Transcriptome analysis of Taenia solium cysticerci using Open reading Frame ESTS (ORESTES)

<p>Abstract</p> <p>Background</p> <p>Human infection by the pork tapeworm <it>Taenia solium </it>affects more than 50 million people worldwide, particularly in underdeveloped and developing countries. Cysticercosis which arises from larval encystation can be life threatening and difficult to treat. Here, we investigate for the first time the transcriptome of the clinically relevant cysticerci larval form.</p> <p>Results</p> <p>Using Expressed Sequence Tags (ESTs) produced by the ORESTES method, a total of 1,520 high quality ESTs were generated from 20 ORESTES cDNA mini-libraries and its analysis revealed fragments of genes with promising applications including 51 ESTs matching antigens previously described in other species, as well as 113 sequences representing proteins with potential extracellular localization, with obvious applications for immune-diagnosis or vaccine development.</p> <p>Conclusion</p> <p>The set of sequences described here will contribute to deciphering the expression profile of this important parasite and will be informative for the genome assembly and annotation, as well as for studies of intra- and inter-specific sequence variability. Genes of interest for developing new diagnostic and therapeutic tools are described and discussed.</p

Transcriptome analysis of Taenia solium cysticerci using Open Reading Frama ESTs (ORESTES)

Background: Human infection by the pork tapeworm Taenia solium affects more than 50 million people worldwide, particularly in underdeveloped and developing countries. Cysticercosis which arises from larval encystation can be life threatening and difficult to treat. Here, we investigate for the first time the transcriptome of the clinically relevant cysticerci larval form. Results: Using Expressed Sequence Tags (ESTs) produced by the ORESTES method, a total of 1,520 high quality ESTs were generated from 20 ORESTES cDNA mini-libraries and its analysis revealed fragments of genes with promising applications including 51 ESTs matching antigens previously described in other species, as well as 113 sequences representing proteins with potential extracellular localization, with obvious applications for immune-diagnosis or vaccine development. Conclusion: The set of sequences described here will contribute to deciphering the expression profile of this important parasite and will be informative for the genome assembly and annotation, as well as for studies of intra- and inter-specific sequence variability. Genes of interest for developing new diagnostic and therapeutic tools are described and discussed

Transcriptome analysis of Taenia solium cysticerci using Open Reading Frama ESTs (ORESTES)

Background: Human infection by the pork tapeworm Taenia solium affects more than 50 million people worldwide, particularly in underdeveloped and developing countries. Cysticercosis which arises from larval encystation can be life threatening and difficult to treat. Here, we investigate for the first time the transcriptome of the clinically relevant cysticerci larval form. Results: Using Expressed Sequence Tags (ESTs) produced by the ORESTES method, a total of 1,520 high quality ESTs were generated from 20 ORESTES cDNA mini-libraries and its analysis revealed fragments of genes with promising applications including 51 ESTs matching antigens previously described in other species, as well as 113 sequences representing proteins with potential extracellular localization, with obvious applications for immune-diagnosis or vaccine development. Conclusion: The set of sequences described here will contribute to deciphering the expression profile of this important parasite and will be informative for the genome assembly and annotation, as well as for studies of intra- and inter-specific sequence variability. Genes of interest for developing new diagnostic and therapeutic tools are described and discussed

Extraction of PMN from the air-pouch.

<p>Absolute number of PMN at extraction time for both irradiated and non-irradiated mice of <i>Pb</i> and Zymosan (Zy) groups; p = 0.0001 (*) between non-irradiated and irradiated PMN of <i>Pb</i> groups; p = 0.0001 (*) between non-irradiated and irradiated PMN of Zy groups; p = 0.0001 (*) between PMN from non-irradiated <i>Pb</i> and Zy groups; p = 0.0399 (*) between PMN from irradiated <i>Pb</i> and Zy groups.</p

PMN protein production.

<p>Protein production kinetics of irradiated and non-irradiated PMN. <b>A</b>—<i>Pb</i> infected mice (non-irradiated and irradiated); <b>B</b>—Zymosan (Zy) inoculated mice (non-irradiated and irradiated); <b>C—</b>PMN recruited by <i>Pb</i> infection and co-cultivated with <i>Pb</i>; and <b>D—</b>recruited by Zy inoculation and co-cultivated with <i>Pb</i>. <b>A:</b> p = 0.002 (*) between 2 and 6 hours of <i>Pb</i> non-irradiated; p = 0.0001 (*) between <i>Pb</i> non-irradiated and <i>Pb</i> irradiated at 2 hours; p = 0.001 (*) between 2 and 18 hours of <i>Pb</i> irradiated PMN; p = 0.001 (*) between 6 and 18 hours of <i>Pb</i> irradiated PMN; and p = 0.001 (*) between the <i>Pb</i> irradiated and non-irradiated PMN at 18 hours; <b>B</b>: p = 0.009 (*) between Zy non-irradiated and Zy irradiated at 2 hours; and p = 0.003 (*) between Zy non-irradiated and Z irradiated at 6 hours; <b>C:</b> p = 0.005 (*) between the non-irradiated PMN of the <i>Pb</i> group at 2 and 18 hours of co-cultivation; p = 0.0481 (*) between the non-irradiated PMN of the <i>Pb</i> group at 6 and 18 hours of co-cultivation with <i>Pb</i>; and p = 0.002 (*) between the non-irradiated and irradiated PMN at 18 hours of co-cultivation; <b>D:</b> p = 0.0043 (*) between the non-irradiated and irradiated PMN at 2 hours of co-cultivation; p = 0.0069 (*) between the non-irradiated PMN at 2 and 18 hours of co-cultivation.</p

PMN mitochondrial activity after co-cultivation.

<p>Mitochondrial activity of light irradiated and non-irradiated PMN recruited by either <i>Pb</i> or Zymosan stimuli and co-cultivated with <i>Pb</i> in vitro. p = 0.0029 (*) between the irradiated and non-irradiated PMN of the <i>Pb</i> group; p = 0.0012 (*) between the irradiated PMN of <i>Pb</i> group and irradiated PMN of the Zy group; p = 0.0004 (*) between the irradiated and non-irradiated PMN of the Zy group; p = 0.0001 (*) between the non-irradiated PMN of the <i>Pb</i> group and the non-irradiated PMN of the Zy group.</p

Kinetic behavior of PMN viability.

<p>Kinetics of PMN cell viability in irradiated and non-irradiated mice after stimuli with: <b>A</b>—<i>Pb</i>: higher viability for the irradiated PMN at 6 hours—p = 0.0278 (*); <b>B</b>—Zymosan: higher viability for the irradiated PMN at 2 hours—p = 0.0274 (*); <b>C</b>—<i>Pb</i> stimulated and co-cultivated with <i>Pb</i>; <b>D—</b>Zymosan stimulated and co-cultivated with <i>Pb</i>.</p

Fungicidal capacity of PMN.

<p><b>A—</b>Colony forming units (CFU) of <i>Pb</i> at 7 and 12 days evaluation for non-irradiated and irradiated PMN. p = 0.0002 (*) between non-irradiated and irradiated PMN after 12 days of fungal growth; p = 0.0118 (*) between the irradiated groups at 7 and 12 days of fungal growth and p = 0.0003 (*) between the non-irradiated groups at 7 and 12 days of fungal growth. The colony forming units of <i>Pb</i> after co-cultivation with PMN recruited by <i>Pb</i> infection or Zy inoculation are shown after: <b>B—</b>7 days: p = 0.0369 (*) between the irradiated and non-irradiated PMN of the <i>Pb</i> group; p = 0.0232 (*) between the irradiated and non-irradiated PMN of the Zy group; or <b>C—</b>12 days: p = 0.0193 (*) between the irradiated and non-irradiated PMN of the <i>Pb</i> group; p = 0.0492 (*) between the irradiated and non-irradiated PMN of the Zy group.</p

Subcutaneous air-pouch.

<p>Clinical appearance of the air-pouch on the dorsum of the mouse after skin flap procedure.</p