Animal research of the inhibition effect on acute graft-versus-host disease by rapamycin-induced dendritic cells
HE Chulin;ZHOU Qianqian;ZHANG Yulong;ZHAO Man;MA Cong;ZHAN Linsheng;WANG Xiaohui;Guangxi Medical University;Institute of Transfusion Medicine,Academy of Military Medical Sciences;
Objective To evaluate the therapeutic effects of rapamycin-induced dendritic cells on acute graft-versushost disease( a GVHD). Methods We established an MHC mismatched mouse a GVHD model via allogeneic hematopoietic stem cell( HSC) transplantation where donor cells from 15 BALB/C mouse( MHC is H-2 Kd) were transferred to 30 recipient C57 BL/6 J mouse( MHC is H-2 Kb) processed by 8. 5 Gy total body irradiation( TBI); to verify the a GVHD model,C57 BL/6 J mice were divided into 3 groups: group MHC mismatch transplantation,group MHC match transplantation and group irradiation only( n = 10). The 3 groups of mouse processed by 8. 5 Gy TBI were intravenously infused with C57 BL/6 Jderived bone marrow cells/spleen cells,BALB/C-derived bone marrow cells/spleen cells and plain PBS solution,respectively. The transplantation success ratio of BALB/C mouse in the recipients were detected by flow cytometry and the survival status of them was observed. In vitro generated dendritic cells( DCs) were treated by rapamycin( RAPA-t DCs) and PBS( PBS-DCs) as control. a GVHD model mice were randomly divided into 3 groups( n = 10) : group RAPA-t DCs( intravenously injected with RAPA-t DCs 4 106/mice),group PBS-DCs( intravenously injected with PBS-DCs 4 106/mice) and group PBS( intravenously injected with PBS). The in vivo distribution and migration of DCs were monitored by bioluminescence imaging;the changes of mice weight and survival period of the 3 groups were examined to evaluate the inhibiting effect of RAPA-t DCs on a GVHD. Results Compared with PBS-DCs,RAPA-t DCs exhibited a suppressed expression of allostimulatory markers of CD80,CD86 and MHCII( P0. 05),the mean fluorescence intensity of CCR1 and CCR5 of RAPA-t DCs and PBS-DCs were14. 5±1. 4 vs 10. 0±2. 1 and 12. 7±2. 3 vs 7. 2±1. 2( P0. 05) while that of CCR7 was 7. 8±1. 3 vs 6. 2±2. 5( P0. 05). The bioluminescence imaging analysis showed that rapamycin induction did not affect DCs' in vivo distribution in the GVHD mice,and rapamycin-conditioned DCs can also mobilize to lymphonodus or spleen. Compared with the mice of group PBSDCs,the weight( g) of RAPA-t DCs decreased slower-4. 3±0. 2 vs-3. 2±0. 6( P0. 05) on day 19 after the transplantation.The Survival time( days) of RAPA-t DCs was longer 10. 1 ± 5. 5 vs 16. 6 ± 6. 7( P 0. 05),but all mice dead eventually,which means no difference in the overall survival rates. Conclusion The rapamycin could induce DCs towards the tolerant state by suppression the expressions of certain surface markers. The adoptively infused RAPA induced tolerant DCs( RAPAt DCs) preserved certain capability of homing to the T cell-abundant region. In addition,the adoptively infused RAPA-t DCs could attenuate a GVHD to some extent but could not improve the overall survival rate.
【Fund】： 国家自然科学基金面上项目(81770196)及青年项目(81701583);; 军队十二五重大专项(AWS14C014);; 军事医学科学院创新基金(2017CXJJ32);; 后勤科研项目(17QNP038)
【CateGory Index】： R457.7
【CateGory Index】： R457.7