Donor lymphocytes expressing the herpes simplex virus thymidine kinase suicide gene: detailed immunological function following add-back after haplo-identical transplantation

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• 作者：Hisayoshi Hashimoto¹ ; Shigehisa Kitano² ; ³ ; Shizuka Yamagata⁴ ; Akiko Miyagi Maeshima⁵ ; Ryosuke Ueda¹ ; Ayumu Ito⁴ ; Kohei Tada³ ; ⁴ ; Shigeo Fuji⁴ ; Takuya Yamashita⁴ ; Daisuke Tomura⁶ ; Ikuei Nukaya⁶ ; Junichi Mineno⁶ ; Takahiro Fukuda⁴ ; Shinichiro Mori⁷ ; Yoichi Takaue⁸ ; Yuji Heike⁹ ; ¹⁰ ; ^{heiyuji@luke.ac.jp" class="auth_mail" title="E-mail the corresponding author}
• 关键词：haplo-identical hematopoietic stem cell transplantation ; herpes simplex virus thymidine kinase suicide gene ; T-cell function ; T-cell repertoire ; insertional oncogenesis
• 刊名：Cytotherapy
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：17
• 期：12
• 页码：1820-1830
• 全文大小：1385 K

文摘

Haplo-identical hematopoietic stem cell transplantation (HSCT) with add-back of donor lymphocytes expressing the herpes simplex virus thymidine kinase suicide gene (TK cells) is one of the most widely applied promising new gene therapy approaches. However, the immunological status of added-back TK cells after HSCT has yet to be well characterized.

Methods

We investigated TK cells through the use of flow cytometry, T-cell receptor (TCR) Vβ repertoire spectratyping and linear amplification-mediated polymerase chain reaction followed by insertion site analysis in a patient enrolled in our clinical trial.

Results

A comparison of onset with remission of acute graft-versus-host disease confirmed that TK cells were predominantly eliminated and that proliferative CD8⁺ non-TK cells were also depleted in response to ganciclovir administration. The TCR Vβ-chain repertoire of both TK cells and non–TK cells markedly changed after administration of ganciclovir, and, whereas the TCR repertoire of non–TK cells returned to a normal spectratype long after transplantation, that of TK cells remained skewed. With the long-term prophylactic administration of acyclovir, TK cells oligoclonally expanded and the frequency of spliced variants of TK cells increased. Known cancer-associated genes were not evident near the oligoclonally expanded herpes simplex virus (HSV)-TK insertion sites.

Conclusions

We demonstrate obvious differences in immunological status between TK cells and non-TK cells. In addition, we speculate that long-term prophylactic administration of acyclovir increases the risk of oligoclonal expansion of spliced forms of TK cells.