A Rapid Embryonic Stem Cell-Based Mouse Model for B-cell Lymphomas Driven by Epstein-Barr Virus Protein LMP1.
A Rapid Embryonic Stem Cell-Based Mouse Model for B-cell Lymphomas Driven by Epstein-Barr Virus Protein LMP1.
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DOI：
10.1158/2326-6066.cir-15-0058
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发表时间：
2015-06
影响因子：
10.1
通讯作者：
Alt FW
中科院分区：
文献类型：
--
作者：
Ba Z;Meng FL;Gostissa M;Huang PY;Ke Q;Wang Z;Dao MN;Fujiwara Y;Rajewsky K;Zhang B;Alt FW
EB病毒（EBV）潜伏膜蛋白1（LMP 1）有助于致癌人类B细胞转化。条件性表达LMP 1的小鼠B细胞不倾向于B细胞恶性肿瘤，因为表达LMP 1的B细胞被T细胞消除。然而，具有条件性B细胞LMP 1表达和α/β和γ/δ T细胞的遗传消除的小鼠（“CLT”小鼠）与B细胞淋巴细胞增殖和淋巴瘤发生相关地早期死亡。CLT小鼠的产生涉及近交多个独立分离的等位基因。因此，虽然在CLT模型中引入额外的激活或敲除突变对于进一步的B细胞扩增和免疫监视研究是理想的，但通过种系繁殖进行此类实验耗时、昂贵，有时甚至不可行。为了产生更易处理的模型，我们从CLT胚胎产生克隆CLT ES细胞，并将其注射到RAG 2缺陷的胚泡中以产生嵌合小鼠，其像种系CLT小鼠一样具有脾CLT B细胞而缺乏T细胞。通过这种RAG 2缺陷型胚泡互补（“RDBC”）方法产生的CLT嵌合小鼠与B细胞淋巴增殖和淋巴瘤相关地快速死亡。由于CLT淋巴瘤通常表达激活诱导的胞苷脱氨酶（AID）抗体多样化剂，我们通过消除CLT ES细胞中的AID基因并通过RDBC测试它们来测试潜在的AID作用。我们发现，CLT和艾滋病缺陷CLT ES嵌合体有不可区分的表型，表明艾滋病是不是LMP 1诱导的淋巴瘤的发生是必不可少的。除了扩大CLT小鼠作为癌症免疫治疗模型的可及性和实用性之外，我们的研究还提供了一种新的方法来促进遗传复杂的小鼠癌症模型的产生。
The Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) contributes to oncogenic human B-cell transformation. Mouse B cells conditionally expressing LMP1 are not predisposed to B-cell malignancies, as LMP1-expressing B cells are eliminated by T cells. However, mice with conditional B-cell LMP1 expression and genetic elimination of α/β and γ/δ T cells (“CLT” mice) die early in association with B-cell lymphoproliferation and lymphomagenesis. Generation of CLT mice involves in-breeding multiple independently segregating alleles. Thus, while introduction of additional activating or knock-out mutations into the CLT model is desirable for further B-cell expansion and immunosurveillance studies, doing such experiments by germline breeding is time-consuming, expensive and sometimes unfeasible. To generate a more tractable model, we generated clonal CLT ES cells from CLT embryos and injected them into RAG2-deficient blastocysts to generate chimeric mice, which like germline CLT mice harbor splenic CLT B cells and lack T cells. CLT chimeric mice generated by this RAG2-deficient blastocyst complementation (“RDBC”) approach die rapidly in association with B-cell lymphoproliferation and lymphoma. As CLT lymphomas routinely express the Activation-Induced Cytidine Deaminase (AID) antibody diversifier, we tested potential AID roles by eliminating the AID gene in CLT ES cells and testing them via RDBC. We found that CLT and AID-deficient CLT ES chimeras had indistinguishable phenotypes, showing that AID is not essential for LMP1-induced lymphomagenesis. Beyond expanding accessibility and utility of CLT mice as a cancer immunotherapy model, our studies provide a new approach for facilitating generation of genetically complex mouse cancer models.