Study Of Donor Lymphocyte Infusion On The Treatment Of Mouse H₂₂ Transplanted Hepatocarcinoma

Objective To explore the effect of donor lymphocyte infusion and which combined with chemotherapy (flurouracil, 5-FU) on mouse H22 transplanted hepatocarcinoma.Methods H22 transplanted hepatocarcinoma model were established in KM mice by oxter hypodermic injection of H22 carcinoma cells and divided randomly into four groups:control group (group A):without any treatment;5-FU group(group B):received daily i.p. 5-FU(20mg/kg) for 5 days;DLI group (group C):select BALB/C mice as donors, KM mice were given an intravenous infusion of 5×107 donor spleen cells (donor specific transfusion, DST)on day 0,and intraperitoneal infusion of cyclophosphamide (CP,200 mg/ kg) on day 2.On day 3,3×107 donor spleen cells were given to KM mice intravenously again to induce the specific immune tolerance status in KM mice to BALB/C mice.Afterwards these recipient mice were infused donor spleen cells on day 14,21,28 and the cells were 0.5×107, 0.5×107, 1×107, respectively.5-FU plus DLI group (group D):KM mice were induced the specific immune tolerance status to BALB/C mice by the method of DST+CP+DST like group C,then received 5-FU as group B and DLI like group C.Mixed lymphocyte reaction (MLR) was examined to confirm the tolerance status.Observe survival time, tumor inhibitory rates, histopathologic changes and investigate T-lymphocytes subsets in peripheral blood and serum Interleukin-2 levels by using Flow Cytometer (FCM) and ELISA, respectively. Detect the CD34+ hematopoietic stem cells in normal BALB/C mice with immunohistochemical staining method.Results The KM mice splenocytes of group C and D exhibited a significant reduction of MLR to BALB/C mice.The survival time of group A,B,C,D were (16.4±2.5)d,(26.3±3.1)d,(23.3±3.2)d and (33.7±5.3)d,respectively.The survival time of group B,C were longer than group A(P<0.01) and there was a significant difference between group D and other groups(P<0.01).The tumor inhibitory rates of group B,C,D were 61.3%,34.7% and 89.5%, respectively.The T-lymphocytes subpopulation of CD3+ and CD4+ were higher of group C and D than that of group A and B(P<0.05).The ratio of CD4+/CD8+ and serous interleukin-2 levels were significantly improved in group C and D(P<0.05).A lot of inflammatory cells could be seen under optic microscopy in group C and D.There were some cluster of CD34+ hematopoietic stem cells at the edge of splenic corpuscles of normal BALB/C mice.Conclusion The method of DST+CP+DST can induce specific tolerance status in KM mice to BALB/C mice. DLI, combined with chemotherapy, has evident inhibition on mouse H22 transplanted hepatic cancer and life prolongation in bearing mice, which suggests DLI has synergetic effect with chemotherapy. It also can enhance the immune abilities of H22 transplanted hepatocarcinoma bearing mice. Donor spleen cells including lots of allogeneic lymphocytes and some stem cells can induce effective graft-vesus-tumor effect to eliminate the residual cancer cells after infused into bearing bodies.It is a potential and feasible adoptive cellular immunotherapy of solid tumors.