Wang et al.

BMC Veterinary Research (2015) 11:119

DOI 10.1186/s12917-015-0423-8

RESEARCH ARTICLE Open Access

Proteomic analysis of the excretory/secretory

products and antigenic proteins of Echinococcus
granulosus adult worms from infected dogs
Ying Wang1, Di Xiao2, Yujuan Shen1, Xiuming Han3, Fei Zhao2, Xiaohong Li1, Weiping Wu1, Hejun Zhou1,
Jianzhong Zhang2* and Jianping Cao1*

Abstract
Background: Cystic echinococcosis, which is caused by Echinococcus granulosus, is one of the most widespread
zoonotic helminth diseases that affects humans and livestock. Dogs, which harbor adult worms in their small
intestines, are a pivotal source of E. granulosus infection in humans and domestic animals. Therefore, novel
molecular approaches for the prevention and diagnosis of this parasite infection in dogs need to be developed.
Results: In this study, we performed proteomic analysis to identify excretory/secretory products (ES) and antigenic
proteins of E. granulosus adult worms using two-dimensional electrophoresis, tandem matrix-assisted laser desorption/
ionization time-of-flight (MALDI-TOF/TOF), and Western blotting of sera from infected dogs. This study identified 33 ES
product spots corresponding to 9 different proteins and 21 antigenic protein spots corresponding to 13 different
proteins. Six antigenic proteins were identified for the first time.
Conclusions: The present study extended the existing proteomic data of E. granulosus and provides further
information regarding host-parasite interactions and survival mechanisms. The results of this study contribute to
vaccination and immunodiagnoses for E. granulosus infections.
Keywords: Echinococcus granulosus, Adult worm, Excretory/secretory products, Antigenic protein, 2-dimensional gel
electrophoresis, MALDI-TOF/TOF

Background through the ingestion of parasite eggs released in the feces

Cystic echinococcosis (CE) is a type of zoonosis caused by of definitive hosts or through consumption of foods con-
Echinococcus granulosus, a canine tapeworm [1]. Accord- taminated with the parasite eggs [5]. Dogs, as definitive
ing to recent estimates, 4 million individuals are affected hosts, are therefore pivotal in the transmission of CE.
with CE and 40 million individuals are at risk [2–4]. The One of the strategies to reduce the risk of infection is to
life cycle of E. granulosus is complex and involves two interrupt the transmission of CE. Vaccination of the de-
hosts: definitive and intermediate hosts. The definitive finitive hosts is an effective method. As there are far fewer
hosts are primarily dogs, which harbor adult worms in dogs than sheep and cattle in endemic areas, far fewer ani-
their small intestines. The intermediate hosts, e.g., humans mals consequently need to be vaccinated. However, a limi-
and herbivores, particularly sheep and cattle, get infected tation of vaccination is that the immune modulatory
mechanisms of E. granulosus are not fully understood.
* Correspondence: [email protected]; [email protected] The immune modulatory mechanisms of the parasite
2
Collaborative Innovation Center for Diagnosis and Treatment of Infectious might involve antigenic proteins and excretory-secretory
Diseases, State Key Laboratory for Infectious Disease Prevention and Control,
National Institute for Communicable Disease Control and Prevention,
(ES) products released by the parasite [6, 7]. The surveil-
Chinese Center for Disease Control and Prevention, Beijing 102206, China lance of E. granulosus infections in definitive hosts
1
National Institute of Parasitic Diseases, Chinese Center for Disease Control through sensitive diagnostic procedures is of paramount
and Prevention; Laboratory of Parasite and Vector Biology, MOH, China; WHO
Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai
importance; coproantigen detection and arecoline purga-
200025, China tion are not sufficiently sensitive. Therefore, there is an
Full list of author information is available at the end of the article

© 2015 Wang et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain
Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article,
unless otherwise stated.
Wang et al. BMC Veterinary Research (2015) 11:119 Page 2 of 7

urgent need to identify immune markers that can be Sample preparation

used in diagnosis and vaccine development. The identi- The concentrated supernatant was precipitated over-
fication of ES products and antigenic proteins could night at −20 °C with five volumes of ice-cold acetone
provide valuable insights into host-parasite interactions containing 0.2 % dithiothreitol (DTT; w/v) and 20 %
and improve the repertoire of candidate proteins used trichloroacetic acid (TCA; w/v). Protein precipitates
in immunodiagnoses, vaccination, and therapy. How- were collected by centrifugation (10,000 rpm, 4 °C,
ever, E. granulosus adult worms have received little at- 10 min) and washed three times with ice-cold acetone
tention; most studies have focused on the pre-adult containing 0.2 % DTT (w/v). The resulting pellet was
stage of the parasite [8–11]. The proteomic profile of freeze-dried, suspended in lysate buffer (6 M urea, 2 M
ES products and antigenic proteins from adult E. gran- thiourea, 4.0 % CHARPS, 40 mM DTT, and 0.5 % IPG;
ulosus remain to be elucidated. pH 3–10), and sonicated in ice until the suspension be-
In this study, we investigated the ES products and came clear. The homogenate was centrifuged at
antigenic proteins of E. granulosus adult worms from 12,000 rpm for 15 min at 4 °C. The resulting super-
infected dogs using two-dimensional polyacrylamide natant contained ES products.
gel electrophoresis (2DE) and tandem matrix-assisted E. granulosus adult worms were washed in PBS, sus-
laser desorption/ionization time-of-flight (MALDI-TOF/ pended in lysate buffer, and sonicated in ice until the
TOF) mass spectroscopy. The results obtained from suspension became clear. The homogenate was centri-
this study are crucial for understanding the survival fuged at 12,000 rpm at 4 °C for 15 min. The supernatant
mechanisms of E. granulosus and host-parasite inte- contained adult worms.
ractions. Furthermore, the results could assist in the The samples were cleaned up using the 2D Clean-up
development of vaccine antigens, drug targets, and im- kit (Amersham Biosciences), quantified using the 2D
munodiagnosis markers. Quant Kit (Amersham Biosciences), and subjected to
2DE.
Methods
Ethics statement Two-dimensional electrophoresis
This study was performd in strict accordance with the rec- One-dimensional isoelectric focusing (IEF) and two-
ommendations of the Guide for the Care and Use of dimensional sodium dodecyl sulfate polyacrylamide gel
Laboratory Animals of the National Institute of Parasitic electrophoresis (SDS-PAGE) were performed (Amersham
Diseases, Chinese Center for Disease Control and Preven- Biosciences). Briefly, 100 μg of the ES sample in 130 μl
tion. The protocol was approved by the Laboratory Ani- rehydration buffer was loaded onto a 7-cm immobiline
mal Welfare & Ethics Committee (LAWEC), National IPG drystrip (pH3-10NL; Amersham Biosciences);
Institute of Parasitic Diseases, Chinese Center for Disease 800 μg of adult worm protein sample in 450 μl of rehy-
Control and Prevention (Permit Number: IPD 2010–007). dration buffer was loaded onto a 24-cm immobiline
Three dogs used in the research belonged to local farmers. IPG drystrip (pH3-10NL; Amersham Biosciences). IEF
The owners of the all dogs have oral consent for the use of was performed at 80,000 Vh using Ettan™IPGphorII
their dogs in this research by explaining the purpose of (Amersham Biosciences). The drystrips were equili-
the research and ensuring the welfare of animals. brated twice (15 min each time) in equilibration buffer
(50 mM Tris–HCl [pH 8.8], 6 M urea, 30 % glycerol, 2
Collection and culture of parasites % SDS, and 1 % DTT for the first equilibration; 50 mM
All dogs were infected with E. granulosus protoscoleces Tris–HCl [pH 8.8], 6 M urea, 30 % glycerol, 2 % SDS,
from sheep after dewormed with praziquantel, and and 4.8 % iodacetamide for the second equilibration).
sacrificed to obtain intestines at 40 d post-infection. Two-dimensional SDS-PAGE was performed on a 12 %
All surgery was performed under sodium pentobarbital polyacrylamide gel in the Ettan™DALTsix and Ettan™VE
anesthesia, and all efforts were made to minimize suf- systems (Amersham Biosciences). Two replicates were
fering. Worms were released by soaking the intestinal performed per protein sample.
contents in phosphate-buffered saline (PBS, Gibco,
California, USA), washed in sterile PBS containing 100 Protein identification by MALDI-TOF/TOF analysis
U/ml penicillin G and 100 μg/ml streptomycin (Gibco), The gels were stained with Coomassie brilliant blue
and cultured for 24 h at 37 °C at 500 worms/ml in (CBB) G-250 (BioRad). Protein spots were excised by
serum-free RPMI 1640 medium (Gibco) supplemented an Ettan Spot Picker (Amersham Biosciences). In-gel
with 2 % glucose (Sigma, St.Louis, USA) and antibi- digestion was performed as previously described [12].
otics. The supernatant was harvested and concentrated Protein identification was carried out using MALDI-
using a micro-concentrator with a 3-kDa cut-off (Millipore, TOF/TOF mass spectrometry (4700 MALDI-TOF/TOF
Massachusetts, USA). mass spectrometer; Applied Biosystems, California, USA).
Wang et al. BMC Veterinary Research (2015) 11:119 Page 3 of 7

Immunoblot analysis of antigenic proteins

Adult worm proteins were electro-transferred from the
2DE gels to polyvinylidene fluoride (PVDF) membranes
(Amersham Biosciences) at 300 mA for 2 h using a
TE77 semi-dry transfer unit (Amersham Biosciences).
Membranes were blocked for 1 h in 5 % nonfat milk
powder in TBS (20 mM Tris, 500 mM NaCl, pH 7.5) at
room temperature and incubated overnight with serum
from E. granulosus-infected dogs (1:500 dilution) at 4 °C.
After three washes with TBST (TBS containing 0.05 %
v/v Tween-20), the membranes were incubated at room
temperature with HRP-labeled anti-dog IgG (1:10000 di-
lution; Sigma-Aldrich) for 1 h. Bound antibodies were
revealed using the DAB reagent. Antigenic spots on the
2DE gels were identified based on matches with the 2DE
proteomic map.

Results
ES products of adult E. granulosus
Fig. 1 Representative 2DE gels of the excretory/secretory products Approximately 50 spots representative of the ES products
of E. granulosus adult worms. Proteins (100 μg) were separated on a were resolved on Coomassie-stained 2DE gels (Fig. 1). A
linear pH range of 3–10 using IEF in the first dimension and 12 %
SDS-PAGE in the second dimension. The proteins were stained with
total of 48 spots were subjected to MALDI-TOF/TOF
Coomassie blue. Molecular weight markers are shown on the left. analysis; 33 protein spots corresponding to 8 different pro-
The proteins identified are shown in detail in Table 1 teins were identified (Table 1).
The identified proteins were subjected to functional an-
notation based on the GO and KOG classification systems.
33, 30, and 20 proteins had GO terms for molecular func-
The spectra were processed and analyzed by the Global tions, cellular components, and biological processes, re-
Protein Server Workstation (GPS; Applied Biosystems), spectively. A summary of the GO and KOG annotations is
which uses the internal Mascot v2.1 software (Matrix provided in Fig. 2. Three proteins could not be classified
Science, London, UK) to search for peptide mass fin- by KOG.
gerprints and MS/MS data based on the NCBI non-
redundant protein database. Identification with a GPS Antigenic proteins of E. granulosus adult worms
confidence interval >95 % was accepted. Gene ontology To identify adult worm antigenic proteins, we per-
(GO) terms were applied to the identified proteins; pie formed 2DE immunoblot analyses using serum from E.
charts of the GO terms for molecular functions, cel- granulosus-infected dogs. In this study, 36 antigen
lular components, and biological processes were ge- spots were detected (Fig. 3). Using the corresponding
nerated. Additionally, Eukaryotic Orthologous Group 2DE proteomic map, 21 antigen spots were identified,
(KOG) annotation was assigned (http://genome.jgi.doe.gov/ corresponding to 12 different proteins (Table 2), three
Tutorial/tutorial/kog.html). of which were identified for the first time: severin,

Table 1 ES products of E. granulosus adult worms identified by MALDI-TOF/TOF analysis

GI number Protein name Species MW pI Protein score C.I.%
547974 Paramyosin E. granulosus 98681.8 5.21 100
262192839 Enolase E. granulosus 46531.9 6.48 100
576695773 Actin E. granulosus 41717.8 5.3 100
576698524 Small heat shock protein p36 E. granulosus 35821.2 5.92 99.995
6016537 Malate dehydrogenase, cytoplasmic E. granulosus 36627.8 8.11 100
2316076 Glutathione S-transferase E. granulosus 25536.8 7.51 100
189016336 Thioredoxin peroxidase E. granulosus 21419.7 5.78 100
31077167 Cyclophilin E. granulosus 17343.4 6.41 99.003
Wang et al. BMC Veterinary Research (2015) 11:119 Page 4 of 7

Fig. 2 Functional analysis of the excretory/secretory proteins identified in E. granulosus adult worms. Gene ontology terms for the subcategories
molecular function (a), cellular component (b), and biological process (c) were assigned to the proteins identified in the adult worms. KOG
functional categories (d) were assigned to the proteins identified in the adult worms. The percentage and number (in parentheses) of proteins
identified in each functional category are indicated in each sector. KOG functional categories: (Z) cytoskeleton; (O) posttranslational modification,
protein turnover, chaperones; (G) carbohydrate transport and metabolism; (C) energy production and conversion; (noKOG) protein not assigned
to any KOG category. The number of proteins in the graph might exceed the total number of identified proteins because some were grouped
into more than one functional category

hypothetical protein EGR_06319, and triosephosphate Discussion

isomerase. In this study, we performed the first proteomic analysis of
A total of 61 adult worm protein spots corresponding ES products from adult E. granulosus. Enolase was the
to 26 different proteins were identified (Table 3); 2 of most abundant ES product Enolase has been described as
these proteins were identified in E. granulosus adult a multifunctional surface protein and an ES product of
worms for the first time. parasites; it exhibits host-interacting properties and is

Fig. 3 Antigenic proteins on corresponding representative 2DE gels of the proteins expressed by E. granulosus adult worms. Proteins (800 μg)
were separated on a linear pH range of 3–10 by using IEF in the first dimension and 12 % SDS-PAGE in the second dimension. Proteins were
electrotransferred to PVDF membranes and probed with sera from E. granulosus-infected dogs. Antigenic protein spots are indicated by red
circles. Molecular weight markers are shown on the left. The proteins identified are shown in detail in Table 2
Wang et al. BMC Veterinary Research (2015) 11:119 Page 5 of 7

Table 2 Antigenic proteins of E. granulosus adult worms identified by immunoblot analysis

GI number Protein name Species MW pI Protein score C.I.%
547974 Paramyosin E. granulosus 98681.8 5.21 100
148613837 Calreticulin E. granulosus 42199.2 4.47 100
256274460 HSP60 E. granulosus
674568928 HSP70 E. granulosus 70640 5.47 100
26399708 Severina E. granulosus 41681.9 5.74 100
262192839 Enolase E. granulosus 46531.9 6.48 100
576695773 Actin E. granulosus 41717.8 5.3 100
a
576695197 Hypothetical protein EGR_06319 E. granulosus 23119 4.77 100
6016537 Malate dehydrogenase, cytoplasmic E. granulosus 36627.8 8.11 100
a
576692880 Triosephosphate isomerase E. granulosus 27149.8 6.6 100
576691284 Superoxide dismutase E. granulosus 15442.8 6.15 99.992
31077167 Cyclophilin E. granulosus 17343.4 6.41 99.003
a
Antigenic proteins identified for the first time in adult worms are shown in bold

Table 3 Proteins of E. granulosus adult worms identified by MALDI-TOF/TOF analysis

GI number Protein name Species MW pI Protein score C.I.%
547974 Paramyosin E. granulosus 98681.8 5.21 100
148613837 Calreticulin E. granulosus 42199.2 4.47 100
256274460 HSP60 E. granulosus
674568928 HSP70 E. granulosus 70640 5.47 100
576695773 Actin E. granulosus 41745.8 5.39 100
26399708 Severin E. granulosus 41681.9 5.74 100
262192839 Enolase E. granulosus 46531.9 6.48 100
576695773 Actina E. granulosus 41717.8 5.3 100
576695197 Hypothetical protein EGR_06319 E. granulosus 23119 4.77 100
6016537 Malate dehydrogenase, cytoplasmic E. granulosus 36627.8 8.11 100
576692880 Triosephosphate isomerase E. granulosus 27149.8 6.6 100
576691284 Superoxide dismutase E. granulosus 15442.8 6.15 99.992
31077167 Cyclophilin E. granulosus 17343.4 6.41 99.003
29336624 78-kDa glucose-regulated protein E. granulosus 71876.1 5.16 100
674566315 Transitional endoplasmic reticulum atpase E. granulosus 71581.6 5.17 100
110558962 Ferritin E. granulosus 16676.2 5.24 100
189016336 Thioredoxin peroxidase E. granulosus 21419.7 5.78 100
674565853 Succinate dehydrogenase ubiquinone E. granulosus 54812.5 7.62 100
576691476 Transketolase E. granulosus 72574.6 6.53 100
576693013 T-complex protein 1 subunit zetaa E. granulosus 26703 7.77 100
182676451 Tropomyosin E. granulosus 32249.2 4.6 100
576698524 Small heat shock protein p36 E. granulosus 35821.2 5.92 99.995
674567794 Oncosphere protein Tso22e E. granulosus 31416.3 6.61 100
2316076 Glutathione S-transferase E. granulosus 25536.8 7.51 100
193213138 Phosphoglycerate mutase Chlorobaculum parvum 28366.6 5.76 95.745
576700988 Fructose-bisphosphate aldolase E. granulosus 39702.4 8.03 100
a
Proteins identified for the first time in adult worms are shown in bold
Wang et al. BMC Veterinary Research (2015) 11:119 Page 6 of 7

involved in invasion [13–17]. Enolase is likely to play Some of these proteins have potential in vaccine devel-
an important role in E. granulosus-host interactions opment, such as paramyosin. Paramyosins, which are
and parasite evasion mechanisms via the immunomo- multifunctional modulators of the host immune re-
dulation of the host immune system. The second most sponse, play a role in binding complement components,
abundant ES product was cyclophilin, which plays a immunoglobulins, and secreted components of cellular
role in parasite development and host-parasite interac- immune response [31–36]. Paramyosin has been pro-
tions [18]. Furthermore, cyclophilin is a target com- posed by WHO/TDR as a vaccine candidate against
pound for the immunosuppressive agent, cyclosporine schistosomiasis and has shown some protection, which
A [19–21]. may suggest a potential for this protein in the treat-
Actin and paramyosin, cytoskeletal proteins, were other ment of E. granulosus infections [37–39].
ES products. Similar results have been obtained in other Recently, the protein profile of E. granulosus adult
parasites [22–25]. The presence of these proteins may in- worms has been analyzed by two dimensional LC-MS
dicate parasite damage or death; however, it is more likely [40]. In addition to proteins that had been reported in
that these proteins are products of tegument shedding. that work, we identified two novel proteins of this para-
The continuous shedding of the parasite tegument is site, which has increased the known repertoire of the
thought to release components that aid the parasite in parasitic protein profile and demonstrates the value of
evading an effective immune response [26]. our experimental approaches for the proteomic analysis
An antioxidant defense mechanism is another form of of E. granulosus proteins.
parasite survival. Among the ES products identified, only
thioredoxin peroxidase was involved in redox homeosta- Conclusions
sis. This enzyme plays a major role in protecting the The present study investigated the ES products and anti-
adult worms from oxidative damage. This result is in ac- genic proteins of E. granulosus adult worms from in-
cordance with the findings of a study performed of fected dogs. The results obtained from this study extend
hydatid cyst fluid (HCF) that contained ES products the existing proteomic data regarding E. granulosus and
from E. granulosus protoscoleces [9]. could assist in the development of vaccine antigens,
Metacestodes (hydatid cysts), protoscoleces, and adult drug targets, and immunodiagnostic markers.
worms are different developmental stages of E. granulo-
sus. The HCF is a reservoir of ES products from both Abbreviations
2DE: Ttwo-dimensional polyacrylamide gel electrophoresis; CE: Cystic
the germinal layer and protoscoleces. Compared with echinococcosis; ES: Excretory-secretory products; GO: Gene ontology;
HCF [9] and ES products from protoscoleces [11], the KOG: Eukaryotic Orthologous Group (KOG); MALDI-TOF/TOF: Tandem
ES products from adult worms comprise different pro- matrix-assisted laser desorption/ionization time-of-flight.
teins. Some of the proteins in this study have not been
Competing interests
identified in HCF, including enolase, malate dehydrogen- The authors declare that they have no competing interests.
ase, GST, and cyclophilin. Similarly, some of the proteins
have not been identified in the ES products from proto- Authors’ contributions
scoleces, including paramyosin, small HSP p36, and GST. In YW designed the study and experiments, interpreted the experimental data
and drafted the manuscript; DX, FZ and HZ performed the experiments; XH,
another study, we observed that the ES products from E. XL and WW contributed to helminthes collection; YS helped to data analysis;
granulosus adult worms failed to induce dendritic cell JZ conducted the experiments perform; JC conducted the study design and
maturation and cytokine production (data not pub- perform, and delivered the manuscript draft. All authors read and approved
the final manuscript.
lished); this result, however, is in contrast with the find-
ing that HCF induces dendritic cell activation [27]. Acknowledgments
These differences could be attributed to the different This work was supported by grants from the National Natural Science
proteins in ES products and HCF; however, this needs Foundation (No. 81371841 to J.C., 81371842 to Y.S.), and the National
S & T Major Program (No.2012ZX10004-201 to J.C.).
to be confirmed.
The search for antigenic proteins by 2DE immunoblots Author details
1
contributed to the identification of 12 parasite proteins National Institute of Parasitic Diseases, Chinese Center for Disease Control
and Prevention; Laboratory of Parasite and Vector Biology, MOH, China; WHO
that were recognized in the serum of E. granulosus-in- Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai
fected dogs. Nine of these proteins (malate dehydrogenase, 200025, China. 2Collaborative Innovation Center for Diagnosis and Treatment
HSP60, HSP70, calreticulin, enolase, actin, cyclophilin, of Infectious Diseases, State Key Laboratory for Infectious Disease Prevention
and Control, National Institute for Communicable Disease Control and
paramyosin and superoxide dismutase) were previously Prevention, Chinese Center for Disease Control and Prevention, Beijing
identified in E. granulosus and E. multilocularis human in- 102206, China. 3Qinghai Institute for Endemic Disease Prevention and
fections [9, 18, 28–30]. The other three proteins (severin, Control, Xining, 811602, China.
hypothetical protein EGR_06319 and triosephosphate Received: 23 November 2014 Accepted: 29 April 2015
isomerase,) were identified in this study for the first time.
Wang et al. BMC Veterinary Research (2015) 11:119 Page 7 of 7

References 25. Bernal D, Carpena I, Espert AM, De la Rubia JE, Esteban JG, Toledo R, et al.
1. Moro P, Schantz PM. Echinococcosis: a review. Int J Infect Dis. 2009;13:125–33. Identification of proteins in excretory/secretory extracts of Echinostoma friedi
2. Craig PS, McManus DP, Lightowlers MW, Chabalgoity JA, Garcia HH, Gavidia (Trematoda) from chronic and acute infections. Proteomics. 2006;6:2835–43.
CM, et al. Prevention and control of cystic echinococcosis. Lancet Infect Dis. 26. Van Hellemond JJ, Retra K, Brouwers JF, van Balkom BW, Yazdanbakhsh M,
2007;7:385–94. Shoemaker CB, et al. Functions of the tegument of schistosomes: clues from
3. Eckert J, Conraths FJ, Tackmann K. Echinococcosis: an emerging or the proteome and lipidome. Int J Parasitol. 2006;36:691–9.
re-emerging zoonosis? Int J Parasitol. 2000;30:1283–94. 27. Kanan JH, Chain BM. Modulation of dendritic cell differentiation and
4. McManus DP, Zhang W, Li J, Bartley PB. Echinococcosis. Lancet. cytokine secretion by the hydatid cyst fluid of Echinococcus granulosus.
2003;362:1295–304. Immunology. 2006;118:271–8.
5. McManus DP. Echinococcosis with particular reference to Southeast Asia. 28. Wang YH, Cheng Z, Lu XF, Tang CT. Echinococcus multilocularis: proteomic
Adv Parasitol. 2010;72:267–303. analysis of the protoscoleces by two-dimensional electrophoresis and mass
6. Casaravilla C, Pittini A, Ruickerl D, Seoane PI, Jenkins SJ, MacDonald AS, et al. spectrometry. Exp Parasitol. 2009;123:162–7.
Unconventional maturation of dendritic cells induced by particles from the 29. Virginio VG, Hernandez A, Rott MB, Monteiro KM, Zandonai AF, Nieto A,
laminated layer of larval Echinococcus granulosus. Infect Immun. et al. A set of recombinant antigens from Echinococcus granulosus with
2014;82:3164–76. potential for use in the immunodiagnosis of human cystic hydatid disease.
7. Nono JK, Pletinckx K, Lutz MB, Brehm K. Excretory/secretory-products of Clin Exp Immunol. 2003;132:309–15.
Echinococcus multilocularis larvae induce apoptosis and tolerogenic 30. Colebrook AL, Lightowlers MW. Serological reactivity to heat shock protein
properties in dendritic cells in vitro. PLoS Negl Trop Dis. 2012;6:e1516. 70 in patients with hydatid disease. Parasite Immunol. 1997;19:41–6.
8. Chemale G, van Rossum AJ, Jefferies JR, Barrett J, Brophy PM, Ferreira HB, 31. Gobert GN, McManus DP. Update on paramyosin in parasitic worms.
et al. Proteomic analysis of the larval stage of the parasite Echinococcus Parasitaol Int. 2005;54:101–7.
granulosus: Causative agent of cystic hydatid disease. Proteomics. 32. Wei J, Gu Y, Yang Y, Wang S, Cui S, Zhu X. Identification and
2003;3:1633–6. characterization of protective epitope of Trichinella spiralis paramyosin.
9. Monteiro KM, de Carvalho MO, Zaha A, Ferreira HB. Proteomic analysis of Vaccine. 2011;29:3162–8.
the Echinococcus granulosus metacestode during infection of its 33. Jiz M, Wu HW, Meng R, Pond-Tor S, Reynolds M, Friedman JF, et al. Pilot-
intermediate host. Proteomics. 2010;10:1985–99. scale production and characterization of paramyosin, a vaccine candidate
10. Aziz A, Zhang W, Li J, Loukas A, McManus DP, Mulvenna J. Proteomic for schistosomiasis japonica. Infect Immun. 2008;76:3164–9.
characterization of Echinococcus granulosus hydatid cyst fluid from sheep, 34. Zhao QP, Moon SU, Na BK, Kim SH, Cho SH, Lee HW, et al. Paragonimus
cattle and humans. J Proteomics. 2011;74:1560–72. westermani: biochemical and immunological characterization of paramyosin.
11. Virginio VG, Monteiro KM, Drumond F, de Carvalho MO, Vargas DM, Zaha A, Exp Parasitol. 2007;115:9–18.
et al. Excretory/secretory products from in vitro-cultured Echinococcus 35. Park TJ, Kang JM, Na BK, Sohn WM. Molecular cloning and characterization
granulosus protoscoleces. Mol Biochem Parasitol. 2012;183:15–22. of a paramyosin from Clonorchis sinensis. Korean J Parasitol. 2009;47:359–67.
12. Zou Q, Yan X, Li B, Zeng X, Zhou J, Zhang J. Proteome analysis of sorbitol 36. Jiz M, Fridman JF, Leenstra T, Jarilla B, Pablo A, Langdon G, et al.
fermentation specific protein in Vibrio cholerae by 2-DE and MS. Proteomics. Immunoglobulin E (IgE) responses to paramyosin predict resistance to
2006;6:1848–55. reinfection with Schistosoma japonicum and are attenuated by IgG4. Infect
13. Perez-Sanchez R, Ramajo-Hernandez A, Ramajo-Martin V, Oleaga A. Proteomic Immun. 2009;77:2051–8.
analysis of the tegument and excretory-secretory products of adult 37. Hemandez MG, Hafalla JC, Acosta LP, Aligui FF, Aligui GD, Ramirez BL, et al.
Schistosoma bovis worms. Proteomics. 2006;6:226–36. Paramyosin is a major target of the human IgA response against
14. Bernal D, de la Rubia JE, Carrasco-Abad AM, Toledo R, Mas-Coma S, Marcilla A. Schistosoma japonicum. Parasite Immunol. 1999;21:641–7.
Identification of enolase as a plasminogen-binding protein in excretory- 38. Zhang DM, Pan WQ, Qian L, Duke M, Shen LH, McManus DP. Investigation
secretory products of Fasciola hepatica. FEBS let. 2004;563:203–6. of recombinant Schistosoma japonicum paramyosin fragments for
15. Seweryn E, Pietkiewicz J, Szamborska A, Gamian A. Enolase on the surface immunogenicity and vaccine efficacy in mice. Parasite Immunol.
of prokaryotic and eukaryotic cells is a receptor for human plasminogen. 2006;28:77–84.
Postepy Hig Med Dosw. 2007;61:672–82. 39. Chen G, Dai Y, Chen J, Wang X, Tang B, Zhu Y, et al. Oral delivery of the
Sj23LHD-GST antigen by Salmonella typhimurium type III secretion system
16. Ramajo-Hernandez A, Perez-Sanchez R, Ramajo-Martin V, Oleaga A.
protects against Schistosoma japonicum infection in mice. PLoS Negl Trop
Schistosoma bovis: plasminogen binding in adults and the identification of
Dis. 2011;5:e1313.
plasminogen-bingding proteins from the worm tegument. Exp Parasitol.
40. Cui SJ, Xu LL, Zhang T, Xu M, Yao J, Fang CY, et al. Proteomic
2007;115:83–91.
characterization of larval and adult developmental stages in Echinococcus
17. Antunez K, Anido M, Arredondo D, Evans JD, Zunino P. Paenibacillus larvae
granulosus reveals novel insight into host-parasite interactions. J Proteomics.
enolase as a virulence factor in honeybee larvae infection. Vet Microbiol.
2013;84:158–75.
2011;10:83–9.
18. Ortona E, Vaccari S, Margutti P, Delunardo F, Rigano R, Profumo E, et al.
Immunological characterization of Echinococcus granulosus cyclophilin, an
allergen reactive with IgE and IgG4 from patients with cystic
echinococcosis. Clin Exp Immunol. 2002;128:124–30.
19. Galat A. Function-dependent clustering of orthologues and paralogues of
cyclophilins. Proteins. 2004;56:808–20.
20. Bell A, Monaghan P, Page AP. Peptidyl-prolyl cis-trans isomerases (immunophilins)
and their roles in parasite biochemistry, host-parasite interaction and antiparasitic Submit your next manuscript to BioMed Central
drug action. Int J Parasitol. 2006;36:261–76. and take full advantage of:
21. Kameyama K, Nishimura M, Punsantsogvoo M, Ibrahim HM, Xuan X, Furuoka
H, et al. Immunological characterization of Neospora caninum cyclophilin.
• Convenient online submission
Parasitology. 2012;139:294–301.
22. Mulvenna J, Sripa B, Brindley PJ, Gorman J, Jones MK, Colgrave ML, et al. • Thorough peer review
The secreted and surface proteomes of adult stage of the carcinogenic • No space constraints or color figure charges
human liver fluke Opisthorchis viverrini. Proteomics. 2010;10:1063–78.
• Immediate publication on acceptance
23. Kundsen GM, Medzihradszky KF, Lim KC, Hansell E, McKerrow JH. Proteomic
analysis of Schistosoma mansoni cercarial secretions. Mol Cell Proteomics. • Inclusion in PubMed, CAS, Scopus and Google Scholar
2005;4:1862–75. • Research which is freely available for redistribution
24. Liu F, Cui SJ, Hu W, Feng Z, Wang ZQ, Han ZG. Excretory/secretory
proteome of the adult developmental stage of human blood fluke,
Schistosoma japonicum. Mol Cell Proteomics. 2009;8:1236–51. Submit your manuscript at
www.biomedcentral.com/submit