表型之间是否存在相关性。为了实现这一目标，该科帮助组织了一个大型联盟，该联盟能够积累227个家庭的262名患者的分子和临床数据。本研究提供了有关遗传缺陷在患者中发生的谱和频率以及AS的临床谱以及遗传缺陷与临床疾病之间的相关性的有价值的信息。为了进一步推进这些研究，我们使用抗WASP的单抗开发了一种策略，利用多色细胞表面和细胞内流式细胞术在单细胞水平上测量WASP蛋白的表达。我们能够测量WASP在正常人外周血细胞中的表达，并证明在IS患者的细胞中WASP表达不足。此外，这种缺陷的表达可以通过异基因骨髓移植来纠正。自20世纪70年代末以来，骨髓移植一直被用作AS的最终治疗方法。主要组织相容性复合体(MHC)相合的、MHC半相合的和匹配无关的造血干细胞捐献者使用了一种有点经验性的移植前条件处理方案，并取得了不同的成功。虽然这些移植获得了不同程度的成功，但关于什么是必要的或足够的植入才能获得良好的临床结果，数据很少。此外，关于植入的持久性或植入的细胞谱系的数据很少。使用现有的WASP表达的流式细胞术检测，我们研究了12例异基因骨髓移植后的患者。6个完全植入供体细胞，6个是不同细胞系中供体细胞和宿主细胞(嵌合体)的混合体。这些结果表明，在美国对骨髓移植进行更多的研究将有助于确定骨髓移植的最佳条件。

The goal of this project is to identify and characterize the genetic basis for certain of the primary immunodeficiency disorders, which are associated with an increased incidence of cancer. While these disorders are rare, their study has been extremely instructive in defining previously unsuspected elements of importance in human immune function. The Immunophysiology Section conducts a clinical translational research program. Basic observations in the laboratory on recombinant immunostimulatory molecules have led to two clinical trials. A clinical trial (00-I-006) of recombinant CD40 ligand (CD40L) in patients with CD40L deficiency (XHIM, OMIM 308230) and a clinical trial of BLySTM in patients with selective IgA deficiency (OMIM 137100) have both been successfully completed with full patient accrual. Further clinical trials in patients with XHIM and selective IgA deficiency are currently being planned. Studies of the Wiskott-Aldrich syndrome (WAS) and the responsible gene (WASP) have continued. The hypothesis is that an understanding of WASP protein function(s) would provide insight into the molecular mechanisms underlying the WAS and provide important clues for innovative diagnostic and therapeutic strategies. Using monoclonal antibodies to WASP produced in the laboratory, we identified several proteins, which interacted with the WASP protein and played critical roles in cytoskeletal organization and function. We took this knowledge to the clinic to see what insights the patients might provide which could yield unique insights into a molecular understanding of the WASP protein. One of the difficulties in understanding the WAS is that no one center could accumulate sufficient patients to define the full genetic and clinical spectrum of the disease. We were interested in whether there was a correlation between the genetic mutations, which occurred in the patients, and the clinical phenotype that was observed. To accomplish this goal, the Section helped organize a large consortium, which was able to accumulate molecular and clinical data on 262 patients from 227 families. This study provided valuable information about the spectrum and frequency of the genetic defects occurring in patients and the clinical spectrum of the WAS as well as correlations between the genetic defects and the clinical disease. To further advance these studies, we used a monoclonal anti-WASP antibody to develop a strategy to measure WASP protein expression at the single cell level using multicolor cell surface and intracellular flow cytometry. We were able to measure WASP expression in the peripheral blood cells of normal individuals and document deficient WASP expression in cells from WAS patients. Moreover, this deficient expression could be corrected by allogeneic bone marrow transplantation. Bone marrow transplantation has been used as the definitive treatment of the WAS since the late 1970s. Using a somewhat empiric pre-transplant conditioning regimen, Major Histocompatibility Complex (MHC)-matched, MHC- haploidentical, and matched-unrelated hematopoetic stem cell donors have been used with varying success. While these transplants were successful to varying degrees, there was sparse data regarding what was necessary or sufficient in terms of engraftment to obtain a good clinical outcome. Moreover, there was little data about the durability of engraftment or the cell lineages engrafted. Using the available flow cytometric test for WASP expression, we studied 12 patients following allogeneic bone marrow transplantation. Six were fully engrafted with donor cells while six were a mixture of donor and host cells (chimeric) in various cell lineages. These results suggested that additional studies of bone marrow transplants in the WAS would be useful in defining the optimal conditions for bone marrow transplantation.