Ya Zhiya, Hailemichael Yared, Overwijk Willem, Restifo Nicholas P
National Cancer Institute, Surgery Branch, Bethesda, Maryland.
Department of Melanoma Medical Oncology-Research, Division of Cancer Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas.
Curr Protoc Immunol. 2015 Feb 2;108:20.1.1-20.1.43. doi: 10.1002/0471142735.im2001s108.
This unit describes protocols for developing tumors in mice, including subcutaneous growth, pulmonary metastases of B16 melanoma, and spontaneous melanoma in B-Raf V600E/PTEN deletion transgenic mouse models. Two immunization methods to prevent B16 tumor growth are described using B16.GM-CSF and recombinant vaccinia virus. A therapeutic approach is also included that uses adoptive transfer of tumor antigen-specific T cells. Methods including CTL induction, isolation, testing, and genetic modification of mouse T cells for adoptive transfer by using retrovirus-expressing genes of interest are provided. Additional sections, including growing B16 melanoma, enumerating pulmonary metastases, tumor imaging technique, and use of recombinant viruses for vaccination, are discussed together with safety concerns.
本单元描述了在小鼠体内诱导肿瘤的实验方案，包括皮下肿瘤生长、B16黑色素瘤的肺转移以及B-Raf V600E/PTEN缺失转基因小鼠模型中的自发性黑色素瘤。还描述了两种使用B16.GM-CSF和重组痘苗病毒预防B16肿瘤生长的免疫方法。此外，还包括一种使用肿瘤抗原特异性T细胞过继转移的治疗方法。提供了相关方法，包括用于过继转移的小鼠T细胞的CTL诱导、分离、检测以及利用表达感兴趣基因的逆转录病毒进行基因改造。还讨论了其他部分内容，包括培养B16黑色素瘤、计数肺转移灶、肿瘤成像技术以及使用重组病毒进行疫苗接种，并提及了安全问题。
