Microarray-based analysis of differential gene expression between infective and noninfective larvae of Strongyloides stercoralis.

Authors

Roshan Ramanathan, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland
Sudhir Varma, Bioinformatics and Computational Biosciences Branch, National Institutes of Health, Bethesda, Maryland
José M C Ribeiro, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases National Institutes of Health, Bethesda, Maryland
Timothy G Myers, Microarray Research Facility, National Institutes of Health, Bethesda, Maryland
Thomas J Nolan, Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania
David Abraham, Department of Microbiology and Immunology, Thomas Jefferson University, Philadelphia, PennsylvaniaFollow
James B Lok, Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania
Thomas B Nutman, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland

Document Type

Article

Publication Date

5-1-2011

Comments

This article has been peer reviewed and is published in PLoS Neglected Tropical Diseases 2011, 5(5). The published version is available at DOI: 10.1371/journal.pntd.0001039. © Public Library of Science

Abstract

BACKGROUND: Differences between noninfective first-stage (L1) and infective third-stage (L3i) larvae of parasitic nematode Strongyloides stercoralis at 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 3,571 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 935 differentially expressed genes (469 L3i-biased; 466 L1-biased) having two-fold expression differences or greater and microarray signals with a p value<0.01. Based on a functional analysis, L1 larvae have a larger number of genes putatively involved in transcription (p = 0.004), and L3i larvae have biased expression of putative heat shock proteins (such as hsp-90). Genes with products known to be immunoreactive in S. stercoralis-infected humans (such as SsIR and NIE) had L3i biased expression. Abundantly expressed L3i contigs of interest included S. stercoralis orthologs of cytochrome oxidase ucr 2.1 and hsp-90, 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 25 most highly expressed L3i genes. The sperm-containing glycoprotein domain, utilized in a vaccine against the nematode Cooperia punctata, was exclusively found in L3i biased genes 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.

Recommended Citation

Ramanathan, Roshan; Varma, Sudhir; Ribeiro, José M C; Myers, Timothy G; Nolan, Thomas J; Abraham, David; Lok, James B; and Nutman, Thomas B, "Microarray-based analysis of differential gene expression between infective and noninfective larvae of Strongyloides stercoralis." (2011). Department of Microbiology and Immunology Faculty Papers. Paper 22.
https://jdc.jefferson.edu/mifp/22

PubMed ID

21572524