Mattsson J, Uzunel M, Remberger M, Tammik L, Omazic B, Levitsky V, Zou J Z, Hentschke P, Ringdén O
Centre for Allogeneic Stem Cell Transplantation, Department of Clinical Immunology and Surgery, Karolinska Institute and Huddinge Hospital, Huddinge, Sweden.
Clin Exp Immunol. 2001 Jan;123(1):162-9. doi: 10.1046/j.1365-2249.2001.01429.x.
Two adults with primary liver cancer underwent liver transplantation from 5/6 and 4/6 major HLA-antigen mismatched unrelated donors. They were then conditioned with 4 x 2 Gy of total lymphoid irradiation, 120 mg/kg cyclophosphamide, 7.5 Gy total body irradiation and anti-T cell antibodies. Thereafter, the patients received T cell-depleted autologous: unrelated mismatched bone marrow in a proportion of 0.5:3.0 and 0.35:1.1 x 10(6) CD34+ cells/kg, respectively. After allogeneic stem cell transplantation (ASCT), both became mixed chimeras, as determined with polymerase chain reaction amplification of variable number tandem repeats from DNA obtained from CD3+, CD19+ and CD45+ magnetic bead-separated cells. Due to a reduction in donor T cells, the first patient was given 10(5) donor T cells/kg and became a complete donor chimera within 3 months. The second patient rejected all donor cells within 1 month after ASCT. Leucocytes normalized in both patients within 1 month. CD8+ cells normalized after 4 and 2 months in the two patients, respectively. However, CD4+, CD56+ and CD19+ cells remained low, except for a transient increase in patient 2. Lymphocyte responses to mitogens were negative in patient 1 from 1 to 5 months after ASCT. This patient also showed an oligoclonal pattern of the B cell repertoire, performed by CDR3 spectratyping. Epstein-Barr virus DNA in lymphocytes increased by 4-5 log in both patients. Prior to ASCT, recipients and donors were mutually reactive in mixed lymphocyte cultures (MLC). In the first patient, who became a complete donor chimera, the chimera cells showed no response to recipient or donor, but a positive response to third party. In the other patient, recipient cells reacted vigorously against donor lymphocytes at the time of rejection. Both patients suffered from overwhelming bacterial, fungal and viral infections, and died of pneumonia 5 and 3 months after ASCT, respectively. To conclude, with a major HLA-mismatch barrier, stable mixed chimerism seems difficult to achieve. The first patient became a full donor chimera and the second one rejected the graft. Both suffered from immune incompetence.
两名原发性肝癌成年患者接受了来自5/6和4/6主要HLA抗原错配的无关供体的肝移植。然后他们接受了4×2 Gy的全身淋巴照射、120 mg/kg环磷酰胺、7.5 Gy全身照射以及抗T细胞抗体预处理。此后，患者分别以0.5:3.0和0.35:1.1×10(6)个CD34+细胞/kg的比例接受了去除T细胞的自体：无关错配骨髓。异基因干细胞移植（ASCT）后，通过对从CD3+、CD19+和CD45+磁珠分离细胞获得的DNA进行可变数目串联重复序列的聚合酶链反应扩增确定，两人均成为混合嵌合体。由于供体T细胞减少，第一名患者接受了10(5)个供体T细胞/kg，3个月内成为完全供体嵌合体。第二名患者在ASCT后1个月内排斥了所有供体细胞。两名患者的白细胞均在1个月内恢复正常。两名患者的CD8+细胞分别在4个月和2个月后恢复正常。然而，除了第二名患者有短暂升高外，CD4+、CD56+和CD19+细胞仍然偏低。ASCT后1至5个月，第一名患者对丝裂原的淋巴细胞反应为阴性。该患者通过CDR3谱型分析还显示了B细胞库的寡克隆模式。两名患者淋巴细胞中的爱泼斯坦-巴尔病毒DNA均增加了4至5个对数。ASCT前，受体和供体在混合淋巴细胞培养（MLC）中相互反应。在成为完全供体嵌合体的第一名患者中，嵌合体细胞对受体或供体无反应，但对第三方有阳性反应。在另一名患者中，排斥时受体细胞对供体淋巴细胞有强烈反应。两名患者均遭受严重的细菌、真菌和病毒感染，分别在ASCT后5个月和3个月死于肺炎。总之，存在主要HLA错配障碍时，似乎难以实现稳定的混合嵌合状态。第一名患者成为完全供体嵌合体，第二名患者排斥了移植物。两人均存在免疫功能不全。
