Mass cytometry identifies characteristic immune cell subsets in bronchoalveolar lavage fluid from interstitial lung diseases

Abstract

Immune cells have been implicated in interstitial lung diseases (ILDs), although their phenotypes and effector mechanisms remain poorly understood. To better understand these cells, we conducted an exploratory mass cytometry analysis of immune cell subsets in bronchoalveolar lavage fluid (BALF) from patients with idiopathic pulmonary fibrosis (IPF), connective-tissue disease (CTD)-related ILD, and sarcoidosis, using two panels including 64 markers. Among myeloid cells, we observed the expansion of CD14+ CD36hi CD84hiCCR2– monocyte populations in IPF. These CD14+ CD36hi CD84hi CCR2– subsets were also increased in ILDs with a progressive phenotype, particularly in a case of acute exacerbation (AEx) of IPF. Analysis of B cells revealed the presence of cells at various stages of differentiation in BALF, with a higher percentage of IgG memory B cells in CTD-ILDs and a trend toward more FCRL5+ B cells. These FCRL5+ B cells were also present in the patient with AEx-IPF and sarcoidosis with advanced lung lesions. Among T cells, we found increased levels of IL-2R+ TIGIT+ LAG3+ CD4+ T cells in IPF, increased levels of CXCR3+ CD226+ CD4+ T cells in sarcoidosis, and increased levels of PD1+ TIGIT+ CD57+ CD8+ T cells in CTD-ILDs. Together, these findings underscore the diverse immunopathogenesis of ILDs.

Exploratory mass cytometry analysis reveals immunophenotypes of cancer treatment-related pneumonitis

“…In both rheumatoid arthritis (Owczarczyk et al, 2011) and granulomatosis with polyangiitis and microscopic polyangiitis (Owczarczyk et al, 2020), increased expression of FCRL5 was indicative of a positive response to rituximab. Additionally, our recent research revealed an increase in FCLR5 + B cells in connective-tissue disease-related ILD (Hata et al, 2023). These findings suggest a potential pathological role of FCRL5 in autoimmunity.…”

“…Metal-tagged antibodies were obtained from Standard Biotools or purchased in a purified form (Supplementary Table 1) and then labeled with metals using the Maxpar Antibody Labeling Kit (Standard Biotools) as instructed by the manufacturer. The cell labeling was performed as previously described (Hata et al, 2023). Upon collection, BALF samples were immediately centrifuged at 300 g for 5 minutes to pellet the cells.…”

“…This technique provides a unique advantage over traditional flow cytometry, which is limited by spectral overlap and the number of parameters that can be analyzed simultaneously. Mass cytometry has been used to study a variety of biological systems, including immune-mediated diseases, such as cancer and autoimmune conditions (Matsubara et al, 2021) (Hata et al, 2023) (Couloume et al, 2021) (Chedid et al, 2022). Through high-throughput analysis of large numbers of single cells, mass cytometry can provide a more comprehensive understanding of the cellular heterogeneity and signaling pathways involved in these diseases, allowing for the identification of potential therapeutic targets and biomarkers.…”

Shared genetic etiology between hypothyroidism and complex diseases: a large-scale genome-wide cross-trait analysis

“…CD226, a member of the immunoglobulin superfamily, is a functional protein originally produced on natural killer and T cells [54]. A recent study has reported that CD226 expression in T cells from sarcoidosis [55]. Furthermore, it was proposed that CD4 T cells moving to the lungs via CXCR3 could be activated by CD226, potentially contributing to the pathogenesis of sarcoidosis.…”

Expansion of ST2-expressing macrophages in a patient with bronchiolitis obliterans syndrome

“…The proportion of cell populations in CD45 + gated BALF cells as determined by tSNE analysis. The methodology has been previously described [9].…”