Investigation Title Sm29 is a protective surface protein from the tegument of lung-stage Schistosoma mansoni
Comment[Submitted Name] Sm29 is a protective surface protein from the tegument of lung-stage Schistosoma mansoni
Experimental Design transcription profiling by array
Experimental Design Term Source REF EFO
Comment[AEMIAMESCORE] 3
Comment[SecondaryAccession] GSE10777
Comment[ArrayExpressReleaseDate] 2010-05-15
Comment[Publication DOI] 10.1371/journal.pntd.0000308
Comment[ArrayExpressAccession] E-GEOD-10777
Comment[MAGETAB TimeStamp_Version] 2010-07-29 22:17:42 Last Changed Rev: 13058
Experimental Factor Name
Experimental Factor Type
Experimental Factor Term Source REF
Person Last Name Oliveira Venancio Caliari Melo Almeida Cardoso Kitten Verjovski-Almeida Mati Macedo Gava Venancio Cardoso
Person First Name Sergio Thiago Marcelo Alan Giulliana Luciana Gregory Sergio Vitor Gilson Elisandra Thiago Fernanda
Person Mid Initials C M V L T S T L C Motta C
Person Email thiago.venancio@gmail.com
Person Phone
Person Fax
Person Address NCBI-NLM-NIH, 8600 Rockville Pike, Bethesda, MD, USA
Person Affiliation NCBI-NLM-NIH
Person Roles submitter
Person Roles Term Source REF The MGED Ontology
Quality Control Type
Quality Control Term Source REF
Replicate Type
Replicate Term Source REF
Normalization Type
Normalization Term Source REF
Date of Experiment
Public Release Date 2010-05-15
PubMed ID 18827884
Publication DOI 18827884
Publication Author List Cardoso FC, Macedo GC, Gava E, Kitten GT, Mati VL, de Melo AL, Caliari MV, Almeida GT, Venancio TM, Verjovski-Almeida S, Oliveira SC
Publication Title Schistosoma mansoni tegument protein Sm29 is able to induce a Th1-type of immune response and protection against parasite infection.
Publication Status journal_article
Publication Status Term Source REF The MGED Ontology
Experiment Description Schistosomiasis continues to be a significant public health problem1. Although vaccine development against this disease has experienced more failures than successes, encouraging results have recently been obtained using membrane-spanning protein antigens from the tegument of S. mansoni. Our group recently identified Sm29, an antigen that is predominantly recognized by IgG1 and IgG3 antibodies of resistant patients2. In the present study, we show that Sm29 is located on the surface of adult worms and lung-stage schistosomula. Immunization of mice with recombinant (r) Sm29 engendered 51% reduction in adult worm burdens, 60% reduction in intestinal eggs and 55% reduction in liver granulomas. Protective immunity in mice was associated with high titers of specific IgG1 and IgG2a and elevated production of IFN-γ, TNF-α and IL-12. Further, cellular responses of infected schistosomiasis patients to rSm29 consisted of elevated IFN-γ and an absence of IL-5. Gene expression analysis of worms recovered from rSm29 vaccinated mice relative to control mice revealed a significant (q< 0.01) down-regulation of 498 genes, while no up-regulation was detected. Among down-regulated genes, many of them encode surface antigens and proteins associated with immune signals suggesting that under immune attack schistosomes reduce the expression of critical surface proteins. This study demonstrates that the membrane-bound Sm29 protein is a new molecule that has great potential as a vaccine candidate against schistosomiasis. Keywords: Schistosoma mansoni gene expression in vaccinated mice Two biological samples were used in the study, each used in two arrays (technical replicates with dye swap) and each array containing two replicates of each spot. Overall, there are eight data points data points for each spot. Only cases with more than 4 of 8 points are valid ratios were considered. Value and its respective dye swap value [after changing the log(ratio) signal] were averaged, reducing the data point to four. The average values were than used in the significance testing using SAM (Tusher et al., 2001). The samples were used to get average values using the following combination: 3000139790L with 3000139792L; 3000139790R with 3000139792R; 3000139791L with 3000139795L; 3000139791R with 3000139795R The normalized, averaged dyeswap values are provided in a supplementary file at the foot of the record.
Protocol Name P-GSE10777-2 P-GSE10777-3 P-GSE10777-4 P-GSE10777-5 P-GSE10777-6 P-GSE10777-1
Protocol Type nucleic_acid_extraction labeling hybridization image_aquisition feature_extraction bioassay_data_transformation
Protocol Description Total RNA was obtained using Trizol reagent (Invitrogen) according to the manufacturer's instructions. The RNA was quantified using the Nanodrop ND-1000 UV/Vis spectrophotometer and RNA integrity was checked by electrophoresis with an Agilent 2100 Bioanalyzer. One microgram of total RNA from each sample was amplified using the T7-RNA polymerase-based linear amplification protocol. Three micrograms of aminoallyl-modified amplified RNA was labeled with Cy3 or Cy5 by indirect labeling (GE Healthcare). The Cy3- and Cy5-labeled samples were then combined, dried and re-suspended in hybridization buffer (50 % formamide, 25 % RNase free water and 25 % Microarray Hybridization buffer 4x from GE Healthcare). The Cy3- and Cy5-labeled samples were then combined, dried and re-suspended in hybridization buffer (50 % formamide, 25 % RNase free water and 25 % Microarray Hybridization buffer 4x from GE Healthcare). Slides were air dried and scanned using a microarray dual channel laser scanner (GenePix 4000B, Molecular Devices, USA) at 5 Ãm resolution, 100 % laser power and PMT levels were adjusted in order to obtain similar average intensities of red and green signal. Data was extracted from images using the Array Vision 8.0 program. Unreliable spots, having intensities too similar to local background (ie, low intensity) or saturated, were filtered out. Saturated signals [intensity greater than >990 fluorescence units] were also discarded. Normalization was carried out by LOWESS fitting on a M versus A plot ID_REF =
CY3_RAW = C1. Median-based Trimmed Mean density value for each spot. The reported value represents the mean of all the pixels remaining in a target, after first removing pixels with density values that exceed four median absolute deviations (MADs) above or below the median. This measure removes the influence of image artifacts (e.g., dust particles) on density estimation.
CY3_BKG = C1. Median of a region outside of the spot
CY3_MTM = C1. Subtracted MTM Density value. MTM Density value of the spot, minus the background Density value.
CY5_RAW = C2. Median-based Trimmed Mean density value for each spot. The reported value represents the mean of all the pixels remaining in a target, after first removing pixels with density values that exceed four median absolute deviations (MADs) above or below the median. This measure removes the influence of image artifacts (e.g., dust particles) on density estimation.
CY5_BKG = C2. Median of a region outside of the spot
CY5_MTM = C2. Subtracted MTM Density value. MTM Density value of the spot, minus the background Density value.
VALUE = normalized, log2 (test/control) ratio; flagged VALUEs labeled as 'null'
Protocol Parameters
Protocol Hardware
Protocol Software
Protocol Contact
Protocol Term Source REF The MGED Ontology The MGED Ontology
SDRF File E-GEOD-10777.sdrf.txt
Term Source Name The MGED Ontology ArrayExpress EFO The MGED Ontology
Term Source File http://mged.sourceforge.net/ontologies/MGEDontology.php http://www.ebi.ac.uk/arrayexpress http://www.ebi.ac.uk/efo/ http://mged.sourceforge.net/ontologies/MGEDontology.php
Term Source Version