目的通过预测分析严重急性呼吸综合征冠状病毒（SARS-CoV）与SARS-CoV-2结构蛋白中潜在的辅助性T细胞（Th细胞）表位,为新型冠状病毒疫苗的开发设计寻找合适的靶点。方法在美国国家生物技术信息中心的GenBank数据库中检索SARS-CoV与SARS-CoV-2病毒的参考序列,利用序列对比软件MEGA7和GeneDoc对2种病毒的刺突蛋白（S蛋白）、包膜蛋白（E蛋白）、膜蛋白（M蛋白）和核衣壳蛋白（N蛋白）的氨基酸序列进行比较分析。利用SYFPEITHI、IEDB和NetMHCIIpan在线生物信息学工具预测筛选SARS-CoV和SARS-CoV-2可能的Th细胞表位,并在此基础上寻找2种病毒同源的潜在表位。最后利用ProtScale对含同源表位的蛋白质进行疏水性分析比较。结果 2种病毒S蛋白同源性为75%,其他蛋白同源性分别为E蛋白94%、M蛋白90%及N蛋白90%。在线软件预测到SARS-CoV-2有22个潜在的Th细胞表位,SARS-CoV有25个潜在的Th细胞表位。进一步对比获得2对完全同源表位和6对高度同源表位,位于3对蛋白中,且ProtScale对比分析2种病毒相应蛋白的疏水性差异微小。结论对比分析得到8对完全同源或高度同源表位,且含差异氨基酸的对应肽段疏水性差异较小,同源性较高的Th细胞表位可考虑作为SARS-CoV-2疫苗设计的候选表位。

Luteolin, a naturally found dietary flavonoid, has a wide range of beneficial biological effects, including effects against tumors and oxidants. Studies proved that luteolin can modulate immune responses. In this study, we investigated the function of luteolin as an antitumor vaccine adjuvant (to treat malignant melanoma) in vitro and in vivo. We found that Luteolin may activated the PI3K-Akt pathways in APCs (Antigen Presenting Cells), induced the activation of APCs, enhanced CTL (Cytotoxic T Lymphocyte) responses, and inhibited tolerogenic T cells. To prove the role of CD8+T cells in immune process, we sorted the CD8+T cells from the immunized mice and transferred them to the B16F10 tumor-bearing mice, the result showed that the survival rate was improved. We also observed that in the mice immunized with Luteolin as an adjuvant, the tumor growth was significantly reduced. Taken together, the result demonstrated that luteolin showed promising properties as a vaccine adjuvant for treating malignant melanoma.

目的 利用生物信息学方法分析预测人巨细胞病毒(HCMV)膜表面糖蛋白gH以及磷酸化蛋白65(PP65)的B细胞、T细胞优势抗原表位,为HCMV亚单位疫苗的研发提供理论依据.方法 从美国国立生物技术信息中心(NCBI)数据库中获取HCMV的gH和PP65蛋白的氨基酸序列.通过ABCpred、BCPred和BepiPred软件预测gH和PP65蛋白的B细胞优势抗原表位;利用NetMHCⅡ2.3 Server和NetMHCⅡpan 4.0 Server软件预测gH和PP65蛋白的CD4+T细胞优势抗原表位;使用NetMHC 4.0 Server和IEDB软件预测gH和PP65蛋白的CD8+T细胞优势抗原表位;最后由VaxiJen v.2.0 Server软件对上述得到的所有抗原表位进行抗原性分析.结果 gH和PP65蛋白的二级结构中无规卷曲均占比最高,分别为40.51％、52.94％;综合所有软件的预测结果并进行抗原性分析后,获得g H蛋白CD8+T细胞优势抗原表位20个,CD4+T细胞优势抗原表位15个,B细胞优势抗原表位1个,获得PP65蛋白CD8+T细胞优势抗原表位8个,CD4+T细胞优势抗原表位8个,B细胞优势抗原表位3个.结论 HCMV的gH和PP65蛋白均含有丰富的B、T细胞优势抗原表位,可为HCMV亚单位疫苗的抗原肽选择提供理论依据.

Hesperetin (HES) is a dihydroflavone with the molecular formula of C16H14O6. It has been reported that Hesperetin has antioxidant and anticancer effects. Recent studies showed that it can also regulate immune responses. To assess its potential function as a vaccine adjuvant, we formulated HES with inactivated B16F10 melanoma cells and determined whether it would enhance the activation of antigen‐presenting cells by experiments in vivo and in vitro. We found that HES activated the PI3K‐Akt signalling pathway in antigen‐presenting cells (APCs), enhanced cytotoxic T lymphocyte (CTL) responses and deactivated tolerogenic T cells. We also observed that inactivated B16F10 cells in combination with HES vaccine inhibited the growth of mice tumours, resulting in improved overall survival compared to the effects of inactivated B16F10 cell vaccine. To verify that CD8 + T cells play a key role in inhibiting the development of melanoma, we transferred the sorted CD8 + T cells from immunized mice to B16F10 challenged models and found that the survival rate of tumour‐bearing mice was significantly prolonged. Taken together, these results suggest that hesperetin can be used as a potential adjuvant to improve tumour immune responses and antigen immunogenicity.

Gastrodin (GAS) is a Chinese medicine with wide application for the treatment of nervous system disease. Previous studies reported that GAS exhibited non-specific immunomodulatory activities. To explore the effects of GAS as a vaccine adjuvant, the expression levels of CD80, CD86, MHCI and MHCII activated markers were detected after GAS treatment in vitro and in vivo, and the expression levels of IL-2 and TNF-α in splenocytes were detected after GAS treatment in vivo. Besides, the expression levels of IL-2 and IFN-γ in CD4+T cells and perforin, TNF-α and IFN-γ in CD8+T cells were detected. The effects of GAS on the survival rate and tumor size of tumor-challenged mice and the effect of cytotoxicity on CD8+T cells were also investigated. Our data showed that GAS ameliorated CD8+T cell mediated immune response and significantly improved protection of tumor-challenged animals. The results demonstrated that GAS is a potential adjuvant contributing to anticancer immunomodulation.

Colorectal cancer (CRC) is the third most prevalent cancer in the world. Although great progress has been made, the specific molecular mechanism remains unclear. This study aimed to explore the differentially expressed genes (DEGs) and underlying mechanisms of CRC using bioinformatics analysis. In this study, we identified a total of 1353 DEGs in the database of GSE113513, including 715 up- and 638 downregulated genes. Gene ontology analysis results showed that upregulated DEGs were significantly enriched in cell division, cell proliferation, and DNA replication. The downregulated DEGs were enriched in immune response, relation of cell growth and inflammatory response. The Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that upregulated DEGs were enriched in cell cycle and p53 signaling pathway, whereas the downregulated DEGs were enriched in drug metabolism, metabolism of xenobiotics by cytochrome P450, and nitrogen metabolism. A total of 124 up-key genes and 35 down-key genes were identified from the protein-protein interaction networks. Furthermore, we identified five up-modules (up-A, up-B, up-C, up-D, and up-E) and three down-modules (d-A, d-B, and d-C) by module analysis. The module up-A was enriched in sister chromatid cohesion, cell division, and mitotic nuclear division. Pathways associated with cell cycle, progesterone-mediated oocyte maturation, oocyte meiosis, and p53 signaling pathway. Whereas the d-A was mainly enriched in G-protein coupled receptor signaling pathway, cell chemotaxis, and chemokine-mediated signaling pathway. The pathways enriched in chemokine signaling pathway, cytokine-cytokine receptor interaction, and alcoholism. These key genes and pathways might be used as molecular targets and diagnostic biomarkers for the treatment of CRC.

T lymphocytes synergize with the cellular immune system to promote hepatocyte regeneration. The T‐cell receptor (TCR) immune repertoire is closely associated with the host immune response and regenerative proliferation. High‐throughput sequencing of TCR provides deep insight into monitoring the immune microenvironment. Here, we aimed to determine the role of the TCRβ immune repertoire in liver regeneration (LR). We investigated hepatic regeneration in TCRβ chain‐deficient (tcrb–/–) mice by two‐thirds partial hepatectomy (PHx) method. Our results demonstrated that tcrb–/– mice revealed a reduced capacity for LR, which was characterized by impaired hepatocyte proliferation and enhanced hepatocyte apoptosis. Dysregulation of inflammatory signaling activation and inflammatory factors was observed in regenerated tcrb–/– livers. Simultaneously, significantly altered immunocyte levels and aberrant cytokine levels were observed during hepatic regeneration. In addition, we first determined the profile of the TCRβ immune repertoire during LR, indicating that PHx resulted in remarkably lower TCRβ diversity in intrahepatic T lymphocytes. Conclusion: Taken together, our data suggest that TCRβ deficiency gives a rise to aberrant intrahepatic immune microenvironment that impairs LR, and the TCRβ reconstitution is required for hepatic immunocyte recruitment and activation during LR.

Recently, increasing evidences show that procyanidin (PC) modulate immune responses in human. To evaluate adjuvant effects of PC on vaccine immune modulation and anti-tumor activity, we formulated PC with B16F10 tumor antigen as tumor vaccine to immune C57BL/6 mice and used intramuscular injection before challenge with tumor B16F10 cells. Our results revealed that PC enhanced T cell-mediated immune responses both in vitro and in vivo. Moreover, the B16F10 tumor vaccine induced some degree of anti-tumor effects as evaluated by the inhibition of tumor growth and the prolongation of survival. The tumor-bearing mice showed a high level of specific cytotoxic activity and had activated CD8 T cells that secreted perforin, IFN-γ and TNF-α in response to the stimulation with antigen in vitro. Taken together, current study presents evidence that PC may be used as a promising vaccine adjuvant.

Gastrodin (GAS) is one of the rare traditional Chinese medicinal materials in our country and it has been widely used in the treatment of cardiovascular and cerebrovascular diseases. Baicalein is a traditional Chinese herbal medicine, the active ingredient of Scutellaria baicalensis Georgi has a wide range of pharmacological effects. in recent years Fan Jinghui et al found that baicalein has a significant anti-tumor effect. In this study, we used LPS as a positive control, different concentrations of gastrodin and baicalein respectively stimulated DC2.4 cell line and RAW264.7 cell line. After 48 hours, flow cytometry was used to detect the expression of CD80, CD86, MHCI, MHCII expression changes.

目的 从体外和体内2个方面检测黄芩素对抗原提呈细胞的免疫增强活性,为促进临床肿瘤免疫治疗的发展提供实验基础.方法 在体外实验中,以LPS作为阳性对照,不同浓度的黄芩素分别刺激DC2.4细胞系和RAW264.7细胞系,应用流式细胞仪检测2种细胞表面CD80、CD86、MHC-LMHC-Ⅱ表达量变化;在体内试验中,以弗式完全佐剂与灭活的肿瘤抗原作为阳性对照,黄芩素与灭活的肿瘤抗原作为实验组对小鼠进行免疫,免疫3d后对其免疫效果进行评价.结果 在体外实验中,不同浓度的黄芩素分别刺激DC2.4和RAW264.7细胞后,细胞表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ都有不同程度表达上调,在体内实验中,不同浓度的黄芩素可使抗原提呈细胞表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ都有不同程度表达上调,同时使细胞因子IL-12和TNF-α有不同程度表达上调,而使IL-10表达下调.结论 黄芩素在体外和体内具有较强的激活抗原提呈细胞的作用,并且使抗原提呈细胞表面标志和细胞因子上调,黄芩素有可能成为具有潜在应用价值的肿瘤细胞疫苗佐剂.

Objective:To detect immunological effects of procyanidin ,gastrodin,baicalein and apigenin on antigen presenting cells of RAW264.7 and DC2.4 and search for new tumor vaccine adjuvant.Methods: After different concentrations of four kinds of Chinese herbal monomer and LPS were used to stimulate RAW 264.7 or DC2.4 cells for 48 hours,cells were obtained and stained with CD80,CD86,MHCⅠand MHCⅡ antibody.Then protein expression levels of CD80,CD86,MHCⅠ and MHCⅡ on RAW264.7 and DC2.4 cells were analyzed by flow cytometry.Results: Results showed that after 48 hours stimulated with different concentrations of four kinds of Chinese herbal monomer on RAW 264.7 or DC2.4 cells,the expression levels of CD80,CD86,MHCⅠ,MHCⅡ were all up-regulated in comparison with control , procyanidin and baicalein exhibited stronger immunological stimulating activity on a cellular level.Conclusion:Procyanidin and baicalein were found having potential application value in clinical treatment as tumor vaccine adju -vants.