TY - JOUR
T1 - Systems Biology Studies of Adult Paragonimus Lung Flukes Facilitate the Identification of Immunodominant Parasite Antigens
AU - McNulty, Samantha N.
AU - Fischer, Peter U.
AU - Townsend, R. Reid
AU - Curtis, Kurt C.
AU - Weil, Gary J.
AU - Mitreva, Makedonka
N1 - Funding Information:
The authors would like to thank Dr. Patricia Willkins, Dr. Scott Folk, and Washington University physicians for providing sera from P. kellicotti patients. We thank John Martin, Phillip Ozersky, Rahul Tyagi, and Dr. Bruce Rosa from the Mitreva Lab for assistance with similarity searching and figure preparation, and the Genome Institute production team for RNAseq data generation. The RNAseq data was generated as part of the Foodborne trematode genome project funded by NHGRI (P. kellicotti NCBI BioProject Accession: PRJNA179523). The proteomic sample processing and LC-MS data acquisition by Petra Gilmore, Anne Kettler and Rose Connors at the Washington University Proteomics Core Laboratory is gratefully acknowledged. We thank Jeanne Rumsey for expert proteomic data processing and protein database searching.
Publisher Copyright:
© 2014 McNulty et al.
PY - 2014
Y1 - 2014
N2 - Background:Paragonimiasis is a food-borne trematode infection acquired by eating raw or undercooked crustaceans. It is a major public health problem in the far East, but it also occurs in South Asia, Africa, and in the Americas. Paragonimus worms cause chronic lung disease with cough, fever and hemoptysis that can be confused with tuberculosis or other non-parasitic diseases. Treatment is straightforward, but diagnosis is often delayed due to a lack of reliable parasitological or serodiagnostic tests. Hence, the purpose of this study was to use a systems biology approach to identify key parasite proteins that may be useful for development of improved diagnostic tests.Methodology/Principal Findings:The transcriptome of adult Paragonimus kellicotti was sequenced with Illumina technology. Raw reads were pre-processed and assembled into 78,674 unique transcripts derived from 54,622 genetic loci, and 77,123 unique protein translations were predicted. A total of 2,555 predicted proteins (from 1,863 genetic loci) were verified by mass spectrometric analysis of total worm homogenate, including 63 proteins lacking homology to previously characterized sequences. Parasite proteins encoded by 321 transcripts (227 genetic loci) were reactive with antibodies from infected patients, as demonstrated by immunoaffinity purification and high-resolution liquid chromatography-mass spectrometry. Serodiagnostic candidates were prioritized based on several criteria, especially low conservation with proteins in other trematodes. Cysteine proteases, MFP6 proteins and myoglobins were abundant among the immunoreactive proteins, and these warrant further study as diagnostic candidates.Conclusions:The transcriptome, proteome and immunolome of adult P. kellicotti represent a major advance in the study of Paragonimus species. These data provide a powerful foundation for translational research to develop improved diagnostic tests. Similar integrated approaches may be useful for identifying novel targets for drugs and vaccines in the future.
AB - Background:Paragonimiasis is a food-borne trematode infection acquired by eating raw or undercooked crustaceans. It is a major public health problem in the far East, but it also occurs in South Asia, Africa, and in the Americas. Paragonimus worms cause chronic lung disease with cough, fever and hemoptysis that can be confused with tuberculosis or other non-parasitic diseases. Treatment is straightforward, but diagnosis is often delayed due to a lack of reliable parasitological or serodiagnostic tests. Hence, the purpose of this study was to use a systems biology approach to identify key parasite proteins that may be useful for development of improved diagnostic tests.Methodology/Principal Findings:The transcriptome of adult Paragonimus kellicotti was sequenced with Illumina technology. Raw reads were pre-processed and assembled into 78,674 unique transcripts derived from 54,622 genetic loci, and 77,123 unique protein translations were predicted. A total of 2,555 predicted proteins (from 1,863 genetic loci) were verified by mass spectrometric analysis of total worm homogenate, including 63 proteins lacking homology to previously characterized sequences. Parasite proteins encoded by 321 transcripts (227 genetic loci) were reactive with antibodies from infected patients, as demonstrated by immunoaffinity purification and high-resolution liquid chromatography-mass spectrometry. Serodiagnostic candidates were prioritized based on several criteria, especially low conservation with proteins in other trematodes. Cysteine proteases, MFP6 proteins and myoglobins were abundant among the immunoreactive proteins, and these warrant further study as diagnostic candidates.Conclusions:The transcriptome, proteome and immunolome of adult P. kellicotti represent a major advance in the study of Paragonimus species. These data provide a powerful foundation for translational research to develop improved diagnostic tests. Similar integrated approaches may be useful for identifying novel targets for drugs and vaccines in the future.
UR - http://www.scopus.com/inward/record.url?scp=84920527872&partnerID=8YFLogxK
U2 - 10.1371/journal.pntd.0003242
DO - 10.1371/journal.pntd.0003242
M3 - Article
C2 - 25329661
AN - SCOPUS:84920527872
VL - 8
JO - PLoS Neglected Tropical Diseases
JF - PLoS Neglected Tropical Diseases
SN - 1935-2727
IS - 10
ER -