Cryopreserved-skin grafting in Rats under administration of anti-CD2 menochlonal antibody

应用免疫抑制剂进行心脏、肺、肝、肾等重要器官的同种异体移植已被广泛接受。然而，这些药物确实有副作用，而且很难证明将这些药物用于其他非重要器官或组织的转移是合理的。不使用这些药物的组织移植的发展将扩大非重要器官或组织的适应症，用于严重创伤或恶性肿瘤切除后复杂缺损的重建。我们使用抗cd2单克隆抗体（mab）进行大鼠大鼠大失配异体皮肤移植。材料与方法：采用Lewis （LEW）大鼠和Brown Norway （BN）大鼠的大失配模型。用培养的杂交瘤细胞OX34腹腔注射制备大鼠抗cd2单克隆抗体。用流式细胞术测定制备抗体的有效剂量为2mg /kg。通过手术前后两次给予相同剂量的抗体，证实了两例BN大鼠异位心脏移植成功。将BN大鼠全层腹部皮肤（2张1.5x2 cm皮肤）移植到LEW背部。创建了三个组。A组（12个移植物）：对照组（12个移植物）：从移植前2天开始，连续2天腹腔注射抗cd2单克隆抗体2 mg/kg。C组（12个）：从移植前一天开始，连续9天注射相同剂量。以PVG为第三方进行混合淋巴细胞反应试验，评价受体大鼠淋巴细胞功能。结果：未处理大鼠和处理2 d大鼠的皮肤移植在术后10 d内均出现排斥反应。相比之下，移植皮肤处理9天的大鼠存活20天以上（平均生存期51天，最长生存期98天）。混合淋巴细胞反应试验显示，A组对BN的反应强于PVG， B组对BN和PVG均表现出类似的强烈反应，提示单抗给药2周后淋巴细胞功能恢复。C组对BN和PVG的反应小于A组和B组，对PVG的反应明显更强，表明连续给药9周后对淋巴细胞功能的抑制更广泛。讨论：本报告显示抗cd2单克隆抗体治疗可显著延缓大鼠同种异体皮肤移植的排斥反应。考虑到同种异体皮肤移植的困难，使用抗cd2单克隆抗体进行全肢、关节或神经的其他同种异体组织移植可能更成功。少

Introduction : It is well accepted to use immuno-suppressing agents for the allografting of the important organs such as heart, lung, liver or kidney. However, these agents do have side effects and it is difficult to justify to use these drugs for the transfer of other non-vital organs or tissues. Development of tissue transfer without using these agents would extend indications for non-vital organs or tissues for the reconstruction of complicated defect after sever trauma or malignant tumor resection. We used anti-CD2 monoclonal antibodies (mAbs) for allo-skin grafting in rat major mismatch model.Materials and Metods : Major mismatch model of Lewis (LEW) and Brown Norway (BN) rats was used. Rat anti-CD2 monoclonal antibiesantibodies was made by intra-peritoneal injection of cultured hybridoma cell (OX34) to mouse. The effective dose of created antibodies was determined as 2 mg/kg by flow-cytometry. Two successful heterotopic heart transplantation from BN to LEW rat were demonstrated b … More y the twice administration of the same dose antibodies before and after surgery. Full thickness abdominal skin (2 sheets of 1.5x2 cm skin) of BN rats was grafted onto the back of LEW.Three groups were created. Group A (12 grafts) : control Group B (12 grafts) : Anti-CD2 mAbs of 2 mg/kg were injected intraperitoneally to LEW rats for 2 consecutive days beginning two days before grafting. Group C (12 grafts) : same dose injection for 9 consecutive days beginning one day before grafting. Mixed lymphocyte reaction test using PVG as the third party was performed to evaluate lymphocyte function of the recipient rat.Results : Skin grafts of untreated rats or those of 2 day treated rats were all rejected within 10 days postoperatively. In contrast, grafted skin of rats treated for 9 days survived for than 20 days (mean survival period : 51 days, longest : 98 days). Mixed lymphocyte reaction test showed that group A demonstrated more reaction to BN than PVG and group B showed similar strong reaction to both BN and PVG suggesting recovery of lymphocyte function 2 weeks after mAbs administration. In group C,reaction to BN and PVG was less than that in group A or B and significantly stronger reaction to PVG suggesting more generalized suppression to lymphocyte function 2 weeks after 9 consecutive administration of mAbs.Discussion : This report shows that anti-CD2 monoclonal antibody treatment may significantly delay rejection of allo-skin grafting in major mismatched rat model. Considering the difficulty in allo-skin grafting, other allo-tissue transfer using anti-CD2 monoclonal antibodies for total limb, joint or nerve may be more successful. Less