Background

Human type 2 innate lymphoid cells (ILC2s) are identified by coupled detection of CRTH2 and IL7Rα on lineage negative (Lin^–) cells. Type 2 cytokine production by CRTH2^–IL7Rα^– innate lymphoid cells (ILCs) is unknown.

Objective

We sought to identify CRTH2^–IL7Rα^– type 2 cytokine–producing ILCs and their disease relevance.

Methods

We studied human blood and lung ILCs from asthmatic and control subjects by flow cytometry, ELISA, RNA sequencing, quantitative PCR, adoptive transfer to mice, and measurement of airway hyperreactivity by Flexivent.

Results

We found that IL-5 and IL-13 were expressed not only by CRTH2⁺ but also by CRTH2^–IL7Rα⁺ and CRTH2^–IL7Rα^– (double-negative [DN]) human blood and lung cells. All 3 ILC populations expressed type 2 genes and induced airway hyperreactivity when adoptively transferred to mice. The frequency of type 2 cytokine–positive IL7Rα and DN ILCs were similar to that of CRTH2 ILCs in the blood and lung. Their frequency was higher in asthmatic patients than in disease controls. Transcriptomic analysis of CRTH2, IL7Rα, and DN ILCs confirmed the expression of mRNA for type 2 transcription factors in all 3 populations. Unexpectedly, the mRNA for GATA3 and IL-5 correlated better with mRNA for CD30, TNFR2, ICOS, CCR4, and CD200R1 than for CRTH2. By using a combination of these surface markers, especially CD30/TNFR2, we identified a previously unrecognized ILC2 population.

Conclusions

The commonly used surface markers for human ILC2s leave a majority of type 2 cytokine–producing ILC2s unaccounted for. We identified top GATA3-correlated cell surface–expressed genes in human ILCs by RNA sequencing. These new surface markers, such as CD30 and TNFR2, identified a previously unrecognized human ILC2 population. This ILC2 population is likely to contribute to asthma.

中文翻译：

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使用额外的表面标记对人类常规和非常规 (CRTH2-IL7Rα-) ILC2s 进行最佳识别。

背景

人类 2 型先天淋巴细胞 (ILC2) 通过在谱系阴性 (Lin ^– ) 细胞上对 CRTH2 和 IL7Rα 进行耦合检测来识别。CRTH2 ^– IL7Rα ^–先天淋巴细胞 (ILC)产生的 2 型细胞因子是未知的。

客观的

我们试图确定 CRTH2 ^– IL7Rα ^–产生 2 型细胞因子的 ILC 及其疾病相关性。

方法

我们通过流式细胞术、ELISA、RNA 测序、定量 PCR、对小鼠的过继转移以及通过 Flexivent 测量气道高反应性研究了来自哮喘和对照受试者的人血液和肺 ILC。

结果

我们发现，IL-5和IL-13不仅被用CRTH2表达⁺也受到CRTH2 ^- IL7Rα ⁺和CRTH2 ^- IL7Rα ^-（双阴性 [DN]）人血和肺细胞。当过继转移到小鼠时，所有 3 个 ILC 群体都表达 2 型基因并诱导气道高反应性。2 型细胞因子阳性 IL7Rα 和 DN ILC 的频率与血液和肺中 CRTH2 ILC 的频率相似。哮喘患者的频率高于疾病对照组。CRTH2、IL7Rα 和 DN ILC 的转录组学分析证实了所有 3 个群体中 2 型转录因子的 mRNA 表达。出乎意料的是，与 CRTH2 相比，GATA3 和 IL-5 的 mRNA 与 CD30、TNFR2、ICOS、CCR4 和 CD200R1 的 mRNA 的相关性更好。通过使用这些表面标记的组合，尤其是 CD30/TNFR2，我们确定了一个以前无法识别的 ILC2 群体。

结论

人类 ILC2 的常用表面标记使大多数产生 2 型细胞因子的 ILC2 无法解释。我们通过 RNA 测序鉴定了人类 ILC 中与 GATA3 相关的细胞表面表达基因。这些新的表面标志物，如 CD30 和 TNFR2，确定了以前未被识别的人类 ILC2 群体。该 ILC2 群体可能会导致哮喘。