Figure 1.
The APACHE-II and PMEWS scores and Ct value in patients.
High APACHE-II scores (A) and the high PMEWS scores (B) were observed in severe patients compared with mild patients; (C) The ages of patients versus APACHE-II score with Spearman's correlation coefficients. Each point represents an individual patient. Results are shown for patients with severe H7N9 infection (red symbols) with mild H7N9 infection (blue symbols). (D) The Ct values on the 10th day after symptom onset in mild and severe patients. Data represent mean ±SD.
Table 1.
Epidemiological and clinical features of patients with H7N9 virus infection.
Figure 2.
Leucocyte subset counts and their correlation with the severity of H7N9 infection.
(A) The total number of leucocytes and the percentages of lymphocytes in patients during disease progression. Each line represents an individual patient. Results are shown for patients with severe H7N9 infection (red tones symbols and lines) with mild H7N9 infection (blue tone symbols and lines). (B) The percentages of neutrophils, lymphocytes and monocytes in patients on the 7th day after symptom onset. Data represent mean ±SD. (C) the lowest percentages for neutrophils, lymphocytes and monocytes vs. APACHE-II score with Spearman's rank correlation coefficients. Each point represents an individual patient. Results are shown for patients with severe H7N9 infection (red tones symbols and lines) with mild H7N9 infection (blue tone symbols and lines).
Figure 3.
Levels of plasma cytokines and chemokines over the course of H7N9 infection.
(A) Plasma levels of IFN-α, IL-2 and IL-12; (B) IFN-γ, IL-17A and IL-10; (C) IP-10, MCP-1 and MIP-1β in patients with severe versus mild H7N9 infection on different days after the onset of symptoms. Data represent mean ±SD for patients with severe H7N9 infection (red tones symbols and lines) and with mild H7N9 infection (blue tone symbols and lines). Experiments were performed in duplicates and repeated at least three times.
Figure 4.
The percentages of lymphocyte subsets in peripheral blood.
The distribution of CD3+ cells, CD4+ cells, CD8+ cells(A), B cells (B), NK cells (C), dendritic cells (D), CD14+ cells (E) and the percentages of HLA-DR expression on CD14+ monocytes (F and G) in patients during acute and recovery phase were detected by flowcytometry. Data represent mean ±SD. (H) The absolute numbers of HLA-DR+CD14+ monocytes in patients during acute and recovery phase were detected by flowcytometry. Data represent mean ±SD. (I) The lowest percentages for HLA-DR expression level on CD14+ monocytes versus APACHE-II score with Spearman's correlation coefficients. Each point represents an individual patient. Results are shown for patients with severe H7N9 infection (red symbols) with mild H7N9 infection (blue symbols).
Figure 5.
Phagocytosis and antigen presentation capacity of CD14+ cells.
(A) Percentages of HLA-DR expressing on E.coli+ CD14+ cells following RFP-labeled E. coli stimulation. Data represent mean ±SD. (B) HLA-DR (stained by green) expressions on phagocytic internalization of E. coli stained by DAPI (blue) in PBMC (shown by red arrows in merged image) was confirmed by fluorescence microscopy. Original magnification: ×400 & ×600. (C) Intracellular IFN-γ expression in CD4+ T cells from health control (HC) after co-cultured with CD14+ cells from patients with severe and mild H7N9 infection (the ratio of CD14+ cells/CD4+ T cells is 1:1) following polyI:C stimulation (20 ng/ml) (left); intracellular IFN-γ expression in CD4+ T cells from patients with severe and mild H7N9 infection after co-cultured with CD14+ cells from HC following polyI:C stimulation middle); intracellular IFN-γ expression in CD4+ T cells from HC after co-cultured with CD14+ cells from HC following polyI:C stimulation (right).
Figure 6.
In vitro analysis of cytokine and chemokine production by patients PBMC.
(A) The levels of cytokines and chemokines secreted by inactivated H7N9 virus (1Moi) stimulated PBMC from patients with severe and mild H7N9 infection (5 μl/ml). Data represent mean ±SD. (B) ELISPOT analysis of IFN-γ and IL-17 secreting cells following polyI:C stimulation of PBMC from patients with severe and mild H7N9 infection (20 ng/ml). All ELISPOT assays were performed in duplicate. Intracellular IFN-γ expression in CD8+ T (C), CD4+T (D) cells and intracellular IL-17A expression in CD4+T (E) cells after heat inactivated H7N9 virus stimulation was analyzed by flow cytometry. Data represent mean ± SD.