Figure 1.
Alignment of amino acids sequences of the SmTAL family.
Helix-loop-helix EF-hand domains (Pfam00036) are indicated. The canonical aspartic acid residues at the start of the loop are denoted D on black background. Residues in the dynein light chain domain (EMBL/EBI IPR001372) are shaded grey. Alignment was performed using Clustal W.
Figure 2.
Transcription profiles of SmTALs.
Profiles from the S. mansoni lifecycle microarray data available via Array express (31) under the experimental accession number E-MEXP-2094. Values are mean normalized fluorescence units ± sem. In primate infections, larvae remain in the skin for 2–5 days (Wilson et al. 1990). In the figure cercariae to 3 d schistosomula are denoted “skin stage”.
Figure 3.
Electrophoresis of purified SmTAL proteins.
2 µg of each of the indicated proteins was run under reducing conditions on a 4–12% gradient SDS-PAGE gel and stained with Coomassie blue.
Figure 4.
Antibody responses to SmTAL1, SmTAL3 and SmTAL5 in the S. mansoni infected cohort.
Recombinant SmTAL1, 3 and 5 were used in ELISA to measure antigen-specific IgE (A) or IgG4 (B) before and after praziquantel treatment in 200 males infected with S. mansoni. Only individuals whose levels exceeded the seropositive threshold (mean+3xSD uninfected controls) for each response are graphed. For the whole cohort, the prevalence of each response (C) is shown in 5 age groups, 7–9 (n = 36), 10–14 (n = 43), 15–24 (n = 35), 25–34 (n = 43) and 35–60 (n = 43) Shown is % seropositive for each group after treatment +95% confidence intervals.
Figure 5.
Antibody responses to SmTAL2, SmTAL4 and SmTAL13 in the S. mansoni infected cohort.
Recombinant antigens were used in ELISA to measure IgE (A) or IgG4 (B) before and after treatment in infected cohort. Only individuals whose levels exceeded the seropositive threshold (mean+3xSD uninfected controls) for each response are graphed. The prevalence of each response (C) is shown (% seropositive for each group after treatment +95% confidence intervals).
Figure 6.
Expression of SmTAL4 in cercarial tail only.
For PCR analysis (A), total RNA from isolated from heads (H) and tails (T) after mechanical separation and used to prepare cDNA for use with specific primers to generate amplicons for Sm ß-actin (203 bp), SmTAL4 (152 bp), SmTAL5(228 bp) and SmTAL13(206 bp). Products were separated on a 2% agarose gel and detected with ethidium bromide. For immunostaining (B) 8 µ sections of frozen sections of whole cercariae were fixed and stained with rat anti-SmTAL4 antiserum and TRITC -anti-rat antibody (red). Nuclei were counterstained with DAPI (blue). In the negative control (insert) anti- SmTAL4 antiserum was pre-absorbed with recombinant SmTAL4.