Identification of tumor antigen derived peptides having the potential to promise candidates for cancer vaccine therapy from prostate cancer patients

鉴定肿瘤抗原衍生肽有可能成为前列腺癌患者癌症疫苗治疗的候选者

• 批准号: 20791130
• 负责人: MINAMI Takafumi
• 金额: $ 2.83万
• 依托单位: Kinki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20791130/
• 关键词: 前立腺癌; 癌ワクチン; HLA-A3; supertype; alleles; HLA-A3 supertype alleles; ペプチドワクチン; 潜伏感染

Peripheral blood mononuclear cells(PBMCs) were obtained from prostate cancer patients who had provided a written informed consent. Fifty milliliter of peripheral blood was obtained, and the PBMCs were prepared by Ficoll-Conray density gradient centrifugation. The expression of HLA-A11,-A31,and -A33 molecules on the PBMCs was determined by flow cytometry using these antibodies-anti-HLA-A11, anti-HLA-A31, anti-HLA-A33-and FITC-conjugated anti-mouse immunoglobulin G(IgG) monoclonal antibodies.7 prostate cancer-related antigens(SART2, CypB, PTH-rP, HER2neu, EGFR, MRP3, EZH2) prepared based on the binding motifs to the MHC-class I alleles.To generate LNCaP sublines expressing each of the HLA-A11, -A31, and-A33 molecules, an HLA-A1101, -A3101, or. A3303 plasmid cDNA was inserted into the eukaryotic expression vector pCR3.1.20 patient's peripheral blood mononuclear cells(PBMCs) were obtained from prostate cancer patients. The expression of HLA-A11, -A31, -A33 molecules on the PBMCs was determined by flow cytometry using these antibodies.

提供书面知情同意书的前列腺癌患者外周血单个核细胞（PBMCs）。取外周血50 ml，采用Ficoll-Conray密度梯度离心法制备pbmc。利用抗HLA-A11、抗hla -A31、抗hla -A33和fitc偶联抗小鼠免疫球蛋白G（IgG）单克隆抗体，流式细胞术检测HLA-A11、-A31和-A33分子在PBMCs上的表达。基于mhc - I类等位基因结合基序制备7种前列腺癌相关抗原（SART2、CypB、PTH-rP、HER2neu、EGFR、MRP3、EZH2）。为了生成表达HLA-A11， -A31和a33分子的LNCaP子序列，HLA-A1101， -A3101或。将A3303质粒cDNA插入真核表达载体pCR3.1.20例前列腺癌患者外周血单核细胞（PBMCs）。利用这些抗体，流式细胞术检测HLA-A11、-A31、-A33分子在PBMCs上的表达。