Impact of malaria on shaping immunity to EBV in the etiology of Burkitt lymphoma

疟疾对伯基特淋巴瘤病因中 EBV 免疫力的影响

• 批准号: 10655570
• 负责人: ANN M MOORMANN
• 金额: $ 56.02万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-10 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655570
• 关键词: 1 year old; 6 year old; 9 year old; Activities of Daily Living; Acute; Adult; Africa; African; African Burkitt' s lymphoma; Antigens; Area; B-Lymphocytes; Biological Assay; Blood; Burkitt Lymphoma; CD8-Positive T-Lymphocytes; CD8B1 gene; CTLA4 gene; Cell Line; Cell physiology; Cells; Child; Childhood; Chronic; Coculture Techniques; Cytotoxic T-Lymphocytes; Development; Diagnosis; EBV specific T-cells; Enrollment; Environment; Epstein-Barr Virus Infections; Epstein-Barr Virus-Related Malignant Neoplasm; Etiology; Exposure to; FOXP3 gene; Falciparum Malaria; Family member; Fever; Flow Cytometry; Frequencies; Granzyme; Herpesviridae Infections; Heterogeneity; Homeostasis; Human; Human Herpesvirus 4; IL24 gene; IL2RA gene; Immune; Immunity; Immunocompetent; Immunologic Surveillance; Immunologics; Impairment; In Vitro; Individual; Infection; Inflammatory; Interferon Type II; Interleukin-10; Intervention; Lead; Ligands; Link; Malaria; Malignant Childhood Neoplasm; Malignant Neoplasms; Measures; Mediating; Mediator; Memory; Mus; Natural Killer Cells; OX40; Parasites; Pathogenesis; Pathologic; Pathway interactions; Patients; Plasmodium falciparum; Recording of previous events; Regulatory T-Lymphocyte; Risk; Role; Schedule; Shapes; Signal Transduction; Site; T cell response; T memory cell; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Time; Viral; Viral load measurement; Virus Replication; Visit; cancer cell; chronic infection; co-infection; cohort; cytokine; cytotoxicity; density; design; effector T cell; exhaustion; experience; experimental study; immunopathology; immunoregulation; improved; in vitro testing; infected B cell; interleukin 20; interleukin-19; interleukin-22; lymphoblastoid cell line; malaria infection; permissiveness; prevent; programmed cell death protein 1; receptor; receptor expression; restraint; single-cell RNA sequencing; transcriptome sequencing; transforming virus; tumor; tumorigenesis

Plasmodium falciparum (Pf) malaria and Epstein-Barr Virus (EBV) co-infections in children residing in malaria holoendemic areas have been linked to an increased risk of an EBV-associated cancer called endemic Burkitt lymphoma (eBL). Most African children are infected with EBV before 1 year of age, yet this B-cell cancer does not occur until years later. It has been postulated that repeated episodes of malaria ‘suppress’ immunity to EBV, creating a permissive environment for eBL pathogenesis. However, the mechanisms responsible are not fully understood. Our prior studies found that malaria-exposed children had pathologically high EBV loads; naïve-like EBV-specific CD8+ T cells with diminished effector functions; unconventional, innate-like CD8+ T cells that expressed Granzyme B in lieu of IFN-γ; and an expansion of ‘chronic-infection induced’ CD56neg Natural Killer (NK) cells with impaired cytotoxicity. Thus, we have identified proximate immunologic alterations that allow unrestrained EBV replication and eBL tumorigenesis. In this renewal application, we will our central hypothesis that malaria-induced immunoregulatory mechanisms restrain T cell cytotoxicity against EBV-infected B cells and eBL tumors. This will be tested by the following Specific Aims. Aim 1. Determine if repeated Pf-malaria infections, known to induce EBV reactivation, lead to increased inhibitory co- receptor expression on EBV-specific CD8+ αβ T cells. Expression of TIGIT, PD1, CTLA4, LAG3, TIM3, CD160, 2B4, KLRG1, BTLA, on T cell subsets will be measured by flow cytometry. Exhaustion versus cytotoxicity signatures will be further defined with single cell RNA sequencing, and functional capacity tested in vitro by cytotoxic T lymphocyte (CTL) assays using EBV-transformed lymphoblastoid cell lines (LCLs). Aim 2. Determine if repeated Pf-malaria infections induce IL-10 producing CD4+ or CD8+ T cells that exert an immune-regulatory effect on EBV-specific T cells. The frequency of IL-10 secreting Foxp3neg regulatory CD25+, CD4+, Tr1 cells (CD49b+, LAG3+, CD226+/DNAM1+), Treg-of-B cells (LAG3+, ICOS+, PD1+, GITR+, OX40+) and CD8+ CD25neg Foxp3neg T cells will be measured by flow cytometry and RNAseq to distinguish them from classical CD4+Fox+p3+ regulatory T cells (Tregs). CTL assays will determine the impact of IL-10 cytokine family members on CD8+ T cell cytotoxicity, in vitro. Aim 3. Determine if repeated Pf-malaria infections influence the frequency of γδT to NK cell subsets and how their relative ratios impact cytotoxicity to eBL tumors. The frequency of γδT and NK cell subsets will be evaluated by flow cytometry and associated with malaria exposure. Cytotoxicity of γδ T and NK cell subsets will be quantified in vitro against BL tumors, including our newly established patient-derived eBL cell lines. Ligand-receptor blocking experiments will evaluate the relative contribution of each subset to overall cytotoxicity. Understanding how malaria influences the human immunologic landscape, especially in children, will allow us to explore interventions that modulate regulatory mechanisms while maintaining protective immunity to EBV.

疟疾儿童中恶性疟原虫 (Pf) 疟疾和 EB 病毒 (EBV) 双重感染 全流行地区与 EB 病毒相关癌症（称为地方性伯基特病）的风险增加有关 大多数非洲儿童在一岁之前感染 EBV，但这种 B 细胞癌却会感染。 据推测，疟疾的反复发作会“抑制”对人体的免疫力。 EBV，为 eBL 发病机制创造了宽松的环境，但其机制却并非如此。 我们之前的研究发现，接触过疟疾的儿童具有病理性的高 EBV 载量； 幼稚样 EBV 特异性 CD8+ T 细胞，效应功能减弱； 表达颗粒酶 B 代替 IFN-γ 的细胞以及“慢性感染诱导的”CD56neg 的扩增； 细胞毒性受损的自然杀伤（NK）细胞因此，我们已经确定了直接的免疫学改变。 允许不受限制的 EBV 复制和 eBL 肿瘤发生 在这个更新应用中，我们将成为我们的核心。 假设疟疾诱导的免疫调节机制抑制 T 细胞的细胞毒性 EBV 感染的 B 细胞和 eBL 肿瘤将通过以下具体目标进行测试。 1. 确定。 如果重复的 Pf-疟疾感染​​（已知会诱导 EBV 重新激活）导致抑制性协同增加 EBV 特异性 CD8+ αβ T 细胞上的受体表达 TIGIT、PD1、CTLA4、LAG3、TIM3、 T 细胞亚群上的 CD160、2B4、KLRG1、BTLA 将通过流式细胞术测量。 将通过单细胞 RNA 测序进一步定义细胞毒性特征，并测试功能能力 使用 EBV 转化的淋巴母细胞系 (LCL) 进行体外细胞毒性 T 淋巴细胞 (CTL) 测定。 目标 2. 确定重复的 Pf-疟疾感染​​是否诱导产生 IL-10 的 CD4+ 或 CD8+ T 细胞 对 EBV 特异性 T 细胞发挥免疫调节作用 IL-10 分泌 Foxp3neg 的频率。 调节性 CD25+、CD4+、Tr1 细胞（CD49b+、LAG3+、CD226+/DNAM1+）、Treg-of-B 细胞（LAG3+、ICOS+、PD1+、 GITR+、OX40+) 和 CD8+ CD25neg Foxp3neg T 细胞将通过流式细胞术和 RNAseq 进行测量 将它们与经典的 CD4+Fox+p3+ 调节性 T 细胞 (Treg) 区分开来，以确定其影响。 IL-10 细胞因子家族成员对 CD8+ T 细胞细胞毒性的体外作用 目标 3. 确定 Pf-疟疾是否重复。 感染影响 NK 细胞亚群的 γδT 频率以及它们的相对比率如何影响 对 eBL 肿瘤的细胞毒性将通过流式细胞术评估 γδT 和 NK 细胞亚群的频率。 以及与疟疾暴露相关的 γδ T 和 NK 细胞亚群的细胞毒性将在体外进行量化。 对抗 BL 肿瘤，包括我们新建立的源自患者的 eBL 细胞系。 实验将评估每个子集对总体细胞毒性的相对贡献。 疟疾影响人类免疫状况，尤其是儿童的免疫状况，这将使我们能够探索 调节调节机制，同时维持对 EBV 的保护性免疫力的干预措施。

项目成果

Profiling genome-wide recombination in Epstein Barr virus reveals type-specific patterns and associations with endemic-Burkitt lymphoma.

分析 Epstein Barr 病毒的全基因组重组揭示了类型特异性模式以及与地方性伯基特淋巴瘤的关联。

• 发表时间: 2022-12-08
• 影响因子: 4.8
• 作者: Agwati, Eddy O;Oduor, Cliff I;Ayieko, Cyrus;Ong'echa, John Michael;Moormann, Ann M;Bailey, Jeffrey A
• 通讯作者: Bailey, Jeffrey A

High pathogen burden in childhood promotes the development of unconventional innate-like CD8+ T cells.

儿童时期的高病原体负担促进了非常规先天样 CD8 T 细胞的发育。

• 发表时间: 2017-08-03
• 影响因子: 8
• 作者: Falanga, Yves T;Frascoli, Michela;Kaymaz, Yasin;Forconi, Catherine;Ong'echa, John Michael;Bailey, Jeffrey A;Berg, Leslie J;Moormann, Ann M
• 通讯作者: Moormann, Ann M

Viral and host factors drive a type 1 Epstein-Barr virus spontaneous lytic phenotype.

病毒和宿主因素驱动 1 型 Epstein-Barr 病毒自发裂解表型。

• 发表时间: 2023-11-16
• 影响因子: 6.4
• 作者: Willard, Katherine A;Barry, Ashley P;Oduor, Cliff I;Ong'echa, John Michael;Bailey, Jeffrey A;Moormann, Ann M;Luftig, Micah A
• 通讯作者: Luftig, Micah A

Modeling of EBV Infection and Antibody Responses in Kenyan Infants With Different Levels of Malaria Exposure Shows Maternal Antibody Decay is a Major Determinant of Early EBV Infection.

对不同疟疾暴露水平的肯尼亚婴儿的 EBV 感染和抗体反应进行建模表明，母体抗体衰减是早期 EBV 感染的主要决定因素。

• DOI: 10.1093/infdis/jiw396
• 发表时间: 2016-11-01
• 期刊: The Journal of infectious diseases
• 作者: A. Reynaldi;T. Schlub;E. Piriou;S. Ogolla;O. Sumba;A. Moormann;R. Rochford;M. Davenport
• 通讯作者: M. Davenport

Regulatory T Cells in Endemic Burkitt Lymphoma Patients Are Associated with Poor Outcomes: A Prospective, Longitudinal Study.

地方性伯基特淋巴瘤患者的调节性 T 细胞与不良预后相关：一项前瞻性纵向研究。

• 发表时间: 2016
• 影响因子: 3.7
• 作者: Parsons, Emily;Otieno, Juliana A;Ong'echa, John Michael;Nixon, Christina E;Vulule, John;Münz, Christian;Stewart, V Ann;Moormann, Ann M
• 通讯作者: Moormann, Ann M