WT1多肽乳佐剂疫苗的免疫增强效应及其对急性髓系白血病抗肿瘤效应研究

叶演; 张泽珑; 朱宝行; 刘姝琳; 宋振; 袁庆鹏; 杨赟; 李海波; 邹全明; 曾浩; 孙红武

doi:10.16016/j.2097-0927.202401037

您当前的位置：

首页 >

文章列表页 >

WT1多肽乳佐剂疫苗的免疫增强效应及其对急性髓系白血病抗肿瘤效应研究

药学 | 更新时间：2026-01-09

- WT1多肽乳佐剂疫苗的免疫增强效应及其对急性髓系白血病抗肿瘤效应研究
- Improved immune response and anti-tumor effect of WT1 peptide emulsion adjuvant vaccine for acute myeloid leukemia
- 陆军军医大学学报 2024年46卷第9期页码：1024-1033
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系微生物与生化药学教研室，国家免疫生物制品工程技术研究中心
- 作者简介：
  
  [孙红武，E-mail：sunhongwu2001@163.comSUN Hongwu, E-mail: sunhongwu2001@163.com
- 基金信息：
  
  ‍基‍金‍项‍目‍］‍ ‍国家重点研发计划(2021YFC2302603);国家自然科学基金面上项目(32070924);国家自然科学基金面上项目(31970078);重庆市自然科学基金面上项目(2019jcyjA-msxmx0159)
- DOI：10.16016/j.2097-0927.202401037
  中图分类号： R392.7;R730.51;R733.71
- 收稿：2024-01-10，
  
  修回：2024-02-26，
  
  纸质出版：2024-05-15
- 稿件说明：
移动端阅览
叶演, 张泽珑, 朱宝行, 等. WT1多肽乳佐剂疫苗的免疫增强效应及其对急性髓系白血病抗肿瘤效应研究[J]. 陆军军医大学学报, 2024,46(9):1024-1033.

Yan YE, Zelong ZHANG, Baohang ZHU, et al. Improved immune response and anti-tumor effect of WT1 peptide emulsion adjuvant vaccine for acute myeloid leukemia[J]. Journal of Army Medical University, 2024, 46(9): 1024-1033.
叶演, 张泽珑, 朱宝行, 等. WT1多肽乳佐剂疫苗的免疫增强效应及其对急性髓系白血病抗肿瘤效应研究[J]. 陆军军医大学学报, 2024,46(9):1024-1033. DOI： 10.16016/j.2097-0927.202401037.

Yan YE, Zelong ZHANG, Baohang ZHU, et al. Improved immune response and anti-tumor effect of WT1 peptide emulsion adjuvant vaccine for acute myeloid leukemia[J]. Journal of Army Medical University, 2024, 46(9): 1024-1033. DOI： 10.16016/j.2097-0927.202401037.

摘要

目的

评价Wilm瘤基因1(Wilms’ tumor gene1

WT1)多肽联合AddaVax

乳佐剂疫苗的稳定性、安全性、免疫增强效应及其对急性髓系白血病抗肿瘤效应。

方法

用MALDI-TOF-MS飞行时间质谱评价该佐剂疫苗中WT1多肽的稳定性；将C57BL/6雌性小鼠按照随机数字表法分成3组：PBS组、WT1组和WT1+AddaVax

组，使用免疫剂量为抗原50 μg/只、佐剂50 μg/只，按照第0、14、28天肌肉注射免疫程序进行免疫。用HE染色评价疫苗肌肉注射小鼠脏器组织毒性；用ELISA检测细胞因子水平；ELISpot检测脾细胞分泌IFN-γ细胞数量；流式细胞术检测疫苗促体外骨髓来源树突状细胞(bone marrow-derived dendritic cells，BMDCs)成熟和促淋巴细胞活化及H-2Db WT1四聚体检测特异性CD8

T细胞比例；用C1498-mWT1稳转细胞株构建小鼠白血病肿瘤模型后，评价其预防与治疗的抗肿瘤效应。

结果

WT1多肽可在疫苗中稳定存在，对小鼠肌肉注射未发生明显的脏器组织变化；对小鼠肌肉注射WT1+AddaVax

疫苗后，与WT1组相比，WT1+AddaVax

组可诱导高水平的Th1细胞免疫应答γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)及其Th17免疫应答白介素17A(IL-17A)(

0.05)；与WT1组相比，WT1+AddaVax

组促进脾淋巴细胞分泌IFN-γ的细胞数量增加(

0.01)，促进BMDCs成熟的标志物CD40

/CD11c

、CD86

CD80

/CD11c

细胞比例增加(

0.05)，促进淋巴细胞中CD4

/CD3

T、CD69

/CD8

T细胞比例增加(

0.05)及其特异性CD8

T细胞比例增加(

0.05)；利用C1498-mWT1小鼠急性髓系白血病皮下荷瘤模型的抗肿瘤效应中，WT1+AddaVax

组中位生存期相比于WT1组小鼠延长了6 d。在第50天时，WT1多肽+AddaVax

组小鼠生存率为28.5%，其他组小鼠全部死亡(

0.05)。

结论

WT1多肽联合AddaVax

乳佐剂疫苗具有良好的免疫学和明显抗肿瘤效果。

Abstract

Objective

To evaluate the stability

safety and immune enhancement and anti-tumor effects of Wilms' tumor gene 1 (WT1) peptide combined with AddaVax

emulsion vaccine for acute myeloid leukemia.

Methods

The stability of WT1 peptide in the adjuvant vaccine was evaluated using MALDI-TOF-MS time-o

f-flight mass spectrometry. Female C57BL/6 mice were randomly divided into PBS group

WT1 peptide group

and WT1 peptide+AddaVax

emulsion adjuvant vaccine group. The immunization was performed at a dose of 50 μg/mouse for antigen and 50 μg/mouse for adjuvant

with intramuscular injection on days 0

and 28. HE staining was used to assess the toxicity of intramuscular vaccination on mouse organ tissues. Cytokine levels were detected by ELISA

and the number of IFN-γ-secreting splenocytes was measured by ELISpot. Flow cytometry was employed to detect the maturation of bone marrow-derived dendritic cells (BMDCs) promoted by the vaccine

in vitro

and the promotion for lymphocyte activation

and H-2Db WT1 tetramer was utilized to detect the proportion of specific CD8

T cells. After establishing a mouse leukemia tumor model using the C1498-mWT1 stable cell line

the anti-tumor effects of the vaccine for prevention and treatment were evaluated.

Results

The WT1 peptide stably existed in the vaccine without causing significant organ tissue changes in mice after intramuscular injection. Compared to the mice immunized with WT1 aqueous solution

the mice after intramuscular injection of the WT1 peptide emulsion adjuvant vaccine showed stronger immune responses of Th1 cells

including IFN-γ and TNF-α

as well as Th17 cells of IL-17A (

0.05)

and the mice had not only promoted number of IFN-γ secreting splenocytes (

0.01) but also enhanced maturation of BMDCs

as indicated by an increase in the proportions of CD40

/CD11c

and CD86

CD80

/CD11c

cells (

0.05). Additionally

there were increases in both the proportion of CD4

/CD3

T and CD69

/CD8

T cells (

0.05) and the proportion of specific CD8

T cells (

0.05). In the anti-tumor effect study

using the C1498-mWT1 mouse model

the median survival time of the WT1+AddaVax

group was extended by 6 d compared to the WT1 aqueous solution group. At day 50

the survival rate of mice in the WT1+AddaVax

group was still 28.5%

while all mice in the other groups had died (

0.05).

Conclusion

The vaccine with the WT1 peptide and AddaVax

emulsion adjuvant exhibits good immunological and anti-tumor effects.

关键词

Keywords

references

李秀钰, 胡绍雯, 朱惠芳. 白血病干细胞在急性髓系白血病治疗中的研究进展[J]. 中国现代医学杂志, 2023, 33(21): 49-57. DOI: 10.3969/j.issn.1005-8982.2023.21.008.

LI X Y, HU S W, ZHU H F. The research progress of therapeutic strategies on acute myeloid leukemia by targeting leukemic stem cells[J]. China J Mod Med, 2023, 33(21): 49-57. DOI: 10.3969/j.issn.1005-8982.2023.21.008.

王婉頔, 常桃, 江思远, 等. 急性髓系白血病新型预后分子标志物NKX2-3的发现[J]. 中山大学学报(医学科学版), 2024, 45(1): 63-68. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).20240004.014.

WANG W D, CHANG T, JIANG S Y, et al . Discovery of A new prognostic molecular marker NKX2-3 for acute myeloid leukemia[J ] . J Sun Yat Sen Univ Med Sci, 2024, 45(1): 63-68. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).20240004.014.

KUMAI T, FAN A, HARABUCHI Y, et al . Cancer immunotherapy: moving forward with peptide T cell vaccines[J ] . Curr Opin Im munol, 2017, 47: 57-63. DOI: 10.1016/j.coi.2017.07.003.

OUE T, UEHARA S, YAMANAKA H, et al . Expression of Wilms tumor 1 gene in a varietyof pediatric tumors[J ] . J Pediatr Surg, 2011, 46(12): 2233-2238. DOI: 10.1016/j.jpedsurg.2011.09.004.

XU N, CHEN W M, LI L D, et al . High WT1 expression predicted induction chemotherapy failure in acute myeloid leukemia patients with non-favorable cytogenetic risk[J ] . Clin Exp Med, 2023, 23(6): 2629-2638. DOI: 10.1007/s10238-023-00995-5.

CLARKE J M, MORSE M A, LYERLY H K, et al . Adenovirus vaccine immunotherapy targeting WT1-expressing tumors[J ] . Expert Opin Biol Ther, 2010, 10(6): 875-883. DOI: 10.1517/14712591003798278.

OKA Y, TSUBOI A, KAWAKAMI M, et al . Development of WT1 peptide cancer vaccine against hematopoietic malignancies and solid cancers[J ] . Curr Med Chem, 2006, 13(20): 2345-2352. DOI: 10.2174/092986706777935104.

OKA Y, TSUBOI A, OJI Y, et al . WT1 peptide vaccine for the treatment of cancer[J ] . Curr Opin Immunol, 2008, 20(2): 211-220. DOI: 10.1016/j.coi.2008.04.009.

ACHEAMPON G D O, ADOKOH C K, ASANTE D B, et al . Immunotherapy for acute myeloid leukemia (AML): a potent alternative therapy[J ] . Biomed Pharmacother, 2018, 97: 225-232. DOI: 10.1016/j.biopha.2017.10.100.

AUGSBERGER C, HÄNEL G, XU W, et al . Targeting intracellular WT1 in AML with a novel RMF-peptide-MHC-specific T-cell bispecific antibody[J ] . Blood, 2021, 138(25): 2655-2669. DOI: 10.1182/blood.2020010477.

O'HAGAN D T, OTT G S, NEST G V, et al . Thehistory of MF59 ® adjuvant: a phoenix that arose from the ashes[J ] . Expert Rev Vaccines, 2013, 12(1): 13-30. DOI: 10.1586/erv.12.140..

ALZUA G P, PIHL A F, OFFERSGAARD A, et al . Inactivated genotype 1a, 2a and 3a HCV vaccine candidates induced broadly neutralising antibodies in mice[J ] . Gut, 2023, 72(3): 560-572. DOI: 10.1136/gutjnl-2021-326323.

KIM E H, WOODRUFF M C, GRIGORYAN L, et al . Squalene emulsion-based vaccine adjuvants stimulate CD8 T cell, but not antibody responses, through a RIPK3-dependent pathway[J ] . eLife, 2020, 9: e52687. DOI: 10.7554/eLife.52687.

KITAGAWA K, ODA T, SAITO H, et al . Development of oral cancer vaccine using recombinant Bifidobacterium displaying Wilms' tumor 1 pr otein[J ] . Cancer Immunol Immunother, 2017, 66(6): 787-798. DOI: 10.1007/s00262-017-1984-0..

NAKAGAWA N, HASHⅡ Y, KAYAMA H, et al . An oral WT1 protein vaccine composed of WT1-anchored, genetically engineered Bifidobacterium longum allows for intestinal immunity in mice with acute myeloid leukemia[J ] . Cancer Immunol Immunother, 2023, 72(1): 39-53. DOI: 10.1007/s00262-022-03214-4..

FUJITA T, TERAMOTO K, OZAKI Y, et al . Inhibition of transforming growth factor-beta-mediated immunosuppression in tumor-draining lymph nodes augments antitumor responses by various immunologic cell types[J ] . Cancer Res, 2009, 69(12): 5142-5150. DOI: 10.1158/0008-5472.CAN-08-2499.

JEANBART L, BALLESTER M, TITTA A D, et al . Enhancing efficacy of anticancer vaccines by targeted delivery to tumor-draining lymph nodes[J ] . Cancer Immunol Res, 2014, 2(5): 436-447. DOI: 10.1158/2326-6066.CIR-14-0019-T.

PENAFUERTE C, NG S, BAUTISTA-LOPEZ N, et al . B effector cells activated by a chimeric protein consisting of IL-2 and the ectodomain of TGF-β receptor Ⅱ induce potent antitumor immunity[J ] . Cancer Res, 2012, 72(5): 1210-1220. DOI: 10.1158/0008-5472.CAN-11-1659.

YUBA E, KONO Y, HARADA A, et al . The application of pH-sensitive polymer-lipids to antigen delivery for cancer immunotherapy[J ] . Biomaterials, 2013, 34(22): 5711-5721. DOI: 10.1016/j.biomaterials.2013.04.007.

VADAKEKOLATHU J, MINDEN M D, HOOD T, et al . Immune landscapes predict chemotherapy resistance and immunotherapy response in acute myeloid leukemia[J ] . Sci Transl Med, 2020, 12(546): eaaz0463. DOI: 10.1126/scitranslmed.aaz0463.

REZVANI K. Peptide vaccine therapy for leukemia[J]. Int J Hematol, 2011, 93(3): 274-280. DOI: 10.1007/s12185-011-0781-3.

WANG F. Identification of tumor antigens and immune subtypes of acute myeloid leukemia for mRNA vaccine development[J]. Clin Transl Oncol, 2023, 25(7): 2204-2223. DOI: 10.1007/s12094-023-03108-6.

O'HAGAN D T, OTT G S, GREGORIO E D, et al . The mechanism of action of MF59—an innately attractive adjuvant formulation[J ] . Vaccine, 2012, 30(29): 4341-4348. DOI: 10.1016/j.vaccine.2011.09.061.

JIE J, LIU G M, FENG J Y, et al . MF59 promoted the combination of CpG ODN1826 and MUC1-MBP vaccine-induced antitumor activity involved in the enhancement of DC maturation by prolonging the local retention time of antigen and down-regulating of IL-6/STAT3[J ] . Int J Mol Sci, 2022, 23(18): 10887 . DOI: 10.3390/ijms231810887.

SON M H, JUNG M Y, CHOI S, et al . IL-32γ induces chemotaxis of activated T cells via dendritic cell-derived CCL5[J ] . Biochem Biophys Res Commun, 2014, 450(1): 30-35. DOI: 10.1016/j.bbrc.2014.05.052.

LUO R, ONYSHCHENKO K, WANG L, et al . Necroptosis-dependent immunogenicity of cisplatin: implications for enhancing the radiation-induced abscopal effect[J ] . Clin Cancer Res, 2023, 29(3): 667-683. DOI: 10.1158/1078-0432.CCR-22-1591.

O'HAGAN D T, FRIEDLAND L R, HANON E, et al . Towards an evidence based approach for the development of adjuvanted vaccines[J ] . Curr Opin Immunol, 2017, 47: 93-102. DOI: 10.1016/j.coi.2017.07.010.

浏览量

182

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

核仁磷酸蛋白1突变的分子异质性对急性髓系白血病预后的影响

植物源性免疫刺激分子麦冬皂苷的免疫增强效应及初步机制

高原低氧改善肿瘤免疫微环境并抑制结直肠癌皮下肿瘤生长

Auer小体对初诊非M3型急性髓细胞白血病患者预后的影响

miRNA-495靶向STYX调节MLL-AF9的急性髓系白血病细胞的增殖和凋亡