Research Of Cytotherapy By DCs Vaccine Combined With ADPT For Breast Cancer Model Of Mammal

Cancer is a deadly disease which has been a challenge of bio-medical field in the worldwide because of the infinite proliferation and malignant metastasis of cancer cells. Breast cancer is a malignant tumor which occurs in epithelial tissue of mammary gland. Breast cancer has been a common cancer attacking women. The incidence of breast cancer ranks first in all female disease. Of all patients with cancer, women take 99% among them. Recently, breast cancer has been a common tumor threatening physical and mental health of women. Breast cancer cells lose the characteristics of normal cells. The adhesiveness of breast cancer cells decrease which leads to scatter and transfer in the body. The rapid growth rate and high mortality of breast cancer incidence leads to great attention of government on a global scale, so it is a major task to perform effective treatment for breast cancer.Dendritic cells (DCs), the most powerful antigen presenting cells (APC) yet discovered in vivo, play an important role in antigen capture, processing and presenting. DCs modulate the immune response by presenting antigens to surface of T lymphocyte cells. DCs vaccine is created by loading tumor antigens to DCs which are induced by mononuclear cells in vitro by adding cytokines. Once infused into the body, DCs vaccine can stimulate great proliferation of lymphocytes and destroy cancer by long-term monitoring of tumor cells. The technical of DCs production and DCs vaccine manufacture has been industrialized gradually. DCs vaccine has become focus of cancer therapy and thus can be wisely utilized as an immunotherapeutic strategy for cancer regimens. Vaccine for cancer therapy can strengthen the immune system of patients. DCs vaccine could modulate the immune response, so it is potential for cancer regimens. The development of DCs vaccine on biology and functions promotes its development against tumor.DCs is cultured in vitro with rmGM-CSF and rmIL-4. rmGM-CSF can induce that precursors of DCs divided into DCs and promote proliferation of DCs. rmGM-CSF also can promote the formation of clusters from individual DC to DCs. rmIL-4 mainly inhibit the differentiation in the direction of macrophages. The combined utilization of rmGM-CSF and rmIL-4 leads to immature DCs. DCs vaccine is made by loading tumor antigens to immature DCs. In order to induce immune response, MHC molecules of mature DCs must be loaded with corresponding tumor antigens. The most widely used method of DCs antigen loading is tumor antigen cracking. In fact, tumor antigens are beneficial to mature of DCs. Next, we apply rmTNF-a in order to induce the mature of DCs. In the meanwhile, rmTNF-a can increase the number of mature DCs and lead to loss of antigen presentation ability. rmTNF-a plays an important role in stimulating naive lymphocytes. DCs vaccine, DCs after sensitization, can induce strong T cell-mediated immune response and have the strong targeting ability. DCs vaccine can make antigen presenting cells produce a large amount of IL-12, a naive adjuvant dependent on T cells. DCs vaccine not only capture antigens, but also can be a target of vaccine due to its identity of immune stimulation cells. So far, the majority of studies on breast cancer vaccine take steps of tumor antigen cracking to sensitize DCs.Immunologic adjuvant is a kind of non-specific immune agents which is lack of antigenicity. When infused into organism individually or with antigen jointly, adjuvant can enhance the immunogenicity or change the type of immune response. Research shows that DCs can enhance immune response and immune memory with assistance of adjuvant. Besides, DCs act as guards of immune system by monitoring immune environment and recognizing antigens. When danger signal in immune environment activates DCs, DCs then migrate to lymph gland and T-lymphocytes response. Some adjuvant can promote cytokine secretion and enhance immune level of people. Absorbed diphtheria-pertussis-tetanus vaccine (ADPT) is mixed with diphtheria, pertussis and tetanus in a proper proportion. It is used to prevent diphtheria, pertussis and tetanus. Adjuvant for this experiment is ADPT.DCs vaccine, a new vaccine for immunotherapy, is regard as a most potential treatment. Studies of DCs vaccine for breast cancer treatment are rare both in China and abroad, especially for early treatment of breast cancer. We set up tumor models of breast cancer by means of subcutaneously injecting 4T1 beneath the right axillary mammary gland fat pad of the female Balb/c mice. DCs were induced by cytokines and loaded with antigens to mature, then we analyzed therapeutic effects on breast cancer of BALB/c mice. Besides, we tested joint antitumor effects with adjuvant simultaneously and prematurely.In the research, We collected bone marrow cells under sterile conditions and generated in follwing cytokines——GM-CSF, IL-4 and TNF-a. Next, DCs were cultured by 4T1 tumor lysates in vitro in order to prepare DCs vaccine. We injected 4T1 in situ of the female Balb/C mice and divided them into six groups——the PBS group, DCs group,the ADPT group, DCs combined ADPT group, the ADPT memory response group, DCs combined ADPT group. On one hand, we can test whethere ADPT individually has better effects on breast cancer than DCs combined ADPT group. On the other hand, we injected ADPT in advance so that mice can provoke immunological memory. When injected ADPT combined with DCs, the tumor-burdened mice were checked in order to detect treatment effects. The results showed that the treatment effect on breast cancer by using DCs vaccine alone is better than ADPT alone regardless of ADPT injection in advance. In the meanwhile, ADPT vaccine and DCs vaccine have synergy effect on breast cancer. DCs vaccine combined with ADPT has significant effects on breast cancer which is superior to DCs vaccine alone or ADPT alone. The mice can form immune memory after injection of ADPT vaccine in advance, then we injected DCs vaccine combined with ADPT which work best. The experiment results provide a basis for early-stage breast cancer treatment and delay the malignant development of tumor effectively. Besides, it lays the foundation for tumor biological immune therapy in clinic application and provides multi-dimension and comprehensive ideas for clinic breast cancer treatment.