Research On The Clinical Efficacy And Immunological Mechanisms Of Chemotherapy Combined With Loading Autoantigen Sensitization Of DC-CIK Treatment On The Multiple Myeloma

Objective:To study the immunoloregulation and molecular mechanisms ofchemotherapy (Epirubicin, EPI) on multiple myeloma, and provide atheoretical basis for chemotherapy combined with adoptive immunotherapy(loading autoantigen sensitization of DC-CIK) on myeloma.Methods:1. Basic Research:CD95expression on myeloma cell strain KM3at different time pointsafter the same concentration of EPI treatment were analyzed through FCM.Same method was used to assay the CD95expression on KM3cells afterdifferent concentration of EPI treatment.2. Clinical research Ⅰ:①Enzyme linked immunosorbent assay was used to detect the changesof peripheral blood cytokines Interleukin-6, Interleukin-17, Transforminggrowth factor-beta and Hsp70of two groups of patients with multiplemyeloma before and after the chemotherapy.②Flow cytometry was used to detect the changes of peripheral blood Tcell subgroups （CD3~+CD4~+, CD3~+CD8~+and CD4~+CD25~+regulatory T cells）of two groups of patients with multiple myeloma before and after thechemotherapy. 3. Clinical research Ⅱ:Of43patients eligible for enrollment were selected in our study,20cases were in the chemotherapy group (VAD chemotherapy group),23ones inthe immunochemotherapy group (VAD chemotherapy combined withDC-CIK treatment group). Clinical indicators including ALB, ESR, LDH,β2-MG, content of urine light chain within24h, Scr, and PS score werecompared after the chemotherapy and the immunochemotherapy after fivecycles of treatment. The objective response rate, safety and tolerability in bothgroups were also compared.Results:1. Basic Research:The expression level of cell surface molecule CD95in KM3myelomawere up-regulated after treated with the chemotherapy, EPI. And itsexpression level gradually was increasing with drug concentration and thetreatment time.2. Clinical researchⅠ:①The Elisa detection showed that IL-6、IL-17、TGF-β in serumincreased in the newly diagnosed myeloma patients compare with the healthycontrols, and the difference was statistically significant（P＜0.05）；And thesecytokines obviously decreased after the chemotherapy, also the difference wasstatistically significant (P＜0.05). There was no statistically significantdifference on serum Hsp70levels between the patients newly diagnosedmyeloma and healthy controls, the difference was no statistically significant（P＞0.05）；But level of HSP70in serum increased after the chemotherapy,the difference was statistically significant (P＜0.05). ②Compared with healthy controls by Flow cytometry, patients withnewly diagnosed myeloma had the same level of peripheral blood CD3~+CD4~+cells but the higher level of CD3~+CD8~+cells.So the CD3~+CD4~+/CD3~+CD8~+ratio decreased, the difference was statistically significant (P＜0.05). TheCD3~+CD4~+/CD3~+CD8~+ratio was increased in myeloma patient afterchemotherapy, and the difference was statistically significant (P＜0.05).Number of peripheral blood CD4~+CD25~+regulatory T cells in newlydiagnosed myeloma patients was significantly higher（P＜0.05） than that inhealthy controls. However its number significantly reduced (P＜0.05) afterchemotherapy in myeloma patients.3. Clinical researchⅡ:①After5cycles of treatment, the levels of Albumin, Hemoglobin,β2-MG, Serum creatinine, Performance status score in the combined treatmentgroup (loading autoantigen sensitization of DC-CIK combined withchemotherapy group) were higher than in the VAD chemotherapy alone group（P＜0.05）; the levels of urine light chain within24h, ErythrocyteSedimentation Rate, Lactate dehydrogenase were no significant differencebetween the two groups（P＞0.05）。②The objective response rate after the combined treatment mentionedabove was higher than that in the VAD chemotherapy alone group（P＜0.05）.③The combination group had gentler adverse reaction, better tolerancecompared with the VAD group.Conclusion:1. The chemotherapeutic drug EPI can effectively increase theexpression of FAS on the surface of myeloma cells, which contributes directly to the CIK-induced apoptosis of myeloma cells through the Fas/FasLpathway.2. After the chemotherapy, the levels of IL-6, IL-17, TGF-β1andCD4~+CD25~+regulatory T cells in multiple myeloma patients significantlydecrease, which indicates the chemotherapy effectively breakes the tumorimmunosuppression and promotes the immunological effect of DC-CIK.3. Chemotherapy combined with loading autoantigen sensitization ofDC-CIK has better clinical effect on multiple myeloma than VADchemotherapy alone. It may be a new direction in the treatment on multiplemyeloma in the future.