Volume 54, Issue 5, Pages 1463-1469 (November 1998) Calcitriol modulates in vivo and in vitro cytokine production: A role for intracellular calcium  Vincenzo Panichi, Stefano De Pietro, Brunella Andreini, Anna Maria Bianchi, Massimiliano Migliori, Daniele Taccola, Luca Giovannini, Ciro Tetta, Roberto Palla  Kidney International  Volume 54, Issue 5, Pages 1463-1469 (November 1998) DOI: 10.1046/j.1523-1755.1998.00152.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 In vitro studies. TNF-α production in PBMC stimulated by LPS (100ng/ml for 12hours). The simultaneous addition of different doses of calcitriol to these cultures induced a significantly dose-dependent inhibition of TNF-α production in all groups. Symbols are: (□) normal subjects; (□) chronic predialytic; (▪) hemodialysis patients. Bars are the means ± 1 sd of fifteen experiments performed in triplicate (*P < 0.05) Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 In vitro studies. IL-1β production in PBMC stimulated by LPS (100ng/ml for 12hours). The simultaneous addition of different doses of calcitriol to these cultures induced a significantly dose-dependent inhibition of IL-1β production in all groups. Symbols are: (□) normal subjects; (□) chronic predialytic; (▪) hemodialysis patients. Bars are the means ± 1 sd of fifteen experiments performed in triplicate (*P < 0.05). Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 In vivo studies. TNF-α production by PBMC of 12 hemodialytic patients assayed at the start (□) and at the end (□) of a single midweek hemodialysis session, before and after three months of calcitriol therapy. The basal intradialytic increase was significantly reduced (*P < 0.05). Bars are the means ± 1 sd of 12 dialyzed uremic patients. Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 In vivo studies. IL-1β production by PBMC of 12 hemodialytic patients assayed at the start (□) and at the end (□) of a single midweek hemodialysis session, before and after three months of calcitriol therapy. The basal intradialytic increase was significantly reduced (*P < 0.05). Bars are the means ± 1 sd of 12 dialyzed uremic patients. Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 In vitro studies. Intracellular calcium ([Ca++]i) concentration in PBMC of normal subjects (□), chronic predialytic (□) and hemodialytic patients (▪) as the control. Intracellular calcium was significantly increased following LPS stimulation (100ng/ml for 12hours;*P < 0.05) in all groups. The simultaneous addition of calcitriol (at a dose of 2.5 × 10-10m) was able to prevent [Ca++]i increase. Bars are the means ± 1 sd of ten experiments performed in duplicate in each group of subjects. Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 6 In vivo studies. (A) Significant (P< 0.001) correlation between IL-1β levels and [Ca++]i on PBMC of 12 hemodialytic patients assayed at the start of a single midweek hemodialysis session before calcitriol therapy (basal). (B) Significant (P< 0.001) correlation between IL-1β levels and [Ca++]i on PBMC of 12 hemodialytic patients assayed at the start of a single midweek hemodialysis session after three months of calcitriol therapy. Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Kidney International 1998 54, 1463-1469DOI: (10. 1046/j. 1523-1755 Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Kidney International 1998 54, 1463-1469DOI: (10. 1046/j. 1523-1755 Kidney International 1998 54, 1463-1469DOI: (10.1046/j.1523-1755.1998.00152.x) Copyright © 1998 International Society of Nephrology Terms and Conditions