Targeting the tissue factor coagulation initiation complex prevents antiphospholipid antibody development
Antiphospholipid antibodies (aPL) in primary or secondary antiphospholipid syndrome (APS) areis a major cause for acquired thrombophilia. Here we test the effect of specific inhibition of the TF coagulation initiation complex with nematode anticoagulant protein c2 (NAPc2) on the activation of monocytes by aPL. NGS data show that aPL-induced proinflammatory and prothrombotic activation of monocytes, but not interferon regulated gene induction is prevent by NAPc2. Overall design: Monocytic MM1 cells were stimulated with 400 ng/ml aPL HL5B without (n=4) and with 200 nM NAPc2 (n=3) or HL7G without (n=4) or with (n=4) NAPc2, 10 ng/ml LPS (n=3), control IgG (n=4), no addition (n=4), NAPc2 (n=4). RNA isolated from MM1 cells was sequenced at the MDC Berlin and NGS data were mapped to GRCh38.p13 with STAR on the fly.  NGS sequencing of splenic DCs used spleens digested with collagenase II and DNase I followed by dissociated cell enrichment on an Optiprep density gradient and FACS of live DCs identified by MHCII and CD11c in the CD3/CD19/NK1.1 negative population. DC RNA of 4 saline and 4 NAPc2 treated mice was sequenced by Novogene and mapped to the GRCm39 mouse genome with STAR on the fly.