, #FCSTM18-22). For the Human Magnetic Luminex Assay (CD163, ST2, and CD14, LBP) (R&D Systems, LXSAHM-02), plates were prepared according to the manufacturer's recommendations and read on a Bio-Plex, Quantification of serum analytes. Twenty-nine parameters were analyzed in serum samples, by magnetic bead assays or ELISA: Human XL Cyt Disc Premixed Mag Luminex Perf Assay Kit (CCL2/MCP-1, CCL4/MIP-1?, RANTES/CCL5, CCL11/Eotaxin, CCL19/MIP-3?, CCL20/MIP-3?, CD40Ligand, Fractalkine/ CX3CL1, CXCL10/IP-10, EGF, Flt-3 Ligand, granzyme B, IL-1RA

, Multiparametric flow cytometry panel was performed using a battery of antibodies: anti-CD38 FITC #340909 (dilution: 1/10), Cell phenotyping. Immune phenotyping was performed with an LSRII Fortessa 4-laser (488, 640, 561, and 405 nm) flow cytometer (BD Biosciences), and Diva software version 6.2. FlowJo software version 9, vol.9, pp.8-15

, #560179 (dilution: 1/20), anti-CD3 Alexa 700 #557943 (dilution: 1/100), p.7

, CD21 PE #555422 (dilution: 1/10), CD27 APC #337169 (dilution: 1/100), CD45 Alexa 700 #560566 (dilution: 1/100), anti-CD56 PECF594 #564849 (dilution: 1/500), anti-HLA-DR BV605 #562845 (dilution: 1/50), anti-CD33 BV421 #562854 (dilution: 1/20), anti-CD141 BV711 #563155 (dilution: 1/20), anti-CD45RA PerCpCy5.5 #563429 (dilution: 1/20), anti-HLA-ABC BV786 #740982 (dilution: 1/50), anti-CD86 PECF594 #562390 (dilution: 1/20) (all from BD Biosciences), vol.647, p.38

, anti-IgM Pacific Blue #314514 (dilution: 1/ 100), anti-CD71 BV650 #334116 (dilution: 1/20), anti-CD20 APC Cy7 #302314 (dilution: 1/20), anti-CD16 APC Cy7 #302018

, All antibodies were commercially available. See the corresponding manufacturer datasheets on webpages for reference and validation CD4 + and CD8 + T cells were analyzed for CD45RA and CCR7 expression, to identify the naive, memory, and effector cell subsets, and for co-expression of the activation markers HLA-DR and CD38. CD19 + B cell subsets were analyzed for the CD21 and CD27 markers. Antibody-secreting cells (plasmablasts) were identified as CD19 + cells expressing CD38 and CD27. We used CD16 and CD56 to identify NK cell subsets, For intracellular cytokine staining (ICS) analyses, cells were stained with surface monoclonal antibodies: anti-CD4 PE PECF594 #562281 (dilution: 1/33), anti-IFN? FITC #557718 (dilution: 1/20), anti-TNF? PE-Cy7 #557647 (dilution: 1/20), anti-MIP-1? PE #550078 (dilution: 1/200), and anti-IL-2 BV421 #564164 (dilution: 1/20) (all from BD Biosciences), vol.46

, Libraries were prepared with the TruSeq® Stranded mRNA Kit, according to the Illumina protocol. Libraries were sequenced on an Illumina HiSeq 2500 V4 system. The single read sequencing depth was was about 50 million reads, with a fragment length of 101 bp. Sequencing quality control was performed with Sequence Analysis Viewer. FastQ files were generated from.bcl files on Illumina BaseSpace sequence hub. After trimming (QPhred score ?25), reads were aligned to the hg19 human reference genome, using STAR -2.5.3ar, and quantified relative to annotation model hg19 -GENCODE Genes -release 19, Characterization of EBOV-specific immune responses. Cellular responses to EBOV peptides were assessed with EpiMax technology 47 . Briefly, PBMCs were stimulated in vitro with 158 overlapping 15-mer peptides (11-amino acid overlaps), covering the Ebola virus Mayinga variant GP, in two pools of 77 (EBOV1) and 81 peptides (EBOV2) (JPT Technologies). Cell functionality was assessed by ICS, with Boolean gating (Fig. 5b, Supplementary Appendix), vol.22, 2019.

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