Selective Immunomodulation of Inflammatory Pathways in Keratinocytes by the Janus Kinase (JAK) Inhibitor Tofacitinib: Implications for the Employment of JAK-Targeting Drugs in Psoriasis

Table 2

Tofacitinib effects on the expression of inflammatory molecules induced by IL-22 in keratinocytes.

	Untreated	TOF	IL-22	IL-22 + TOF
Membrane molecules
ICAM1	2.4 ± 0.2	2.0 ± 0.18	2.5 ± 0.24	1.8 ± 0.16
HLA-DR	1.4 ± 0.15	1.4 ± 0.12	1.4 ± 0.12	1.4 ± 0.11
MHC-I	72 ± 5.2	63 ± 4.5	57 ± 4.8	54 ± 5.2
Chemokines
CX3CL1	0.40 ± 0.05	0.36 ± 0.08	46.72 ± 4.57	19.72 ± 1.87†
CXCL1	0.80 ± 0.12	0.80 ± 0.07	0.80 ± 0.09	0.40 ± 0.035
CXCL8	0.92 ± 0.11	2.60 ± 0.32	83.04 ± 9.43	49.04 ± 4.85†
CXCL10	29.08 ± 2.2	25.04 ± 0.14	36.60 ± 3.57	43.28 ± 3.19
CXCL12	107.20 ± 9.72	110.20 ± 2.43	119.12 ± 12.91	111.16 ± 5.76
CXCL16	2.92 ± 0.32	5.28 ± 0.68	12.92 ± 1.39	10.52 ± 1.15
CCL1	0.80 ± 0.1	0.80 ± 0.14	10.60 ± 1.08	10.60 ± 1.10
CCL2	0.80 ± 0.95	0.76 ± 0.67	1.72 ± 0.08	0.72 ± 0.62
CCL5	2.0 ± 0.4	1.9 ± 0.24	1.95 ± 0.16	1.73 ± 0.13
MIF	337.08 ± 35.71	637.04 ± 52.47	1800.12 ± 105.9	1807.00 ± 135.7
Cytokines, AMPs, SOCS
IL-6	5.92 ± .0.79	5.28 ± 0.48	7.24 ± 0.84	6.24 ± 0.54
IL-20	1.00 ± 0.12	0.90 ± 0.10	3.17 ± 0.17	3.16 ± 0.05
LL-37	1.00 ± 0.11	0.89 ± 0.09	0.84 ± 0.11	1.51 ± 0.13
HBD2	1.00 ± 0.12	0.66 ± 0.08	3.62 ± 0.39	3.96 ± 0.56
S100A7	1.00 ± 0.14	1.11 ± 0.2	1.73 ± 0.16	1.71 ± 0.18
SOCS3	1.00 ± 0.12	1.13 ± 0.10	8.12 ± 0.95	0.64 ± 0.05

Note: IL: interleukin; TOF: tofacitinib; ICAM: intercellular adhesion molecule; HLA-DR: human leukocyte antigen-antigen D related; MHC: major histocompatibility complex; CXCL: CXC-chemokine ligand; CL: chemokine ligand; MIF: macrophage migration inhibitory factor; LL37: antimicrobial peptide; HBD: human-defensin; S100: S100 calcium-binding protein; SOCS: suppressor of cytokine signaling. Keratinocyte cultures were left untreated or treated with 5 μM of tofacitinib and stimulated or not with 75 ng of IL-22. After 6 hours, IL-20, LL-37, HBD2, S100A7, and SOCS3 mRNA levels were analysed by real-time PCR and normalized to β-actin mRNA levels. Results are expressed as mean 2^−ΔΔCT ± SD. After 24 hours, cells were stained with ICAM1, HLA-DR, and MHC-I mAb followed by FITC-conjugated anti-mouse IgG and then analysed by flow cytometry. Data are expressed as mean ΔMFI ± SD. At the same time, supernatants were collected and, chemokines and IL-6 were measured by Bioplex, except for CCL5 which has been evaluated by ELISA. Results are expressed as mean pg/ml ± SD. compared to untreated or stimulated keratinocytes.