摘要
目的观察供体脾细胞及其输注途径以及联合抗CD154单克隆抗体（anti CD154 mAb，AH．F5）对非清髓性骨髓移植预处理方案建立异基因嵌合体的影响。方法供、受体分别为Lewis（Rt1^u）人鼠及Wistar（Rt1^u）大鼠，SD大鼠为无关第3品系。受鼠随机分为4组（n=20）：A组：第0天经阴茎背静脉输注2×10^8脾细胞（Sc）；B组：第0天经门静脉输注同量Sc；C组：在B组基础上，第3天再经阴茎背静脉输注1×10^8骨髓细胞（BMc）；D组：在B组基础上2h后腹腔注射AH．F53ug，第3天经阴茎背静脉输注1×10^8BMc。第20、40天检测受鼠、胸腺Rt1^1 ＋嵌合体水平，第100天对D组加做此项检查；通过第20、40天混合淋巴细胞反应（MLR）实验对受鼠耐受程度进行评估。结果单向MLR中第20、40天D组刺激指数（SI）分别为4．43±0．19及4．97±0．25，均显著低于其余各组（P〈0．01）。嵌合体检测结果表明A组在脾和胸腺中均不能形成嵌合体；B和C组在第20天能建立较低水平嵌合体；而D组第20天[脾（32．40±3．29）％、胸腺（31．08±2．60）％]，第40天[脾（23．06±3．77）％、胸腺（24．52±3．22）％]均可形成稳定的高水平嵌合，并持续超过100d。结论单纯2×10^8 Sc门静脉输注只能诱导轻度免疫耐受，建立短暂（〈20d）、低水平（〈20％）的外周性嵌合体；在此基础上联合1×10^8个BMc输注可延长嵌合体水平至40d，但仍小稳定；而加入AH．F5后叮显著增强BMc植入率，三者具有协同作用。外周嵌合体〉30％、中枢嵌合体〉20％是保证骨髓细胞发挥强大致耐受作用的基础。
Objective To explore the role of donor spleen cells （Scs） （including transfusion pathway） combined with anti CD154 monoclonal antibody （mAb） on bone marrow transplantation in non- myeloablative pre-conditioning regimen for induction of allogeneic mixed chimerism and donor-specific immune tolerance. Methods Lewis rats （Rtlt） were used as donors,Wistar-Furth rats （Wistar,Rt1^u） as recipients,and Sprague-Dawley （SD） rats as the third party donors. Scs and bone marrow cells （BMCs） were harvested from donor Lewis rats. Recipient Wistar rats were randomly divided into 4 groups ,20 rats in each group. In group A,dorsum penis vein （i. v. ） injection of Lewis rats 2 × 10^8 Scs on day 0 alone. In group B,portal vein （P. V. ） injection of Lewis rats 2 × 10^8 Scs on day 0 alone. In group C,P. V. injection of Lewis rats 2 × 10^8 Scs on day 0,in combination with i. v. injection of Lewis rats 1 × 10^8 BMCs on day 3. In group D,P. V. injection of Lewis rats 2 × 10^8 Scs on day 0,intraperitoneal injection of 3ug AH. F5 （ hamster anti-rat CD154 mAb） 2 h later,in combination with i. v. injection of Lewis rats 1 × 10^8 BMCs on day 3. Tolerance was assessed by stimulation index （SI） of one-way mixed lymphocyte reaction （MLR）. Rtl^1 positive chimersim in recipient spleen and thymus was measured by flow cytometric （FACS） analysis on the day 20,40. In rats in group D were additionally subjected to chimerism detection on the day 100. Results SI in group D on the day 20, and 40 was significantly lower than other groups （4.43 ± 0. 19, 4.97 ±0.25 ,P 〈 0.01 ）. The results of Rt1^＋ assay revealed that there was no chimersim in group A. While chimersim was transient and lower in groups B and C on the day 20, and rapidly decreased. Only group D had stable and higher level chimerism in spleen （32.40 ± 3.29 ） %, （ 23.06 ±3.77 ） % and thymus （31.08 ± 2.60） %, （24.52 ± 3.22 ） % on the day 20, and 40. Furthmore, long-term chimerism （ 〉 100days） was observed in the group D. Conclusion High level of chimerism （ 〉 30% in peripheral spleen and 〉 20% in central thymus） provides the foundation to BMCs to play a critical role in the induction or maintenance of tolerance. P. V. injection of Scs and anti CD154 mAb, plus i. v. injection of BMCs had synergistic effect to induce donor-specific tolerance and establish stable allogeneic mixed chimersim, without participation of megadose BMCs, myleosuppressive drugs and irradiation.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第9期1182-1184,共3页
Chinese Journal of Experimental Surgery
关键词
骨髓移植
脾细胞
CD154
嵌合体
免疫耐受
Bone marrow transplantation
Spleen ceils
CD154
Chimerism
Immune tolerance
