Figure 1
Figure 1. Human monocytes cocultured in vitro with BCP-ALL cells show increased expression of chemokine CXCL10. (A) Human monocytes (Mo) were cocultured with CD19+ cALL blasts isolated from cALL patients in a transwell system for 18 hours. Thereafter, the cALL-containing transwells were removed, the monocyte monolayers were washed, lysed, and use for accessing the expression of a panel of cytokine, chemokine, and tumor promotion (tumor angiogenesis and invasion) related genes. Mo indicates monocytes alone; and Mo + cALL, monocytes cocultured with cALL tumor blasts. A Mo/cALL ratio of 1:5 was used in this experiment. *P < .04, versus Mo. (B) Monocytes were cultured alone or cocultured with cALL cells as described in panel A. Thereafter, the cALL-containing transwells were removed; the monocyte monolayers were washed, incubated in fresh media for 1 hour, and stimulated with LPA (100 ng/mL) for 4 hours. Expression of the same panel of genes as described in panel A was assessed by quantitative PCR. Mo + LPA indicates monocytes cultured alone stimulated with LPA; and Mo + cALL + LPA, monocytes cocultured with cALL tumor blasts followed by LPA stimulation. *P < .05, versus Mo + LPA. (A-B) Data are mean ± SEM (n = 4). (C) LPA-induced expression of CXCL10 gene was assessed in monocytes cocultured with the cALL tumor blasts at different ratios as indicated in the figure. The procedure for coculturing and LPA stimulation remains the same as indicated for panel A. cALL + LPA indicates cALL blasts stimulated with 100 ng/mL LPA for 4 hours. Data are mean ± SEM (n = 3). *P < .05, versus Mo (1:0). **P < .05, versus Mo (1:0) + LPA. (D) Expression of CXCL10 protein by monocytes cocultured with cALL blasts, pre-B ALL blasts, or B cells from healthy donors (normal B), treated with or without 100 ng/mL LPA overnight. The procedure for coculturing remains the same as indicated for panel A. CXCL10 was measured by ELISA in the cell-free culture supernatant of the cells. Data are mean ± SEM (n = 3). *P < .03, versus Mo + LPA. (E) Monocytes were cultured alone or cocultured with a panel of human BCP-ALL cell lines for 18 hours using the same protocol as described in panel A. Thereafter, cells were stimulated or not with LPA (100 ng/mL) for 4 hours and CXCL10 gene expression assessed. Data are mean ± SD from a representative experiment. *P < .05, versus Mo. **P < .05, versus Mo + LPA.

Human monocytes cocultured in vitro with BCP-ALL cells show increased expression of chemokine CXCL10. (A) Human monocytes (Mo) were cocultured with CD19+ cALL blasts isolated from cALL patients in a transwell system for 18 hours. Thereafter, the cALL-containing transwells were removed, the monocyte monolayers were washed, lysed, and use for accessing the expression of a panel of cytokine, chemokine, and tumor promotion (tumor angiogenesis and invasion) related genes. Mo indicates monocytes alone; and Mo + cALL, monocytes cocultured with cALL tumor blasts. A Mo/cALL ratio of 1:5 was used in this experiment. *P < .04, versus Mo. (B) Monocytes were cultured alone or cocultured with cALL cells as described in panel A. Thereafter, the cALL-containing transwells were removed; the monocyte monolayers were washed, incubated in fresh media for 1 hour, and stimulated with LPA (100 ng/mL) for 4 hours. Expression of the same panel of genes as described in panel A was assessed by quantitative PCR. Mo + LPA indicates monocytes cultured alone stimulated with LPA; and Mo + cALL + LPA, monocytes cocultured with cALL tumor blasts followed by LPA stimulation. *P < .05, versus Mo + LPA. (A-B) Data are mean ± SEM (n = 4). (C) LPA-induced expression of CXCL10 gene was assessed in monocytes cocultured with the cALL tumor blasts at different ratios as indicated in the figure. The procedure for coculturing and LPA stimulation remains the same as indicated for panel A. cALL + LPA indicates cALL blasts stimulated with 100 ng/mL LPA for 4 hours. Data are mean ± SEM (n = 3). *P < .05, versus Mo (1:0). **P < .05, versus Mo (1:0) + LPA. (D) Expression of CXCL10 protein by monocytes cocultured with cALL blasts, pre-B ALL blasts, or B cells from healthy donors (normal B), treated with or without 100 ng/mL LPA overnight. The procedure for coculturing remains the same as indicated for panel A. CXCL10 was measured by ELISA in the cell-free culture supernatant of the cells. Data are mean ± SEM (n = 3). *P < .03, versus Mo + LPA. (E) Monocytes were cultured alone or cocultured with a panel of human BCP-ALL cell lines for 18 hours using the same protocol as described in panel A. Thereafter, cells were stimulated or not with LPA (100 ng/mL) for 4 hours and CXCL10 gene expression assessed. Data are mean ± SD from a representative experiment. *P < .05, versus Mo. **P < .05, versus Mo + LPA.

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