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Progress of Listeria monocy togenes as tumor vaccine vector

Feifei DUAN ; Yuelan YIN ; Meiqin KANG ; Weijun TAN ; Chengwu TAO ; Zhiming PAN ; Jinlin HUANG ; Xinan JIAO

Chinese Journal of Zoonoses.2014;(7):743-746,752. doi:10.3969/cjz.j.issn.1002-2694.2014.07.016

Listeriamonocytogenes is a facultative intracellular bacterium that enters professional antigen presenting cells , presents passenger antigens to the major histocompatibility complex class I and II pathways ,then elicits CD+4 and CD+8 T-cell-mediated immune responses .It was demonstrated that attenuated Listeriamonocytogenes as a novel live vaccine vector in deliv-ering tumor antigens of cervical cancer and melanoma etc .,could induce strong protective immune response ,and shows effec-tive antitumor immunotherapeutics .This review discussed the characteristics of immune responses elicited by Listeria monocy-togenes ,and the progress of its antitumor immunotherapeutics as delivery vaccine vector .

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Progress of microRNA-related research in tuberculosis

Shuaili CAO ; Li YUAN

Chinese Journal of Zoonoses.2014;(7):757-760,771. doi:10.3969/cjz.j.issn.1002-2694.2014.07.019

The miRNA is a class of small molecule non-coding RNA with a length of about 20-24 bp ,which is transcrip-tion from the endogenous gene .The hairpin structure and the post-transcriptional regulation of gene expression are its typical characteristics .It is suggested that the miRNA play an important control and management role in latent infection and active in-fection in tuberculosis ,and the expression situation of miRNA is also different between latent infection and active infection in tuberculosis .Some studies also found that there are differences in the expression of miRNAs such as miR-155 ,miR-29a ,miR-361-5p ,miR-889 ,miR-576-3p and so on ,as a marker of TB diagnose .

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Age impact in mouse model of secondary hepatic alveolar echinococcus

Haiyang XIA ; Jiefeng ZHAO ; Qikun WANG ; Li LI ; Jingxia TANG ; Shijie ZHANG ; Xinyu PENG ; Hongqiang YANG

Chinese Journal of Zoonoses.2014;(7):723-726,732. doi:10.3969/cjz.j.issn.1002-2694.2014.07.012

Age impact in mouse model of secondary hepatic alveolar echinococcus was investigated in this research . Twenty-nine 8-week-old ,twenty-five 18-week-old and twenty-five 28-week-old female mice were anesthetized with 20% ure-thane by intraperitoneal injection and then transhepatically injected by Echinococcus multilocularis (E .m) tissue suspension through skin incision and abdominal muscle to liver in all three groups to establish mouse model of secondary hepatic alveolar e-chinococcus .Results showed that the survival rates for the three groups of mice were 62 .1% ,84% and 68% ,respectively (P>0 .05) .The E .m infection rates in liver were 72 .2% ,71 .4% and 76 .5% ,respectively (P>0 .05) .The diameter of E .m cysts in liver were 0 .915 ± 0 .103 cm ,1 .247 ± 0 .112 cm and 1 .215 ± 0 .197 cm ,respectively (P>0 .05) .The mass of E .m cysts in liver were 0 .332 ± 0 .035 g ,0 .532 ± 0 .155 g and 0 .382 ± 0 .085 g ,respectively (P> 0 .05) .HE stain showed no difference in pathology .Results indicated that the establishment of secondary hepatic alveolar echinococcus model by using transhepatic injection through skin incision and abdominal muscle of 18-week-old mice was capable of simplifying operation and improving the survival rate of the mice .

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Case study of Yersinia enterocolitica infection

Yi LIU ; Jianyuan FENG ; Yanchun LI ; Lili LIU ; Teng QI ; Ziren ZHOU

Chinese Journal of Zoonoses.2014;(7):769-771. doi:10.3969/cjz.j.issn.1002-2694.2014.07.022

Yersinia enterocolitica infection in humans and animals was investigated in this study to explore the source of infection .We collected samples from patients and pigs ,and then the strains were conducted to undergo serotyping ,virulence gene detection and PFGE molecular typing .Of the 150 patients samples ,3 were tested positive for Yersinia enterocolitica ,and one of them was O∶3 .Of the 222 pigs samples ,14 were tested positive for Yersinia enterocolitica ,and all of them were O∶3 .PFGE molecular typing showed that the Yersinia enterocolitica from patient and pig had high homology .Our study con-firmed that Yersinia enterocolitica from pigs could infect people and find that it could exist in the cured patient for at least two months .

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Assembly and components of the type III secretion system of Salmonel l a

Long CHEN ; Qingfeng MENG ; Yuanhuan KANG ; Xiaofeng SHAN ; Liang CAO ; Aidong QIAN

Chinese Journal of Zoonoses.2014;(7):753-756. doi:10.3969/cjz.j.issn.1002-2694.2014.07.018

Salmonella is the main food-borne pathogen that causes food poisoning and gastroenteritis in human and ani-mals .The type III secretion system in Salmonella has played an important role in the invasion of host cell .In recent years ,the research of the composition ,assembly and related pathogenesis of Salmonella T3SS have made some progress .This article re-views the composition and assembly of Salmonella type III secretion system ,which could provide further study the pathogene-sis of Salmonella and also the new strategies and methods toward the treatment and prevention of Salmonella infections .

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Research progress of molecular examination and genotyping method for Entamoeba

Haiju DONG ; Rongjun WANG ; Longxian ZHANG

Chinese Journal of Zoonoses.2014;(7):747-752. doi:10.3969/cjz.j.issn.1002-2694.2014.07.017

Entamoeba is a zoonotic protozoan that can parasitize in the intestine and other organs in human and animals . There are many species in this genus ,but only Entamoeba histolytica can cause amebic colitis and liver abscess ,and even death .The cysts and trophozoits are difficult to be discriminated for pathogenic and nonpathogenic Entamoeba .And there is al-so difference in the pathogenicity ,infectivity etc .,even in the same species .Therefore ,it is difficult to confirm the Entamoeba species according to traditional method ,thus affecting the use of medicine .In this article ,the research progress of molecular examination and genotyping method about entamoeba are reviewed .

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Fine-scale population genetic structure of Oncomelania hupensis based on microsatellite DNA markers

Bin CUI ; Wei GUAN ; Ping YOU ; Shizhu LI

Chinese Journal of Zoonoses.2014;(7):701-708. doi:10.3969/cjz.j.issn.1002-2694.2014.07.009

The genetic structure of small-scale landscape groups of Oncomelania hupensis in Songzi City ,Hubei Province was identify in this study .O .hupensis snails were collected from 10 habitats in Songzi City ,of which 6 polymorphic microsat-ellite DNA loci (T6-17 ,P101 ,D11 ,B14 ,T4-33 ,and C22) were carried out with GeneScan .The number of alleles (Na) ,het-erozygosity (H) ,fixation index (FST) of snails in each group ,genetic distance between groups ,and the polymorphic informa-tion content (PIC) were calculated .Cluster analysis was then carried out based on genetic distance ,and hierarchical AMOVA calculation was conducted .By certified the shells of snails ,10 groups were divided into light and ribbed shell (including shallow rib and deep rib) .There were 141 alleles in total detection on 10 populations and 20-34 alleles in each locus ,which were detec-ted for 23 .5 on average .The average number of alleles in 6 loci was 1 .575 and the number of alleles in each locus was uneven , showing large numerical differences ranged from 0 .445 to 3 .060 .The average observed heterozygosity (Ho) ranged from 0 .438 ue between paired populations was from -0 .015 64 to 0 .252 47 ,and the polymorphic information content in the population ranged from 0 .528 -0 .857 ,showing a high polymorphism .Hierarchical AMOVA calculations showed that inter-individual variation of the snails occupied 88 .4% of the total variations .Cluster analysis revealed that the three ribbed shell population in Munu Kou Village ,Hengti Village and Yixing Village first clustered to the three light shell population in Mashizizu Village , Mingzhu Village and Tuqiao Village ,then clustered to the light shell population in Tuanshan Village and Jiama Cao Village with the two shallow rib population in Desheng Village and Tianmu He Village .Under the different landscape environment of Songzi Area ,there were different shells presenting on the morphology of O .hupensis .Although there was a rich diversity on O .hupensis of Songzi City ,the genetic differences mostly present in individuals .Different groups didn’t show the significant genetic differentiation among the different shell morphology of O .hupensis .

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Potency of a novel multi-epitope vaccine against foot-and-mouth disease type Asia 1 in guinea pigs

Junjun SHAO ; Jingfeng WANG ; Shandian GAO ; Tong LIN ; Huiyun CHANG

Chinese Journal of Zoonoses.2014;(7):692-697. doi:10.3969/cjz.j.issn.1002-2694.2014.07.007

The potency of an improved recombinant multi-epitope vaccine against FMDV type Asia1 was evaluated in this study .A multi-epitope gene based on FMDV type Asia1 was designed and a recombinant expression plasmid (pRE-oIgG) was constructed .The proteins ,RE-oIgG and 3D were expressed in E .coli cells and purified with Ni-NTA agarose resin by affinity chromatography .The proteins ,RE-oIgG ,3D and RE-oIgG plus 3D ,were emulsified in an oil adjuvant ISA 206 .Twenty-five female guinea pigs were randomly divided into five groups and intramuscularly vaccinated for with RE-oIgG ,3D ,RE-oIgG plus 3D ,an inactivated FMDV vaccine (type Asia1) ,and PBS .All animals were vaccinated for two times .Anti-FMDV specific an-tibodies ,neutralization antibodies ,protection potency ,and lymphoproliferation assay were detected by ELISA ,virus neutrali-zation assay ,challenge test ,and flow cytometry ,respectively .Results showed that RE-oIgG plus 3D elicited significant high-level anti-FMDV specific antibodies compared to RE-oIgG alone (P<0 .05) .All the vaccinated animals induced higher level lymphoproliferation responses in vitro except PBS .Both 3D alone and PBS produced the negligible neutralizing antibodies and anti-FMDV specific antibodies .RE-oIgG plus FMDV 3D not only elicited high levels of anti-FMDV neutralizing antibodies ,but also induced significant lymphoproliferation responses .More importantly ,RE-oIgG plus 3D conferred complete protection to guinea pigs against challenge with 1 000 GPID50 .Interestingly ,two of five vaccinated animals with 3D alone were full protected against challenge ,and other three animals significantly showed a delay of 2-3 days in the onset of clinical signs .Therefore ,we considered that RE-oIgG plus 3D induces strong humoral and cellular immune responses ,which may be used for control and prevention of FMD in the future .

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Rapid identification of bacteriological negative pulmonary TB and lung disease by proteomic fingerprinting technology

Lin WANG ; Lizhen WENG ; Xiaohong CHEN ; Mingxiang HUANG ; Xueling LI ; Jiandong LIN ; Zhiping GUO ; Lijun XIONG ; Tanye LIU

Chinese Journal of Zoonoses.2014;(7):688-691. doi:10.3969/cjz.j.issn.1002-2694.2014.07.006

To explore the application of protein fingerprint technique and differential diagnosis in bacteriological negative pulmonary tuberculosis and pneumonia ,60 patients with bacteriological negative pulmonary tuberculosis ,60 patients with pneumonia ,and 60 healthy volunteers were selected from known clinical cases .Surface strengthening laser desorption ioniza-tion time of flight mass spectrometry (SELDI ToF Ms) and protein chip technology were applied to detect serum proteins ,and analyze their protein peaks by Ciphergen protein chip 3 .1 .1 software .Comparison of the serum protein fingerprinting data from the pool of 180 patients and healthy volunteers showed significant difference in 5 protein peaks (1 028 .49 ,4 796 .56 ,7 564 .77 , 8 048 .02 ,and 11 526 .75 m/z) identified between pulmonary tuberculosis and pneumonia (P<0 .01) .The total effective rate of the 5 protein peaks as a diagnosis model for differential diagnosis of bacteriological negative pulmonary tuberculosis and pneumonia was 84 .2% (101/120) ,the specificity was 82 .5% (52/63) ,the sensitivity was 85 .9% (49/57) ,the positive pre-dictive value was 86 .7% (52/60) ,and the negative predictive value was 81 .7% (49/60) .The total effective rate of the diagno-sis model for differential diagnosis of bacteriological negative pulmonary tuberculosis ,pneumonia and healthy volunteers was 89 .4% (161/180) .The specificity was 100% (60/60) ,the sensitivity was 84 .2% (101/120) ,the positive predictive value was 100% (101/101) ,and the negative predictive value was 75 .9% (60/79) .Protein fingerprinting technology is advanta-geous of being a simple method ,quick detection ,and requires less amount of sample .It is an effective means to screening the tuberculosis specific markers .We found the good diagnosis model through the detection of serum protein by protein fingerprint-ing technology .

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Protective function of S jcb2 DNA vaccine in mice infected with Schistosoma j aponicum

Angui FENG ; Yongqiang GAO ; Yu LIANG ; Yuan WU ; Hu CHEN ; Yukuai ZHENG ; Kegeng WANG ; Shanshan CHEN ; Jianhua XIAO

Chinese Journal of Zoonoses.2014;(7):673-678. doi:10.3969/cjz.j.issn.1002-2694.2014.07.003

The protective effect and mechanism of Schistosoma japonicum cathepsin B (Sjcb2) DNA vaccine in the mouse model of schistosomiasis were studied through construction pcDNA3 .1 (+ ) / Sjcb2 DNA recombinant vector ,which provided effective candidate antigen for anti-schistosome vaccine .The 6-week-old female BALB/c mice were randomly divided into pcDNA3 .1(+ )/Sjcb2 DNA vaccine group ,pcDNA3 .1(+ ) plasmid group and normal saline group ,respectively .Each group was composed of 35 mice ,and 100 μg of S jcb2 plasmid DNA was injected in the hind leg quadriceps of mice once every two weeks .PCR and immunohistochemistry assay were used to detect the expression and stability of Sjcb2 gene in mice .MTT assay was used for testing the specific proliferation response of mice spleen lymphocytes .The level of Sjcb2 antibodies in mouse serum and the IFN-γand IL-4 levels in mice spleen lymphocyte culture supernatant before and after schistosome infection were assayed by ELISA .At last ,we counted load of Schistosome adult worms in mouse and eggs in liver of mouse .The results showed that the Sjcb2 gene was detected in all mice of the Sjcb2 DNA vaccine group ,and Sjcb2 gene expression was positive in the muscle cells in Sjcb2 DNA immunized mice by IHC assay .MTT assay showed that T-cell proliferation rate was in-creased significantly in S jcb2 DNA vaccinated group .ELISA results showed that the IFN-γlevels were increased significantly in the vaccinated group ,while the IL-4 levels were significantly increased after Schistosoma japonicum infection in all mice of every group .The load of worms and eggs in Sjcb2 DNA vaccinated group was reduced significantly than that of control group (P<0 .05) ,the reduction rates of adult worms and eggs were 36 .32% and 60 .61% respectively .In conclusion ,the Sjcb2 gene was stably expressed in muscle cells of mice after injection of S jcb2 recombinant plasmid ,and S jcb2 produced protective effects of anti-schistosoma infection in mice possibly by mean of regulating Th1 cell subgroups through increasing the IFN-γ level and decreasing IL-4 levels .

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