Journal: Scientific Reports
Article Title: Growth arrested live-attenuated Leishmania infantum KHARON1 null mutants display cytokinesis defect and protective immunity in mice
Figure Lengend Snippet: Humoral and cellular immune responses induced by different schemes of vaccination with Δ Likh1 . Humoral immune response was evaluated by conventional ELISA measurement of serum levels of different isotypes of Li WT SLA-specific antibodies (IgG total , IgG 1 and IgG 2a ). ( a ) Seroconversion of mice 20 days post-immunization with Δ Likh1 . ( b ) IgG2a:IgG1 ratio between IV and SC prime-boosted groups. ( c – h ) Levels of cytokines in pg/mL in the supernatant of Li WT SLA-stimulated splenocytes. The concentration of Li WT SLA in the experiments was 25 μg/mL and splenocytes were stimulated for 72 h. CpG ODN at 5 μg/mL was used as a positive control of a Th1-response and α-MEM medium with 20% of FBS was used as negative control. Cytokines were measured by flow cytometry with the BD Cytometric Bead Array (CBA) Mouse Th1/Th2/Th17 Cytokine Kit (BD Biosciences, San Jose, CA, USA) following manufacturer instructions. The acquisition and data analyses were made in BD FACSVerse Instrument and FCAP Array Software v3. The detection limit of IL-2, IL-4, IL-6, IFN-γ, TNF, IL-17A and IL-10 cytokines were respectively 0.1; 0.03; 1.4; 0.5; 0.9; 0.8 and 16.8 pg/mL as well as cytokines secreted by Li WT SLA-stimulated splenocytes from immunized mice. Data are shown as means ± SE. Statistical analyses used One-way-ANOVA followed by Bonferroni’s multiple comparison test. *** p
Article Snippet: Promastigote forms of L. infantum (MHOM/MA/67/ITMAP-263) were cultured in α-MEM medium (Gibco/Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum, 5 μg/mL of hemin and 5 μM of biopterin, in pH 7 at 26 °C.
Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay, Concentration Assay, Positive Control, Negative Control, Flow Cytometry, Cytometry, Crocin Bleaching Assay, FCAP Assay, Software