Determining the role of NLRP3 on the placement and protective potential of CD73+ BRMs post influenza infection
Determining the role of NLRP3 on the placement and protective potential of CD73+ BRMs post influenza infection
批准号:
10393080
负责人:
S Rameeza Allie
金额:
$20.27万
依托单位国家:
美国
项目类别:
财政年份:
2020
资助国家:
美国
项目状态:
已结题
起止时间:
2020-03-01 至 2022-02-28
关键词:
AdenosineAffinityAntibodiesAntibody FormationAntibody ResponseAntigensAreaB cell differentiationB-LymphocytesBiological AssayCell CommunicationCell Differentiation processCellsCenters for Disease Control and Prevention (U.S.)Cessation of lifeComplexConsequentialismDataDevelopmentDiseaseEmigrantFutureGenesGrantHemagglutininHistologyHomeImageImmuneImmunityImmunoglobulin-Secreting CellsIn SituIn VitroIndividualInfectionInflammasomeInfluenzaInterleukin-6InterleukinsInvadedInvestigationKnockout MiceLightLocationLungLymphoid TissueMaintenanceMemoryMemory B-LymphocyteMolecular TargetMorbidity - disease rateMusMutationOutcomePhenotypePopulationProductionPropertyPublic HealthReactionReporterReportingResearch PersonnelResistanceResolutionRoleSentinelSignal TransductionStructure of germinal center of lymph nodeT cell responseT-LymphocyteTNFRSF5 geneTarget PopulationsTestingTissuesVaccinationVaccine DesignVaccinesViralVirusbasecross reactivitycytokineeffector T cellfunctional outcomesin vitro Assayinfluenza infectionlymph nodeslymphoid organlymphoid structuresmacrophagemortalityneutralizing antibodynovelpathogenplasma cell differentiationpopulation basedpulmonary functionreceptorresponsetranscriptometranscriptome sequencingvaccine accessvaccine immunotherapy
中文摘要
项目摘要
关于抗原(Ag)特异性记忆B(BMEM)细胞,在公共卫生相关的背景下知之甚少。
病原体我们的研究清楚地解决了非循环、流感特异性、常驻记忆B的子集
BRM细胞(BRM)。BRM的建立依赖于早期CD 40信号传导,并维持CD 73表达。
淋巴结(LN)中CD 73-表达下降,而淋巴结(LN)中CD 73-表达下降。
人口这就提出了CD 73+和CD 73-在形成和功能上的内在差异的问题。
BRM。CD 73是LN中的生发中心(GC)迁移物的标志物,并且LN中的记忆细胞驻留在
靠近捕获和转移抗原的巨噬细胞和可以提供帮助的T细胞;因此,
我们假设CD 73 + BRM将归巢于肺中有组织的淋巴样结构区域,
依赖于T细胞的帮助我们的RNASeq数据显示NLRP 3(NOD样受体蛋白3),一种先天危险
传感复合物,是最上调的基因在CD 73 + BRM。据我们所知,NLRP 3活性不是
与B细胞分化相关,这使得这一发现新颖,并可能开创先天性研究。
在适应性反应中类似于B细胞的功能。NLRP 3活性产生白细胞介素B(IL-1 B),IL-1 B是
一种众所周知的白细胞介素6(IL-6)上调剂,可促进GC和浆细胞分化。作为我们
CD 73 + BRM可能有更高的NLRP 3表达,我们假设NLRP 3在回忆过程中是活跃的,
产生GC和ASC,而CD 73-BRM可能是肺中的哨兵,
ASCs,以应对返回的病毒。
为了验证这些假设,我们提出以下问题:1)CD 73+和CD 73- BRM细胞是否依赖于GC?如果
因此,GC特异性阻断应减少肺中的CD 73 + BRM并产生低亲和力BCR。2)做CD 73 +
BRM优先驻留在IBALT中?如果是这样,使用组织学,我们应该观察到它们聚集在
而CD 73-BRM分散在实质中。3)CD 73+和CD 73- BRM细胞是否应答
在功能分析中的不同?我们将通过对BRM进行体外测定来回答这个问题
使用细胞因子混合物和分选的T细胞将细胞分化为ASC。4)NLRP 3复合体是否会成为
在BRM中被激活如果是这样,使用NLRP 3炎性小体报告小鼠,我们预计,
CD 73 + BRM细胞,而不是CD 73- BRM细胞,以增加NLRP 3活性。5)NLRP 3是否监管
BRM的形成或功能?如果是这样的话,B细胞特异性NLRP 3缺乏应该改变以下比例:
CD 73+和CD 73-BRM及其保护功能,在发病率和死亡率方面存在明显差异。
这项研究的结果,尽管结果将是极其重要的,以允许解决一个
用于有效疫苗设计靶群体和潜在分子靶。
英文摘要
Project Summary
Little is known about antigen (Ag) specific memory B (BMEM) cells, in the context of a public health relevant
pathogen. Our studies clearly resolve the subsets of non-circulating, influenza specific, resident memory B
cells (BRM) in the lung. BRM establishment relied on early CD40 signaling and maintained a CD73
(ectoenzyme)+ and CD73- population, whereas the lymph node (LN) showed a decline in the CD73-
population. This raised the question of the inherent differences in formation and function of CD73+ and CD73-
BRMs. CD73 is a marker of germinal center (GC) emigrants in the LN and memory cells in the LN reside in
niches close to macrophages that capture and transfer antigen and T cells which can provide help; therefore,
we hypothesized that the CD73+BRMs will home to areas of organized lymphoid structures in the lung and be
dependent on T cell help. Our RNASeq data shows NLRP3 (NOD-like receptor protein 3), an innate danger
sensing complex, to be the most upregulated gene in CD73+BRMs. To our knowledge NLRP3 activity is not
associated with B cell differentiation, which makes this finding novel and may inaugurate studies into innate
like functions in B cells during adaptive responses. NLRP3 activity produces interleukin B (IL-1B) and IL-1B is
a well-known up regulator of interleukin 6 (IL-6), which promotes GCs and plasma cell differentiation. As our
CD73+BRMs may have higher NLRP3 expression, we hypothesize that NLRP3 will be active during recall and
produce GCs and ASCs, while the CD73-BRMs may be sentinels in the lung which spontaneously convert to
ASCs, in response to the returning virus.
To test these hypotheses, we ask the following: 1) Are CD73+ and CD73- BRM cells dependent on GCs? If
so, GC-specific blockade should reduce CD73+ BRMs in the lung and produce low affinity BCRs. 2) Do CD73+
BRMs preferentially reside in iBALT? If so, using histology, we should observe them congregating in the
iBALT while the CD73-BRMs are scattered in the parenchyma. 3) Do CD73+ and CD73- BRM cells respond
differently in functional assays? We will answer this question by performing in vitro assays for BRM
differentiation to ASCs, using cytokine cocktails and sorted T cells. 4) Does the NLRP3 complex become
activated in BRMs upon viral challenge? If so, using NLRP3 inflammasome reporter mice, we expect that
CD73+ BRM cells, but not CD73- BRM cells to increase NLRP3 activity. 5) Does NLRP3 regulate the
formation or function of BRMs? If so, B cell specific NLRP3 deficiency should alter the proportions of
CD73+ and CD73-BRMs and their protective function with observable differences in morbidity and mortality.
The findings from this study despite the outcome will be extremely important to allow for the resolution of a
target population and potential molecular targets for an effective vaccine design.
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会议论文
Establishment and metabolic control of influenza-specific lung-resident memory B cells
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批准号:8982505
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项目类别:
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资助金额:$5.34万
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财政年份:2015
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负责人:S Rameeza Allie
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依托单位:
Establishment and metabolic control of influenza-specific lung-resident memory B cells
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批准号:9120660
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项目类别:
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资助金额:$5.69万
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财政年份:2015
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负责人:S Rameeza Allie
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依托单位:
海外基金