Absolute numbers of LDGs, CD14⁺ cells and white blood cells.

<p>The white blood cell counts were determined after Ficoll using trypan blue exclusion and are expressed as cell number per ml of blood.</p

L-arginine levels and CD3ζ expression in CD4⁺ and CD8⁺ T cells.

<p>Plasma was isolated by centrifugation of the blood of controls (n = 11), VL patients (n = 11) and treated VL patients (n = 11) and (A) the levels of L-arginine were measured by HPLC. PBMCs were isolated by density gradient from the blood of controls (n = 10), VL patients (n = 10) and treated VL patients (n = 10) and the mean fluorescence intensity of CD3ζ was determined in CD4⁺ T cells (B) and CD8⁺ T cells (C). Box = interquartile range and median; whiskers = range. Statistical significance was determined by a two-tailed Mann-Whitney test. NS = not significant. Patients = VL patients before treatment; treated patients = VL patients after 3–4 weeks treatment.</p

Phenotype and frequency of arginase-expressing cells in PBMCs.

<p>PBMCs were isolated by density gradient from the blood of controls (n = 14), VL patients (n = 17) and treated VL patients (n = 12). The phenotype of arginase-expressing cells was determined by flow cytometry. (A) Dot plot profile of CD15⁺ arginase⁺ cells, data show the results of one representative experiment out of 17 independent experiments; (B) dot plot profile of CD14⁺ arginase⁺ cells, data show the results of one representative experiment out of 17 independent experiments. (C) Percentages of CD15⁺arginase⁺ cells in the PBMCs of controls (n = 14), VL patients (n = 17) and treated VL patients (n = 12). Box  =  interquartile range and median; whiskers = range. Statistical significance was determined by a two-tailed Mann-Whitney test. NS = not significant. Patients = VL patients before treatment; treated patients = VL patients after 3–4 weeks treatment.</p

Arginase activity in PBMCs.

<p>PBMCs were isolated by density gradient from the blood of controls (n = 10), VL patients (n = 14) and treated VL patients (n = 12) and the activity of arginase was measured by enzymatic assay. Box = interquartile range and median; whiskers = range. Statistical significance was determined by a two-tailed Mann-Whitney test. NS = not significant. Patients = VL patients before treatment; treated patients = VL patients after 3–4 weeks treatment.</p

Frequency of LDGs and monocytes in PBMCs.

<p>PBMCs were isolated by density gradient from the blood of controls (n = 14), VL patients (n = 17) and treated VL patients (n = 12), (A) the frequencies of monocytes were determined by flow cytometry and (B) the ratio of low-density granulocytes (LDGs) to monocytes was calculated by dividing the percentages of LDGs by the percentages of monocytes. Box = interquartile range and median; whiskers = range. Statistical significance was determined by a two-tailed Mann-Whitney test. NS = not significant. Patients = VL patients before treatment; treated patients = VL patients after 3–4 weeks treatment.</p

Haematological data.

<p>Hct = hematocrit; Hb = haemoglobin.</p><p>Normal range: platelets (×10³) = 150–450; white blood cells (×10³) = 4.5–10.5; Hct (%) = 35–60; hb (g/dl) = 11–18.</p><p>The white blood cells were counted in total blood before Ficoll using a COULTER Ac•T diff Hematology Analyzer and are expressed as number of WBC (×10³) per µl of blood.</p

Clinical data.

<p>Spleen and liver size = measurement below left costal margin and right subcostal margin (respectively).</p