Infection-induced TIMP1 in BM stromal cells enhances myeloid cell production. (A) Cytokine array assay. Serum samples were pooled from three to five sham or CL&P mice at 10 wk after surgery. (1) G-CSF, (2) GM-CSF, (3) IL-1α, (4) IL-1β, (5) IL-6, (6) TNFα, (7) TIMP1. (B) ELISA measurement of circulating TIMP1 levels in mice at the indicated time points after sham surgery (n = 10) or CL&P surgery (n = 14). ** P < 0.01; *** P < 0.001; **** P < 0.0001. (C) Correlation between serum levels of TIMP1 and the number of BM granulocytes in mice at 12 wk after sham surgery (n = 9) or CL&P surgery (n = 9). Each symbol represents an individual mouse, and the data were pooled from three independent experiments. Bars represent the mean value ± SD. The Spearman correlation coefficient (R) and its associated P value (P) are indicated. (D) Numbers of colonies formed in the methylcellulose medium after a 10-day culture. The cultures were supplied with serum from sham or CL&P mice at 10 wk after surgery or with serum of CL&P mice plus anti-TIMP1 antibody. Symbols represent data of independent experiments with bars showing the mean value ± SD. * P < 0.05; ns, P > 0.05. (E) FACS analysis of cells collected from the culture described in Fig. 5 D. Percentages of Ly6G+ granulocytes are shown with the mean value ± SD. Data were collected from three independent cultures. (F) Representative FACS plots show the frequencies of CD11b+ and CD11b− cells in the peripheral blood of Timp1−/− host mice transferred with TIMP1-expressing BM cells or control BM cells. The right panel shows the percentages of CD11b+ cells. Data were collected from five mice/group. Bars represent the mean value ± SD. * P < 0.05. (G) qRT-PCR analysis of Timp1 transcripts in CD45+ cells, CD45−CD31+ endothelial cells and CD45−CD31− stromal cells purified from sham BM (n = 6) and CL&P BM (n = 6). Symbols represent data of individual mice compiled from two independent experiments, with the mean value ± SD. * P < 0.05; ns, P > 0.05 (unpaired two-tailed Student’s t test). Source data are available for this figure: SourceData F5.
Figure 5.

Infection-induced TIMP1 in BM stromal cells enhances myeloid cell production. (A) Cytokine array assay. Serum samples were pooled from three to five sham or CL&P mice at 10 wk after surgery. (1) G-CSF, (2) GM-CSF, (3) IL-1α, (4) IL-1β, (5) IL-6, (6) TNFα, (7) TIMP1. (B) ELISA measurement of circulating TIMP1 levels in mice at the indicated time points after sham surgery (n = 10) or CL&P surgery (n = 14). ** P < 0.01; *** P < 0.001; **** P < 0.0001. (C) Correlation between serum levels of TIMP1 and the number of BM granulocytes in mice at 12 wk after sham surgery (n = 9) or CL&P surgery (n = 9). Each symbol represents an individual mouse, and the data were pooled from three independent experiments. Bars represent the mean value ± SD. The Spearman correlation coefficient (R) and its associated P value (P) are indicated. (D) Numbers of colonies formed in the methylcellulose medium after a 10-day culture. The cultures were supplied with serum from sham or CL&P mice at 10 wk after surgery or with serum of CL&P mice plus anti-TIMP1 antibody. Symbols represent data of independent experiments with bars showing the mean value ± SD. * P < 0.05; ns, P > 0.05. (E) FACS analysis of cells collected from the culture described in Fig. 5 D. Percentages of Ly6G+ granulocytes are shown with the mean value ± SD. Data were collected from three independent cultures. (F) Representative FACS plots show the frequencies of CD11b+ and CD11b− cells in the peripheral blood of Timp1−/− host mice transferred with TIMP1-expressing BM cells or control BM cells. The right panel shows the percentages of CD11b+ cells. Data were collected from five mice/group. Bars represent the mean value ± SD. * P < 0.05. (G) qRT-PCR analysis of Timp1 transcripts in CD45+ cells, CD45−CD31+ endothelial cells and CD45−CD31− stromal cells purified from sham BM (n = 6) and CL&P BM (n = 6). Symbols represent data of individual mice compiled from two independent experiments, with the mean value ± SD. * P < 0.05; ns, P > 0.05 (unpaired two-tailed Student’s t test). Source data are available for this figure: SourceData F5.

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