Fig 2: Increased HPSE concentration in HTNV–infected podocytes. Podocytes were infected with live HTNV (MOI 10 or 1), UV–inactivated HTNV (MOI 10) or remained uninfected (mock). Supernatants were collected and cells either fixed for immunofluorescence analysis or subjected to RNA extraction at 2 and 4 dpi. (A) Fixed and permeabilized podocytes were stained with rabbit serum against viral nucleocapsid N protein followed by AF488–conjugated secondary antibody (green) or Hoechst 33,420 to detect nuclei (blue). (B) HPSE levels measured from podocyte supernatants. (C) Isolated RNA was subjected to multiplex RT–qPCR with primers and probes detecting HPSE and GAPDH mRNA. The signal for HPSE mRNA was normalized based on GAPDH mRNA levels and fold change calculated in reference to mock–infected podocytes. Statistically significant differences were assessed with Tukey’s multiple comparisons test. ****, *** and * indicate p < 0.0001, p < 0.001 and p < 0.05, respectively.

Fig 3: Heparanase and syndecan–1 levels during PUUV–HFRS. (A) Urinary syndecan–1:creatinine ratio (B), HPSE:creatinine ratio and (C) albumin:creatinine ratio. (D) HPSE concentration in plasma. Concentrations were measured from sequential samples of 49 (55 for plasma) patients hospitalized due to PUUV–HFRS (with a total of 159 urine and 262 plasma samples). Days after onset of fever (aof) 3–9 represent the acute stage, 20–30 the convalescent and 182–365 the controls. Differences between individual time points and the last time point (365 aof), which was considered to represent full recovery and used as controls, were assessed by GEE and significant differences indicated as *** <0.001, ** <0.01 and * < 0.05. Red lines represent mean ± standard deviation.

Fig 4: Spearman’s rank correlation coefficient matrix. The colors of the round circles indicate the level of the correlation coefficient and increasing size with a lower p–value. Statistical significance is specified as * = p < 0.05, **= p < 0.01, ***= p < 0.001, ****= p < 0.0001. AOF = after onset of fever, LOS = length of stay, eGFR = estimated glomerular filtration rate, P = plasma, U = urine, crea = creatinine, alb = albumin, HPSE = heparanase, Synd–1 = syndecan–1, IL = interleukin, IP–10 = interferon gamma–induced protein 10, MCP–1 = monocyte chemoattractant protein 1, min MAP = minimum mean arterial pressure, WBC = white blood cells, MPO = plasma myeloperoxidase, CRP = plasma C–reactive protein, tromb = blood thrombocytes.

Fig 5: HPSE2 protein and peptides treatment have beneficial effects in LPS-induced glomerulonephritis (A). BUN, (B). plasma creatinine, and (C). Urinary albumin/creatinine ratio of mice injected with LPS (2 days) and treated with different concentrations of HPSE2 protein or HPSE2 peptide mix (D). Cortical HPSE1, (E). cortical TNF-a, (F). cortical IL-6 mRNA expression of mice injected with LPS (2 days) and treated with different concentrations of HPSE2 protein or HPSE2 peptide mix of 381–410, 391–420, and 401–430. Data are expressed as mean ± SEM. n = 4 *p < 0.05, **p < 0.01, ***p < 0.001. HPSE2, heparanase-2; HPSE1, heparanase-1; Prt, His-mHPSE2 protein; pep, HPSE2 peptide mix of 381–410, 391–420, and 401–430; BUN, Blood urea nitrogen plasma level.