Immunomodulatory Effects of 1,25-Dihydroxyvitamin D₃ on Dendritic Cells Promote Induction of T Cell Hyporesponsiveness to Myelin-Derived Antigens

Table 3

Immunophenotypic analysis of in vitro differentiated mo-DC from healthy controls before and after cryopreservation.

Healthy controls () cc-mDC
Marker	Type	Fold change (cc-mDC/iDC)	MFI ± SD	value	value
CD86	Conventional DC	1.03	637 ± 98	n.s.	< 0.05
	1,25(OH)2D3 DC	1.19	209 ± 155	n.s.
CD83	Conventional DC	1.43	45 ± 10	< 0.05	< 0.05
	1,25(OH)2D3 DC	0.90	22 ± 6	n.s.
CD80	Conventional DC	1.24	23 ± 7	n.s.	< 0.05
	1,25(OH)2D3 DC	1.00	15 ± 4	n.s.
HLA-DR	Conventional DC	0.89	103 ± 21	n.s.	n.s.
	1,25(OH)2D3 DC	0.92	61 ± 36	n.s.

CD14+ monocytes were cultured for 7 days in the presence of IL-4 and GM-CSF or in the presence of IL-4, GM-CSF, and 1,25(OH)₂D₃ to obtain immature conventional DC or 1,25(OH)₂D₃-treated DC, respectively. On day 7, iDC were frozen and stored at −80°C. Next, cryopreserved iDC were thawed, rested for 2 h at 37°C, and stimulated with a proinflammatory cytokine cocktail for 24 h (i.e., cc-mDC). The mean fluorescent intensity (MFI) of costimulatory molecules, CD80 and CD86, of maturation marker, CD83, and of HLA-DR by various DC subsets of healthy controls () was determined Results are expressed as fold change, calculated as the ratio between the MFI value after maturation following a freeze-thaw cycle and the MFI value obtained at immature stage following a freeze-thaw cycle.
The values indicated are calculated for cc-mDC DC after cryopreservation versus iDC after cryopreservation.
The values indicated are calculated for conventional cc-mDC after cryopreservation versus 1,25(OH)₂D₃-treated cc-mDC after cryopreservation.
MFI, mean fluorescence intensity; cc-mDC, cytokine cocktail-matured DC; iDC, immature DC; and n.s., nonsignificant.