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基于硝基化T细胞表位的PD-L1疫苗佐剂的筛选

周虹佑, 高向东, 姚文兵, 田浤

周虹佑,高向东,姚文兵,等. 基于硝基化T细胞表位的PD-L1疫苗佐剂的筛选[J]. 中国药科大学学报,2024,55(3):397 − 403. DOI: 10.11665/j.issn.1000-5048.2023032801
引用本文: 周虹佑,高向东,姚文兵,等. 基于硝基化T细胞表位的PD-L1疫苗佐剂的筛选[J]. 中国药科大学学报,2024,55(3):397 − 403. DOI: 10.11665/j.issn.1000-5048.2023032801
ZHOU Hongyou, GAO Xiangdong, YAO Wenbing, et al. Screening of adjuvant for PD-L1 vaccine based on nitrated T cell epitope[J]. J China Pharm Univ, 2024, 55(3): 397 − 403. DOI: 10.11665/j.issn.1000-5048.2023032801
Citation: ZHOU Hongyou, GAO Xiangdong, YAO Wenbing, et al. Screening of adjuvant for PD-L1 vaccine based on nitrated T cell epitope[J]. J China Pharm Univ, 2024, 55(3): 397 − 403. DOI: 10.11665/j.issn.1000-5048.2023032801

基于硝基化T细胞表位的PD-L1疫苗佐剂的筛选

基金项目: 国家自然科学基金项目(No.81973222,No.82073754);新疆维吾尔自治区重点研发计划项目(No.2020B03003)
详细信息
    通讯作者:

    田浤: Tel:025-83271543 E-mail:tinahew@139.com

  • 中图分类号: R186;R967

Screening of adjuvant for PD-L1 vaccine based on nitrated T cell epitope

Funds: This study was supported by the National Natural Science Foundation of China (No.81973222,No.82073754) and the Key Research and Development Program of Xinjiang Uygur Autonomous Region (No.2020B03003)
  • 摘要:

    为了充分发挥本课题组前期设计的基于硝基化T表位的靶向PD-L1的肿瘤疫苗(PD-L1-NitraTh)的抑瘤活性,选择了作用机制不同的几种佐剂进行比较,以期筛选出最适用于此类疫苗的佐剂。研究结果显示,Poly(I:C)、CPG1018、肿节风多糖SGP2及GM-CSF等佐剂均可以提高PD-L1-NitraTh疫苗的免疫原性,其中,Poly(I:C)组诱导产生的抗体滴度最高。对T细胞分化相关转录因子的qPCR检测结果显示,Poly(I:C)减少了GATA3和FoxP3的表达,提示对CD4+T细胞分化有较强的影响。同时,相比其他佐剂,Poly(I:C)可以辅助PD-L1-NitraTh增加肿瘤内T淋巴细胞以及CD11b+细胞浸润,提示 Poly(I:C)佐剂可能适用于以硝基化T表位为基础的肿瘤疫苗。

    Abstract:

    To enhance the anti-tumor activity of tumor vaccine targeting PD-L1 based on the nitrated T-epitope (PD-L1-NitraTh), this research compared several adjuvants with different mechanisms to screen out the adjuvant most suitable for PD-L1-NitraTh. The results showed that Poly(I:C), CPG1018, swollen knotted polysaccharide SGP2 and GM-CSF could enhance the immunogenicity of PD-L1-NitraTh when used as adjuvants, with the Poly(I:C) group inducing the highest antibody titer. The results of qPCR for T cell differentiation-related cytokines showed that Poly(I:C) reduced the expression of GATA3 and FoxP3, indicating a strong effect on CD4+ T cell differentiation. Besides, compared with other adjuvants, Poly(I:C) could assist PD-L1-NitraTh to increase the infiltration of T cells as well as CD11b+ cells within tumor, suggesting that Poly(I:C) may be the suitable adjuvant for tumor vaccines based on the nitrated T epitopes.

  • Figure  1.   Immunogenicity of PD-L1-NitraTh with different adjuvants ($ \bar{x}\pm s $, n =5)

    A: Determination of anti-PD-L1 antibody titer in mouse serum; B: Number of IFN-γ ELISPOT spots of mouse spleen cells; C: IFN-γ ELISPOT speckle pattern of mouse spleen cells **P < 0.01,***P < 0.001, ****P < 0.0001

    Figure  2.   Antitumor activity of PD-L1-NitraTh with different adjuvants ($ \bar{x}\pm s $, n =5)

    A:Tumor volume measured from the beginning of PD-L1-NatriTh treatment; B: Volume of the tumors excised after the mice were sacrificed (circle indicates tumor regression; triangle represents the death of the mouse); C: Tumor volume of mice at the end of the experiment**P < 0.01

    Figure  3.   Detection of relevant proteins relative expression levels in lymph node cells by qRT-PCR($ \bar{x}\pm s $, n =5)

    A: GATA3; B: Tbet; C: Foxp3**P < 0.01

    Figure  4.   Detection of tumor-infiltrating immune cells in mouse tumor tissues($ \bar{x}\pm s $, n =5)

    A: Frequency of CD11b+ cells in tumor tissues after immunization; B: Frequency of CD3+ cells in tumor tissues after immunization *P < 0.05, ****P < 0.0001

    Figure  5.   Detection of relevant proteins relative expression levels in tumor cells by qRT-PCR($ \bar{x}\pm s $, n =5)

    A:Tbet; B:IFN-γ; C: Foxp3

    Table  1   Grouping situation and immune dose of mice

    GroupDose of
    PD-L1-NitraTh/μg
    AdjuvantDose of
    adjuvant/μg
    Blank---
    PD-L150--
    CPG101850CPG101835
    Poly(I:C)50Poly(I:C)50
    GM-CSF50GM-CSF0.2
    SGP250SGP2200
    Blank group was inoculated with saline, PD-L1 group was inoculated with PD-L1-NitraTh only, and the other groups were inoculated with PD-L1-NitraTh mixed with the corresponding adjuvant respectively
    GM-CSF:Granulocyte-macrophage colony-stimulating factor ;SGP2:
    An acidic polysaccharide from Sarcandra glabra
    下载: 导出CSV

    Table  2   Primer sequence for PCR

    GeneForward primer sequence(5′→3′)Reverse primer sequence(5′→3′)
    TNF-αCCCTCACACTCAGATCATCTTCTGCTACGACGTGGGCTACAG
    INF-γATGAACGCTACACACTGCATCCCATCCTTTTGCCAGTTCCTC
    GATA3CTCGGCCATTCGTACATGGAAGGATACCTCTGCACCGTAGC
    CD11cCTGGATAGCCTTTCTTCTGCTGGCACACTGTGTCCGAACTCA
    CD206CTCTGTTCAGCTATTGGACGCCGGAATTTCTGGGATTCAGCTTC
    Foxp3CCCATCCCCAGGAGTCTTGACCATGACTAGGGGCACTGTA
    β-actinACTCCTATGTGGGTGACGAGCATCTTTTCACGGTTGGCCTTAG
    下载: 导出CSV
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出版历程
  • 收稿日期:  2023-03-27
  • 网络出版日期:  2024-06-24
  • 刊出日期:  2024-06-24

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