During visceral leishmaniasis (VL), T helper 1 (Th1)-based inflammation is induced to control intracellular parasites. Inflammation-based pathology has been shown to be dampened by interleukin 10 and eventual Programmed Death1 (PD1)-mediated T cell exhaustion. Cell type(s) responsible for the initiation of T cell-produced IL-10 during VL are unknown. CD19+, CD5−, CD1d−, IgDhi regulatory B cells from healthy controls produced IL-10 in absence of infection or stimulation in contrast to IgDlo/neg B cells. IgDhi B cells may have a de novo vs. induced regulatory program. IgDhi B cells increased three-fold in population size as VL progressed. B cells from VL dogs were necessary and sufficient to suppress T helper 1 (Th1) cell effector function. IgDhi B cells induced T cell and IgDlo B cell IL-10 production. Blockage of B cell-specific PD-L1 restored Th1 responses. IgDhi regulatory B cells represent a novel regulatory B cell which may precipitate T cell exhaustion during VL.

Immunopathological studies have contributed to the characterization of in situ inflammatory infiltrates in cutaneous leishmaniasis (CL). However, little is known about the T-cell antigen reactivity of these lesions. Our objective was to analyze the responsiveness of lymphocytes from CL lesions to leishmanial and nonrelated antigens in terms of proliferation and the production of cytokines. Mononuclear cells were extracted from lesions, and blood from CL patients infected with Leishmania (Viannia) braziliensis. Activated cells accounted for 35-45% of lesions T-cell subsets. Elevated levels of C1.7/CD244(+)CD8(+) T cells suggest in situ cytotoxic effector function. Lymphocytes isolated from the leishmaniasis lesions proliferated and produced IFN-gamma in response to leishmanial antigens as well as to irrelevant antigens such as Toxoplasma gondii (Tg). Patients presenting with larger lesions had the highest lymphocyte proliferation indexes. A high frequency of Tg-specific cells was detected in the lesions by limiting dilution assay, similar to the frequency of Leishmania-specific cells. Importantly, Tg-reactive cells were not found in lesions of patients without a history of toxoplasmosis. The proportion of Leishmania-reactive CD4(+) and CD8(+) T cells in the lesions was quite variable. Overall, these data suggest that T cells reactive to nonrelevant antigens can migrate to leishmanial lesions and possibly influence the pathogenesis of the disease.

Leishmania (Viannia) braziliensis control and tissue damage relate to the effector immune response, which in turn affects clinical outcome. Leishmania reactive CD4+ and CD8+ T cells are expanded in long-term healed cutaneous leishmaniasis (hCL) patients but their functional characteristics remain to be determined. This study investigates antigen-specific recall in long-term healed CL caused by L. braziliensis infection. Healed CL subjects were grouped according to the time elapsed since the end of therapy: less than two years and two to five years. Activation phenotype (CD69+ or CD25+) and subpopulations of memory T cell phenotypes [central memory (Tcm): CD45RO+ CCR7+ or effector memory (Tem): CD45RO+ CCR7-] were quantified in ex vivo blood mononuclear cells and after Leishmania antigens stimuli. A reduction in the percentage of activated Leishmania-responder CD4+ and CD8+ T cells in hCL was associated with the time elapsed since clinical cure. Percentage of CD69+ in TCD4+ and TCD8+ cells were negatively correlated with IL-10 levels. Ex vivo analyses showed contracted Tem CD4+ and Tem CD8+ compartments from hCL with long time elapsed since clinical cure, although renewal of these compartments was observed following in vitro exposure to leishmanial stimuli. Our results show that healed L. braziliensis infected patients exhibit a recall response to Leishmania antigens with evident expansion of effector memory T cells. Regulated leishmanial-specific response seems to emerge only about two years after initial contact with the parasite antigens.

Parasitic diseases are a human health problem mainly in low-income areas. The drugs available for the treatment of these diseases are far from satisfactory due to high costs, toxicity, and drug resistance. Sulfated polysaccharides are a complex group of bioactive polymers and can be obtained from seaweeds. The heterofucans from Sargassum filipendula (SF) present strong antiproliferative and antioxidant activities. However, their immunomodulatory and antimicrobial capacity have not been evaluated until now. In this study, five sulfated fucose-rich fractions were isolated (named SF0.5V, SF0.7V, SF1.0V, SF1.5V, and SF2.0V). The chemical composition showed slight differences among polysaccharides and, consequently, biological activity of these polymers. Three fractions (SF0.5V, SF0.7V, and SF1.0V) showed a strong immunomodulatory activity enhancing the release of nitric oxide (NO) by murine macrophages (RAW 264.7), though only SF0.5V was able to induce interleukin-6 (IL-6) and TNF-α release from RAW cells. The sugar to sulfate ratio was not correlated with these activities. Meanwhile, the contents of xylose (P = 0.98 for NO; P = 0.98 for IL-6; P = 0.96 for TNF-α) and glucuronic acid (P = 0.91 for NO; P = 0.9190 for IL-6; P = 0.79 for TNF-α) were strongly positively correlated. SF0.7V and SF1.0V inhibited biofilm formation by Klebsiella pneumoniae (4.2 and 6.8%, respectively), whereas SF0.5V