Observing Efficacy Of DC-CIK Celluar Immunotherapy In Malignant Solid Tumor And Monitoring Levels Of Treg And Ts

ObjectivesObserving short-term effect and immune function, analysing the changes ofRegulatory T cell (Treg) and Suppressor T cell (Ts) in peripheral blood pre-treatmentand post-treatment of malignant and solid tumor patients treated by DC-CIK.Toprovide immune monitoring index for judging the efficacy of DC-CIK.Methods①We collected and separated patients peripheral blood mononuclear cells,andthen cultured in vitro to obtain mature DC, CIK cells. Mature DC, CIK cells wereinfused to the patient by intravenous.②43cases of cancer in patients received1-2courses that4doses of DC (1x107)and5doses of CIK (1x109) were infused by intravenous. Patients who received DCand CIK cell therapy combine with chemotherapy infused cells at3days afterchemotherapy.③Immunophenotype of DC and CIK were analyzed by flow cytometry tocontrol quality of DC and CIK. Lymphocyte subsets, CD4+CD25+Foxp3+T, CD8+CD28-T, CD8+CD28+T were analyzed by flow cytometry. While the12casesof healthy people of physical examination (45-65years old) as a healthy controlgroup.④The treatment group were divided into CR+PR+SD group and PD group，andthen compare Treg and Ts in peripheral blood of two groups.⑤Clinical observation included imaging, tumor markers, the degree of pain andside effects of patients before and after treatment.Results(1)The surface of mature DC expressed high CD83(83.42±13.53), CD86(93.97±8.45),CD40(78.18±17.28),HLADR(92±9.09),and CIK CD3(96.58±2.54),CD3+CD4+(30.21±11.51), CD3+CD8+(58.08±9.48), CD3+CD56+(15.37±1.23).CD3ratio on CIK surface after culturing in vitro was significantly improved thanbefore (72.98±10.71VS96.58±2.54, P <0.05).(2)In43patients, the ratio of CD3+,CD3+CD4+,CD3+CD8+,CD3+CD56+ratios were higher than before treatment, and the improve of CD3had statisticalsignificance.(3)The percentage of Treg and Ts in peripheral blood of43case of patientsbefore treatment were higer than healthy control group, and CTL were lower, thedifference was statistically significant（P<0.05）.(4)The percentage of Treg and Ts in peripheral blood of43case of patients aftertreatment were lower than pre-treatment, but only the change of Treg had statisticallysignificant（P<0.05）.And the CTL ratio were higer than pre-treatment, the differencewas statistically significant（P<0.05）.(5)The percentage of Treg and Ts in peripheral blood of43case of patients aftertreatment were higer than healthy control group, And the change of Ts was no longerstatistically significant (P>0.05),that Treg remain had statistically significant. andthat CTL were lower, but the difference was no statistically significant（P<0.05）.(6) According to RECIST, there were3complete response cases,8partialresponse cases,26stable disease cases and6cases with progressed disease. Noserious adverse reaction was found in all patients. The effective rate (CR+PR) was23.63%, disease control rate (CR+PR+SD) was88.37%. (7)The treatment group was divided into (CR+PR+SD) and PD group. In the(CR+PR+SD) group, the ratio of Treg increased in6cases, decreased in31cases thanbefore treatment. In the SD group, there were4case increase and2cases reduced.The difference had statistically significant. So as to Treg, the difference of Ts between(CR+PR+SD) and PD group was no statistically significant.(8)Molecular tumor markers including CA199, CEA, CA125,CA724and AFPwere decreased in the treat group after treatment, but the difference was no statisticalsignificance（P>0.05）.(9)There were no Ⅲ°and the above adverse reaction. There were3cases patientswith fever in the process of cell reinfusion, the highest temperature reached39℃,continuing3-5h, temperature returned to normal after physical cooling therapy; Therewere no other adverse reactions, such as allergies, shock, bone marrow suppression,liver and kidney dysfunction.(10)We observed16cases of digestive cancer patients. We found that comparedto the control group Treg and Ts ratios before treatment were significantly higher andCTL ratio was significantly lower (P <0.05). After treatment, Treg and Ts ratio werehigher and CTL ratio was lower than the healthy control group. But the differencewas no longer statistically significant (P>0.05). Treg ratio after treatment wassignificantly lower than before treatment, CTL ratio was significantly higher thanbefore treatment (P <0.05).Conclusions：1.The ratio of CD86, CD83, CD40, HLA-DR were>75%.The ratio ofCD3+CD56+T was15.37%. Trypan blue staining activity was>90%.2.The levels of Treg and Ts in cancer patients were higher that in health people.The immune function in cancer patients lower that in health people.3.DC combined CIK cell therapy could improve the level of CTL and immunityof tumer patients, effectively reduced the level of Treg cells in peripheral blood oftumer patients, made the level of Ts had downward trend.4.DC combined CIK cell therapy could effectively improve the level of CTL andreduced the level of Treg in peripheral blood of digestive cancer patients.