摘要A204:一种基于gd3的保护性疫苗增加了C57BL/6小鼠模型中的nkt细胞

R. Milner, B. Sahay, Matt Cascio, M. Salute
{"title":"摘要A204:一种基于gd3的保护性疫苗增加了C57BL/6小鼠模型中的nkt细胞","authors":"R. Milner, B. Sahay, Matt Cascio, M. Salute","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-A204","DOIUrl":null,"url":null,"abstract":"The disialyl gangliosides GD2/GD3 have been implicated in the enhancement of malignancy in a number of human and animal cancers and as a tumor antigen target for immunotherapy. Earlier we reported on a GD3-based vaccine, which protects canine melanoma patients when provided as an adjunct to standard of care (1). Despite a significant increase in the median survival time, the mechanism by which vaccine works remains unclear; therefore, we vaccinated C57BL/6 mice with the GD3-vaccine formulation and appropriate controls and evaluated changes in the immune cells. Methods: C57BL/6 mice were vaccinated weekly for 4 weeks followed by a week of rest. Mice were then euthanized to collect blood, liver, and spleen for evaluation of NKT-cells. Liver lymphocytes were collected by making mono-cellular suspension of liver obtained after enzymatic digestion. Collected lymphocytes were stained for CD3, CD4, TCRβ, NK1.1, CD49b and dead cells. In these experiments untreated and α-galactosylceramide (α-GalCer) treated mice were used as controls. Results: Mice treated with the subcutaneous injections of the GD3 vaccine showed an increase in NKT-cells (NK1.1+CD49b+CD3+CD4+TCRb+) in the liver but failed to show an increase in the spleen and blood when compared with the untreated mice. The mice treated with α-GalCer also showed an accumulation of NKT-cells in the liver; however, the increase in the livers obtained from vaccine-treated mice was 5-10-fold higher compared to the α-GalCer treated mice. Discussion: NKT-cells are the Swiss army knife of the immune system which are capable of producing different cytokines and chemokines to regulate the overall immune system. After their discovery two decades ago, activation of NKT-cells have shown to have a crucial protective role in various infectious, and non-infectious diseases. These cells form a bridge between the innate and adaptive immune cells. The activation and maintenance of these cells are dependent upon the presentation of lipid molecules on CD1 receptors by dendritic cells. Since the target antigen in the vaccine is a lipid (GD3) it is very likely it would be presented on CD1 receptors for the activation of NKT-cells. Due to the lack of validated and suitable canine CD1 receptor reagents, we were unable to define the mechanism behind the hypothetical protection found in the canine GD3 based vaccine (1). In the murine model NKT-cells are well characterized, and provided us with an opportunity to understand the possible selective activation of NKT-cells by the GD3 based vaccine. Our data found an increase in NKT-cells in the liver for mice vaccinated with GD3 and α-GalCer, but no discernible differences were found in blood and spleen between vaccinated mice and normal controls. Further investigation using the B16 melanoma cell line in C57BL/6 mice vaccinated with the GD3 vaccine and suitable controls may identify the NKT response in the tumor microenvironment. Conclusions: While the GD3-based vaccine and α-GalCer both increased NKT-cells in the murine liver, GD3 increased NKT counts 5-10 fold over α-GalCer. In addition, monitoring changes in NKT numbers in the peripheral blood may not be of benefit due to low cell counts. Acknowledgment: The study is funded by a grant from the American Kennel Club Health Foundation and The UF CVM Foundation. Reference: 1. Milner RJ. The immune response to disialoganglioside GD3 vaccination in dogs with spontaneous occurring melanoma: A large animal translational model. Poster # 123628_1 presented at: Proceeding American Association for Cancer Research, Tumor Immunology: Basic and Clinical Advances; 2010 Dec 30; Miami Beach, Florida. Citation Format: Rowan J. Milner, Bikash Sahay, Matthew Cascio, Marc Salute. A protective GD3-based vaccine increases NKT-cells in a C57BL/6 murine model [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A204.","PeriodicalId":170885,"journal":{"name":"Regulating T-cells and Their Response to Cancer","volume":"23 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Abstract A204: A protective GD3-based vaccine increases NKT-cells in a C57BL/6 murine model\",\"authors\":\"R. Milner, B. Sahay, Matt Cascio, M. Salute\",\"doi\":\"10.1158/2326-6074.CRICIMTEATIAACR18-A204\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The disialyl gangliosides GD2/GD3 have been implicated in the enhancement of malignancy in a number of human and animal cancers and as a tumor antigen target for immunotherapy. Earlier we reported on a GD3-based vaccine, which protects canine melanoma patients when provided as an adjunct to standard of care (1). Despite a significant increase in the median survival time, the mechanism by which vaccine works remains unclear; therefore, we vaccinated C57BL/6 mice with the GD3-vaccine formulation and appropriate controls and evaluated changes in the immune cells. Methods: C57BL/6 mice were vaccinated weekly for 4 weeks followed by a week of rest. Mice were then euthanized to collect blood, liver, and spleen for evaluation of NKT-cells. Liver lymphocytes were collected by making mono-cellular suspension of liver obtained after enzymatic digestion. Collected lymphocytes were stained for CD3, CD4, TCRβ, NK1.1, CD49b and dead cells. In these experiments untreated and α-galactosylceramide (α-GalCer) treated mice were used as controls. Results: Mice treated with the subcutaneous injections of the GD3 vaccine showed an increase in NKT-cells (NK1.1+CD49b+CD3+CD4+TCRb+) in the liver but failed to show an increase in the spleen and blood when compared with the untreated mice. The mice treated with α-GalCer also showed an accumulation of NKT-cells in the liver; however, the increase in the livers obtained from vaccine-treated mice was 5-10-fold higher compared to the α-GalCer treated mice. Discussion: NKT-cells are the Swiss army knife of the immune system which are capable of producing different cytokines and chemokines to regulate the overall immune system. After their discovery two decades ago, activation of NKT-cells have shown to have a crucial protective role in various infectious, and non-infectious diseases. These cells form a bridge between the innate and adaptive immune cells. The activation and maintenance of these cells are dependent upon the presentation of lipid molecules on CD1 receptors by dendritic cells. Since the target antigen in the vaccine is a lipid (GD3) it is very likely it would be presented on CD1 receptors for the activation of NKT-cells. Due to the lack of validated and suitable canine CD1 receptor reagents, we were unable to define the mechanism behind the hypothetical protection found in the canine GD3 based vaccine (1). In the murine model NKT-cells are well characterized, and provided us with an opportunity to understand the possible selective activation of NKT-cells by the GD3 based vaccine. Our data found an increase in NKT-cells in the liver for mice vaccinated with GD3 and α-GalCer, but no discernible differences were found in blood and spleen between vaccinated mice and normal controls. Further investigation using the B16 melanoma cell line in C57BL/6 mice vaccinated with the GD3 vaccine and suitable controls may identify the NKT response in the tumor microenvironment. Conclusions: While the GD3-based vaccine and α-GalCer both increased NKT-cells in the murine liver, GD3 increased NKT counts 5-10 fold over α-GalCer. In addition, monitoring changes in NKT numbers in the peripheral blood may not be of benefit due to low cell counts. Acknowledgment: The study is funded by a grant from the American Kennel Club Health Foundation and The UF CVM Foundation. Reference: 1. Milner RJ. The immune response to disialoganglioside GD3 vaccination in dogs with spontaneous occurring melanoma: A large animal translational model. Poster # 123628_1 presented at: Proceeding American Association for Cancer Research, Tumor Immunology: Basic and Clinical Advances; 2010 Dec 30; Miami Beach, Florida. Citation Format: Rowan J. Milner, Bikash Sahay, Matthew Cascio, Marc Salute. A protective GD3-based vaccine increases NKT-cells in a C57BL/6 murine model [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A204.\",\"PeriodicalId\":170885,\"journal\":{\"name\":\"Regulating T-cells and Their Response to Cancer\",\"volume\":\"23 1\",\"pages\":\"0\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2019-02-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Regulating T-cells and Their Response to Cancer\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-A204\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Regulating T-cells and Their Response to Cancer","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-A204","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

双醚神经节苷脂GD2/GD3与许多人类和动物癌症的恶性增强有关,并作为免疫治疗的肿瘤抗原靶点。早些时候,我们报道了一种基于gd3的疫苗,当作为标准护理的辅助手段提供时,它可以保护犬黑色素瘤患者(1)。尽管中位生存时间显着增加,但疫苗起作用的机制尚不清楚;因此,我们用gd3疫苗制剂和适当的对照接种C57BL/6小鼠,并评估免疫细胞的变化。方法:C57BL/6小鼠每周接种4周,休息1周。然后对小鼠实施安乐死,收集血液、肝脏和脾脏用于评估nkt细胞。酶解后制备肝脏单细胞悬液,收集肝淋巴细胞。收集淋巴细胞进行CD3、CD4、TCRβ、NK1.1、CD49b和死细胞染色。实验以未处理和α-半乳糖神经酰胺(α-GalCer)处理的小鼠为对照。结果:与未注射GD3疫苗的小鼠相比,皮下注射GD3疫苗的小鼠肝脏中nkt细胞(NK1.1+CD49b+CD3+CD4+TCRb+)增加,但脾脏和血液中未显示出增加。α-GalCer处理的小鼠肝脏中也出现了nkt细胞的积累;然而,与α-GalCer处理的小鼠相比,疫苗处理小鼠肝脏的增加是α-GalCer处理小鼠的5-10倍。讨论:nkt细胞是免疫系统的瑞士军刀,能够产生不同的细胞因子和趋化因子来调节整个免疫系统。在二十年前发现nkt细胞后,nkt细胞的激活已被证明在各种传染性和非传染性疾病中具有至关重要的保护作用。这些细胞在先天免疫细胞和适应性免疫细胞之间形成了一座桥梁。这些细胞的激活和维持依赖于树突状细胞在CD1受体上的脂质分子的呈现。由于疫苗中的靶抗原是脂质(GD3),因此它很可能会在CD1受体上呈现以激活nkt细胞。由于缺乏经过验证和合适的犬CD1受体试剂,我们无法确定犬GD3疫苗中假设的保护机制(1)。在小鼠模型中,nkt细胞被很好地表征,这为我们提供了一个了解基于GD3的疫苗可能选择性激活nkt细胞的机会。我们的数据发现,接种GD3和α-GalCer的小鼠肝脏中的nkt细胞增加,但在接种小鼠和正常对照组之间的血液和脾脏中未发现明显差异。在接种GD3疫苗和适当对照的C57BL/6小鼠中进一步研究B16黑色素瘤细胞系,可能会确定NKT在肿瘤微环境中的反应。结论:虽然基于GD3的疫苗和α-GalCer均能增加小鼠肝脏中NKT细胞,但GD3比α-GalCer能使NKT细胞数量增加5-10倍。此外,由于细胞计数低,监测外周血中NKT数的变化可能没有好处。致谢:本研究由美国养犬俱乐部健康基金会和UF CVM基金会资助。参考:1。米尔纳RJ。自发性黑色素瘤犬对二联神经节苷脂GD3疫苗的免疫反应:大型动物转化模型海报# 123628_1发表于:美国癌症研究协会会刊,肿瘤免疫学:基础和临床进展;2010年12月30日;迈阿密海滩,佛罗里达州。引用格式:Rowan J. Milner, Bikash Sahay, Matthew Cascio, Marc Salute。在C57BL/6小鼠模型中,基于gd3的保护性疫苗增加了nkt细胞[摘要]。第四届CRI-CIMT-EATI-AACR国际癌症免疫治疗会议:将科学转化为生存;2018年9月30日至10月3日;纽约,纽约。费城(PA): AACR;癌症免疫学杂志,2019;7(2增刊):摘要nr A204。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Abstract A204: A protective GD3-based vaccine increases NKT-cells in a C57BL/6 murine model
The disialyl gangliosides GD2/GD3 have been implicated in the enhancement of malignancy in a number of human and animal cancers and as a tumor antigen target for immunotherapy. Earlier we reported on a GD3-based vaccine, which protects canine melanoma patients when provided as an adjunct to standard of care (1). Despite a significant increase in the median survival time, the mechanism by which vaccine works remains unclear; therefore, we vaccinated C57BL/6 mice with the GD3-vaccine formulation and appropriate controls and evaluated changes in the immune cells. Methods: C57BL/6 mice were vaccinated weekly for 4 weeks followed by a week of rest. Mice were then euthanized to collect blood, liver, and spleen for evaluation of NKT-cells. Liver lymphocytes were collected by making mono-cellular suspension of liver obtained after enzymatic digestion. Collected lymphocytes were stained for CD3, CD4, TCRβ, NK1.1, CD49b and dead cells. In these experiments untreated and α-galactosylceramide (α-GalCer) treated mice were used as controls. Results: Mice treated with the subcutaneous injections of the GD3 vaccine showed an increase in NKT-cells (NK1.1+CD49b+CD3+CD4+TCRb+) in the liver but failed to show an increase in the spleen and blood when compared with the untreated mice. The mice treated with α-GalCer also showed an accumulation of NKT-cells in the liver; however, the increase in the livers obtained from vaccine-treated mice was 5-10-fold higher compared to the α-GalCer treated mice. Discussion: NKT-cells are the Swiss army knife of the immune system which are capable of producing different cytokines and chemokines to regulate the overall immune system. After their discovery two decades ago, activation of NKT-cells have shown to have a crucial protective role in various infectious, and non-infectious diseases. These cells form a bridge between the innate and adaptive immune cells. The activation and maintenance of these cells are dependent upon the presentation of lipid molecules on CD1 receptors by dendritic cells. Since the target antigen in the vaccine is a lipid (GD3) it is very likely it would be presented on CD1 receptors for the activation of NKT-cells. Due to the lack of validated and suitable canine CD1 receptor reagents, we were unable to define the mechanism behind the hypothetical protection found in the canine GD3 based vaccine (1). In the murine model NKT-cells are well characterized, and provided us with an opportunity to understand the possible selective activation of NKT-cells by the GD3 based vaccine. Our data found an increase in NKT-cells in the liver for mice vaccinated with GD3 and α-GalCer, but no discernible differences were found in blood and spleen between vaccinated mice and normal controls. Further investigation using the B16 melanoma cell line in C57BL/6 mice vaccinated with the GD3 vaccine and suitable controls may identify the NKT response in the tumor microenvironment. Conclusions: While the GD3-based vaccine and α-GalCer both increased NKT-cells in the murine liver, GD3 increased NKT counts 5-10 fold over α-GalCer. In addition, monitoring changes in NKT numbers in the peripheral blood may not be of benefit due to low cell counts. Acknowledgment: The study is funded by a grant from the American Kennel Club Health Foundation and The UF CVM Foundation. Reference: 1. Milner RJ. The immune response to disialoganglioside GD3 vaccination in dogs with spontaneous occurring melanoma: A large animal translational model. Poster # 123628_1 presented at: Proceeding American Association for Cancer Research, Tumor Immunology: Basic and Clinical Advances; 2010 Dec 30; Miami Beach, Florida. Citation Format: Rowan J. Milner, Bikash Sahay, Matthew Cascio, Marc Salute. A protective GD3-based vaccine increases NKT-cells in a C57BL/6 murine model [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A204.
求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信