Fig 1: Immunoassay-based validation of differentially expressed proteins.Immunoturbidimetric measurement of serum haptoglobin (A) and Apo A-1 (B) levels in healthy subjects (n = 20), falciparum malaria (n = 20), vivax malaria (n = 17) and febrile controls (n = 6). Histograms depicting the haptoglobin and Apo A-1 concentrations determined by immunoturbidimetry. Patients with malaria infection found to have lower serum level of haptoglobin and Apo A-1 compared to the HC and FC (p<0.0001; Mann-Whitney test). (C) Retinol-binding protein 4 (RBP4) level was measured in sera derived from malaria patients (n = 24), healthy subjects (n = 20) and febrile controls (n = 6) by ELISA and results are shown as bar-diagrams (mean ± SE), which indicates down-regulation of RBP4 in FM and VM patients (p<0.01) compared to HC, while serum level of RBP4 found to be unaltered in FC (leptospirosis patients) (D &E) Western blot analysis of haptoglobin (HP) serum amyloid A (SAA), clusterin (CLU) and retinol-binding protein (RBP) from serum samples of HC (n = 12), FM (n = 12),VM (n = 12) and leptospirosis patients (n = 6). Representative blots of the target proteins (D) are depicted along with their respective relative intensities (X 104) (E). Western blot analysis revealed up-regulation of serum amyloid A and down-regulation of haptoglobin, clusterin and retinol-binding protein in FM and VM patients (p<0.01) compared to the controls (HC and FC). All data are represented as mean ± SE.
Fig 2: Validation cohort using CD4+ T-cell count gain as a marker of immune recovery. (A) Flow chart illustrating cases (n = 101) categorized according to CD4+ T-cell count gain after 144 weeks of ART initiation instead of 250 cell/µL CD4+ T-cell count cut-off: A group for those who gained = 100 cells/µL, and B group for those who gained < 100 cells/µL. (B) Circulating ZAG, RBP4, and APLNR concentrations at baseline (B,C) at 144 weeks of ART. p-value indicates significant differences among indicated groups by nonparametric Mann–Whitney test.
Fig 3: Difference in RBP4 levels after 6-weeks of chemotherapy. Data were analyzed using a Wilcoxon test. RBP4, retinol-binding protein 4.
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