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Prevalence of antifolate drug resistance markers in Plasmodium vivax in China

Frontiers of Medicine 2022年第1期16卷 83-92页

作者： Fang Huang Yanwen Cui He Yan Hui Liu Xiangrui Guo Guangze Wang Shuisen Zhou Zhigui Xia National Institute of Parasitic Diseases Chinese Center for Disease Control and PreventionChinese Center for Tropical Diseases ResearchWHO Collaborating Center for Tropical DiseasesNational Centre for International Research on Tropical DiseasesNHC Key Laboratory of Parasite and Vector Biology(National Institute of Parasitic DiseasesChinese Center for Disease Control and Prevention)Shanghai 200025China Yunnan Institute of Parasitic Diseases Puer 665000China Yingjiang County for Disease Control and Prevention Yingjiang 679300China Hainan Center for Disease Control&Prevention Haikou 570203China

The dihydrofolate reductase(dhfr)and dihydropteroate synthetase(dhps)genes of Plasmodium vivax,as antifolate resistance-associated genes were used for drug resistance surveillance.A total of 375 P.vivax isolates collected from different geographical locations in China in 2009–2019 were used to sequence Pvdhfr and Pvdhps.The majority of the isolates harbored a mutant type allele for Pvdhfr(94.5%)and Pvdhps(68.2%).The most predominant point mutations were S117T/N(77.7%)in Pvdhfr and A383G(66.8%)in Pvdhps.Amino acid changes were identified at nine residues in Pvdhfr.A quadruple-mutant haplotype at 57,58,61,and 117 was the most frequent(57.4%)among 16 distinct Pvdhfr haplotypes.Mutations in Pvdhps were detected at six codons,and the double-mutant A383G/A553G was the most prevalent(39.3%).Pvdhfr exhibited a higher mutation prevalence and greater diversity than Pvdhps in China.Most isolates from Yunnan carried multiple mutant haplotypes,while the majority of samples from temperate regions and Hainan Island harbored the wild type or single mutant type.This study indicated that the antifolate resistance levels of P.vivax parasites were different across China and molecular markers could be used to rapidly monitor drug resistance.Results provided evidence for updating national drug policy and treatment guidelines.

关键词： drug resistance antifolates molecular markers Plasmodium vivax China

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Polymorphisms of potential drug resistant molecular markers in Plasmodium vivax from China-Myanmar border during 2008-2017

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Infectious Diseases of Poverty 2022年第2期11卷 93-94页

作者： Zhensheng Wang Chunyan Wei Yunchun Pan Zhihua Wang Xin Ji Qianqian Chen Lianhui Zhang Zenglei Wang Heng Wang Department of Microbiology and Parasitology Institute of Basic Medical Sciences Chinese Academy of Medical SciencesSchool of Basic Medicine Peking Union Medical College5#Dong Dan San TiaoBeijing 100005People’s Republic of China NHC Key Laboratory of Systems Biology of Pathogens Institute of Pathogen BiologyChinese Academy of Medical Sciences and Peking Union Medical College9#Dong Dan San TiaoBeijingPeople’s Republic of China 不详

Background:Plasmodium vivax remains the predominant species at the China–Myanmar border,imposing a major challenge to the recent gains in regional malaria elimination.To closely supervise the emerging of drug resistance in this area,we surveyed the variations in genes potentially correlated with drug resistance in P.vivax parasite and the possible drug selection with time.Methods:A total of 235 P.vivax samples were collected from patients sufering uncomplicated malaria at Yingjiang,Tengchong,and Longling counties,and Nabang port in China,Yunnan province,and Laiza sub-township in Myanmar,from 2008 to 2017.Five potential drug resistance genes were amplifed utilizing nested-PCR and analyzed,including pvdhfr,pvdhps,pvmdr1,pvcrt-o,and pvk12.The Pearson’s Chi-squared test or Fisher’s exact test were applied to determine the statistical frequency diferences of mutations between categorical data.Results:The pvdhfr F57I/L,S58R,T61M and S117T/N presented in 40.6%,56.7%,40.1%,and 56.0% of the sequenced P.vivax isolates,and these mutations signifcantly decreased with years.The haplotype formed by these quadruple mutations predominated in Yingjiang,Tengchong,Longling and Nabang.While a mutation H99S/R(56.6%)dominated in Laiza and increased with time.In pvdhps,the A383G prevailed in 69.2% of the samples,which remained the most prevalent haplotype.However,a signifcant decrease of its occurrence was also noticed over the time.The S382A/C and A553G existed in 8.4% and 30.8% of the isolates,respectively.In pvmdr1,the mutation Y976F occurred at a low frequency in 5/232(2.2%),while T958M was fxed and F1076L was approaching fxed(72.4%).The K10 insertion was detected at an occurrence of 33.2% in pvcrt-o,whereas there was no signifcant diference among the sites or over the time.No mutation was identifed in pvk12.Conclusions:Mutations related with resistance to antifolate drugs are prevalent in this area,while their frequencies decrease signifcantly with time,suggestive of increased susceptibility of P.vivax parasite to antifolate drugs

关键词： Plasmodium vivax Antimalarial drug Drug resistance Molecular marker China-Myanmar border

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Genetic diversity of Plasmodium vivax revealed by the merozoite surface protein-1 icb5-6 fragment

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Infectious Diseases of Poverty 2017年第1期6卷 801-809页

作者： Wei Ruan Ling-ling Zhang Yan Feng Xuan Zhang Hua-liang Chen Qiao-yi Lu Li-nong Yao Wei Hu Department of Communicable Diseases of Control and Prevention Zhejiang Provincial Centre for Disease Control and PreventionHangzhouChina School of Life Sciences FuDan UniversityShanghaiChina

Background:Plasmodium vivax remains a potential cause of morbidity and mortality for people living in its endemic areas.Understanding the genetic diversity of P.vivax from different regions is valuable for studying population dynamics and tracing the origins of parasites.The PvMSP-1 gene is highly polymorphic and has been used as a marker in many P.vivax population studies.The aim of this study was to investigate the genetic diversity of the PvMSP-1 gene icb5-6 fragment and to provide more genetic polymorphism data for further studies on P.vivax population structure and tracking of the origin of clinical cases.Methods:Nested PCR and sequencing of the PvMSP-1 icb5-6 marker were performed to obtain the nucleotide sequences of 95 P.vivax isolates collected from Zhejiang province,China.To investigate the genetic diversity of PvMSP-1,the 95 nucleotide sequences of the PvMSP-1 icb5-6 fragment were genotyped and analyzed using DnaSP v5,MEGA software.Results:The 95 P.vivax isolates collected from Zhejiang province were either indigenous cases or imported cases from different regions around the world.A total of 95 sequences ranging from 390 to 460 bp were obtained.The 95 sequences were genotyped into four allele-types(Sal I,Belem,R-III and R-IV)and 17 unique haplotypes.R-III and Sal I were the predominant allele-types.The haplotype diversity(Hd)and nucleotide diversity(Pi)were estimated to be 0.729 and 0.062,indicating that the PvMSP-1 icb5-6 fragment had the highest level of polymorphism due to frequent recombination processes and single nucleotide polymorphism.The values of dN/dS and Tajima’s D both suggested neutral selection for the PvMSP-1icb5-6 fragment.In addition,a rare recombinant style of R-IV type was identified.Conclusions:This study presented high genetic diversity in the PvMSP-1 marker among P.vivax strains from around the world.The genetic data is valuable for expanding the polymorphism information on P.vivax,which could be helpful for further study on population dynamics and tracking the origin of P.vivax.

关键词： Plasmodium vivax PvMSP-1 Genetic diversity Malaria

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Epidemiological characterization of imported recurrent Plasmodium vivax and plasmodium ovale in China,2013–2020

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Infectious Diseases of Poverty 2021年第4期10卷 104-104页

作者： Yanwen Cui Li Zhang Zhigui Xia Hejun Zhou Fang Huang National Institute of Parasitic Diseases Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research)Shanghai 200025China

Background China has reached important milestones in the elimination of malaria.However,the numbers of imported recurrent cases of Plasmodium vivax and P.ovale are gradually increasing,which increases the risk of malaria re-establishment in locations where Anopheles mosquitoes exist.The aim of this study is to characterize the epidemiological profiles of imported recurrent P.vivax and P.ovale cases,quantifying the recurrence burden and guiding the development of appropriate public health intervention strategies.Methods Individual-level data of imported recurrent P.vivax and P.ovale cases were collected from 2013 to 2020 in China via the Parasitic Diseases Information Reporting Management System.Demographic characteristics,temporal and spatial distributions,and the interval from previous infection to recurrence were analyzed by SAS,ArcGIS and GraphPad Prism software,respectively,to explore the epidemiological profiles of imported recurrent cases.Results A total of 307 imported recurrent cases,including 179 P.vivax and 128 P.ovale cases,were recorded.The majority of cases occurred in males(P.vivax 91.1%,P.ovale 93.8%)and migrant workers(P.vivax 43.2%,P.ovale 44.7%).Individuals aged 30–39 years had the highest P.vivax and P.ovale recurrent infection rates,respectively.The number of imported recurrent cases of infection by these two malaria species increased from 2013 to 2018,and P.vivax infection showed well-defined seasonality,with two peaks in February and June,respectively.More than 90%of patients with recurrent cases did not receive radical treatment for previous infection.Most imported recurrent P.vivax cases were reported in Yunnan Province and were imported from Myanmar,Ethiopia,and Pakistan,while most recurrent P.ovale cases were reported in southern China and primarily imported from Cameroon,Ghana,and Nigeria.The intervals from previous malaria infection to recurrence among different continents were significantly different(P=0.0016)for P.vivax malaria but not for P.ovale malaria(P=0.2373).Conclusions The la

关键词： Recurrence Imported malaria Plasmodium vivax plasmodium ovale China

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Plasmodium vivax Merozoite surface protein 8 is a highly immunogenic protein that localizes to the food vacuole of the malaria parasite

Plasmodium vivax Merozoite surface protein 8 is a highly imm...

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第十一届全国免疫学学术大会

作者： Cheng Yang Wang Bo Sattabongkot Jetsumon Tsuboi Takafumi Han Eun Taek Department of Parasitology Wuxi Medical School Jiangnan University Department of Medical Environmental Biology and Tropical Medicine School of Medicine Kangwon National University Chuncheon Mahidol vivax Research Unit Faculty of Tropical Medicine Mahidol University Division of Malaria Research Proteo-Science Center Ehime University

Plasmodium vivax merozoite surface protein 8(Pv MSP8)contains GPI motif and double EGF-like domains,which are considered potential targets of protective immunity.In our previous screening,recombinant Pv MSP8(r Pv MSP8)has been highly detected for recognizing with vivax-infected patient sera.However,the vaccine potential of Pv MSP8 needs adequately evaluated.In present study,purified r Pv MSP8 protein possessed potent antigenicity

关键词： Plasmodium vivax malaria MSP8 food vacuole Immunity

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Plasmodium vivax GPI-anchored micronemal antigen(PvGAMA) binds human erythrocytes independent of Duffy antigen status

Plasmodium vivax GPI-anchored micronemal antigen(PvGAMA) bin...

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中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工...

作者： Yang Cheng Feng Lu Bo Wang Jian Li Jin-Hee Han Daisuke Ito Deok-Hoon Kong Lubin Jiang Jian Wu Kwon-Soo Ha Eizo Takashima Jetsumon Sattabongkot Won Sun Park Seok-Ho Hong Sanjay A.Desai Louis H.Miller Takafumi Tsuboi Eun-Taek Han Department of Medical Environmental Biology and Tropical Medicine School of MedicineKangwon National University Department of Parasitology Wuxi Medical School Jiangnan University Laboratory of Malaria and Vector Research (LMVR) National Institute of Allergy and Infectious Diseases (NIAID) National Institutes of Health (NIH) Division of Malaria Research Proteo-Science Center Ehime University Department of Molecular and Cellular Biochemistry School of Medicine Kangwon National University Key Laboratory of Molecular Virology & Immunology Unit of Human Parasite Molecular and Cell Biology Institut Pasteur of Shanghai Mahidol vivax Research Unit Faculty of Tropical Medicine Mahidol University Department of Physiology School of Medicine Kangwon National University Department of Internal Medicine School of Medicine Kangwon National University

Plasmodium vivax, a major agent of malaria in both temperate and tropical climates, has a limited capacity for infecting humans lacking the Duffy(Fy) blood group antigen as this receptor is critical for erythrocyte invasion. Recent surveys in various endemic regions, however, have reported P. vivax infections in Duffy-negative individuals, suggesting that the parasite may utilize alternative receptor-ligand pairs to complete erythrocyte invasion. Here, we identified and characterized a novel parasite ligand, GPI-anchored micronemal antigen(Pv GAMA), that can bind human erythrocytes regardless of Duffy antigen status. Specific antibodies raised against PvG AMA localize the protein to the parasite microneme, consistent with a role in invasion. These antibodies also inhibited Pv GAMA binding to erythrocytes in a dose-dependent manner. Pv GAMA appears to play a role in a sialic acid-independent, chymotrypsin-sensitive invasion pathway. Thus, Pv GAMA may be an adhesion molecule for invasion of Duffy positive and negative human erythrocytes and is an important P. vivax vaccine candidate.

关键词： Plasmodium vivax GAMA erythrocyte binding Duffy negative sialic acid-independent vaccine candidate

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Identification and characterization of a novel Plasmodium vivax exported protein PvEXP100

Identification and characterization of a novel Plasmodium vi...

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第十一届全国寄生虫学青年工作者学术研讨会

作者： Xinxin Zhang Ruilin Chu Haitian Fu Xiaodan Shi Yinghua Xuan Yang Cheng Laboratory of Pathogen Infection and Immunity Department of Public Health and Preventive MedicineWuxi School of MedicineJiangnan University

Malaria is one of the serious infectious diseases of humans in the world,and vivax malaria is the most geographically widespread of the human malaria.It is widely accepted that during parasite invasion and development within host cells,P.vivax exported protein specifically cytoadhere to human spleen fibroblast(HSF) cells where they are protected from macrophage clearance and upon disruption of barrier cells during crisis,free merozoites will immediately encounter.However,the detailed mechanism is still unclear.We investigated a hypothetic P.vivax protein Pv EXP100 in blood stage.Immunofluorescence assay data reveals that Pv EXP100 is a novel protein which exported to infected red blood cell surface,and anti-Pv EXP100 causes partial immune response(sensitivity 42%,specificity 96.9%).Additionally,the data showed Pv EXP100 drove HSF cells migration,and Pv EXP100 could bind HSF cells.In summary,Pv EXP100 is a novel exported protein that may be a potential molecule involved in the immune evasion of P.vivax.

关键词： Plasmodium vivax export protein human spleen fibroblast immune evasion

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Production of Plasmodium vivax Enolase in Escherichia coli and its Protective Properties

Production of Plasmodium vivax Enolase in Escherichia coli a...

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中国动物学会寄生虫学专业委员会第十届全国寄生虫学青年工...

作者： Chao Zhang Yaping Gu Jianxia Tang Feng Lu Yuanyuan Cao Huayun Zhou Guoding Zhu Jun Cao Qi Gao Key Laboratory on Technology for Disease Prevention and Control(Ministry of Health) Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology Jiangsu Institute of Parasitic Diseases Public Health Research Center Jiangnan University

Objective: Plasmodium vivax predominates in South-East Asia and the American continent, causes significant morbidity and inflicts a huge socioeconomic burden. Sequencing completion of the Plasmodium vivax genome and transcriptome provides the chance to identify antigens. Enolase is the eighth enzyme in the glycolytic pathway, which, apart from its glycolytic function, also possess antigenic properties and is present on the cell wall of many invasive organisms, such as Candida albicans. In order to assess whether enolase of Plasmodium vivax is also antigenic, in this study, we first reported the expression and purification of recombinant Plasmodium vivax enolase(r-Pven) in Escherichia coli, using prokaryotic expression vector. Methods and results: The r-Pven was expressed in soluble form in E. coli, and the expression was verified by SDS-PAGE and western blotting analysis. The r-Pven was purified to 90% purity by nickel–nitrilotriacetic acid(Ni–NTA) resin chromatography. For reactivity with r-Pven, compared with the average values of the reactivity of control serum samples, the average values of the reactivity of 99 individual serums from vivax malaria patients appeared higher, and there was significant difference between them(p=0.0117plasmodium berghei. The antibodies raised against r-Pven were specific for plasmodium and did not react to the host tissues. Conclusion: These observations established Plasmodium vivax enolase to be a potential protective antigen.

关键词： Malaria Plasmodium vivax Enolase Prokaryotic expression Vaccine candidate

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Plasmodium vivax subtelomeric transmembrane protein 1(PvSTP1), a homolog of P. falciparum SURFIN, is highly polymorphic

Plasmodium vivax subtelomeric transmembrane protein 1(PvSTP1...

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第八届全国免疫学学术大会

作者： Xiaotong Zhu Morakot Kaewthamasorn Kazuhide Yahata Ya-Ming Cao Osamu Kaneko Department of Immunology China Medical University Department of Protozoology Institute of Tropical Medicine (NEKKEN) Nagasaki University

Background and Objective:Plamodium vivax subtelomeric transmembrane protein 1(PvSTP1) is a predicted type I transmembrane protein, which shares homology with the P.falciparum SURFIN proteins that are located on the surface of infected erythrocytes.The structure of SURFIN–PvSTP1 family members incorporate features of several erythrocyte surface parasite-encoded proteins from human, rodent, and monkey malaria parasites;such as Pf EMP-1, pir, and SICAvar.Parasite proteins that are located on the surface of erythrocytes are often highly polymorphic in nature, possibly reflecting the action of positive diversifying selection driven by host immune factors.Methods:We analyzed the level of polymorphism in an 1182 bp nucleotide(nt) region encoding the predicted extracellular domain of PvSTP1 using 70 sequences originating from 52 P.vivax Chinese field isolates.Results:We found that the extracellular domain of PvSTP1 is highly polymorphic;a total of 307 polymorphic sites were observed with an average nucleotide diversity of 0.0999.Based on the predicted protein structure, we divided the sequenced region into two parts:a Cysteine-rich domain(nt 67 – 597) and a variable domain(nt 598 – 1248).Both domains were highly polymorphic with nucleotide diversities of 0.0995 and 0.1010, respectively.A significant excess of nonsynonymous substitutions over synonymous substitutions was detected in the variable domain.Tajima’s D test and Fu & Li’s D* and F* tests all detected the signature of positive selection on extracellular region.Conclusion:Thus, the extracellular region of PvSTP1 appears to have evolved under positive diversifying selection, possibly mediated by host immune pressure.

关键词： Plasmodium vivax Diversity polymorphism selection

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Insights into the elimination of vivax malaria in China

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Infectious Diseases of Poverty 2023年第2期12卷 101-105页

作者： Fang Huang Li Zhang Zhi-Gui Xia Shanghai Municipal Center for Disease Control and Prevention Shanghai200336China National Institute of Parasitic Diseases Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research)NHC Key Laboratory of Parasite and Vector BiologyWHO Collaborating Center for Tropical DiseasesNational Center for International Research on Tropical DiseasesShanghai200025China

Background Malaria is caused by multiple parasitic species of the genus plasmodium.Plasmodium vivax is the most geographically widespread and poses challenges in elimination due to its unique biological and epidemiological characteristics.The aim of study was to highlight the practices and experience targeting vivax malaria control and elimination in China.Main body P.vivax malaria was historically endemic in more than 70%of counties in China,with reported vivax malaria cases as high as 26 million a year.After around 70 years of effort,China was certified as malaria-free in June of 2021.The key insights into China’s vivax malaria control and elimination were offered,including radical cure strategies,comprehensive but adaptive strategies targeting species of plasmodium and Anopheles,mass drug administration,and case-/focus-centred surveillance and response systems.Conclusion The complete global eradication of P.vivax and eventually malaria will be more difficult,and China’s practices and experience could be a valuable reference in this campaign.

关键词： Malaria Plasmodium vivax Elimination China

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