Serum Markers Associated with Disease Severity in a Bosnian Hemorrhagic Fever with Renal Syndrome Cohort

Abstract

Puumala orthohantavirus (PUUV) is endemic in Europe and can cause hemorrhagic fever with renal syndrome (nephropathia epidemica). Disease features include fever, thrombocytopenia, and acute kidney injury (AKI). This retrospective cohort study of forty PUUV patients aims to characterize associations of serum immunological, hemostatic or kidney injury markers to disease severity. While interleukin-18 (IL-18) was significantly increased in severely thrombocytopenic patients (<100 × 109 platelets/L) compared to patients with higher platelet counts, RANTES was significantly decreased in these patients. These data suggest that patients with significant thrombocytopenia might have experienced pronounced Th1 immune responses. When kidney dysfunction was used as the primary disease outcome, recently identified AKI biomarkers (Cystatin C, insulin-like growth factor-binding protein 7, Nephrin, and trefoil factor 3) were significantly upregulated in patients with severe PUUV infection, defined as the estimated glomerular filtration rate (eGFR) below 30 m/min/1.73 m2. The increased expression of these markers specifically indicates pathology in glomeruli and proximal tubuli. Furthermore, E-selectin was significantly higher while interferon gamma-induced protein 10 (IP-10) was significantly lower in PUUV patients with more severe kidney dysfunction compared to patients with higher eGFR-values. Increased E-selectin illustrates the central role of endothelial cell activation, whereas decreased IP-10 could indicate a less important role of this cytokine in the pathogenesis of kidney dysfunction.

A pan-orthohantavirus preclinical human lung xenograft mouse model

“…Anti-ANDV mouse monoclonal antibody KL-AN-5E8 7 and anti-influenza virus human monoclonal antibody CR9114 43 , and murine serum samples from pre-infection, 3, 5 and 10 dpi were tested to determine their capacities to neutralize ANDV infection in vitro. The protocol was adapted from an existing orthohantavirus microneutralization assay for heatinactivated human patient sera 56 . In short, antibodies were ten-fold diluted to a final range from 100 μg/ml to 1.0 x 10 -4 μg/ml.…”

“…Plates were scanned on an Amersham Tyhpoon Biomolecular Imager (channel Cy2, GE Healthcare, Chicago, United States). Neutralizing titers were determined by scoring in which wells infection was inhibited ≥ 95% (MNT95) as described previously 56,57 .…”