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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

疟疾疟原虫（PF）疟疾和Epstein-Barr病毒（EBV）居住在疟疾的儿童中全甘登岩地区与EBV相关的癌症的风险增加了，称为伯基特癌症淋巴瘤（EBL）。大多数非洲儿童在1岁之前感染了EBV，但这种B细胞癌确实有人认为，疟疾的重复发作“抑制”免疫 EBV，为EBL发病机理创造一个宽松的环境。但是，负责的机制不是完全理解。我们先前的研究发现，暴露于疟疾的儿童的EBV负荷高。幼稚的EBV特异性CD8+ T细胞具有下降的效应函数；非常规的，先天的CD8+ T 表达颗粒状B代替IFN-γ的细胞；以及“慢性感染诱导” CD56NEG的扩展天然杀伤（NK）细胞具有细胞毒性受损。这，我们已经确定了接近免疫学的改变这允许不受约束的EBV复制和EBL肿瘤发生。在此续签应用中，我们将中央假设疟疾诱导的免疫调节机制抑制了T细胞毒性针对T细胞毒性 EBV感染的B细胞和EBL肿瘤。这将通过以下特定目的对此进行测试。目标1。确定如果重复的PF-麦乳ari症感染已知会诱导EBV重新激活，则会导致抑制性共同感染 EBV特异性CD8+αβT细胞上的受体表达。 Tigit，pd1，ctla4，lag3，tim3， CD160、2B4，KLRG1，BTLA在T细胞子集上将通过流式细胞仪测量。疲惫与细胞毒性特征将通过单细胞RNA测序进一步定义使用EBV转化的淋巴细胞细胞系（LCLS），通过细胞毒性T淋巴细胞（CTL）测定法。目标2。确定重复的PF-malaria感染是否诱导IL-10产生CD4+或CD8+ T细胞的IL-10 对EBV特异性T细胞产生免疫调节作用。 IL-10分泌foxp3neg的频率调节性CD25+，CD4+，TR1细胞（CD49B+，LAG3+，CD226+/DNAM1+），TREG-B细胞（LAG3+，ICOS+，PD1+， GITR+，OX40+）和CD8+CD25NEG FOXP3NEG T细胞将通过流式细胞仪和RNASEQ进行测量它们与经典的CD4+FOX+P3+调节T细胞（Tregs）不同。 CTL分析将确定在体外，IL-10细胞因子家族成员。 AIM 3。确定是否重复PF-Malaria 感染会影响γδT与NK细胞子集的频率以及它们的相对比率如何影响对EBL肿瘤的细胞毒性。 γδT和NK细胞子集的频率将通过流式细胞仪评估并与疟疾暴露有关。 γδT和NK细胞子集的细胞毒性将在体外进行定量针对BL肿瘤，包括我们新建立的患者衍生的EBL细胞系。配体受体阻塞实验将评估每个子集对总体细胞毒性的相对贡献。了解如何疟疾影响人类的免疫学局势，尤其是在儿童中，将使我们能够探索调节调节机制的干预措施，同时保持对EBV的保护性免疫。

项目成果

期刊论文数量（19）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Evaluation of KIR3DL1/KIR3DS1 allelic polymorphisms in Kenyan children with endemic Burkitt lymphoma.

DOI：
10.1371/journal.pone.0275046
发表时间：
2023
期刊：
PloS one
影响因子：
3.7
作者：
通讯作者：

Integrative microRNA and mRNA deep-sequencing expression profiling in endemic Burkitt lymphoma.

DOI：
10.1186/s12885-017-3711-9
发表时间：
2017-11-13
期刊：
BMC cancer
影响因子：
3.8
作者：
Oduor CI;Kaymaz Y;Chelimo K;Otieno JA;Ong'echa JM;Moormann AM;Bailey JA
通讯作者：
Bailey JA

Endemic Burkitt lymphoma avatar mouse models for exploring inter-patient tumor variation and testing targeted therapies.

DOI：
10.26508/lsa.202101355
发表时间：
2023-05
期刊：
LIFE SCIENCE ALLIANCE
影响因子：
4.4
作者：
Lakshmi, Priya Saikumar;Oduor, Cliff, I;Forconi, Catherine S.;Bana, Viriato M. ';Bly, Courtney;Gerstein, Rachel M.;Otieno, Juliana A.;Muenz, Christian;Luftig, Micah A.;Brehm, Michael A.;Bailey, Jeffrey A.;Moormann, Ann M.;Ong'echa, John M.
通讯作者：
Ong'echa, John M.

Factors influencing survival among Kenyan children diagnosed with endemic Burkitt lymphoma between 2003 and 2011: A historical cohort study.

DOI：
10.1002/ijc.30170
发表时间：
2016-09-15
期刊：
International journal of cancer
影响因子：
6.4
作者：
Buckle G;Maranda L;Skiles J;Ong'echa JM;Foley J;Epstein M;Vik TA;Schroeder A;Lemberger J;Rosmarin A;Remick SC;Bailey JA;Vulule J;Otieno JA;Moormann AM
通讯作者：
Moormann AM

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