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. 2022 Jun 27:13:922000.
doi: 10.3389/fimmu.2022.922000. eCollection 2022.

High-Dimensional Immune Profiling by Mass Cytometry Revealed the Circulating Immune Cell Landscape in Patients With Intracranial Aneurysm

Peicong Ge  1   2   3   4   5 , Chenglong Liu  1   2   3   4   5 , Liujia Chan  6 , Yuheng Pang  6 , Hao Li  1   2   3   4   5 , Qian Zhang  1   2   3   4   5 , Xun Ye  1   2   3   4   5 , Jia Wang  1   2   3   4   5 , Rong Wang  1   2   3   4   5 , Yan Zhang  1   2   3   4   5 , Wenjing Wang  6 , Dong Zhang  1   2   3   4   5   7 , Jizong Zhao  1   2   3   4   5   8
Affiliations

High-Dimensional Immune Profiling by Mass Cytometry Revealed the Circulating Immune Cell Landscape in Patients With Intracranial Aneurysm

Peicong Ge et al. Front Immunol. .

Abstract

Background: Increasing evidence supports a critical role of chronic inflammation in intracranial aneurysm (IA). Understanding how the immunological alterations in IA provides opportunities for targeted treatment. However, there is a lack of comprehensive and detailed characterization of the changes in circulating immune cells in IA.

Objective: To perform a comprehensive and detailed characterization of the changes in circulating immune cells in patients with IA.

Methods: Peripheral blood mononuclear cell samples from IA patients (n = 26) and age-and sex-matched healthy controls (HCs, n = 20) were analyzed using high dimensional mass cytometry, and the frequency and phenotype of immune cell subtypes were assessed.

Results: We identified 28 cell clusters and found that the immune signature of IA consists of cluster changes. IA patients exhibited dysfunction of immunity, with dysregulation of CD4+ T-cell clusters, increased B cells and monocytes, and decreased CD8+ T cells, DNT cells, and DPT cells. Moreover, compared with findings in HC, IA was associated with enhanced lymphocyte and monocyte immune activation, with a higher expression of HLA-DR, CXCR3, and CX3CR1. In addition, the expression of TLR4, p-STAT3, and the exhaustion marker PD1 was increased in T cells, B cells, and NK cells in IA patients.

Conclusions: Our data provide an overview of the circulating immune cell landscape of IA patients, and reveal that the dysfunction of circulating immunity may play a potential role in the development of IA.

Keywords: circulating immune cell; inflammation; intracranial aneurysm; landscape; mass cytometry.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Overview of the imaging mass cytometry study of IA. (A) Flowchart overview of PBMC collection. Twenty-six PBMC samples from IA patients were included in this study, as well as 20 control samples. PBMCs were stained with one antibody panels and acquired on the CyTOF instrument. (B) t-SNE visualization of CD45+ cells from PBMCs. PhenoGraph clusters analysis of manual gated eight-cell lineages, including CD4+ T cells, CD8+ T cells, CD4+CD8+ double-positive T cells (DPT), CD4-CD8- double-negative T cells (DNT), B cells, NK cells, dendritic cells (DCs), and monocytes. (C) Representative t-SNE plot of 28 cell clusters from HC and IA patients:CD4+ T cells, nine clusters (C02, C03, C05, C09, C11, C17, C20, C26, C27); CD8+T cells, six clusters (C06, C10, C12, C13, C19, C23); DPT, one cluster (C15); DNT, three clusters (C14, C22, C24); B cells, three clusters (C04, C08, C28); NK cells, two clusters (C01,C25); monocytes, two clusters (C07, C18); DC, two clusters (C16, C21) (D) Heatmap showing median marker expression across the 8 cell lineages (E) Heatmap of the expression of each protein in 28 cell clusters.
Figure 2
Figure 2
Changes in cell proportions in IA. (A) Percentage of 8 cell lineages in CD45+ cells from HC and IA patients. P values were calculated by the two-sided Wilcoxon test. Significance: ***p < 0.001; **p < 0.01; *p < 0.05; ns p ≥ 0.05. (B) Bar chart of the percentage of 8 cell lineages in CD45+ cells. (C) Percentage of 28 cell lineages in CD45+ cells from HC and IA patients. P values were calculated by the two-sided Wilcoxon test. Significance: ***p < 0.001; **p < 0.01; *p < 0.05; ns p ≥ 0.05. (D) Bar chart of the percentage of 28 cell lineages in CD45+ cells.
Figure 3
Figure 3
Marker expressions in 8 cell lineages. (A) Marker expressions in CD4+ T cells from HC and IA patients. The expression levels of TLR4, PD1, p-STAT3, CD25, CD45RO, HLA-DR, CXCR3, CX3CR1, CCR4, CCR10, and IL-17A in CD4+ T cells were higher in the IA group than in the HC group. The levels of GP130, CD11b, CD27, CD56, CXCR5, CCR6, FOXP3, and IL-10 were lower in the patient groups than in the HC group. (B) Marker expressions in CD8+ T cells from HC and IA patients. The expression levels of TLR4, PD1, p-STAT3, CD25, CD45RA, CD45RO, HLA-DR, CXCR3, CX3CR1, CCR4, and CCR10 in CD8+ T cells were higher in the IA group than in the HC group. However, the expression levels of GP130, CD11b, CD27, CXCR5, CCR6, and IL-10 were lower in the IA group than in the HC group. (C) Marker expressions in DPT cells from HC and IA patients. The expression of PD1, p-STAT3, CD25, CD45RO, HLA-DR, CXCR3, CX3CR1, CCR4, and CCR10 in DPT cells was higher in the IA group than in the HC group. The levels of GP130, CD11b, CD27, CXCR5, CCR6, and IL-10 were lower in the patient groups than in the HC group (D) Marker expressions in DNT cells from HC and IA patients. The expression levels of TLR4, PD1, p-STAT3, CD11c, CD25, CD27, CD45RA, CD45RO, HLA-DR, CXCR3, CX3CR1, CCR4, CCR7, and CCR10 in DNT cells were higher in the IA group than in the HC group. In addition, the levels of CD11b and CCR6 were lower in the patient groups than in the HC group.(E) Marker expressions in B cells from HC and IA patients. The expression levels of TLR4, PD1, p-STAT3, CD27, CD34, CD45RA, CD45RO, CXCR3, CX3CR1, CCR4, CCR7, CCR10, and IL-10 in B cells were higher in the IA group than in the HC group. The levels of CD11b, CCR6, HLA-DR, and CXCR5 were lower in the patient groups than in the HC group. (F) Marker expressions in NK cells from HC and IA patients. The expression levels of TLR4, PD1, p-STAT3, CD45RO, CXCR3, CX3CR1, and HLA-DR in NK cells were higher in the IA group than in the HC group. The levels of CD11b, CD16, and CD56 were lower in the IA group than in the HC group. (G) Marker expressions in monocytes from HC and IA patients. The expression levels of p-STAT3, CD11c, CD14, CD45RA, CD45RO, CXCR3, CX3CR1, and HLA-DR in monocytes were higher in the IA group than in the HC group. However, the levels of GP130, CD11b, CD15, CD16, CD68, CD163, and CXCR5 were lower in the IA group than in the HC group. (H) Marker expressions in DCs from HC and IA patients. The expression levels of TLR4, PD1, p-STAT3, GP130, CD11c, CD45RA, CD45RO, CXCR3, and CX3CR1 in DC cells were higher in the IA group than in the HC group. P values were calculated by the two-sided Wilcoxon test. Significance: ***p < 0.001; **p < 0.01; *p < 0.05; ns p ≥ 0.05.
Figure 4
Figure 4
Marker expressions in Treg cells. The expression levels of TLR4, PD1, p-STAT3, and HLA-DR in Treg cells were higher, while those of GP130, Foxp3, and IL-10 were lower, in IA patients compared with HCs. P values were calculated by the two-sided Wilcoxon test. Significance: ***p < 0.001.
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References

    1. Vlak MH, Algra A, Brandenburg R, Rinkel GJ. Prevalence of Unruptured Intracranial Aneurysms, With Emphasis on Sex, Age, Comorbidity, Country, and Time Period: A Systematic Review and Meta-Analysis. Lancet Neurol (2011) 10(7):626–36. doi: 10.1016/S1474-4422(11)70109-0 - DOI - PubMed
    1. Korja M, Kaprio J. Controversies in Epidemiology of Intracranial Aneurysms and SAH. Nat Rev Neurol (2016) 12(1):50–5. doi: 10.1038/nrneurol.2015.228 - DOI - PubMed
    1. Nieuwkamp DJ, Setz LE, Algra A, Linn FH, de Rooij NK, Rinkel GJ. Changes in Case Fatality of Aneurysmal Subarachnoid Haemorrhage Over Time, According to Age, Sex, and Region: A Meta-Analysis. Lancet Neurol (2009) 8(7):635–42. doi: 10.1016/S1474-4422(09)70126-7 - DOI - PubMed
    1. Bakker MK, van der Spek R, van Rheenen W, Morel S, Bourcier R, Hostettler IC, et al. . Genome-Wide Association Study of Intracranial Aneurysms Identifies 17 Risk Loci and Genetic Overlap With Clinical Risk Factors. Nat Genet (2020) 52(12):1303–13. doi: 10.1038/s41588-020-00725-7 - DOI - PMC - PubMed
    1. Frösen J, Tulamo R, Paetau A, Laaksamo E, Korja M, Laakso A, et al. . Saccular Intracranial Aneurysm: Pathology and Mechanisms. Acta Neuropathol (2012) 123(6):773–86. doi: 10.1007/s00401-011-0939-3 - DOI - PubMed

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