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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in <t>R848-stimulated</t> pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.
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Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in R848-stimulated pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.

Journal: Frontiers in Immunology

Article Title: Integrated analysis of sex differences in human dendritic cell, monocyte, and natural killer cell subsets

doi: 10.3389/fimmu.2026.1750775

Figure Lengend Snippet: Frequency and functions comparisons of DC subsets in females and males. (A) Representative flow cytometry gating strategy for the identification of human DC subsets. Home-made lineage markers included CD3, CD19, CD56. AXL + DC: lineage - HLA-DR + AXL + ; pDCs: lineage - HLA-DR + AXL - CD11c - CD123 + ; CD141 + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD1c - CD141 + ; CD1c + DC: lineage - HLA-DR + AXL - CD123 - CD11c + CD141 - CD1c + . (B) Comparative analysis of the relative frequencies of DC subsets (AXL + DC, pDCs, CD1c + DC, CD141 + DC, and CD1c - CD141 - DC) in PBMCs between males (n = 46) and females (n = 45). (C) Correlation between donor age and the frequency of pDCs (left) and AXL + DCs (right). Each dot represents an individual donor. (D) The UpSet plot illustrating the integrated comparative analysis of differentially expressed genes (DEGs) in DC subsets between females and males. (E) Functional pathway enrichment analysis of DEGs in CD141 + DC, CD1c + DC, and pDCs, as determined by Ingenuity Pathway Analysis (IPA). The color scale represents the activation z-score, with red indicating pathway activation and blue indicating inhibition. Gray indicates that an activation state could not be determined. (F) Predicted upstream regulators of the DEGs in each DC subset from females compared to males. The color scale represents the activation z-score, with red indicating predicted activation and blue indicating predicted inhibition of the regulator in females. (G) Mean Fluorescence Intensity (MFI) of HLA-DR on CD141 + DC, CD1c + DC, and pDCs, comparing male and female donors. (H) Relative expression of IFN-β in R848-stimulated pDCs from females (n=10) and males (n =10), measured by qPCR, and IFN-beta secretion into the supernatant, quantified by ELISA. Data was shown as mean ± SD. Statistical significance for comparisons between two groups was determined using the Mann-Whitney test. Correlations in (C) were assessed by Spearman’s rank correlation. Significance levels are denoted as follows: ns,not significant; * p < 0.05, ** p < 0.01.

Article Snippet: The total pDCs were isolated from PBMCs using EasySep human pDCs isolation kit (Stemcell, Cat. no17977) and 900 pDCs was plated in 96U plate, cells were stimulated with 10ng/ml TLR7/8 agonist R848 compound (TLRL-R848, MCE) for 24hours.

Techniques: Flow Cytometry, Functional Assay, Activation Assay, Inhibition, Fluorescence, Expressing, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY