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Chromosome-level genome assembly of Oncomelania hupensis: the intermediate snail host of schistosoma japonicum

Infectious Diseases of Poverty 2024年第1期13卷 16-26页

作者： Qin Liu Lei Duan Yun-Hai Guo Li-Min Yang Yi Zhang Shi-Zhu Li Shan Lv Wei Hu Nan-Sheng Chen Xiao-Nong Zhou National Institute of Parasitic Diseases at Chinese Center for Disease Control and Prevention(Chinese Centerfor Tropical Diseases Research) NHC Key Laboratory of Parasite and Vector BiologyWHO Collaborating Centre for Tropical DiseasesNational Center for International Research on Tropical DiseasesNational Key Laboratory of Intelligent Tracking and Forecasting for Infectious DiseasesShanghai 200025People's Republic of China School of Global Health Chinese Center for Tropical Diseases ResearchShanghai Jiao Tong University School of MedicineShanghai 200025People's Republic of China School of Life Science Fudan UniversityShanghai 200438People's Republic of China CAS Key Laboratory of Marine Ecology and Environmental Sciences Institute of OceanologyChinese Academy of SciencesQingdaoShandong 266071People'sRepublicofChina

Background schistosoma japonicum is a parasitic flatworm that causes human schistosomiasis,which is a signif-cant cause of morbidity in China,the Philippines and Indonesia.Oncomelania hupensis(Gastropoda:Pomatiopsidae)is the unique intermediate host of S.japonicum.A complete genome sequence of O.hupensis will enable the funda-mental understanding of snail biology as well as its co-evolution with the S.japonicum parasite.Assembling a high-quality reference genome of O.hupehensis will provide data for further research on the snail biology and controlling the spread of S.japonicum.Methods The draft genome was de novo assembly using the long-read sequencing technology(PacBio Sequel I)and corrected with llumina sequencing data.Then,using Hi-C sequencing data,the genome was assembled at the chromosomal level.CAFE was used to do analysis of contraction and expansion of the gene family and CodeML module in PAML was used for positive selection analysis in protein coding sequences.Results A total length of 1.46 Gb high-quality O.hupensis genome with 17 unique full-length chromosomes(2n=34)of the individual including a contig N50 of 1.35 Mb and a scaffold N50 of 75.08 Mb.Additionally,95.03%of these con-tig sequences were anchored in 17 chromosomes.After scanning the assembled genome,a total of 30,604 protein-coding genes were predicted.Among them,86.67%were functionally annotated.Further phylogenetic analysis revealed that O.hupensis was separated from a common ancestor of Pomacea canaliculata and Bellamya purificata approximately 170 million years ago.Comparing the genome of O.hupensis with its most recent common ancestor,it showed 266 significantly expanded and 58 significantly contracted gene families(P<0.05).Functional enrichment of the expanded gene families indicated that they were mainly involved with intracellular,DNA-mediated transposi-tion,DNA integration and transposase activity.Conclusions Integrated use of multiple sequencing technologies,we have successfully constructed the genome at the chromosomal-level of O

关键词： Schistosomiasis schistosoma japonicum Oncomelania hupensis Chromosome-level genome

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schistosoma japonicum: Isolation and Identification of Peptides Mimicking Ferritin Epitopes from Phage Display Library

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Acta Biochimica et Biophysica Sinica 2004年第3期36卷 206-210页

作者： Lian-Fei TANG, Xin-Yuan YI*, Xian-Fang ZENG, Lin-Qian WANG, and Shun-Ke ZHANG Department of Parasitology, Central South University Xiangya School of Medicine, Changsha 410078, China Department of Parasitology Central South University Xiangya School of Medicine Changsha 410078 China

In an attempt to isolate and identify the antigenic epitopes on ferritin of schistosoma japonicum (SjFer) and to test their protective potentiality against schistosoma japonicum (S.j), polyclonal antisera against SjFer was prepared to screen a 12-mer random peptide library. Three rounds of biopan- ning were performed and resulted in an enrichment. Six peptides selected randomly from the third elute were all found to be positive by evaluating the binding to anti-SjFer sera by ELISA and Western blot- ting. Three amino acid sequences were deduced from the six phage clones by sequencing. When they were used to vaccinate mice, the three peptides could induce significant reduction in adult worms (26.7%, 20.4%, and 25.9%) as well as in liver eggs per gram (LEPG) (40.0%, 38.2%, and 40.8%). This result showed that three mimotopes on SjFer were obtained and they could induce significant protective efficacy against S.j.

关键词： phage displayed peptide library epitope schistosoma japonicum ferritin

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schistosoma japonicum attenuates dextran sodium sulfateinduced colitis in mice via reduction of endoplasmic reticulum stress

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World Journal of Gastroenterology 2017年第31期23卷 5700-5712页

作者： Ya Liu Qing Ye Yu-Lan Liu Jian Kang Yan Chen Wei-Guo Dong Department of Gastroenterology Key Laboratory of Hubei Province for Digestive System DiseaseRenmin Hospital of Wuhan University Department of Hospital Infection Renmin Hospital of Wuhan University Departments of Critical Care Medicine Renmin Hospital of Wuhan University

AIM To elucidate the impact of schistosoma(S.) japonicum infection on inflammatory bowel disease by studying the effects of exposure to S. japonicum cercariae on dextran sodium sulfate(DSS)-induced colitis. METHODS Infection was percutaneously established with 20 ± 2 cercariae of S. japonicum, and colitis was induced by administration of 3% DSS at 4 wk post infection. Weight change, colon length, histological score(HS) and disease activity index(DAI) were evaluated. Inflammatory cytokines, such as IL-2, IL-10 and IFN-γ, were tested by a cytometric bead array and real-time quantitative polymerase chain reaction(RT-PCR). Protein and m RNA levels of IRE1α, IRE1β, GRP78, CHOP, P65, P-P65, P-IκBα and IκBα in colon tissues were examined by Western blot and RT-PCR, respectively. Terminal deoxynucleotidyl transferase-mediated d UTP nick-end labeling positive cells, cleaved-caspase 3 expression and Bcl2/Bax were investigated to assess the apoptosis in colon tissues.RESULTS Mice infected with S. japonicum cercariae were less susceptible to DSS. Mice infected with S. japonicum cercariae and treated with DSS showed decreased weight loss, longer colon, and lower HS and DAI compared with mice treated with DSS alone. A substantial decrease in Th1/Th2/Th17 response was observed after infection with S. japonicum. Endoplasmic reticulum(ER) stress and the nuclear factor-kappa B(NF-κB) pathway were reduced in mice infected with S. japonicum cercariae and treated with DSS, along with ameliorated celluar apoptosis, in contrast to mice treated with DSS alone. CONCLUSION Exposure to S. japonicum attenuated inflammatory response in a DSS-induced colitis model. In addition to the Th1/Th2/Th17 pathway and NF-κB pathway, ER stress was shown to be involved in mitigating inflammation and decreasing apoptosis. Thus, ER stress is a new aspect in elucidating the relationship between helminth infection and inflammatory bowel disease(IBD), which may offer new therapeutic methods for IBD.

关键词： Endoplasmic reticulum stress schistosoma japonicum Colitis

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Identification of Immunodominant Th1-type T cell Epitopes from schistosoma japonicum 28 kDa Glutathione-S-transferase,a Vaccine Candidate

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Acta Biochimica et Biophysica Sinica 2005年第11期37卷 751-758页

作者： Guang-Fu LI Yong WANG Zhao-Song ZHANG Xin-Jun WANG Min-Jun JI Xiang ZHU Feng LIU Xiao-Ping CAI Hai-Wei WU Guan-Ling WU Department of Pathogenic Biology,Nanjing Medical University,Nanjing 210029,China Department of Pathogenic Biology Nanjing Medical UniversityNanjing 210029 China

Th1-type cytokines produced by the stimulation of Th1-type epitopes derived from definedschistosome-associated antigens are correlated with the development of resistance to the parasite infection.schistosoma mansoni 28 kDa glutathione-S-transferase(Sm28GST),a major detoxification enzyme,hasbeen recognized as a vaccine candidate and a phase Ⅱ clinical trial has been carried out.Sheep immunizedwith recombinant schistosoma japonicum 28GST(Sj28GST)have shown immune protection against theparasite infection.In the present study,six candidate peptides(P1,P2,P3,P4,P7 and P8)from Sj28GSTwere predicted,using software,to be T cell epitopes,and peptides P5 and P6 were designed by extendingfive amino acids at the N-terminal and C-terminal of P1,respectively.The peptide 190-211 aa in Sj28GSTcorresponding to the Th1-type epitope(190-211 aa)identified from Sm28GST was selected and named P9.The nine candidate peptides were synthesized or produced as the fusion protein with thioredoxin in thepET32c(+)/BL21(DE3)system.Their capacity to induce a Th1-type response in vitro was measured usinglymphocyte proliferation,cytokine detection experiments and flow cytometry.The results showed that P6(73-86 aa)generated the strongest stimulation effect on T cells among the nine candidate peptides,and drovethe highest level of IFN-γ and IL-2.Therefore,P6 is a functional Th1-type T cell epitope that is differentfrom that in Sm28GST,and will be useful for the development of effective vaccines which can triggeracquired immunity against S.japonicum.Moreover,our strategy of identifying the Th1-type epitope by acombination of software prediction and experimental confirmation provides a convenient and cost-savingalternative approach to previous methods.

关键词： schistosoma japonicum epitope Sj28GST Th1 epitope IFN-γ

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Dynamics of CD4~+CD25~+ T Cells in Spleens and Mesenteric Lymph Nodes of Mice Infected with schistosoma japonicum

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Acta Biochimica et Biophysica Sinica 2006年第5期38卷 299-304页

作者： Xiao-Ping CAI Hui ZHANG Yong-Chen ZHANG Yong WANG Chuan SU Min-Jun JI Hai-Wei WU Xiang ZHU Zhao-Song ZHANG Guan-Ling WU Department of Pathogenic Biology,Jiangsu Province Key Laboratoly of Modern Pathogenic Biology,Nanjing Medical University,Nanjing 210029,China Department of Pathogenic Biology Jiangsu Province Key Laboratory of Modern Pathogenic Biology Nanjing Medical UniversityNanjing 210029 China

CD4~+CD25~+T cells play a major role in modulating immune response,but few reports havebeen published about schistosomiasis.Here,we investigated the changes in CD4~+CD25~+T cell populations inspleens and mesenteric lymph nodes of mice infected with schistosoma japonicum.The proportions ofCD4~+CD25~+T cells in total CD4~+T cells were analyzed by flow cytometry.CD25 and Foxp3 expression wasmeasured by real-time quantitative polymerase chain reaction.The suppressive activities of CD4~+CD25~+Tcells were detected by in vitro proliferation of splenocytes.Evidence showed that the percentage of CD4~+CD25~+T cells was the same as controls 3 weeks post-infection.At the acute stage of infection,the percentagedecreased significantly.However,at the chronic stage of infection,it rebounded to normal levels or evenhigher.The expression of the CD25 and Foxp3 showed gradual increase along with the infection progress.Invitro experiment also showed the strong suppressive effect of CD4~+CD25~+T cells,isolated during the chronicstage,on proliferation of the CD25~-splenocytes.This is the first time that the dynamics of CD4~+CD25~+T cellpopulations was demonstrated in mice infected with schistosomiasis.In conclusion,our data indicated thatCD4~+CD25~+cells might be involved in the immune modulation during S.japonicum infection,which en-hances current knowledge of the mechanisms of the immuno-downregulation and re-infection inschistosomiasis.

关键词： schistosoma japonicum CD4~+CD25~+ T cell chronic infection Foxp3

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Vaccination against schistosoma japonicum Infection by DNA Vaccine Encoding Sj22.7 Antigen

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Acta Biochimica et Biophysica Sinica 2007年第1期39卷 27-36页

作者： Gan DAI Shiping WANG Junlong YU Shaorui XU Xianchu PENG Zhuo HE Xueqin LIU Songhua ZHOU and Fen LIU Institute of Schistosomiasis Research.Xiangya School of Medicine,Central South University,Changsha 410078 China Institute of Schistosomiasis Research Xiangya School of Medicine Central South UniversityChangsha 410078 China

To observe the in vitro expression of DNA vaccine pcDNA3-Sj22.7 and its immunologicaleffect in mice,the recombinant plasmid pcDNA3-Sj22.7 was used to transfect HeLa cells with liposome-mediated method and the expression of Sj22.7 mRNA and protein was examined using reversetranscription-polymerase chain reaction, sodium dodecylsulfate-polyacrylamide gel electrophoresis andWestern blot.Then,the ability of pcDNA3-Sj22.7 to protect against schistosoma japonicum challengeinfections was analyzed according to worm reduction rate and egg reduction rate after vaccination of mice.The serum levels of specific IgG antibody and T lymphocyte proliferation response were also determined.After the challenge infection,Sj22.7-driven interferon (IFN)-γand interleukin (IL)-4 was also quantified.Results showed that pcDNA3-Sj22.7 could express Sj22.7 mRNA and protein in vitro.Immunizationresulted in a worm reduction rate of 29.70%,egg reduction rate of 47.25% (liver) and 51.73% (intestine),and egg reduction rate of 25.90% (eggs per female),suggesting induction of significant anti-fecundityin the pcDNA3-Sj22.7 group. Enzyme-linked immunosorbent assay and Western blot analysis indicatedthat immunized mice generated specific IgG against Sj22.7.T lymphocytes from mice immunized withpcDNA3-Sj22.7 showed a significant proliferation response to rSj22.7.The culture of spleen cells showedthat secretion of IFN-γ increased but IL-4 decreased. The results indicate that DNA vaccination bypcDNA3-Sj22.7 is sufficient to elicit significant levels of protective immunity against S.japonicum infection.The DNA vaccine could induce significant cellular and humoral immune response,and display predominantT helper cell type 1 type immune responses,which contribute to the protective immunity against challengeinfection in mice.

关键词： response schistosoma japonicum DNA vaccine Sj22.7 gene expression immune protection immune

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Characterization of CD4~+ T Cell Responses in Mice Infected with schistosoma japonicum

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Acta Biochimica et Biophysica Sinica 2006年第5期38卷 327-334页

作者： Min-Jun JI Chuan SU Yong WANG Hai-Wei WU Xiao-Ping CAI Guang-Fu LI Xiang ZHU Xin-Jun WANG Zhao-Song ZHANG Guan-Ling WU~* Department of Pathogen Biology,Nanjing Medical University,Nanjing 210029,China Department of Pathogen Biology Nanjing Medical UniversityNanjing 210029 China

To better understand the interaction between schistosoma japonicum and its murine host,wecharacterized the immune response of CD4~+ T cells generated during an experimental S.japonicum infectionbased on different key aspects,from gene expression to cell behavior.Mouse oligonucleotide microarrayswere used to compare gene expression profiles of CD4~+ T cells from spleens of mice at 0,3,6 and 13weeks post-infection.Flow cytometry analysis was used to determine type 1 and type 2 cytokine-secretingCD4~+ T cells,to test apoptosis of CD4~+ T cells and to count CD4~+CD25~+ T cells,a kind of regulatorysubpopulation of CD4~+ T cells.The percentage of interleukin-4-producing CD4~+ T cells was found to bemuch higher than that of γ-interferon-producing cells,especially after stimulation with S.japonicum eggantigen,which was consistent with type 1 and type 2 cytokine gene expression in the genechip.Microarraydata also showed that S.japonicum induced the increased expression of Th2 response-related genes,whereassome transcripts related to the Th1 responsive pathway were depressed.Flow cytometry analysis showeda marked increase in the apoptotic CD4~+ T cells from 6 weeks post-infection and in the ratio of CD4~+CD25~+to CD4~+ T cells in infected mice after 13 weeks.We therefore concluded that experimental infection of micewith S.japonicum resulted in a Th2-skewed immune response,which was to a great extent monitored bythe immune regulatory network,including cytokine cross-modulation,cell apoptosis and the subpopulationof regulatory cells.

关键词： schistosoma japonicum CD4~+ T cell immune deviation immune regulatory network

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IFN-inducible p47 GTPases display differential responses to schistosoma japonicum acute infection

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Cellular & Molecular Immunology 2010年第1期7卷 69-76页

作者： Xuesong Chen Xiaotang Du Meijuan Zhang Donghui Zhang Minjun Ji Guanling Wu Department of Pathogen Biology and Immunology Nanjing Medical UniversityNanjingChina Jiangsu Province Key Laboratory of Modern Pathogen Biology NanjingChina

Interferon gamma induced GTPase(IGTP)(also named lrgm3)and interferon gamma inducible protein 47(IRG-47)(also named lIrgd)are interferon(IFN)-inducible p47 GTPases that have been shown to regulate host resistance to intracellular pathogens.Little knowledge has been known about the role of p47 GTPases in host responses against extracellular pathogens.To investigate possible roles of IGTP and IRG-47 in the course of schistosoma japonicum infection,IGTP and IRG-47 knockout and wild-type(WT)mice were challenged with cercariae of S.japonicum,and host responses were analyzed.At the acute stage of S.japonicum infection,mice that lacked IGTP displayed similar parasite burden and pathological damage to WT mice.Importantly,S.japonicum-infected lRG-47-deficient mice,in contrast to IGTP-deficient mice and WT mice,showed significantly reduced worms and lower egg-burden,but intense granulomatous reaction evoked by schistosome eggs in peripheral parts of liver lobes.In addition,upregulation of inflammation-related gene expression was observed in the spleen of IRG-47-deficient mice using oligonucleotide microarrays,in which multiple pathways of cytokine-cytokine receptor interaction,T-cell receptor signaling,complement,coagulation cascades and cell adhesion molecules were highlighted.Taken together,these data suggest that IGTP and IRG-47 might have distinct features that were differentially required for resistance to S.japonicum.

关键词： acute infection immunity p47 GTPase schistosoma japonicum

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Optimisation of the DNA dipstick as a rapid extraction method for schistosoma japonicum in infected mice samples and spiked human clinical samples

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Infectious Diseases of Poverty 2023年第4期12卷 108-108页

作者： Oyime P.Aula Donald P.McManus Malcolm K.Jones Hong You Pengfei Cai Catherine A.Gordon Molecular Parasitology Laboratory QIMR Berghofer Medical Research Institute Brisbane Australia School of Public Health Faculty of Medicine The University of Queensland Brisbane Australia School of Veterinary Science The University of Queensland Brisbane Australia Faculty of Medicine The University of Queensland Brisbane Australia

BackgroundSchistosomiasis remains a public health issue and the need for accurate and affordable diagnostics is crucial in the elimination of the disease. While molecular diagnostics are highly effective, they are expensive, with the main costs been associated with DNA extraction. The DNA dipstick is a rapid, affordable and simple purification method that allows DNA to be extracted from diagnostic samples within 30 s. We aimed to optimise the DNA dipstick method for samples from mice and egg-spiked human samples.MethodsUrine, blood and faeces were collected from mice exposed to schistosoma japonicum infection at weekly intervals from Day 0 to Day 42. Urine and faecal samples were also collected from volunteer, uninfected humans and spiked with S. japonicum eggs. All samples were subject to several optimisation procedures and DNA extracted with the DNA dipstick. Amplification of the target DNA was carried out using LAMP and visualised using agarose gel electrophoresis and flocculation.ResultsThe DNA dipstick successfully identified S. japonicum from infected mice and human clinical samples spiked with cracked eggs or genomic DNA from S. japonicum. Amplification was observed from week 4 post infection in infected mice. For human samples, amplification was observed in sieved faecal samples, filtered urine samples heated at 95 ℃ for 30 min, and sera samples heated at 95℃ for 30 min.ConclusionsThe DNA dipstick combined with LAMP has huge potential in providing cost-effective, simple and accurate detection of schistosomiasis infection in endemic regions. This will allow for rapid treatment, tracking outbreaks—such as occur after typhoons, leading to better health outcomes and contributing to control and eventual elimination of schistosomiasis.

关键词： Schistosomiasis schistosoma japonicum DNA dipstick Loop-mediated isothermal amplification assay Flocculation

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Vaccination of Goats with 31 kDa and 32 kDa schistosoma japonicum Antigens by DNA Priming and Protein Boosting

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Cellular & Molecular Immunology 2007年第2期4卷 153-156页

作者： Lianfei Tang Zhijun Zhou Yuxiao Chen Yonghui Luo Linqian Wang Liyu Chen Fushen Huang Xianfang Zeng Xinyuan Yi Hunan Entry-Exit Inspection and Quarantine Bureau of the People＇s Republic of China Changsha 410007 Hunan China Department of Parasitology Xiangya School of Medicine Central South University Changsha 410078 Hunan China Department of Laboratory Animal Xiangya School of Medicine Central South University Changsha 410078 Hunan China College of Animal Science Hunan Agriculture University Changsha 410128 Hunan China

Two schistosomajaponicum vaccine candidate antigens Sj 31 and Sj 32, which have shown particular promise to induce protective immunity in mice, were used to immunize goats by using a DNA priming-protein boosting strategy in present work. DNA vaccine formulations of the two antigens （VRSj31 and VRSj32） were produced and injected intramuscularly twice at a 2-week interval and then recombinant proteins （rSj31 and rSj32） together with Freund Complete Adjuvant （FCA） were used to boost the goats. The experiment was repeated in different batche cercariae. A strong anamnestic antibody response was induced after boost. A significant reduction of liver egg counts and miracidial hatching was showed in both experiments. Significant protections against challenge infection were elicited with 31.6% of percentage reduction for worm recovery in the second experiment and 20.9% in the first experiment, respectively.

关键词： subunit vaccine DNA vaccine schistosoma japonicum

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