The Cyclophilin-Binding Agent Sanglifehrin A Is a Dendritic Cell Chemokine and Migration Inhibitor

Abstract

Sanglifehrin A (SFA) is a cyclophilin-binding immunosuppressant but the immunobiology of action is poorly understood. We and others have reported that SFA inhibits IL-12 production and antigen uptake in dendritic cells (DC) and exhibits lower activity against lymphocytes. Here we show that SFA suppresses DC chemokine production and migration. Gene expression analysis and subsequent protein level confirmation revealed that SFA suppressed CCL5, CCL17, CCL19, CXCL9 and CXCL10 expression in human monocyte-derived DC (moDC). A systems biology analysis, Onto Express, confirmed that SFA interferes with chemokine-chemokine receptor gene expression with the highest impact. Direct comparison with the related agent cyclosporine A (CsA) and dexamethasone indicated that SFA uniquely suppresses moDC chemokine expression. Competitive experiments with a 100-fold molar excess of CsA and with N-Methyl-Val-4-cyclosporin, representing a nonimmunosuppressive derivative of CsA indicated chemokine suppression through a cyclophilin-A independent pathway. Functional assays confirmed reduced migration of CD4+ Tcells and moDCs to supernatant of SFA-exposed moDCs. Vice versa, SFA-exposed moDC exhibited reduced migration against CCL19. Moreover, SFA suppressed expression of the ectoenzyme CD38 that was reported to regulate DC migration and cytokine production. These results identify SFA as a DC chemokine and migration inhibitor and provide novel insight into the immunobiology of SFA.

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“…SfA ( 40 ) binds in the active site of cyclophilin A, but immunosuppressive activity is not caused by calcineurin binding (Table ). Additionally, there is no inhibition of mTOR or of the purine/pyrimidine de novo synthesis . The mechanism of immunosuppression seems to be connected to dendritic cell (DC) chemokine and migration inhibition but remains to be fully elucidated. , Degradation studies on SfA ( 40 ) revealed that removal of the C24 polyketide side chain (corresponding to R 1 in Table ) eliminated the immunosuppressive properties of the molecule while maintaining the nanomolar binding affinity to hCypA, -B, and -C. ,, One of these nonimmunosuppressive sanglifehrins, AAE931 ( 41 , Table ), showed potent anti-HCV activity in the replicon assay (IC 50 = 130 nM) …”

“…123 SfA (40) binds in the active site of cyclophilin A, 137 but immunosuppressive activity is not caused by calcineurin binding (Table 6). 138 Additionally, there is no inhibition of mTOR or of the purine/pyrimidine de novo synthesis. 139 The mechanism of immunosuppression seems to be connected to dendritic cell (DC) chemokine and migration inhibition but remains to be fully elucidated.…”

“…139 The mechanism of immunosuppression seems to be connected to dendritic cell (DC) chemokine and migration inhibition but remains to be fully elucidated. 123,138 Degradation studies on SfA (40) revealed that removal of the C24 polyketide side chain (corresponding to R 1 in Table 6) eliminated the immunosuppressive properties of the molecule while maintaining the nanomolar binding affinity to hCypA, -B, and -C. 128,136,140 One of these nonimmunosuppressive sanglifehrins, AAE931 (41, Table 6), showed potent anti-HCV activity in the replicon assay (IC 50 = 130 nM). 140 Investigation of the X-ray crystal structure of a nonimmunosuppressive analogue of AAE931 ( 41) complexed with hCypA revealed that the unique piperazic acid functionality occupies the same hydrophobic pocket as the Val11 of CsA (5), while some additional important hydrophobic interactions are maintained between hCypA and the valine group of the sanglifehrin analogue.…”

Insights into peptidyl-prolyl cis–trans isomerase structure and function in immunocytes

“…Immecke et al recently claimed that SfA-mediated suppression of chemokine and CD38 receptor expression in monocyte-derived dendritic cells (moDC) is unique and independent of CyPA [129]. However, they found more of a reduction of chemokines CCL5, CCL17, and CCL19 when moDC were treated with a combination of SfA and CsA, than when treated with SfA or CsA alone.…”

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“…Combined pharmacological approaches are also a feasible possibility. In this regard, treatments with CsA, a cycophilin‐interacting drug, associated to other natural compounds such as sanglifehrin efficiently suppresses chemokine signalling and cell migration . It has been proposed that the use of rapamycin along with metrothrexate and tacrolimus in patients with lymphoma is associated with a significantly decreased risk of disease progression, although a recent clinical trial performed in children with acute lymphatic leukemia also showed increased toxicity and disapproved the therapeutic combination of these drugs.…”