Zhang M X, Guo S, Nadiya Abudukelimu, Xierenguli Alimu, Zhang R, Zeng X J, Zhang L Y, Zhang R R, Qu J H
Hematology Center, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China Xinjiang Uygur Autonomous Region Hematology Research Institute, Urumqi 830001, China.
Zhonghua Xue Ye Xue Za Zhi. 2025 May 14;46(5):445-452. doi: 10.3760/cma.j.cn121090-20241120-00461.
To generate a chronic lymphocytic leukemia (CLL) mouse model with intact immune competence and short latency by adoptively transferring (AT) splenocytes from immunoglobulin heavy-chain enhancer-driven T-cell leukemia/lymphoma 1 (Eμ-TCL1) transgenic donors into wild-type (WT) recipients. Specific pathogen-free C57BL/6J WT mice and H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice were utilized. The H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice were generated using CRISPR/Cas9 technology, and their genotypes were confirmed by PCR. Experimental animals were randomly divided into an adoptive transfer (AT) group and a WT control group (=10 per group). Mice in the AT group received an intraperitoneal injection of splenocytes from H11-Eμ-VH-TCL1-β-globin-PolyA knock-in mice. The weight and general condition of the mice were monitored. Mice were euthanized by cervical dislocation at 9 weeks post-transplantation. The CLL model was validated using key indicators, including pathological manifestations, changes in peripheral blood leukocyte counts, and immunophenotype. AT group mice exhibited significantly increased spleen weight [ (0.92±0.16) g (0.06±0.01) g in WT group, <0.05] and liver weight [ (2.11±0.56) g (1.42±0.13) g in WT group, =0.006], indicative of marked splenomegaly and hepatomegaly. The peripheral blood leukocyte count was significantly higher in the AT group [ (124.33±8.74) ×10(9)/L] compared to the WT group [ (5.55±1.67) ×10(9)/L] (=0.002). Similarly, the percentage of peripheral blood B lymphocytes was markedly increased in the AT group versus the WT group [ (69.13±6.88) % (39.78±5.94) %, <0.05]. Histopathological examination revealed CLL manifestations in the spleen, lymph nodes, and bone marrow of AT group mice, with significant lymphocytic infiltration observed in the liver, lung, and kidney tissues. Flow cytometry analysis showed that the percentages of CD19(+)CD5(+) B lymphocytes among total lymphocytes in peripheral blood, bone marrow, and spleen of the AT group were (61.37±9.92) %, (28.61± 7.08) %, and (86.03±5.78) %, respectively. These were significantly higher (all <0.05) than in the WT group [ (4.51±1.32) %, (5.58±1.46) %, and (14.33±3.20) %]. Furthermore, these CLL-like cells in the AT group were positive for CD43 and CD200, but showed lower expression of CD20, CD22, and CD79b compared to WT B cells. Adoptive transfer of splenocytes from Eμ-TCL1 transgenic mice successfully established a CLL mouse model with a relatively short latency period. This model represents a valuable preclinical tool for investigating CLL and related pathologies.
通过将免疫球蛋白重链增强子驱动的T细胞白血病/淋巴瘤1（Eμ-TCL1）转基因供体的脾细胞过继转移（AT）到野生型（WT）受体中，以生成具有完整免疫能力且潜伏期短的慢性淋巴细胞白血病（CLL）小鼠模型。使用了无特定病原体的C57BL/6J野生型小鼠和H11-Eμ-VH-TCL1-β-珠蛋白-PolyA敲入小鼠。H11-Eμ-VH-TCL1-β-珠蛋白-PolyA敲入小鼠是使用CRISPR/Cas9技术构建的，其基因型通过PCR确认。实验动物被随机分为过继转移（AT）组和野生型对照组（每组=10只）。AT组小鼠接受腹腔注射来自H11-Eμ-VH-TCL1-β-珠蛋白-PolyA敲入小鼠的脾细胞。监测小鼠的体重和一般状况。在移植后9周通过颈椎脱臼法对小鼠实施安乐死。使用包括病理表现、外周血白细胞计数变化和免疫表型等关键指标对CLL模型进行验证。AT组小鼠的脾脏重量显著增加[（0.92±0.16）g，野生型组为（0.06±0.01）g，P<0.05]，肝脏重量也显著增加[（2.11±0.56）g，野生型组为（1.42±0.13）g，P=0.006]，表明有明显的脾肿大和肝肿大。AT组的外周血白细胞计数[（124.33±8.74）×10⁹/L]显著高于野生型组[（5.55±1.67）×10⁹/L]（P=0.002）。同样，与野生型组相比，AT组外周血B淋巴细胞的百分比显著增加[（69.13±6.88）%，野生型组为（39.78±5.94）%，P<0.05]。组织病理学检查显示AT组小鼠的脾脏、淋巴结和骨髓有CLL表现，在肝脏、肺和肾组织中观察到明显的淋巴细胞浸润。流式细胞术分析表明，AT组外周血、骨髓和脾脏中总淋巴细胞中CD19⁺CD5⁺B淋巴细胞的百分比分别为（61.37±9.92）%、（28.61±7.08）%和（86.03±5.78）%。这些均显著高于野生型组[（4.51±1.32）%（5.58±1.46）%和（14.33±3.20）%]（均P<0.05）。此外，AT组中的这些CLL样细胞CD43和CD200呈阳性，但与野生型B细胞相比，CD20、CD22和CD79b的表达较低。从Eμ-TCL1转基因小鼠过继转移脾细胞成功建立了潜伏期相对较短的CLL小鼠模型。该模型是研究CLL及相关病理的有价值的临床前工具。
