PO.IM01.17 · 免疫学
一种基于NSG-SGM3xIL15xDKO的新型PBMC人源化小鼠模型,用于评估免疫疗法的疗效和细胞因子释放
A novel NSG-SGM3xIL15xDKO-based PBMC humanized mouse model to evaluate efficacy and cytokine release of immunotherapies
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摘要 Abstract
中文摘要
我们观察到,在不进行照射的情况下,人外周血单个核细胞(PBMC)在NSG-SGM3xIL15xDKO(SDKO)小鼠中的植入优于NSG-MHC I/II DKO(DKO)小鼠。照射是刺激免疫应答的一种方式。我们检验了照射如何影响PBMC在SDKO小鼠中的植入,并比较了照射后SDKO与DKO小鼠在有无癌细胞情况下的植入。还在照射后的SDKO和DKO的Raji模型中测试了抗CD3xCD19 BiTE的疗效和毒性。方法 对SDKO和DKO小鼠静脉注射PBMC,并伴或不伴照射。通过眼球后(RO)采血用流式细胞术检查植入情况。SDKO和DKO小鼠在照射后静脉注射PBMC之前也接种了癌细胞。收集RO血液和脾脏以检查各类细胞类型的植入。对携带Raji-luc的照射后SDKO和DKO小鼠给予PBS或抗CD3xCD19 BiTE治疗。进行Xenogen成像以测量肿瘤负荷,并收集血浆用Luminex测定测量细胞因子水平。结果 照射加速了人CD45(hCD45)细胞在SDKO小鼠中的植入。在第10天或第14天,照射后的SDKO小鼠hCD45植入量是未接受照射的SDKO小鼠的3倍。与未照射小鼠相比,CD3+ T细胞、CD19+ B细胞和CD56+自然杀伤(NK)细胞在照射后的SDKO小鼠早期时间点的植入量高出2-6倍。与不照射相比,照射显著改善了SDKO小鼠中的B细胞和NK细胞。照射后,PBMC在SDKO中的植入水平高于DKO。这包括血液和脾脏中的CD4+T细胞、CD8+T细胞、B细胞和NK细胞。在第9天,照射后SDKO脾脏中的T细胞、B细胞、NK细胞、树突状细胞(DC)和巨噬细胞比DKO高出2-10倍。B细胞淋巴瘤Raji细胞刺激了PBMC在照射后的植入,包括脾脏中的T细胞、NK细胞和巨噬细胞。更重要的是,在Raji细胞刺激下,SDKO的植入细胞计数显著高于DKO,包括DC和巨噬细胞。最后,我们发现与DKO相比,SDKO在植入三天后支持Raji以更高的肿瘤负荷生长。同时,携带Raji的SDKO对抗CD3xCD19 BiTE的应答比DKO更快,使SDKO成为比DKO更敏感的测试BiTE疗效的模型。此外,在Raji模型中,抗CD3xCD19 BiTE处理后SDKO的细胞因子产生量远高于DKO。结论 照射不仅加速了人PBMC在SDKO小鼠中的植入。在有无癌细胞刺激的照射条件下,包括B细胞、NK细胞、DC和巨噬细胞在内的多种细胞类型以及T细胞在SDKO中的植入细胞计数均高于DKO。此外,接种癌细胞的SDKO是比DKO更敏感的测试药物疗效和毒性的平台。
查看英文原文 English abstract
We have observed better engraftment of human peripheral blood mononuclear cells (PBMCs) in NSG-SGM3xIL15xDKO (SDKO) mice compared to NSG-MHC I/II DKO (DKO) mice without irradiation. Irradiation is a way to stimulate immune responses. We examined how irradiation affects human PBMC engraftment in SDKO mice and compared engraftment in irradiated SDKO and DKO mice with and without cancer cells. Efficacy and toxicity of anti-CD3xCD19 BiTE were also tested in Raji model of irradiated SDKO and DKO. Methods SDKO and DKO mice were intravenously injected with PBMCs with/without irradiation. Retro-orbital (RO) bleeding was performed to check engraftment with flow cytometry. SDKO and DKO mice were also injected with cancer cells prior to intravenous injection of PBMCs after irradiation. RO blood and spleen were collected to check engraftment of various cell types. Raji-luc-bearing SKO and DKO mice with irradiation were treated with PBS or anti-CD3xCD19 BiTE. Xenogen was performed to measure tumor burdens, and plasma was collected to measure cytokine levels with Luminex assay. Results Irradiation accelerated human CD45 (hCD45) cells engraftment in SDKO mice. On day 10 or 14, SDKO mice after irradiation showed 3 folds of hCD45 engraftment compared to SDKO mice that didn't receive irradiation. CD3+ T cells, CD19+ B cells and CD56+ nature killer (NK) cells were engrafted at 2-6 fold higher in irradiated SDKO mice at early timepoints compared to non-irradiated mice. Irradiation significantly improved B cells and NK cells in SDKO mice compared to no irradiation. PBMCs were engrafted at higher levels in SDKO compared to DKO after irradiation. It included CD4+T cells, CD8+T cells, B cells and NK cells in the blood and spleen. On day 9, T cells, B cells, NK cells, dendritic cells (DC) and macrophages in the spleen were 2-10 fold higher in irradiated SDKO compared to DKO. B cell lymphoma Raji cells stimulated PBMCs to engraft after irradiation, including T cells, NKs and macrophages in the spleen. More importantly, SDKO was engrafted at significantly higher cell counts compared to DKO with Raji cells stimulation, including DCs and macrophages. Lastly, we found SDKO supported Raji growth with higher tumor burden on three days after implantation compared to DKO. At the same time, Raji-bearing SDKO responded to anti-CD3xCD19 BiTE faster than DKO, making SDKO a more sensitive model to test BiTE efficacy than DKO. In addition, cytokines production after anti-CD3xCD19 BiTE was much higher in SDKO than DKO in the Raji model. Conclusion Irradiation not only expediates engraftment of human PBMCs in SDKO mice. Various cell types, including B cells, NK cells, DCs and macrophages, plus T cells are engrafted at higher cell counts in SDKO than DKO with/without cancer cells stimulation with irradiation. In addition, SDKO implanted with cancer cells is a more sensitive platform than DKO to test drug efficacy and toxicity.
利益披露 Disclosure
X. Zheng, None..
G. Yang, None..
B. Parry, None..
J. G. Keck, None..
L. Yao, None.