杭州浙大迪迅生物基因工程有限公司Allergy：多维度分析定义多细胞因子T细胞群和IL-21在特应性皮炎中的作用
发布日期：2021-11-19
原标题：高维分析确定了多细胞因子T细胞亚群，并支持IL-21在特应性皮炎中的作用
方法：我们研究了20名中度至重度AD和15名对照者血液中的T细胞极化。使用CyTOF和无监督分析，我们测量了活化的极性CD4+/CD8+ T细胞亚群频率和平均金属强度。使用免疫组织化学、免疫荧光和qRT-PCR分析皮肤样本。
结果：通过检查24个表面、细胞内标记和转录因子，我们确定了6个CD4+和5个 CD8+ T 细胞元簇。与对照组相比，AD中CD4+皮肤归巢IL-13+单细胞因子和新型IL-13+IL-21+多细胞因子元簇增加（p < .01）。虽然 IL-13 特征表征了两个簇，但 IL-21+ 组的水平显着更高。两个集群都与AD严重程度相关（r = 0.49，p = .029）。手动门控证实了这些结果并确定了AD中的其他多细胞因子亚群。通过mRNA表达验证的免疫组织化学和免疫荧光显示，IL-21计数显着增加，并且与AD皮肤中的IL-13/IL-4R共定位。
结论：多细胞因子特征是中重度AD的特征，这可能解释了对一种细胞因子靶向的部分治疗反应，尤其是在重度患者中。血液和皮肤中显着的IL-21特征暗示了IL-21在AD中的潜在致病作用。
延伸阅读
Allergy
DOI: 10.1111/all.14845
Abstract:
Background: Flow cytometry is a well-accepted approach for immune profiling; however, its value is restricted by the limited number of markers that can be analyzed simultaneously. Mass cytometry/CyTOF offers broad-scale immune characterization integrating large number of parameters. While partial blood phenotyping was reported in atopic dermatitis (AD), patients’ comprehensive profiling, critical for leveraging new targeted treatments, is not available. IL-21 may be involved in inflammatory skin diseases but its role in AD is not well established.
Methods: We studied T-cell polarization in the blood of 20 moderate-to-severe AD and 15 controls. Using CyTOF and an unsupervised analysis, we measured the frequencies and mean metal intensities of activated polar CD4+/CD8+ T-cell subsets. Immunohistochemistry, immunofluorescence, and qRT-PCR were used to analyze skin samples.
Results: Examining 24 surface, intracellular markers, and transcription factors, we identified six CD4+ and five CD8+ T-cell metaclusters. A CD4+ skin-homing IL-13+monocytokine and a novel IL-13+IL-21+ multicytokine metaclusters were increased in AD vs. controls (p < .01). While IL-13 signature characterized both clusters, levels were significantly higher in the IL-21+ group. Both clusters correlated with AD severity (r = 0.49, p = .029). Manual gating corroborated these results and identified additional multicytokine subsets in AD. Immunohistochemistry and immunofluorescence, validated by mRNA expression, displayed significantly increasedIL-21 counts and colocalization with IL-13/IL-4R in AD skin.
Conclusion: A multicytokine signature characterizes moderate-to-severe AD, possibly explaining partial therapeutic responses to one cytokine targeting, particularly in severe patients. Prominent IL-21 signature in blood and skin hints for a potential pathogenic role of IL-21 in AD.
First Author:
Tali Czarnowicki
Correspondence:
Emma Guttman-Yassky, The Kimberly and Eric J. Waldman Department of Dermatology, Icahn School of Medicine at Mount Sinai, 1425 Madison Avenue, Box 1047, New York, NY 10029-6574, USA.
2021-10-27 Article
创建过敏性疾病的科研、科普知识交流平台，为过敏患者提供专业诊断、治疗、预防的共享平台。
——浙大迪迅 译
背景：流式细胞术是一种广为接受的免疫分析方法；然而，它的价值受限于可以同时分析的标记物的数量。质谱流式细胞术/CyTOF提供大规模免疫表征整合成大量参数。虽然在特应性皮炎(AD)中报告了部分血液表型，但对利用新的靶向治疗至关重要的患者综合分析尚不可用。IL-21可能与炎症性皮肤病有关，但其在AD中的作用尚未确定。方法：我们研究了20名中度至重度AD和15名对照者血液中的T细胞极化。使用CyTOF和无监督分析，我们测量了活化的极性CD4+/CD8+ T细胞亚群频率和平均金属强度。使用免疫组织化学、免疫荧光和qRT-PCR分析皮肤样本。
结果：通过检查24个表面、细胞内标记和转录因子，我们确定了6个CD4+和5个 CD8+ T 细胞元簇。与对照组相比，AD中CD4+皮肤归巢IL-13+单细胞因子和新型IL-13+IL-21+多细胞因子元簇增加（p < .01）。虽然 IL-13 特征表征了两个簇，但 IL-21+ 组的水平显着更高。两个集群都与AD严重程度相关（r = 0.49，p = .029）。手动门控证实了这些结果并确定了AD中的其他多细胞因子亚群。通过mRNA表达验证的免疫组织化学和免疫荧光显示，IL-21计数显着增加，并且与AD皮肤中的IL-13/IL-4R共定位。
结论：多细胞因子特征是中重度AD的特征，这可能解释了对一种细胞因子靶向的部分治疗反应，尤其是在重度患者中。血液和皮肤中显着的IL-21特征暗示了IL-21在AD中的潜在致病作用。
延伸阅读
Allergy
[IF: 13.146]
High-dimensional analysis defines multicytokine T-cell subsets and supports a role for IL-21 in atopic dermatitisDOI: 10.1111/all.14845
Abstract:
Background: Flow cytometry is a well-accepted approach for immune profiling; however, its value is restricted by the limited number of markers that can be analyzed simultaneously. Mass cytometry/CyTOF offers broad-scale immune characterization integrating large number of parameters. While partial blood phenotyping was reported in atopic dermatitis (AD), patients’ comprehensive profiling, critical for leveraging new targeted treatments, is not available. IL-21 may be involved in inflammatory skin diseases but its role in AD is not well established.
Methods: We studied T-cell polarization in the blood of 20 moderate-to-severe AD and 15 controls. Using CyTOF and an unsupervised analysis, we measured the frequencies and mean metal intensities of activated polar CD4+/CD8+ T-cell subsets. Immunohistochemistry, immunofluorescence, and qRT-PCR were used to analyze skin samples.
Results: Examining 24 surface, intracellular markers, and transcription factors, we identified six CD4+ and five CD8+ T-cell metaclusters. A CD4+ skin-homing IL-13+monocytokine and a novel IL-13+IL-21+ multicytokine metaclusters were increased in AD vs. controls (p < .01). While IL-13 signature characterized both clusters, levels were significantly higher in the IL-21+ group. Both clusters correlated with AD severity (r = 0.49, p = .029). Manual gating corroborated these results and identified additional multicytokine subsets in AD. Immunohistochemistry and immunofluorescence, validated by mRNA expression, displayed significantly increasedIL-21 counts and colocalization with IL-13/IL-4R in AD skin.
Conclusion: A multicytokine signature characterizes moderate-to-severe AD, possibly explaining partial therapeutic responses to one cytokine targeting, particularly in severe patients. Prominent IL-21 signature in blood and skin hints for a potential pathogenic role of IL-21 in AD.
First Author:
Tali Czarnowicki
Correspondence:
Emma Guttman-Yassky, The Kimberly and Eric J. Waldman Department of Dermatology, Icahn School of Medicine at Mount Sinai, 1425 Madison Avenue, Box 1047, New York, NY 10029-6574, USA.
2021-10-27 Article
创建过敏性疾病的科研、科普知识交流平台，为过敏患者提供专业诊断、治疗、预防的共享平台。
