The Experimental Study On The Effect Of GVHD In The Mice Model After Adoptive Haploidentical Tumor-specific DCs-CTLs

Objective:Biological therapy as a new technique of cancertreatment,which has become the fourth therapeutic methed after the surgery,chemotherapy, and radiotherapy.Adoptive immunotherapy is that applyingimmune cells of the patients themselves and feedback to patients afteramplification in vitro,which is a systemic cancer treatment method throughkilling autologous tumours specially. Adoptive immunotherapy has higherefficiency, slight side effects, and lower drug resistance, and has kill functionto both tumer stem cells and stationary phase cells,but has no immunesuppression. Clinically, some patients,who have hepatitis, coagulant functionobstacle or myeloma,or who is valetudinarianism, is unable to tolerate theextraction of mononuclear cells from their peripheral blood or theraputicadoptive cellular transfer. Present experiment aimed to explore the possibilityof graft-versus-host disease after the haploidentical DCs-CTLs feedback totumor-bearing mice, and to provide a theoretical basis and experimental datafor clinical application.Methods: The30health C57BL/6mice were used in present experiment.Mice were subcutaneously inoculated B16cells to establish tumor-bearingmouse model and were randomly divided into three groups: control group,autologous treatment group, and haploidentical treatment group. Three groupswere given7Gy total body irradiation with CO60in tenth day after modling.Theraputics were administered respectively in the blank control group with aninjection of2000microliter normal saline through tail intravenously at thetenth and seventeenth day, in the autologous treatment group with an injectionof C57BL/6origined DCs-CTLs effecter cells with the volumn of1x106/micethrough tail intravenously at the tenth and seventeenth day, and in thehaploidentical treatment group with an injection of haploidentical (BABL/C× C57BL/6cross generation, the son CB6F1) origined DCs-CTLs effecter cellswith the volumn of1x106/mice through tail intravenously at the tenth andseventeenth day. General condition of the mice were observed and clinicalGVHD score was performed according to the weight loss, posture, mobility,fur and skin changes in the integrity of the serious level. Tumor growth andtumor size were observed and tumor inhibition ratio was calculated. The21thday after adoptive cellular transfer, mice were sacrificed, and the skin, spleenand kidney were examined histopathologically.Results:1General conditionNo diarrhea or skin ulcer was observed in any of the three groups aftercellular transfer therapy. As time gose on, weight loss, the reduce of weightgradually and spirit dropping were observed in the control group, and theperformance of arching back and hair warp in the autologous treatment groupwere observed. Accidentally spirit dropping and eating disorders were seen inthe haploidentical treatment group, but as time gose on, it was graduallyimproved.2GVHD score and the pathological examination of the miceThe control group, autologous treatment group, haploidentical treatmentgroup according to the severity of the weight loss, mobility, posture, fur,change, skin ulcer after feedback to do GVHD score respectively in9days,15days and21days to three groups,the9days control group:0.38±0.48;autologous treatment group:0.86±0.35;haploidentical treatment group:0.89±0.73; the15day control group:0.75±0.43; autologous treatment group:1.29±0.49;haploidentical treatment group:1.25±0.46; the21days controlgroup for0.86±0.64;autologous treatment group:1.57±0.79; haploidenticaltreatment group:1.50±0.76.Control group and autologous treatment group, thecontrol group and haploidentical treatment group compared respectively P<0.05, the difference was statistically significant;Autologous treatment groupand autologous treatment group compared P>0.05, the difference was notstatistically significant. Put to death the rats and take its skin, spleen and kidney to do pathological,in light-microscopy case,we observed skin, spleenand kidney and all was not seen obvious GVHD pathological changes inautologous treatment group and control group, haploidentical treatmentgroup.3The growth and the suppress of tumors in different groupsTumor growth curve shows that the control group giving saline can'tsuppress tumor growth; autologous treatment group and haploidenticaltreatment group were significantly inhibit tumor growth.After the end ofexperiment, we weigh the heavy of each group mices, the result shows thatthe weight of control group, autologous treatment group, haploidenticaltreatment group is respectively (3.51±0.05)g and (2.09±0.15)g and (2.11±0.12)g,autologous treatment group and haploidentical treatment group were lowerthan that of the heavy in control group (P<0.05); There was not statisticallysignificant differences between autologous treatment group and haploidenticaltreatment group; the tumor-suppressing rates were44.81%,45.69%respectively in autologous treatment group and haploidentical treatment group,the difference was not statistically significant.Conclusion:1Tumor-bearing mices after throughing haploidentical source DCs-CTLstreatment, does not appear significant GVHD and can better tolerancehaploidentical treatment.2Both haploidentical DCs-CTLs treatment and autologous DCs-CTLstreatment are significantly suppress tumor growth.3Haploidentical source of DCs-CTLs treatment and autologous source ofDCs-CTLs treatment of the tumor-bearing mice is safety and efficacyquite.