IFN-γ levels induced by <i>P</i>. <i>papatasi</i> salivary glands extracts stimulation in the presence of an anti-IL-10 antibody, in individuals living in old or emerging foci of <i>L</i>. <i>major</i> infection.

<p>IFN-γ was quantified by ELISA, in the culture supernatants of PBMC stimulated with <i>P</i>. <i>papatasi</i> salivary gland extracts (1 gland/ml), with or without an anti-IL10 antibody (500ng/ml), during 72h. Analysis of IFN-γ production was performed in individuals from OF and EF (a) and LST+/Scar+ (healed), LST+/Scar- (asymptomatics) and LST-/Scar- (naïve) groups in OF (b) and EF (c). Statistical significance was assigned to a value of p<0.05. Statistically significant differences between stimulated and non-stimulated cultures and between groups are showed. Horizontal lines represent median IFN-γ values.</p

Clinical and immunological parameters in individuals living in various emerging foci of <i>L</i>. <i>major</i> infection.

<p>Clinical and immunological parameters in individuals living in various emerging foci of <i>L</i>. <i>major</i> infection.</p

IgG response against <i>P</i>. <i>papatasi</i> salivary glands extracts in individuals living in old or emerging foci of <i>L</i>. <i>major</i> infection.

<p>Analysis of IgG response was performed in individuals from OF and EF (a) and LST+/Scar+ (healed), LST+/Scar- (asymptomatics) and LST-/Scar- (naïve) groups (b), in each focus. IgG antibodies were measured by ELISA. Results were expressed as relative OD (ROD) defined as the ratio of sample OD/mean OD of sera from negative controls. ROD superior or equal to 2 was considered positive. Negative sera were obtained from healthy controls living outside Tunisia in sand fly-free regions. Statistical significance was assigned to a value of p<0,05. Statistically significant differences between groups are showed for positive responders. Horizontal lines represent median ROD values and dotted lines represent cut-off level. The percentages of positive responders are presented for groups and foci.</p

Study design.

<p>Study design.</p

Proliferative responses to <i>P</i>. <i>papatasi</i> salivary glands extracts in individuals living in old or emerging foci of <i>L</i>. <i>major</i> infection.

<p>PBMC were stimulated for 5 days with <i>P</i>. <i>papatasi</i> salivary gland extracts (1gland/ml). Proliferation was analyzed in individuals from donors residing in an old (OF) or an emerging focus (EF) (a), LST+/Scar+ (healed), LST+/Scar- (asymptomatics) and LST-/Scar- (naïve) groups (b) and LST+, LST- groups (c), in each focus. Results were expressed as stimulation index (SI) obtained by dividing the mean counts of triplicates in antigen-stimulated cultures by the mean counts of triplicates in non-stimulated cultures. The proliferative response was considered positive when SI was superior or equal to 2. Statistical significance was assigned to a value of p<0,05. Horizontal lines represent median SI values and dotted lines represent cut-off level. The percentages of positive responders are presented for groups and foci.</p

Timeline showing the different steps of the prospective study.

<p>790 participants living in endemic areas of ZCL were followed up over 1 year throughout one season of <i>L</i>. <i>major</i> transmission. Parameters such as LST and the presence of typical scars were monitored at the beginning of the study and after the transmission season and the triggering of new cases. Peripheral blood samples were obtained from each donor at the beginning of the study.</p

Proliferative responses to <i>P</i>. <i>papatasi</i> salivary glands extracts in the presence of an anti-IL-10 antibody, in individuals living in old or emerging foci of <i>L</i>. <i>major</i> infection.

<p>PBMC were stimulated with <i>P</i>. <i>papatasi</i> salivary glands extracts (1gland/ml) with or without an anti-IL10 antibody (500ng/ml), for 5 days. Proliferation was analyzed in individuals from donors residing in an old (OF) or an emerging focus (EF) (a) and LST+/Scar+ (healed), LST+/Scar- (asymptomatics) and LST-/Scar- (naïve) groups (b), in each focus. Results were expressed as percentage of positive proliferative responders. The proliferative response was considered positive when SI was superior or equal to 2. Statistical significance was assigned to a value of p<0,05. Statistically significant differences between groups are showed.</p

Clinical and immunological parameters in individuals living in old (OF) or emerging foci (EF) of <i>L</i>. <i>major</i> infection.

<p>Clinical and immunological parameters in individuals living in old (OF) or emerging foci (EF) of <i>L</i>. <i>major</i> infection.</p

IFN-γ levels induced by <i>P</i>. <i>papatasi</i> salivary glands extracts stimulation in individuals living in old or emerging foci of <i>L</i>. <i>major</i> infection.

<p>IFN-γ was quantified by ELISA, in the culture supernatants of PBMC stimulated with <i>P</i>. <i>papatasi</i> salivary gland extracts (1 gland/ml) during 72h. Analysis of IFN-γ production was performed in individuals from OF and EF (a), LST+/Scar+ (healed), LST+/Scar- (asymptomatics) and LST-/Scar- (naïve) groups (b) and LST+, LST- groups (c), in each focus. Statistical significance was assigned to a value of p<0,05. Statistically significant differences between stimulated and non-stimulated cultures and between groups are showed. Horizontal lines represent median IFN-γ values.</p

Sequences of the Tunisian <i>Phlebotomus papatasi</i> 30 kDa salivary protein.

<p>(<b>A</b>) mRNA sequence of PpSP30. (<b>B</b>) Protein sequence of PpSP30. Differences in the nucleotide and protein sequence PpSP30 described in the GenBank (GenBank AF335489.1) are highlighted in pink. Non coding sequence is highlighted in purple.</p