Mixed chimera status in patients with Wiskott-Aldrich syndrome (WAS) after hematopoietic stem cell transplantation ; evaluation by flow cytometric analysis of intracellular WAS protein expression.

Wiskott-Aldrich综合征（WAS）患者造血干细胞移植后的混合嵌合体状态；

• 批准号: 13670776
• 负责人: ARIGA Tadashi
• 金额: $ 1.98万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670776/
• 关键词: Wiskott-Aldrich syndrome; Hematopoietic stem cell transplantation; Flow cytometry; WASP; Mixed chimera

The Wiskott-Aldrich syndrome (WAS) is caused by defects in the WAS protein (WASP) gene on the X-chromosome. Previously we reported that flow cytometric analysis of intracellular WASP expression (FCM-WASP) was useful for the diagnosis of WAS patients and carriers. In this study, we applied FCM-WASP to evaluate the mixed chimera (MC) status of 12 WAS patients who underwent hematopoietic stem cell transplantation (HST). After HST, donor and recipient-derived peripheral blood mononuclear cells (PBMC) could be easily distinguished by this method, as the donor cells are observed to be WASP^<bright>, while the defective recipient cells are WASP^<dim>. Furthermore, by two-color FCM-WASP, the MC status could be characterized by cell lineage. Six of 12 WAS patients were revealed to have the MC status after HST, while others had the complete chimera status. The MC was observed in every cell lineage examined. However, among PBMC, recipient cells were most commonly observed in the monocyte population. Finally, to investigate the naive/memory status of donor and recipient T cells in these patients, three-color FCM-WASP using anti-CD45RA or CD45RO was performed. It was demonstrated that, in contrast to WASP^<bright> T cells, most WASP^<dim> T cells remained naive (CD45RA^+/RO^~) more than one year after HST. No imbalance in the ratio of naive/memory T cells was observed in WAS patients before HST. We conclude that FCM-WASP is a potentially useful method for clinical follow-up of WAS patients who underwent HST. This study may also have significant implications regarding the role of WASP during hematopoietic development.

维斯科特-奥尔德里奇综合征（WAS）是由x染色体上WAS蛋白（WASP）基因的缺陷引起的。以前我们报道过流式细胞术分析细胞内WASP表达（FCM-WASP）对was患者和携带者的诊断有用。在本研究中，我们应用FCM-WASP评估了12例接受造血干细胞移植（HST）的WAS患者的混合嵌合体（MC）状态。HST后，供体和受体来源的外周血单个核细胞（PBMC）可以很容易地区分，供体细胞为WASP^<亮>，而缺陷受体细胞为WASP^<暗>。此外，通过双色FCM-WASP， MC状态可以通过细胞谱系来表征。12例WAS患者中有6例在HST后显示MC状态，而其他患者则具有完全嵌合体状态。在每个细胞系中都观察到MC。然而，在PBMC中，受体细胞最常见于单核细胞群。最后，为了研究这些患者供体和受体T细胞的初始/记忆状态，使用抗cd45ra或CD45RO进行三色FCM-WASP。结果表明，与WASP^<亮> T细胞相比，大多数WASP^<暗> T细胞在HST后一年多仍保持初始状态（CD45RA^+/RO^~）。在HST前，未观察到was患者初始/记忆T细胞比例失衡。我们得出结论，FCM-WASP是一种潜在的有用的方法，用于WAS患者接受HST的临床随访。这项研究也可能对WASP在造血发育中的作用有重要意义。

项目成果

Tadashi Ariga, Tatsuro Kondoh, Koji Yamaguchi, Masahumi Yadama, Satoshi Sasaki, David L. Nelson, Hisami Ikeda, Kunihiko Kobayashi, Hiroyuki Moriuchi, Yukio Sakiyama: "Spontaneous In Vivo Reversion of an Inherited Mutation in the Wiskott-Aldrich Syndrome"T

Tadashi Ariga、Tatsuro Kondoh、Koji Yamaguchi、Masahumi Yadama、Satoshi Sasaki、David L. Nelson、Hisami Ikeda、Kunihiko Kobayashi、Hiroyuki Moriuchi、Yukio Sakiyama：“Wiskott-Aldrich 综合征中遗传性突变的自发体内逆转”T

Ariga T: "Molecular Basis for Paradoxical Carriers of Adenosine Deaminase(ADA) Deficiency That Show Extremely Low Levels of ADA Activity in Peripheral Blood Cells Without Immunodeficiency"J Immmunol. 166. 1698-1702 (2001)

Ariga T：“腺苷脱氨酶 (ADA) 缺乏症的矛盾载体的分子基础，在无免疫缺陷的外周血细胞中显示出极低水平的 ADA 活性”J Immmunol。

Ariga T: "Spontaneous In Vivo Reversion of an Inherited Mutation in the Wiskott-Aldrich Syndrome"Journal of Immunology. 166. 5245-5249 (2001)

Ariga T：“Wiskott-Aldrich 综合征遗传性突变的自发体内逆转”免疫学杂志。

Yamaguchi K: "Mixed chimera status of 12 patients with Wiskott-Aldrich syndrome (WAS) after hematopoietic stem cell transplantation ; evaluation by flow cytometric analysis of intracellular WAS protein expression"Blood. 100. 1208-1214 (2002)

Yamaguchi K：“12 名 Wiskott-Aldrich 综合征 (WAS) 患者造血干细胞移植后的混合嵌合体状态；通过流式细胞术分析细胞内 WAS 蛋白表达进行评估”血液。

Ariga T: "T-cell linesfrom 2 patients with adenosine deaminase(ADA) deficiency showed the restoration of ADA activity resulted from the reversion of an inherited mutation"Blood. 97. 2896-2899 (2001)

Ariga T：“来自 2 名腺苷脱氨酶 (ADA) 缺乏症患者的 T 细胞系显示，由于遗传突变的逆转，ADA 活性得以恢复”Blood.