A Sensitive Whole Blood Assay Detects Antigen-Stimulated Cytokine Release From CD4+ T Cells and Facilitates Immunomonitoring in a Phase 2 Clinical Trial of Nexvax2 in Coeliac Disease.
A Sensitive Whole Blood Assay Detects Antigen-Stimulated Cytokine Release From CD4+ T Cells and Facilitates Immunomonitoring in a Phase 2 Clinical Trial of Nexvax2 in Coeliac Disease.
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DOI：
10.3389/fimmu.2021.661622
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发表时间：
2021
影响因子：
7.3
通讯作者：
Anderson RP
中科院分区：
文献类型：
--
作者：
Hardy MY;Goel G;Russell AK;Chen Yi Mei SLG;Brown GJE;Wang S;Szymczak E;Zhang R;Goldstein KE;Neff KM;Williams LJ;Truitt KE;Dzuris JL;Tye-Din JA;Anderson RP
需要改进评估罕见谷蛋白特异性CD 4 + T细胞功能状态的血液检测，以有效监测乳糜泻（CD）的实验性治疗。我们的目标是开发一种简单但高度敏感的谷蛋白特异性CD 4 + T细胞因子释放试验（CRA），不需要患者事先接受谷蛋白激发，并且在大型多中心临床试验中实用。我们开发了一种增强的CRA，并将其用于Nexvax 2的2期临床试验（“NexCeD”），Nexvax 2是一种基于肽的CD免疫疗法。在一项试点研究中，在3天麸质挑战之前和之后6天，对两名接受治疗的CD参与者进行了评估。检测谷蛋白肽刺激24 h后外周血单个核细胞（PBMC）的增殖反应，并采用多重电化学发光免疫分析法（ECL）检测IL-2、IFN-γ和IL-10的水平。随后，在接受Nexvax 2（维持剂量900 μg，n = 12）或安慰剂（n = 9）的一个亚组中，评估了血液中谷蛋白特异性CD 4 + T细胞。初步研究表明，谷蛋白肽诱导IL-2、IFN-γ和IL-10从PBMC中释放，归因于CD 4 + T细胞，但PBMC CRA的敏感性明显低于全血CRA。仅可检测到适度的谷蛋白肽刺激的IL-2释放，而无需使用PBMC进行先前的谷蛋白激发。相比之下，全血CRA能够在麸质攻击之前和之后检测IL-2和IFN-γ。全血中IL-2和IFN-γ的释放需要超过6小时的孵育。全血孵育从采集开始延迟超过3小时显著降低了抗原刺激的IL-2和IFN-γ分泌。Nexvax 2（而非安慰剂）治疗与谷蛋白肽刺激的全血IL-2和IFN-γ释放以及CD 4 + T细胞增殖的显著降低相关。我们的结论是，使用新鲜的全血，而不是PBMC大大提高了谷蛋白肽刺激的细胞因子分泌，并使罕见的谷蛋白特异性CD 4 + T细胞的评估，而不需要CD患者进行谷蛋白的挑战。全血评估加上超灵敏的细胞因子检测显示出在临床研究中监测罕见抗原特异性T细胞的前景。
Improved blood tests assessing the functional status of rare gluten-specific CD4+ T cells are needed to effectively monitor experimental therapies for coeliac disease (CD). Our aim was to develop a simple, but highly sensitive cytokine release assay (CRA) for gluten-specific CD4+ T cells that did not require patients to undergo a prior gluten challenge, and would be practical in large, multi-centre clinical trials. We developed an enhanced CRA and used it in a phase 2 clinical trial (“RESET CeD”) of Nexvax2, a peptide-based immunotherapy for CD. Two participants with treated CD were assessed in a pilot study prior to and six days after a 3-day gluten challenge. Dye-dilution proliferation in peripheral blood mononuclear cells (PBMC) was assessed, and IL-2, IFN-γ and IL-10 were measured by multiplex electrochemiluminescence immunoassay (ECL) after 24-hour gluten-peptide stimulation of whole blood or matched PBMC. Subsequently, gluten-specific CD4+ T cells in blood were assessed in a subgroup of the RESET CeD Study participants who received Nexvax2 (maintenance dose 900 μg, n = 12) or placebo (n = 9). The pilot study showed that gluten peptides induced IL-2, IFN-γ and IL-10 release from PBMCs attributable to CD4+ T cells, but the PBMC CRA was substantially less sensitive than whole blood CRA. Only modest gluten peptide-stimulated IL-2 release could be detected without prior gluten challenge using PBMC. In contrast, whole blood CRA enabled detection of IL-2 and IFN-γ before and after gluten challenge. IL-2 and IFN-γ release in whole blood required more than 6 hours incubation. Delay in whole blood incubation of more than three hours from collection substantially reduced antigen-stimulated IL-2 and IFN-γ secretion. Nexvax2, but not placebo treatment in the RESET CeD Study was associated with significant reductions in gluten peptide-stimulated whole blood IL-2 and IFN-γ release, and CD4+ T cell proliferation. We conclude that using fresh whole blood instead of PBMC substantially enhances cytokine secretion stimulated by gluten peptides, and enables assessment of rare gluten-specific CD4+ T cells without requiring CD patients to undertake a gluten challenge. Whole blood assessment coupled with ultra-sensitive cytokine detection shows promise in the monitoring of rare antigen-specific T cells in clinical studies.
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影响因子：
7.3
作者：
Ben-Othman R;Cai B;Liu AC;Varankovich N;He D;Blimkie TM;Lee AH;Gill EE;Novotny M;Aevermann B;Drissler S;Shannon CP;McCann S;Marty K;Bjornson G;Edgar RD;Lin DTS;Gladish N;Maclsaac J;Amenyogbe N;Chan Q;Llibre A;Collin J;Landais E;Le K;Reiss SM;Koff WC;Havenar-Daughton C;Heran M;Sangha B;Walt D;Krajden M;Crotty S;Sok D;Briney B;Burton DR;Duffy D;Foster LJ;Mohn WW;Kobor MS;Tebbutt SJ;Brinkman RR;Scheuermann RH;Hancock REW;Kollmann TR;Sadarangani M
通讯作者：
Sadarangani M
影响因子：
4.6
作者：
Ontiveros, N.;Tye-Din, J. A.;Anderson, R. P.
通讯作者：
Anderson, R. P.
影响因子：
4.6
作者：
Anderson, R. P.;Goel, G.;Tye-Din, J. A.
通讯作者：
Tye-Din, J. A.
影响因子：
13.6
作者：
Goel, Gautam;Tye-Din, Jason A.;Anderson, Robert P.
通讯作者：
Anderson, Robert P.
影响因子：
17.1
作者：
Tye-Din, Jason A.;Stewart, Jessica A.;Anderson, Robert P.
通讯作者：
Anderson, Robert P.