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

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

Methods of probing the interactions between small molecules and disordered proteins

It is generally recognized that a large fraction of the human proteome is made up of proteins that remain disordered in their native states. Despite the fact that such proteins play key biological roles and are involved in many major human diseases, they still represent challenging targets for drug discovery. A major bottleneck for the identification of compounds capable of interacting with these proteins and modulating their disease-promoting behaviour is the development of effective techniques to probe such interactions. The difficulties in carrying out binding measurements have resulted in a poor understanding of the mechanisms underlying these interactions. In order to facilitate further methodological advances, here we review the most commonly used techniques to probe three types of interactions involving small molecules: (1) those that disrupt functional interactions between disordered proteins; (2) those that inhibit the aberrant aggregation of disordered proteins, and (3) those that lead to binding disordered proteins in their monomeric states. In discussing these techniques, we also point out directions for future developments.Gabriella T. Heller is supported by the Gates Cambridge Trust Scholarship. Francesco A. Aprile is supported by a Senior Research Fellowship award from the Alzheimer’s Society, UK (grant number 317, AS-SF-16-003)

Planobispora siamensis sp nov., isolated from soil

A novel actinomycete strain, A-T 4600T, which developed cylindrical sporangia containing a longitudinal pair of motile spores forming singly or in bundles on short ramifications of the aerial mycelium, was isolated from soil collected from an evergreen forest in Thailand. The cell-wall peptidoglycan containedmeso-diaminopimelic acid. The whole-cell sugars contained ribose, madurose, mannose and glucose. The predominant menaquinones were MK-9(H2). Mycolic acids were not detected. The diagnostic phospholipids were phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylinositol, phosphatidylglycerol, phosphatidylinositol mannoside, diphosphatidylglycerol and aminophosphoglycolipid. The predominant cellular fatty acids were unsaturated C17 : 1, C18 : 1, saturated C16 : 0, and C17 : 0. The G+C content of the DNA was 70.8 mol%. Phenotypic and chemotaxonomic analyses showed that the isolate had the typical characteristics of members of the genusPlanobispora. Furthermore, 16S rRNA gene sequence analysis also indicated that this strain belonged to the genusPlanobisporabut as a putative novel species. DNA–DNA relatedness values that differentiate the isolate from previously described members of the genusPlanobisporawere significantly below 70 %. Following an evaluation of phenotypic, chemotaxonomic and genotypic studies, it is proposed that the isolate represents a novel species,Planobisporasiamensissp. nov.; the type strain is A-T 4600T( = BCC 39469T = NBRC 107568T).</jats:p