Progress in antitumor effect of tumor-associated macrophages in co-culture system / 中华微生物学和免疫学杂志

Macrophages are important cells of the immune system. Tumor-associated macrophages are enriched macrophages near tumor cells or tissues. Their role is mainly to promote the construction of tumor inflammatory microenvironment and inhibit tumor immune response. Cell co-culture system is a symbiotic culture system formed by mimicking the internal environment of the body in vitro. The co-culture condition is relatively consistent with the environment in vivo, enabling better information exchange and material exchange between cells, which is a supplement to the monolayer cell culture and animal experiments. Tumor-associated macrophages and tumor cells co-exist in the tumor microenvironment. Thus, constructing a co-culture system for tumor-associated macrophages and tumor cells would be conducive to studying the antitumor effect of tumor-associated macrophages and developing new immunotherapy drugs. The co-culture system would provide a new direction for treating malignant tumors. This article mainly reviewed the co-culture patterns of macrophages and the antitumor effects of different phenotypes of macrophages, and highlighted the importance of using immunotherapy to treat malignant tumors in the tumor microenvironment.

TGR5 deficiency activates antitumor immunity in non-small cell lung cancer via restraining M2 macrophage polarization

The bile acid-responsive G-protein-coupled receptor TGR5 is expressed in monocytes and macrophages, and plays a critical role in regulating inflammatory response. Our previous work has shown its role in promoting the progression of non-small cell lung cancer (NSCLC), yet the mechanism remains unclear. Here, using Tgr5-knockout mice, we show that TGR5 is required for M2 polarization of tumor-associated macrophages (TAMs) and suppresses antitumor immunity in NSCLC via involving TAMs-mediated CD8+ T cell suppression. Mechanistically, we demonstrate that TGR5 promotes TAMs into protumorigenic M2-like phenotypes via activating cAMP-STAT3/STAT6 signaling. Induction of cAMP production restores M2-like phenotypes in TGR5-deficient macrophages. In NSCLC tissues from human patients, the expression of TGR5 is associated with the infiltration of TAMs. The co-expression of TGR5 and high TAMs infiltration are associated with the prognosis and overall survival of NSCLC patients. Together, this study provides molecular mechanisms for the protumor function of TGR5 in NSCLC, highlighting its potential as a target for TAMs-centric immunotherapy in NSCLC.

Effects of Bcl3 gene knockout on composition of spleen immune cells and antitumor ability of mice / 中华微生物学和免疫学杂志

Objective:To investigate the effects of Bcl3 gene knockout on the composition of spleen immune cells and antitumor ability of mice.Methods:Bcl3 gene knockout mice (Bcl3 -/-) were established by CRISPR/Cas9 genome editing technology. Blood routine test and flow cytometry were used to detect the immune cell composition in Bcl3 -/- mice. Lung metastasis models were established by injecting mice with B16F10 melanoma cells. The number of tumor nodules in lung and the survival time of mice were used to assess the antitumor ability of wild-type (WT) and Bcl3 -/- mice. Results:Bcl3 -/- mice were successfully bred to a strain with normal growth rate and normal breeding performance. Furthermore, no embryonic death occurred. Compared with WT mice, Bcl3 -/- mice showed splenomegaly and a significant increase in the number of spleen immune cells ( P<0.05). The counts and percentages of platelets and neutrophils in Bcl3 -/- mice were significantly lower than those in WT mice. The proportion of CD19 + B cells showed no significant change, while the proportions of CD3 + T cells and T cell subsets (CD4 + , CD8 + , Treg) increased significantly ( P<0.05). The proportions of NK cells (NK1.1 + ) and neutrophils (Gr1 + ) decreased ( P<0.05), while no significant change in the proportion of DC (CD11b + ) was observed. There were a large number of tumor nodules formed by melanoma cells in the lung of Bcl3 -/- tumor bearing mice, and their survival time was shortened dramatically. Conclusions:Knockout of Bcl3 gene affected the development, differentiation and function of immune cells, thereby reducing the antitumor ability of mice.

Consensus on the perioperative pain management of cancer patients in China (2020 edition) / 中国肿瘤临床

Surgical tumor excision is a cornerstone of primary cancer treatment, and persistent postoperative pain is common after most surgical procedures. Surgical trauma and pain can cause inflammation, hypothalamic-pituitary axis activation, and sympathetic system over-reactivity. These factors, acting either separately or in combination, affect anti-tumor immunity and promote postoperative immunosuppression. Effective management of perioperative pain may help to modulate the unwanted consequences of the stress response on the immune system and improve outcomes. Based on the recent literature and clinical studies, experts from The Society of Oncological Anesthesia and Analgesia, Chinese Anti-Cancer Association have developed this consensus on the perioperative pain management of cancer patients in China (2020 edition). This consensus aims to provide evidence-based clinical recommendations for the acute pain management of cancer patients postoperatively. It does not apply to chronic pain or pain caused by cancer.

knocking out mediated by CRISPR-Cas9 genome editing for PD-L1 attenuation and enhanced antitumor immunity

Blocking the programmed death-ligand 1 (PD-L1) on tumor cells with monoclonal antibody therapy has emerged as powerful weapon in cancer immunotherapy. However, only a minority of patients presented immune responses in clinical trials. To develop an alternative treatment method based on immune checkpoint blockade, we designed a novel and efficient CRISPR-Cas9 genome editing system delivered by cationic copolymer aPBAE to downregulate PD-L1 expression on tumor cells specifically knocking out Cyclin-dependent kinase 5 () gene . The expression of PD-L1 on tumor cells was significantly attenuated by knocking out , leading to effective tumor growth inhibition in murine melanoma and lung metastasis suppression in triple-negative breast cancer. Importantly, we demonstrated that aPBAE/Cas9-Cdk5 treatment elicited strong T cell-mediated immune responses in tumor microenvironment that the population of CD8 T cells was significantly increased while regulatory T cells (Tregs) was decreased. It may be the first case to exhibit direct PD-L1 downregulation CRISPR-Cas9 genome editing technology for cancer therapy. It will provide promising strategy for preclinical antitumor treatment through the combination of nanotechnology and genome engineering.

Primary immunodeficiencies and B-cell lymphomas / Inmunodeficiencias primarias y linfomas de células B

Abstract Introduction: In primary immunodeficiencies there is a failure in the anti-tumor defense. Common variable immunodeficiency (CVID) is one of the most common primary immunodeficiencies characterized by an alteration in the differentiation of B lymphocytes (BL). Epstein-Barr virus (EBV) is an ubiquitous virus that selectively infects the BL. In patients with immunodeficiency, uncontrolled proliferation of infected BL and the action of viral proteins promote the development of lymphomas. Clinical cases: At the University Hospital Sant Joan de Deu, Barcelona, 28 patients were diagnosed with CVID from 2000 to 2013. This paper describes four patients who developed non-Hodgkin's lymphoma (NHL). The lymphoma was associated with EBV in two of the cases. Patients were < 18 years old, diagnosed with lymphoma between 4 and 13 years old. Two patients were treated with rituximab as monotherapy and achieved complete remission. Two patients were treated with CHOP (cyclophosphamide, doxorubicin, vincristine and prednisolone) and radiotherapy or rituximab and achieved complete remission. Conclusions: Early detection of EBV infections and NHL in all patients diagnosed with CVID is recommended, regardless of age at diagnosis.

Resumen Introducción: En las inmunodeficiencias primarias existe un fallo en la defensa antitumoral. La inmunodeficiencia variable común (IDVC) es una de las inmunodeficiencias primarias más frecuentes. Se caracteriza por una alteración en la diferenciación de linfocitos B (LB). El virus de Epstein-Barr (EBV) es un virus ubicuo que infecta de manera selectiva los LB. En pacientes con inmunodeficiencias, la proliferación incontrolada de LB infectados y la acción de proteínas virales promueve la aparición de linfomas. Casos clínicos: En el Hospital Universitario Sant Joan de Déu, Barcelona, se han diagnosticado 28 pacientes con IDVC del 2000 al 2013. En este trabajo se describen cuatro que desarrollaron linfoma no Hodgkin (NHL). El linfoma fue asociado a EBV en dos de ellos. Los pacientes eran menores de 18 años, con el linfoma diagnosticado entre los 4 y 13 años de edad. Dos de los pacientes fueron tratados con rituximab como monoterapia, y lograron la remisión completa. Dos fueron tratados con CHOP (ciclofosfamida, doxorrubicina, vincristina y prednisolona) y radioterapia o rituximab y también alcanzaron la remisión completa. Conclusiones: Se recomienda realizar la detección precoz de las infecciones por EBV y los NHL en todos los pacientes con diagnóstico de IDVC, independientemente de la edad del diagnóstico.

Investigação do papel de SIGIRR/IL-1R8 no crosstalk entre células tumorais e o infiltrado leucocitário / Investigating the role of SIGIRR/IL-1R8 in the crosstalk between tumor cells and the immune system

Células tumorais desenvolvem diversas estratégias para escapar da identificação e eliminação pelo sistema imune. Dessa forma, a investigação dos mecanismos envolvidos na comunicação celular no microambiente tumoral e na desregulação local do sistema imune é crítica para uma melhor compreensão da progressão da doença e para o desenvolvimento de alternativas terapêuticas mais eficazes. Nós aqui demonstramos que SIGIRR/IL-1R8, um importante regulador negativo de receptores de Interleucina-1 (ILRs) e receptores do tipo Toll (TLRs), apresenta expressão aumentada em uma linhagem celular epitelial mamária transformada pela superexpressão do oncogene HER2 e em tumores primários de mama, e promove o crescimento tumoral e metástase através da modulação da inflamação associada ao câncer e da atenuação da resposta imune antitumoral. Observamos que IL-1R8 tem sua expressão correlacionada com HER2 em tecidos mamários e sua alta expressão é fator de pior prognóstico em câncer de mama de baixo grau. Notavelmente, níveis aumentados de IL-1R8 foram observados especialmente nos subtipos HER2+ e Luminais de tumores de mama, e sua expressão aumentada em células epiteliais de mama transformadas por HER2 diminui a ativação da via de NF-κB e a expressão de diferentes citocinas pro-inflamatórias (IL-6, IL-8, TNF, CSF2, CSF3 e IFN-ß1). Meio condicionado de células transformadas por HER2, mas não de variantes celulares com o gene IL-1R8 silenciado, induz a polarização de macrófagos para o fenótipo M2 e inibe a ativação de células NK. Em um modelo murino transgênico de tumorigênese espontânea mediada por HER2, MMTV-neu, verificamos que a deficiência de IL-1R8 (IL-1R8-/-neu) retardou o aparecimento de tumores e reduziu a incidência, a carga tumoral e a disseminação metastática. Contudo, não foram observadas diferenças significativas no crescimento tumoral quando animais IL-1R8-/-neu receberam medula óssea de animais IL-1R8+/+, confirmando um papel importante da expressão de IL-1R8 em células não hematopoiéticas na tumorigênese da mama. Tumores IL-1R8+/+neu apresentaram maiores níveis de citocinas pró-inflamatórias como IL-1ß e VEGF, e menores níveis da citocina imunomodulatória IFN-γ. Além disso, tumores que expressavam IL-1R8 apresentaram menor infiltrado de células NK maduras, células dendríticas (DCs) e linfócitos T-CD8+ e um maior infiltrado de macrófagos M2 e linfócitos T-CD4+. Coletivamente, esses resultados indicam que a expressão de IL-1R8 em tumores de mama pode representar um novo mecanismo de escape da resposta imune e suportam IL-1R8 como potencial alvo terapêutico

Tumor cells develop numerous strategies to fine-tune inflammation and avoid detection and eradication by the immune system. Identification of new players that regulate the cellular crosstalk within the tumor microenvironment and promote local immune dysregulation is critical to understand disease progression and to improve therapeutic strategies. Here, we demonstrate that SIGIRR/IL-1R8, a negative regulator of IL-1R and TLRs, is up-regulated in a HER2-transformed epithelial mammary cell line and in primary breast tumors and promotes tumor growth and metastasis by modulating cancer-related inflammation and impairing anti-tumor immunity. IL-1R8 expression is correlated with HER2 in mammary tissue, and higher tumor IL-1R8 expression is a poor prognostic factor in lower grade breast tumors. Notably, higher levels of IL-1R8 expression were observed in HER2+ and Luminal breast tumor subtypes and IL-1R8 up-regulation in HER2-transformed mammary epithelial cells inhibited NF-κB activation and the expression of pro-inflammatory cytokines (IL-6, IL-8, TNFα, CSF2, CSF3, IFN-ß1). Conditioned medium from HER2-transformed cells, but not from IL-1R8 knockdown variants, induced M2-macrophage polarization and inhibited natural-killer (NK) cell activation. IL-1R8 deficiency in a transgenic mouse model of breast tumorigenesis (MMTV-neu) significantly delayed tumor onset and reduced tumor incidence, burden and metastasis. No significant differences in tumor growth were observed when IL-1R8-/-neu mice were transplanted with bone marrow from IL-1R8+/+ animals, confirming an important role for IL-1R8 expression in non-hematopoietic cells during breast tumorigenesis. IL-1R8+/+neu mammary tumors presented higher levels of pro-inflammatory cytokines such as IL-1ß and VEGF, but lower levels of IFN-γ. Besides, a lower infiltrate of mature NK cells, dendritic cells (DCs) and CD8+ T cells but higher infiltrate of M2-macrophages and CD4+ T cells were present in IL-1R8 expressing tumors. Collectively, our results support IL-1R8 expression as a novel tumor immune escape mechanism in breast cancer and putative target for immunotherapy

Implantation of Liquid Nitrogen Frozen Tumor Tissue after Posterior Decompression and Stabilization for Metastatic Spinal Tumors

STUDY DESIGN: A retrospective study. PURPOSE: To evaluate the immunity-enhancing effect of implantation of a liquid nitrogen-treated tumor. OVERVIEW OF LITERATURE: We have developed a new technique of implanting a tumor frozen in liquid nitrogen after posterior decompression and stabilization, with the aim of enhancing antitumor immunity in order to prolong the survival period of the patient. In the current study, the immunity-enhancing effect of this new technique has been evaluated. METHODS: The subjects were 19 patients in whom we had earlier performed decompression and stabilization between April 2011 and September 2013. The 19 subjects were divided into two groups, namely a frozen autologous tumor tissue implantation group (n=15; "implantation group"), which consisted of patients, who underwent implantation with autologous tumor tissue frozen in liquid nitrogen, and a control group (n=4), which consisted of patients, who did not undergo autologous cancer transplantation. To evaluate the immunity-enhancing effect of the protocol, plasma cytokines (interferon [IFN]-gamma and interleukin [IL]-12) were analyzed before surgery and a month after surgery. RESULTS: The mean rate of increase in IFN-gamma was significantly higher in the implantation group (p=0.03). Regarding IL-12, no significant difference was observed between the groups, although the implantation group exhibited increased levels of IL-12 (p=0.22). CONCLUSIONS: Decompression and stabilization combined with autologous frozen tumor cell implantation can enhance cancer immunity in metastatic spinal tumor patients. It is hypothesized that this procedure might prevent local recurrence and prolong survival period.

Research of antitumor immunity of gastric carcinoma by DC vaccine with AⅣ in vitro / 国际中医中药杂志

Objective Through experiments in vitro,we explored the role of Chinese medicine monomer-Astragaloside Ⅳ with DC vaccine in the body immunologic function.Methods Peripheral blood mononuclear cells (PBMCs)from healthy volunteers were isolated,cultured and generated in vitro,pulsed with tumor antigen from SGC7901 gastric carcinoma cell lysates,produced DC vaccine.Observe T-cell proliferation responses stimulated by DC vaccine with AⅣ group,DC vaccine group and AⅣ group respectively,and the anti-tumor effects on SGC7901 cells in vitro.Results ①The T-cell proliferation rate of DC vaccine with AⅣ group and DC vaccine group were significantly higher than AⅣ group and T-cell group(negative control group)(P=0.000).The stimulating efficacy on T-cell proliferation of DC vaccine with AⅣ group was higher than that of DC vaccine group(S/R 1∶5,1∶10,1∶50,1∶100,P=0.013,0.014,0.017,0.019).Compared with T-cell group,the T-cell proliferation rate of AⅣ group had no statistically significance(P=0.185).②The killing rate of effector cells actived by DC vaccine with AⅣ group and DC vaccine group against SGC7901 gastric carcinoma cells were higher than that of AⅣ group and T-cell group(P=0.000).The Killing power of DC vaccine with AⅣ group was stronger than that of DC vaccine(E/T 5∶1,10∶1,20∶1,50∶1,P=0.023,0.012,0.016,0.011);while the group of AⅣ group and T-cell group cannot killing tumor cells.Both had no statistically significance(P=0.267).Conclusion AⅣ can stimulate T-cell proliferation and enhance the activity of killing tumor cells by DC,which induced specific antitumor response against stomach carcinoma cells effectively.

Therapeutic effect of chemokine SLC combined with immune adjuvant CpG-ODN in treatment of implanted mouse melanoma / 中国肿瘤生物治疗杂志

Objective: To study the therapeutic effect of secondary lymphoid tissue chemokine (SLC) combined with CpG oligodeoxynucleotide (CpG-ODN) in treatment of implanted mouse melanoma and the possible mechanism. Methods: SLC-Fc fusion protein was prepared and its chemotaxis of lymphocytes was detected by chemotaxis assay. Implanted melanoma mouse models were established and randomly divided into 4 groups: control group, SLC-Fc group, CpG-ODN group, and SLC-Fc+CpG-ODN group. The growth of implanted tumors in each group was observed after treatment. Subtype and infiltration of lymphocytes in implanted tumor tissues were examined by flow cytometry. Results: SLC-Fc protein was successfully prepared, and it dose-dependently attracted lymphocytes (0.03, 0.3, and 3 μg/L). Intra-tumor injection SLC-Fc and CpG-ODN alone or in combination significantly inhibited growth of B16-implanted tumors. Tumor size in SLC-Fc+CpG-ODN group was significantly smaller than that in control group (P<0.01), and animals in SLC-Fc+CpG-ODN group survived longer. Tumor-infiltrated CD4~+ T, CD~8+ T, and dendritic cells (DCs) in SLC-Fc+CpG-ODN group were markedly increased as compared with those in control group (P<0.05), and tumor draining lymph nodes were dramatically enlarged. Conclusion: SLC combined with CpG-ODN can inhibit the growth of implanted melanoma by attracting CD4~+ T and CD8~+ T and promoting proliferation of DCs.

Antitumor effect of DC vaccine modified by Hsp70-H22 tumor peptide complex in vitro and in vivo / 中国免疫学杂志

Objective:To study the specific antitumor effect of DC modified by Hsp70 tumor peptide complexes in vitro and in vivo.Methods:The tumor antigen peptides were acquired from H22 liver cancer cells and bound Hsp70 in vitro by using biochemical technique;the mouse marrow cells were cultured with induction of rmGM CSF and rmIL 4 by using cell culture technique;mouse spleen lymphocytes was stimulated.The cultured DC cells were harvested and activation of lymphocytes was detected by MTT test and cytotoxicity of stimulated and proliferated lymphocytes to H22 tumor cells and Ehrilich ascites carcinoma cells was tested;The inhibitation to tumor was observed in vivo,after stimulated DCs were injected in mice inoculated by tumor cells.Results:DCs could become mature with the effect of Hsp70 H22 peptide complexes and secret IL 12?TNF ??IL 1? and effectively activate lymphocyte;The activated and proliferated lymphocytes could specifically kill H22 cells but not Ehrilich ascites carcinoma cells in vitro;DCs modified by Hsp70 H22 peptide complexes could become one useful kind of vaccines to inhibit H22 tumor growth in vivo.Conclusion:DCs orignied from marrow cells can be effectively modified by Hsp70 H22 peptide complexes,these modified DCs can specifically activate lymphocytes in vitro and effectively induce antitumor immune response.

Immune adjuvant-like effect of recombinant human heat shock protein 70D / 第二军医大学学报

Objective: To determine the adjuvant activity of recombinant human HSP70D protein. Methods: Dendritic cells were cultured in vitro , rhHSP70D was added into the culture medium, the cytokine secretion and maturation of dendritic cells(DC) stimulated by rhHSP70D were measured; C57BL/6 mice were immunized with rhHSP70D OVA 257 264 complexs reconstituted in vitro , and the protective antigen specific immune response against tumor challenge were evaluated in the immunized mice. Results: rhHSP70D stimulated human DC to secrete IL 1?, IL 12 p70 and TNF ?, up regulated the HLA DR,CD86 and CD40 expression on DC and improved the antigen presentation function of DC; After immunization with the hybrid of rhHSP70D and OVA 257 264 , the mice were able to resist the challenge of E. G7 OVA, which was a tumor cell line expressing model antigen OVA. Conclusion: rhHSP70D can induce maturation of DC, immunization of rhHSP70D peptide complexs can induce antigen specific immune response, suggesting that rhHSP70D may be a new adjuvant in immunotherapy of the cancer and infection diseases.

Anti-Tumor Effects of Local Injection With Oncolysate Transfected by Recombinant Vaccinia Viruses Encoding Human IL-2 / 中国肿瘤生物治疗杂志

In this experiment, the murine melanoma cell B16-F10 oncolysates transfected by recombinant vaccinia viruses encoding human IL-2(IL-2VBO) were used as vaccine. When the tumor-bearing mice were treated by following injection of IL-2VBO into tumor site, the tumor growth was inhibited and the survival time prolonged. The PBL from IL-2VBO treated mice showed higher cytotoxicity to the wild type B16-F10 but not to YAC-1 cells than that from control groups. The results showed the active specific immunity was induced by the IL-2VBO. On accout of its stronger immuogenicity and some advantages in preparing and storing, the IL-2VBO might be used as a kind of effective vaccine in the cancer active specific immunotherapy.

Antitumor Effects of Adenovirus-Mediated CD/5FC Gene Therapy in Established Colon Adenocarcinoma- Bearing Mice and Its Immunological Mechanisms / 中国肿瘤生物治疗杂志

To further investigate whether the suicide gene therapy affect the immune system, CT26 colon adenocarcinoma of BALB/c mice was adapted as experimental tumor model. The growth of CT26 tumor was suppressed by the adenovirus-mediated CD/5FC system significantly both in vitro and in vivo. But the tumors seemed very difficult to be cured and developed again soon after the end of treatment. After adenovirus-mediated CD/5FC gene therapy, there were about 40%of treated mice were cured from the tumor burden, survived longer and resisted further CT26 cell challenge. These mice also showed improvement of splenic CTL cytotoxicity. Costimulatory molecule B7-2, dendritic cell marker NLDC-145, MHC-I, and F4/80 were present in tumor mass in mice. These data suggested that in vivo suicide gene therapy could induce specific antitumor immunity, but this effect is not strong enough to eradicate established tumors. Further study for more efficient induction of antitumor immunity in CD/5FC gene therapy is warranted.

Experimental study of specific antitumor immunity induced by dendritic cell vaccine transfected with tumor cell total RNA in lung cancer patients / 中国免疫学杂志

Objective:To observe ability of DC vaccine transfected with tumor cell total RNA to induce specific antitumor immunity in lung cancer patients in vitro.Methods:DCs were generated from lung cancer patients' peripheral blood mononuclear cells(PBMC).Total RNA was isolated from lung cancer tumor cell line Calu-6 by Trizol.Autologous DCs transfected with Calu-6 total RNA by liposome were used to induce specific CTL proliferation.Specific cytotoxicity and IFN-? secretion were measured by LDH assay and ELISA method.Results:Transfected DCs exhibited dramatically increased expression of specific membrane markers and function-associated molecules,and were more potent in stimulating allogenous or autologous T cell proliferation than that of control DCs.Specific CTLs induced by transfected DCs showed higher cytotoxicity than LAK against Calu-6 antigens positive target cells.When sensitized lymphocytes were restimulated by transfected DCs again,IFN-? secretion enhanced significantly.Conclusion:Lung cancer patient's autologous DCs transfected with tumor total RNA are effective vaccines in stimulating specific antitumor T-cell immunity in vitro.

Antitumor and immune regulating effect of exosomes cancer vaccine derived from H_(22) in vivo / 重庆医科大学学报

Objective:To investigate the influence ofexosomes tumor vaccine on immunity and grouth ofmouse hepatocellular carcinoma xenograft in mice.Methods:Hepatoma carcinoma H22 cell-derived exosomes were isolated and purified from the culture supernatants by serial ultracentrifugation and sugar density ultracentrifugation.BALB/c mice inoculated with H22 cell were used as tumor models.The mice inoculated with H22 cell were divided into two groups at random:exosomes group and PBS control group.15 days later,the mice were killed;then the tumor inhibitory rate was calculated.The spleens and thymuses were weighed,and the effect of exosomes on spleens and thymus index was determined.The proliferation and cytotoxicity of splenocytes were detected by MTT assay.The level of IL-2 and INF-? in splenocyte culture supernants was detected by enzyme-linked immunoabsorbent assay(ELISA).Results:Exosomes cancer vaccine depressed the growth of tumor remarkably,and its tumor inhibitory rate was 42.38%.Compared with these in PBS control group,the spleen indexes and thymus indexes of mice bearing tumor in experiment group were increased(P