Immunotherapy Mediated By DC Vaccine Plused With Adjuvant Measles, Mumps And Rubella Combined Vaccine For Murine Breast Cancer

Brest cancer is the most widespread female tumor, while its wordwide morbidity present an increasing trend year by year. Breast cancer in situ was not life threatening. But in case of these breast tumor cell transfered to other tissue or organs, would imperil patients’life. With the continually deepening understanding of human breast cancer, the treatment concept of breast cancer updated constantly, and the treatment method of breast cancer was becoming diversification. Among all of cancer therapy, immune therapy is one of the most active area of cancer reaserch in recent years. Immune therapy can invoke the patients’initiative immune system to achieve the goal of getting rid of tumor cells.Dendritic cells is the most powerful antigen presenting cells among all of those discovery so far. And dendritic cell is the only one that able to active the primitive T immune cells. The number of dendritic cells in the human tissue is deficient. And due to the low maturity of dendritic cells, its immune function was largely restrained. In addition, with the research on dendritic cells getting more and more thorough, the technique of culturing and proliferating dendritic cells in vitro has been mature and accessible. Application of dendritic cells vaccine for anti-tumor therapy has become one of the hot spot of current immunotherapy.Immune adjuvant, also known as non-specific immunity, can enhance the immunogenicity of antigen when injected into the body with tumor antigen-plused dendritic cells vaccine. It also can make the body produce efficient, durable and anamnestic immune response.We use the Measles, Mumps and Rubella combined vaccine as an immune adjuvant in this article. Measles, Mumps and Rubella combined vaccine is a widely used vaccine for childhood to prevent from respiratory infections measles, mumps and rubella.Our experiment aims to look for a kind of novel adjuvant to improve the immune efficiency of dendritic cells vaccine loaded with tumor antigen, establishing an experimental basis for clinical immunotherapy for breast cancer.Objective:Explore whether DC vaccine united Measles, Mumps and Rubella combined vaccine can excite a therapeutic effect on breast cancer of experimental animals Balb/c mice.1. We used mice breast cancer cell 4t1 to sensitize DC vaccine, to study the immune therapy efficacy of 4t1 cell lysate acting on DC vaccine.2. We established the in situ breast cancer model of mice, to explor whether DC vaccine united Measles, Mumps and Rubella combined vaccine can play a therapeutic effect on breast cancer of experimental mice.3. We used Measles, Mumps and Rubella combined vaccine to immune mice in different ways, to study the different immune therapy effect of Measles, Mumps and Rubella combined vaccine injected in advance and real time treatment.Method:1. We cultured brest cancer cell 4tlin RPMI-1640 containing 10% FBS, and prepare 4t1 cell lysate use the method of repeated freeze-thaw.2. We extracted marrow bone cell suspension from Balb/c mice, and treated it with red blood cell lysis buffer. Then the monocyte was cultured in RPMI-1640 containing 12% FBS, with the cytokines rmGM-CSF and rmIL-4 in the RPMI-1640 medium, the concentration of these two cytokines were lOng/ml. In the last, we add the cytokines rmTNF-a into the RPMI-1640 medium to stimulate the maturation of Dccell, and the concentration of rmTNF-a was also 10ng/ml.3. We use 4t1 tumor cell lysate to stimulate DC cell, to acquire DC vaccine loaded tumor antigen.4. We inoculated 5 × 104 4tlcells in mice breast subcutaneous pad in situ, to establish a breast tumor model in mice.5. We used the Measles, Mumps and Rubella combined vaccine to immune mice in different ways. Treatment group:7,14 and 21days after inoculate 4tlcells, we inject 50 μl Measles, Mumps and Rubella combined vaccine into mice back subcutaneous, while inoculate DC vaccine at axillary lymph node. Recall and response groups:28 and 14 days prior to inoculate 4tlcells, we inject 50 μl Measles, Mumps and Rubella combined vaccine into mice back subcutaneous in order to make mice produce recall and response, then 7,14 and 21 days after inoculate 4tlcells, we inject 50 u 1 Measles, Mumps and Rubella combined vaccine into mice back subcutaneous, while inoculate DC vaccine at axillary lymph node.6. We collected mice blood by cardiac puncture, then stand the blood in 4℃ refrigerator overnight. The next day, the blood centrifuged 10min by 3000r/min. The we use Elisa kits to test the IL-12and TNF-a level of mice serum.7. We stripped the tumor tissue after execute the mice, and weigh the tumor tissue, calculate the tumor inhibitory rate of each group.Results:1. The DC activated by 4T1 cell lysate present typical dendritic cells morphological characteristics.2. All Balb/c mice develop tumors after inoculate 5 × 104 4tlcells, and the life cycle of mice is 14-16 weeks.3. The mice tumor volume of group Measles, Mumps and Rubella combined vaccine united DC vaccine is significantly lower than the control group, and the life length of group Measles, Mumps and Rubella combined vaccine united DC vaccine is significantly extend.4. The mice tumor weight of group Measles, Mumps and Rubella combined vaccine united DC vaccine is significantly lower than the control group, and the tumor inhibition rate is 46% and 71% respectively.5. The serum cytokine IL-12 and TNF-a of group Measles, Mumps and Rubella combined vaccine united DC vaccine and group Measles, Mumps and Rubella combined vaccine recall united DC vaccine is significantly higher than control group, and the serum cytokine IL-12and TNF-α of group Measles, Mumps and Rubella combined vaccine recall united DC vaccine is highest.Conclusion: The Measles, Mumps and Rubella combined vaccine can promote the secretion of IL-12 and TNF-α when united DCvaccine. In addition, the tumor growth in mice can be retarded and the mice life length also can be extended. All above give us a hint that the Measles, Mumps and Rubella combined vaccine can be considered as a novel adjuvant used in DC immune therapy of breast cancer.