Balance of IL-10 and interferon-gamma plasma levels in human visceral leishmaniasis: implications in the pathogenesis

Abstract

Background: Leishmaniasis remains a serious public health problem in several parts of the developing world. Effective prophylactic measurements are hampered by imprecise comprehension of different aspects of the disease, including its immunoregulation. A better comprehension of immunoregulation in human VL may be useful both for designing and evaluating immunoprophylaxis.

Methods: To explore immunoregulatory mechanisms, 20 visceral leishmaniasis (VL) patients were evaluated during active disease and at different periods up to one year after treatment determining their plasma cytokine levels, clinical parameters (palpable spleen and liver) and antibody levels.

Results: Elevated plasma levels of IFN-gamma and of IL-12 p40 were observed during active disease, significantly decreasing after treatment whereas in vitro Leishmania antigen-stimulated IFN-gamma production by PBMC exhibited an inverse pattern being low during disease and increasing steadily thereafter. Absence of IFN-gamma activity is a hallmark of VL. The main candidate for blunting IFN-gamma activity is IL-10, a cytokine highly elevated in plasma with sharp decrease after treatment. Activity of IL-10 is inferred by high levels of anti-Leishmania specific IgG1 and IgG3. TGF-beta had elevated total, but not of active, levels lessening the likelihood of being the IFN-gamma counterpart. Spleen or liver size presented a steady decrease but return to normal values at only 120 days after treatment. Anti-Leishmania IgG (total and subclasses) levels and DTH or Leishmania-stimulated lymphocyte proliferation conversion to positive also present a slow decrease after treatment. IL-6 plasma levels were elevated in only a few patients.

Conclusion: Taken together our results suggest that IFN-gamma and IL-10 are the molecules most likely involved in determining fate of disease. After treatment, there is a long delay before the immune profile returns to normal what precludes using plasma cytokine levels as criteria of cure as simpler clinical evaluations, as a palpable spleen or liver, can be used.

Figure 1

Clinical and immunological parameters from acute VL at different periods after treatment. Spleen (A) liver (B) size and plasma IgG levels and their subclasses (C) from VL patients during acute VL (day 0) and at different periods after treatment (30, 120 and 210 days) are shown. Stimulation index obtained in proliferation assay from PBMC from VL stimulated with Leishmania antigen (10 μg/ml) are represented in D.

Figure 2

IFN-γ and IL-10 levels in plasma and supernatant from PBMC from active VL and at different periods after treatment. Plasma IFN-γ (A) and IL-10 (C) levels obtained by ELISA from VL patients during acute VL (day 0) and at different periods after treatment (30, 120 and 210 days) are shown. IFN-γ (B) and IL-10 (D) production obtained from PBMC from VL patients during acute VL (day 0) and at different periods after treatment (30, 120 and 210 days) restimulated in vitro for 72 hours with SLA (10 μg/ml) are shown.

Figure 3

Plasma cytokine levels from active VL and at different periods after treatment. Plasma IL-12 p40 (A), total (open symbols) and active (closed symbols) TGF-β (B) and IL-6 (C) levels obtained by ELISA from VL patients during acute VL (day 0) and at different periods after treatment (30, 120 and 210 days) are shown in A, B, C, respectively.