Mechanisms of allergy/immunologyOptimal identification of human conventional and nonconventional (CRTH2–IL7Rα–) ILC2s using additional surface markers
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Human subjects
We studied blood and bronchoalveolar lavage (BAL) ILC2s from patients with allergic asthma, disease controls, and healthy donors. Asthmatic patients and disease controls were recruited from the outpatient clinics at National Jewish Health during the period from July 2015 through December 2019. Healthy donors were recruited from the blood bank of National Jewish Health. Bronchoscopy and BAL were performed as a part of their clinical work-up for poorly controlled asthma, as defined by the Expert
Type 2 cytokines are expressed by CRTH2– and IL7Rα– ILCs
CRTH2 and IL7Rα are commonly used to detect human ILC2s in the Lin–cell population of blood or other tissues (the Lin antibody cocktail contained antibodies against CD3, CD14, CD16, CD19, CD20, CD56, and FcεRI). We asked whether CRTH2 and IL7Rα captured all type 2 cytokine–positive ILCs. To this goal, we examined type 2 cytokine expression by peripheral blood CRTH2+IL7Rα–, CRTH2+IL7Rα+, CRTH2–IL7Rα+, and CRTH2–IL7Rα– ILC populations (Fig 1, A-J) cultured with IL-2/IL-25. The gating strategy is
Discussion
In this article we showed that the commonly used ILC2 surface marker CRTH2 did not identify all type 2 cytokine–positive ILCs. The CRTH2–IL7Rα+ and DN (CRTH2–IL7Rα–) subpopulations also contained type 2 cytokine–positive ILC2s and expressed the type 2 transcription factors. The CRTH2+ ILC population had the highest frequency of type 2 cytokine–positive ILC2s when compared with the IL7Rα+ and DN populations. However, the CRTH2+ population was much smaller in size than the other 2 populations.
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2023, Allergology InternationalCitation Excerpt :However, CRTH2 might be less useful as a marker for ILC2s because some ILC2-like cells lack CRTH2 expression in peripheral blood.25–27 For example, it has been reported that CRTH2– ILCs expressing CD30 and TNFR2 express GATA3 and produce type 2 cytokines.26 It has also been reported that peripheral blood CD117+ CRTH2– ILCs that express another ILC2-related marker, KLRG1, could upregulate CRTH2 expression in vitro upon IL-7 stimulation,27 indicating that CD117+ CRTH2– ILCs are precursors or immature status of ILC2s.
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2021, Journal of Allergy and Clinical ImmunologyCitation Excerpt :Functionally, CCR10− ILC2s showed robust production of type 2 cytokines, whereas CCR10+ ILC2s displayed an ILC1-like phenotype with robust IFN-γ production and little to no IL-5 or IL-13.106 Liu et al107 used IL-5 expression as a functional surrogate for ILC2s and fully characterized ILC2s for their expression of CD127 (IL-7R) and CRTH2 and effector functions in subjects with asthma. CRTH2+CD127+ ILC2s produced type 2 cytokines alone, whereas CRTH2−CD127+ ILC2s and CRTH2−CD127− ILC2s produced IL-17 and IFN-γ, respectively, in addition to type 2 cytokines.
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2021, Annals of Allergy, Asthma and ImmunologyCitation Excerpt :In contrast, other common ILC2 markers such as GATA3, IL-7Ra, and TSLP receptor (cytokine receptor-like factor 2) were expressed more uniformly across all tissues.14 Liu et al15 recently found that some ILC2 subsets in patients with asthma lack cell surface expression of CD127 or CRTH2 but produce type 2 cytokines. Although the heterogeneity of human ILC2s has become clearer, there are no reports about this variability within CRS.
Tissue-Specific Features of Innate Lymphoid Cells
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Supported by National Institutes of Health grants AI102943, AI137970, and HL126895; grants from the Cohen Family Foundation, the NBL Fellowship Foundation, and Colorado Technology; and the Department of Medicine of National Jewish Health.
Disclosure of potential conflict of interest: The authors declare that they have no relevant conflicts of interest.
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These authors contributed equally to this work as first authors.