个体化卵巢癌新抗原的预测和免疫原性验证-现代医学

个体化卵巢癌新抗原的预测和免疫原性验证

单位：湘潭市中心医院生殖与遗传中心, 湖南湘潭 411100

分类号：R737.31

出版年·卷·期（页码）：2022·50·第八期（995-1002）

摘要：

目的：对卵巢癌组织进行高通量测序，联用生物信息学软件和体外试验验证其免疫原性。方法：采集1例晚期卵巢癌患者外周血和肿瘤标本进行全外显子和全转录组测序，结合PSSMHCpan软件和表位呈现综合预测模型(EPIP)对抗原肽与人类白细胞抗原(HLA)Ⅰ类分子结合的亲和力和表位呈递性预测排序，用筛选出的候选抗原肽分别刺激卵巢癌患者和同HLA型的健康人外周血单个核细胞(PBMC)，通过酶联免疫斑点法(Elispot)对免疫反应性进行量化。结果：1.测序结果显示患者具有22个非同义单核苷酸变体、1个插入或缺失。2.根据生物信息学软件，筛选出高亲和力和强呈递性的16条卵巢癌候选抗原肽。3. 16条候选抗原肽经体外试验Elispot验证，其中6个基因组成的10条肽具有成为新抗原的潜力，阳性率62.50%。4.同HLA-A*24：02限制的9条候选抗原肽，在同HLA型的健康志愿者与患者中Elispot结果一致。结论：通过物信息学软件明显提高卵巢癌新抗原的验证率，筛选到的OR2G3、SLC19A2、SPTB、C3、ZBED6CL、LY75成为潜在的卵巢癌新抗原。HLA分型相同的健康人外周血可作为验证抗原免疫原性的一种可能的替代方案。

Objective: To verify immunogenicity of ovarian cancer tissue via high-throughput sequencing in combination with bioinformatics software and in vitro experimentsy.Methods: Peripheral blood and tumor samples from a patient with advanced ovarian cancer were collected for whole exon and whole transcriptomic sequencing,and the affinity and epitope presentation of antigen peptide binding to human leucocyte antigen (HLA) Class Ⅰmolecules were predicted and sequenced using PSSMHCpan software and Epitope Presentation Integrated Prediction (EPIP).The selected candidate antigen peptides were used to stimulate peripheral blood mononuclear cells (PBMC) of patients with ovarian cancer and healthy people with the same HLA type,and the immune reactivity was quantified by enzyme-linked immunospot assay (Elispot).Results: 1.The sequencing results showed that the patients had 22 non-synonymous single-nucleotide variants,1 insertions or deletions.2.According to the bioinformatics software,16 ovarian cancer candidate antigen peptides with high affinity and strong presentation were screened out.3.16 candidate antigenic peptides were verified by Elispot in vitro,of which 10 peptides were composed of 6 genes having the potential to become neoantigens.4.Nine candidate antigen peptides with HLA-A*24:02 limit had consistent Elispot results in healthy volunteers and patients with HLA type.Conclusion: The validation rate of ovarian cancer new antigen is significantly improved using bioinformatics software,the selected OR2G3,SLC19A2,SPTB,C3,ZBED6CL and LY75 become potential neoantigens of ovarian cancer.Peripheral blood of healthy individuals with identical HLA typing can be used as a possible alternative to verify the antigenic immunogenicity.

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