Department of Medicine, Stanford University Medical Center, CA 94305-5623, USA.
Blood. 2010 Mar 11;115(10):2077-87. doi: 10.1182/blood-2009-03-211375. Epub 2010 Jan 6.
Identifying the targets of immune response after allogeneic hematopoietic cell transplantation (HCT) promises to provide relevant immune therapy candidate proteins. We used protein microarrays to serologically identify nucleolar and spindle-associated protein 1 (NuSAP1) and chromatin assembly factor 1, subunit B (p60; CHAF1b) as targets of new antibody responses that developed after allogeneic HCT. Western blots and enzyme-linked immunosorbent assays (ELISA) validated their post-HCT recognition and enabled ELISA testing of 120 other patients with various malignancies who underwent allo-HCT. CHAF1b-specific antibodies were predominantly detected in patients with acute myeloid leukemia (AML), whereas NuSAP1-specific antibodies were exclusively detected in patients with AML 1 year after transplantation (P < .001). Complete genomic exon sequencing failed to identify a nonsynonymous single nucleotide polymorphism (SNP) for NuSAP1 and CHAF1b between the donor and recipient cells. Expression profiles and reverse transcriptase-polymerase chain reaction (RT-PCR) showed NuSAP1 was predominately expressed in the bone marrow CD34(+)CD90(+) hematopoietic stem cells, leukemic cell lines, and B lymphoblasts compared with other tissues or cells. Thus, NuSAP1 is recognized as an immunogenic antigen in 65% of patients with AML following allogeneic HCT and suggests a tumor antigen role.
鉴定异基因造血细胞移植（HCT）后免疫反应的靶标有望提供相关的免疫治疗候选蛋白。我们使用蛋白质微阵列血清学鉴定核仁与纺锤体相关蛋白 1（NuSAP1）和染色质组装因子 1 亚基 B（p60；CHAF1b）作为新抗体反应的靶标，这些抗体在异基因 HCT 后产生。Western blot 和酶联免疫吸附试验（ELISA）验证了它们在 HCT 后的识别，并对 120 名接受 allo-HCT 的其他患有各种恶性肿瘤的患者进行了 ELISA 检测。CHAF1b 特异性抗体主要在急性髓细胞白血病（AML）患者中检测到，而 NuSAP1 特异性抗体仅在移植后 1 年的 AML 患者中检测到（P <.001）。全基因组外显子测序未能在供体和受体细胞之间鉴定出 NuSAP1 和 CHAF1b 的非同义单核苷酸多态性（SNP）。表达谱和逆转录聚合酶链反应（RT-PCR）显示，NuSAP1 在骨髓 CD34(+)CD90(+)造血干细胞、白血病细胞系和 B 淋巴母细胞中表达明显高于其他组织或细胞。因此，NuSAP1 被鉴定为异基因 HCT 后 65%的 AML 患者的免疫原性抗原，并提示其具有肿瘤抗原的作用。
