![Fig. 2. MD-DCs from PBSCs collected in children with NB. / MD-DCs were derived from adherent monocytes and cultured for 7 days in AIM-V medium with 10% FCS supplemented with rhuIL-4 and GM-CSF (1000 IU/mL each). Induction of maturation was performed by adding LPS (20 μg/mL) on day 5 for 48 hours. (A) Immunophenotyping of immature (iDC, LPS−) versus mature (mDC, LPS+) MD-DCs was performed by 3-color flow cytometry by means of the combinations of FITC-, PE-, and PerCP-conjugated mAbs CD40/CD83/HLA-DR, CD86/CD1a/HLA-DR, and CD80/CD14/HLA-DR. One representative experiment is shown. Induction of maturation is evidenced by up-regulation of CD83 (mean fluorescence intensity [MFI], 29.6 versus 3.4); CD86 (MFI, 152.7 versus 60.4); CD80 (MFI, 61.2 versus 18.9); HLA-DR (MFI, 454.1 versus 129.4); and down-regulation of CD1a (MFI, 235.5 versus 187.3). (B) Allostimulatory capacities of immature (LPS−) versus mature (LPS+) MD-DCs. Results are presented as mean ± SEM (n = 9) proliferation of 2 × 105 allogeneic mononuclear cells as a function of decreasing numbers of stimulating MD-DCs. Stars indicate significant statistical differences (P < .01) between values obtained with immature versus mature MD-DCs.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/100/7/10.1182_blood.v100.7.2554/3/h81923173002.jpeg?Expires=1724241475&Signature=z9uaJHNAXb1I2cDvUI0xdnivycXtA7ZSd4FtPNgSf5BbODIyUE9a-M9vFIwfBobT9O54N4-0DTgZDs65x66cNC2FhQGczALnCx5QsXMyfsEY7gs3hoLX1gvBHx8K4A1HsXkHzxO4ij1XCUT371M6phYZeXb5wc2ONql0zU1lPEGWWXHsM-0Djy9I-hHnRWWHANHIJOuDxGxXNbGxsUczmyqjD8A44lTtSJWIKsHqWrSwLlFp1Z7EnMgD~~G-Dt5gfUwrevUZv5LsYfOa-K5bk1yJZQ6~j2hFraAn6AHX5b1TDCjiXFrEY-J99ZtVNyf33~1kS4dnKjpGhfPLpSV1gA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
MD-DCs from PBSCs collected in children with NB.
MD-DCs were derived from adherent monocytes and cultured for 7 days in AIM-V medium with 10% FCS supplemented with rhuIL-4 and GM-CSF (1000 IU/mL each). Induction of maturation was performed by adding LPS (20 μg/mL) on day 5 for 48 hours. (A) Immunophenotyping of immature (iDC, LPS−) versus mature (mDC, LPS+) MD-DCs was performed by 3-color flow cytometry by means of the combinations of FITC-, PE-, and PerCP-conjugated mAbs CD40/CD83/HLA-DR, CD86/CD1a/HLA-DR, and CD80/CD14/HLA-DR. One representative experiment is shown. Induction of maturation is evidenced by up-regulation of CD83 (mean fluorescence intensity [MFI], 29.6 versus 3.4); CD86 (MFI, 152.7 versus 60.4); CD80 (MFI, 61.2 versus 18.9); HLA-DR (MFI, 454.1 versus 129.4); and down-regulation of CD1a (MFI, 235.5 versus 187.3). (B) Allostimulatory capacities of immature (LPS−) versus mature (LPS+) MD-DCs. Results are presented as mean ± SEM (n = 9) proliferation of 2 × 105 allogeneic mononuclear cells as a function of decreasing numbers of stimulating MD-DCs. Stars indicate significant statistical differences (P < .01) between values obtained with immature versus mature MD-DCs.
