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Status |
Public on Feb 09, 2011 |
Title |
Microarray Analysis of Infective and Noninfective Larvae of Strongyloides stercoralis |
Organism |
Strongyloides stercoralis |
Experiment type |
Expression profiling by array
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Summary |
BACKGROUND: Differences between noninfective first-stage (L1) and infective third-stage (L3i) larvae of parasitic nematode Strongyloides stercoralisat the molecular level are relatively uncharacterized. DNA microarrays were developed and utilized for this purpose. METHODS AND FINDINGS: Oligonucleotide hybridization probes for the array were designed to bind 3571 putative mRNA transcripts predicted by analysis of 11,335 expressed sequence tags (ESTs) obtained as part of the Nematode EST project. RNA obtained from S. stercoralis L3i and L1 was co-hybridized to each array after labeling the individual samples with different fluorescent tags. Bioinformatic predictions of gene function were developed using a novel cDNA Annotation System software. We identified 967 differentially expressed genes (457 L3i-biased; 510 L1-biased) having greater than two-fold expression differences and p < 0.01. Based on functional analysis, L1s have a larger number of genes putatively involved in transcription (p = 0.0158), and L3is have higher expression of stress-related genes (such as putative heat shock proteins dnj-12, daf-21, dnj-10). Genes with products that have been shown to be immunoreactive in S. sterocoralis-infected humans (SsIR and NIE) were additionally found to be L3i biased. Unique and abundant L3i contigs of interest included S. stercoralis orthologs of cytochrome oxidase ucr 2.1, daf-12, and daf-21, which may be potential chemotherapeutic targets. The S. stercoralis ortholog of fatty acid and retinol binding protein-1, successfully used in a vaccine against Ancylostoma ceylanicum, was identified among the top 25 most L3i-biased genes. The sperm-containing glycoprotein domain, utilized in a vaccine against the nematode Cooperia punctata, was exclusively found in the L3i group and may be a valuable S. stercoralis target of interest. Conclusions A new DNA microarray tool for the examination of S. stercoralis biology has been developed and provides new and valuable insights regarding differences between infective and noninfective S. stercoralis larvae. Potential therapeutic and vaccine targets were identified for further study.
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Overall design |
For the present study, four technical replicates using pooled samples of L1 stage larvae versus L3i stage larvae were performed, including one dye swap.
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Citation(s) |
21572524 |
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Submission date |
Oct 15, 2010 |
Last update date |
Mar 22, 2012 |
Contact name |
Roshan Ramanathan |
E-mail(s) |
[email protected]
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Phone |
301 402 5969
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Fax |
301 480 3757
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Organization name |
National Institutes of Health
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Department |
NIAID
|
Lab |
Laboratory of Parasitic Diseases
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Street address |
4 Center Drive
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City |
Bethesda |
State/province |
MD |
ZIP/Postal code |
20892 |
Country |
USA |
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Platforms (1) |
GPL11063 |
NIAID Ssca -- Strongyloides stercoralis 4K - mAdb version |
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Samples (4)
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GSM609213 |
L3i vs L1- repeat 2 - mAdbID:103420 |
GSM609214 |
L3i vs L1- repeat 3 - mAdbID:103421 |
GSM609215 |
L3i vs L1- repeat 4 - mAdbID:103422 |
GSM609216 |
L1 vs L3i - repeat 1 - rev. label - mAdbID:103423 |
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Relations |
BioProject |
PRJNA132425 |