In vitro and in vivo functions of T cells produced in complemented thymi of chimeric mice generated by blastocyst complementation.
In vitro and in vivo functions of T cells produced in complemented thymi of chimeric mice generated by blastocyst complementation.
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DOI：
10.1038/s41598-022-07159-7
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发表时间：
2022-02-25
影响因子：
4.6
通讯作者：
Nakauchi H
中科院分区：
文献类型：
--
作者：
Yamazaki K;Kubara K;Ishii S;Li P;Dairiki R;Hihara T;Ishizuka Y;Izumi Y;Kumai M;Kamisako T;Ishizaki H;Sato H;Masaki H;Mizuno N;Mitsuhashi K;Ito M;Hamanaka S;Yamaguchi T;Watanabe M;Sugiyama F;Nakauchi H
囊胚互补是一种有趣的方法，可以产生用于再生医学器官制备的人源化动物，并建立用于药物开发的新模型。证明在嵌合体动物中补充的器官和细胞正常工作对于证明胚泡互补的可行性至关重要。在这里，我们产生了胸腺互补的嵌合小鼠，评估了抗PD-L1抗体在荷瘤嵌合小鼠中的功效，然后研究了T细胞功能。通过将C57 BL/6（B6）胚胎干细胞注射到Foxn 1 nu/nu桑椹胚或胚泡中产生胸腺补充嵌合小鼠。流式细胞仪检测结果表明，嵌合小鼠胸腺上皮细胞（TEC）来源于B6细胞。胸腺外出现T细胞。单细胞RNA测序分析显示TEC基因表达谱与B6小鼠相当。嵌合小鼠的脾T细胞在体外对抗CD 3刺激反应良好; CD 4+和CD 8 + T细胞增殖并产生IFNγ、IL-2和颗粒酶B，与B6小鼠相同。抗PD-L1抗体处理抑制嵌合小鼠中的MC 38肿瘤生长。此外，在嵌合体中，抗PD-L1抗体通过显著降低T细胞上的PD-1表达并增加引流淋巴结和肿瘤中产生IFNγ的T细胞来恢复T细胞活化。因此，由补充胸腺产生的T细胞在体外和体内均正常发挥功能。为了通过胚泡互补成功地产生人源化动物，种间嵌合体中互补器官/细胞的功能和基因表达谱的验证在不久的将来将是重要的。
Blastocyst complementation is an intriguing way of generating humanized animals for organ preparation in regenerative medicine and establishing novel models for drug development. Confirming that complemented organs and cells work normally in chimeric animals is critical to demonstrating the feasibility of blastocyst complementation. Here, we generated thymus-complemented chimeric mice, assessed the efficacy of anti-PD-L1 antibody in tumor-bearing chimeric mice, and then investigated T-cell function. Thymus-complemented chimeric mice were generated by injecting C57BL/6 (B6) embryonic stem cells into Foxn1nu/nu morulae or blastocysts. Flow cytometry data showed that the chimeric mouse thymic epithelial cells (TECs) were derived from the B6 cells. T cells appeared outside the thymi. Single-cell RNA-sequencing analysis revealed that the TEC gene-expression profile was comparable to that in B6 mice. Splenic T cells of chimeric mice responded very well to anti-CD3 stimulation in vitro; CD4+ and CD8+ T cells proliferated and produced IFNγ, IL-2, and granzyme B, as in B6 mice. Anti-PD-L1 antibody treatment inhibited MC38 tumor growth in chimeric mice. Moreover, in the chimeras, anti-PD-L1 antibody restored T-cell activation by significantly decreasing PD-1 expression on T cells and increasing IFNγ-producing T cells in the draining lymph nodes and tumors. T cells produced by complemented thymi thus functioned normally in vitro and in vivo. To successfully generate humanized animals by blastocyst complementation, both verification of the function and gene expression profiling of complemented organs/cells in interspecific chimeras will be important in the near future.
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影响因子：
5.9
作者：
Hamanaka S;Umino A;Sato H;Hayama T;Yanagida A;Mizuno N;Kobayashi T;Kasai M;Suchy FP;Yamazaki S;Masaki H;Yamaguchi T;Nakauchi H
通讯作者：
Nakauchi H
影响因子：
32.4
作者：
Kernfeld EM;Genga RMJ;Neherin K;Magaletta ME;Xu P;Maehr R
通讯作者：
Maehr R
DOI：
10.1073/pnas.93.12.5742
发表时间：
1996-06-11
影响因子：
11.1
作者：
Blackburn, CC;Augustine, CL;Morahan, G
通讯作者：
Morahan, G
影响因子：
64.8
作者：
NEHLS, M;PFEIFER, D;BOEHM, T
通讯作者：
BOEHM, T
影响因子：
4.4
作者：
SEGRE, JA;NEMHAUSER, JL;LANDER, ES
通讯作者：
LANDER, ES