Arthritis Res Ther. 2020;22(1):63.

Reduction of circulating innate lymphoid cell progenitors results in impaired cytokine production by innate lymphoid cells in patients with lupus nephritis.

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제1저자 류승원 · 교신저자 이하정, 김혜영

PMID 32223753 PMC7104540 DOI 10.1186/s13075-020-2114-5 IF 5.4 신장내과

소속 (11)

Laboratory of Mucosal Immunology, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea.
Division of Rheumatology, Department of Internal Medicine, Seoul National University Hospital, Seoul, South Korea.
Division of Nephrology, Department of Internal Medicine, Seoul National University Hospital, Seoul, South Korea.
Kidney Research Institute, Seoul National University College of Medicine, Seoul, 03080, South Korea.
Department of Pathology, Seoul National University College of Medicine, Seoul, South Korea.
Laboratory of Immune Regulation, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, South Korea.
College of Life Sciences and Biotechnology, Korea University, Seoul, South Korea.
Division of Nephrology, Department of Internal Medicine, Seoul National University Hospital, Seoul, South Korea. mdhjlee@gmail.com.
Kidney Research Institute, Seoul National University College of Medicine, Seoul, 03080, South Korea. mdhjlee@gmail.com.
Laboratory of Mucosal Immunology, Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, 03080, South Korea. hykim11@snu.ac.kr.
Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, South Korea. hykim11@snu.ac.kr.

Abstract

BACKGROUND Innate lymphoid cells (ILCs) play an essential role in maintaining homeostasis; however, they can also cause chronic inflammation and autoimmune disease. This study aimed to identify the role of ILCs in the pathogenesis of lupus nephritis (LN).

METHODS The percentage of ILCs within the peripheral blood mononuclear cell (PBMC) population and urine of patients with LN (n = 16), healthy controls (HC; n = 8), and disease controls (ANCA-associated vasculitis (AAV; n = 6), IgA nephropathy (IgAN; n = 9), and other glomerular diseases (n = 5)) was determined by flow cytometry analysis. In addition, ILCs were sorted and cultured with plasma from LN patients or HC to elucidate whether the reduced population of CD117⁺ ILCs observed in LN was due to changes in the ILC progenitor population.

RESULTS The percentage of total ILCs and CD117⁺ ILCs in LN was significantly lower than that in HC. The percentage of cytokine-secreting ILCs was also lower in LN; however, when the disease stabilized, cytokine production was restored to levels similar to those in HC. The increase in the number of exhausted ILCs (cells unable to secrete cytokines) correlated positively with disease activity. When CD117⁺ ILCs were cultured with LN plasma, the number of CD117⁺ ILCs fell, but that of other ILC subsets increased.

CONCLUSIONS The percentage of CD117⁺ ILCs and the capacity of ILCs to secrete cytokines fell as LN severity increased, suggesting that an inflammatory environment of LN induces persistent differentiation and exhaustion of ILCs.

Keywords

Autoimmune kidney diseaseCytokineInnate lymphoid cells (ILCs)Lupus nephritis

Figures

**Fig. 1** Patients with LN show reduced percentages of CD117⁺ ILCs in the circulation and in renal tissue. a Gating strategy for blood ILCs. b Representative flow cytometry plots of ILC subsets within the PBMC. c Percentage of total ILCs (CD45⁺ Lin⁻ CD127⁺), CD117⁺ ILCs, CRTH2⁺ ILCs, and CD117⁻/CRTH2⁻ ILCs within the PBMC from HC (n = 8), LN (n = 12), AAV (n = 5), IgAN (n = 8), and MCD/FSGS (n = 5). d Representative flow cytometry plots of ILC subsets in urine. e Percentage of CD117⁺ ILCs and CRTH2⁺ ILCs in urine from patients with LN (n = 15), AAV (n = 6), IgAN (n = 9), and MCD/FSGS (n = 5). **P <* 0.05; **P < 0.01; ***P < 0.001 (Student’s t test). ^##P < 0.01 (Mann-Whitney test). ILC, innate lymphoid cell; PBMC, peripheral blood mononuclear cell; HC, healthy control; LN, lupus nephritis; AAV, ANCA-associated vasculitis; IgAN, IgA nephropathy; MCD, minimal change disease; FSGS, focal segmental glomerulosclerosis

**Fig. 2** Circulating ILCs in patients with LN are dysfunctional with respect to cytokine production. a Representative flow cytometry plots of IFN-γ-, IL-13-, and IL-17A⁺-producing ILCs within the PBMC. b, c Percentage of IFN-γ-, IL-13-, and IL-17A⁺-producing ILCs in (b) CD45⁺ immune cells and (c) total ILCs from HC (n = 6), LN (n = 11), AAV (n = 5), IgAN (n = 8), and MCD/FSGS (n = 5). d Representative histogram plot of PD-1 expression of circulating ILCs from HC and LN, and percentage of PD-1⁺ ILCs within PBMC compared between HC (n = 5) and LN (n = 6). e Percentage of cytokine-negative ILCs within PBMC from HC (n = 6) and LN (n = 11). **P <* 0.05; **P < 0.01 (Student’s t test)

**Fig. 3** Patients with stable LN have higher numbers of circulating CD117⁺ ILCs and cytokine-producing ILCs. a Differences in clinical parameters of those with active and stable LN. b Changes in total ILCs, CD117⁺ ILCs, CRTH2⁺ ILCs, and CD117⁻/CRTH2⁻ ILCs are shown as min. to max. Box-and-whisker plots (line represents the median). c, d Changes in IFN-γ⁺, IL-13⁺, IL-17A⁺, PD-1⁺ ILCs, and cytokine-negative ILCs in PBMC from those with active LN (n = 12) or stable LN (n = 10). **P <* 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (Student’s t test). ^##P < 0.01 (Mann-Whitney test)

**Fig. 4** Circulating ILCs correlate with clinical parameters that reflect LN disease activity. Correlation between the percentage of (a) total ILCs, (b) CD117⁺ ILCs, and (c) cytokine-negative ILCs within PBMC, and clinical parameters (WBC counts, UPCR, and eGFR). **P < 0.01 (Pearson correlation). ^#P < 0.01 (Spearman correlation). WBC, white blood cell; UPCR, urine protein to creatinine ratio; eGFR, estimated glomerular filtration rate

**Fig. 5** Differentiation of ILC progenitors is affected by LN microenvironment. a PBMCs from HC were cultured for 24 h with plasma from either HC (n = 9) or patients with active LN (n = 12). b Percentages of CD117⁺, CRTH2⁺, and CD117⁻ CRTH2⁻ ILCs in total ILCs were compared under both conditions. c, d Annexin V was stained to compare the apoptosis of ILCs in either conditions (n = 8 for HC; n = 10 for LN). e FACS-sorted CD117⁺ ILCs were cultured for 8 days in plasma from HC (n = 7) or patients with active LN (n = 11). f Percentages of CD117⁺, CRTH2⁺, and CD117⁻ CRTH2⁻ ILCs in CD45⁺ immune cells and total ILCs were compared between two culture systems. g The effect of blocking of IL-1 receptor was compared in HC (n = 9) and LN (n = 12) plasma-cultured condition as in Fig. 5a. h Serum IL-1β level was quantified for HC and LN, presented in optical density at 450 nm (OD_450nm) and concentration (pg/mL). **P <* 0.05 (Student’s t test)

그림 출처: PubMed Central 공개 논문 (PMC7104540)

Conflict of interest statement

The authors declare that they have no competing interests.

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