探讨基因编辑猪-恒河猴异种心脏移植实验免疫抑制方案及其效果，为异种器官移植免疫抑制方案制订提供参考。

经表型鉴定、补体依赖的细胞毒性(CDC)实验以及IgG、IgM结合实验等方式，选取双基因敲除供体猪(耳号4207)和恒河猴(实验动物备案号382350)作为实验动物。手术方式为异位心脏移植，免疫抑制方案包括免疫诱导期与免疫抑制维持期。

于2023年12月16日实施基因编辑猪-恒河猴异位心脏移植术，手术过程顺利，供、受体心脏均成功复搏。受体术后一般情况良好，术后第9天(T＋9)因左下肢缺血坏死行左下肢截肢术。受体猴累计存活46 d，死亡原因可能为慢性排斥反应致供心功能衰退以及T＋46时对受体猴行心肌组织活检前麻醉药物给量过多。受体经免疫抑制后，白细胞、淋巴细胞均处于稳定且较低水平，CDC稳定在4%以下；受体猴流式细胞分析可见T＋46较移植前1 d，以CD19为表面标志物的B细胞和以CD3为表面标志物的T细胞较实验前明显减少。受体猴猪心肌和猴心肌病理检测结果表明，B细胞表面标志物CD19与T细胞表面标志物CD3染色几乎不可见，补体激活标志物C4D染色较浅，巨噬细胞表面标志物CD68、IgG和IgM着色频率偏多。

本实验采用的免疫抑制方案可有效抑制超急性与急性排斥反应，为未来异种器官移植提供了可行的免疫抑制方案参考。

To explore the immunosuppressive protocol and its effect in the gene-edited pig-rhesus monkey xenotransplantation experiment, and to provide a reference for the formulation of immunosuppressive protocol for xenogeneic organ transplantation.

Through preoperative matching experiments including phenotypic identification, complement-dependent cytotoxicity (CDC) experiment, and IgG and IgM binding experiments and so on, a double-gene knockout donor pig (lot number 4207) and a rhesus monkey (experimental animal registration number 382350) were selected as experimental animals. An ectopic heart transplantation surgery was chosen for transplantation. The immunosuppressive protocol was divided into the induction period and the maintenance period of immunosuppression.

On December 16, 2023, the gene-edited pig-rhesus monkey ectopic heart transplantation surgery was performed successfully. The surgical process went on smoothly, and both the donor and recipient hearts successfully resumed beating. The general condition of the recipient was good after the surgery. On the 9th day after the surgery (T+ 9), due to ischemic necrosis of the left lower limb, a left lower limb amputation surgery was performed. The final survival time of the recipient monkey was 46 d. The possible cause of death was chronic rejection-induced decline in donor heart function and excessive dosage of anesthetic drugs administered to the recipient monkey before the myocardial tissue biopsy at T+ 46. After immunosuppression, the white blood cells and lymphocytes of the recipient were in a stable and low level, and the CDC reached steady state, settling below 4%. The flow cytometry analysis results of the recipient monkey showed that compared with 1 d before transplantation, the B cells with CD19 as the surface marker and T cells with CD3 as the surface marker were significantly reduced in T+ 46. The pathological examination results of the pig heart and rhesus monkey heart after the recipient′s death indicated that the staining of B cell surface marker CD19 and T cell surface marker CD3 was almost invisible, and the staining of complement activation marker C4D was relatively shallow; the staining frequency of macrophage surface marker CD68, IgG, and IgM was relatively higher.

The immunosuppressive protocol used in this experiment effectively inhibited hyperacute and acute rejection reactions, providing a feasible immunosuppressive protocol reference for future xenogeneic organ transplantation.