Vaccination with a DNase II recombinant protein against Trichinella spiralis infection in pigs

2020 ◽  
pp. 109069 ◽  
Author(s):  
Daoxiu Xu ◽  
Bin Tang ◽  
Yong Yang ◽  
Xuepeng Cai ◽  
Wanzhong Jia ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Trichinella Spiralis ◽  
Dnase Ii

Molecular cloning of enolase from Trichinella spiralis and the protective immunity in mice

2018 ◽  
Vol 63 (2) ◽  
pp. 252-260 ◽  
Author(s):  
Xuliang Zhang ◽  
Lixin Xu ◽  
Xiaokai Song ◽  
Xiangrui Li ◽  
Ruofeng Yan
Keyword(s):  
Recombinant Protein ◽  
Trichinella Spiralis ◽  
Flow Cytometric Analysis ◽  
Recombinant Plasmids ◽  
Flow Cytometric ◽  
Th2 Immune Response ◽  
Wide Range ◽  
Muscle Larvae ◽  
Ifn Γ ◽  
Negative Controls

Abstract Trichinella spiralis, the main pathogen of trichinosis, infects a wide range of mammalian hosts and is one of the most widespread parasites worldwide. For parasites, glycolysis is the most important way to generate energy. Previous studies showed that some enzymes involved in the glycolytic pathway play roles in regulation the host immunity. In this paper, enolase from T. spiralis was cloned and the protective potentials were studied. One hundred and sixty ICR mice were divided into four groups and vaccinated with recombinant enolase (pET-ENO), eukaryotic recombinant plasmid encoding enolase (pVAX1-ENO) and negative controls (pVAXl and PBS), respectively. Two weeks after the second immunization, each mouse was challenged orally with 200 muscle larvae (MLs) of T. spiralis. Results showed that mice vaccinated with pET-ENO and pVAX1-ENO induced specific antibodies of IgG, IgA, IgM, but no IgE. Subclasses of IgG antibodies showed that mice immunized with recombinant protein and recombinant plasmids induced a Th1/Th2 immune response. Concentrations of serum cytokines were detected and showed significant increase of IFN-γ, IL-4 and TGFβ1, while IL-17 in each group was not significantly different. Flow cytometric analysis showed significant increase of CD4+ and CD8+ T lymphocytes in the groups immunized with recombinant protein and recombinant plasmids. Challenge infection demonstrated that immunized groups had a reduced number of worm burdens. The reductions of larvae per gram muscle (LPG) in pET-ENO and pVAX1-ENO group were 17.7% and 15.8% when compared with PBS control.

scholarly journals Proteomic Analysis ofTrichinella spiralisMuscle Larval Excretory-Secretory Proteins Recognized by Early Infection Sera

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Li Wang ◽  
Zhong Quan Wang ◽  
Dan Dan Hu ◽  
Jing Cui
Keyword(s):  
Molecular Weight ◽  
Serine Proteases ◽  
Trichinella Spiralis ◽  
Early Stage ◽  
False Negative ◽  
Hypothetical Protein ◽  
Secretory Proteins ◽  
Dnase Ii ◽  
Negative Results ◽  
Protective Antibodies

Although the excretory-secretory (ES) proteins ofTrichinella spiralismuscle larvae are the most commonly used diagnostic antigens for trichinellosis, their main disadvantage is the false negative results during the early stage of infection and cross-reaction of their main components (43, 45, 49, and 53 kDa) with sera of patients with other helminthiasis. The aim of this study was to identify early specific diagnostic antigens inT. spiralisES proteins with 30–40 kDa. The ES proteins were analyzed by two-dimensional electrophoresis (2-DE), and a total of approximately 150 proteins spots were detected with isoelectric point (pI) varying from 4 to 7 and molecular weight from 14 to 66 kDa. When probed with sera from infected mice at 18 days postinfection, ten protein spots with molecular weight of 30–40 kDa were recognized and identified by MALDI-TOF/TOF-MS. All of ten spots were successfully identified and characterized to correlate with five different proteins, including two potential serine proteases, one antigen targeted by protective antibodies, one deoxyribonuclease (DNase) II, and one conserved hypothetical protein. These proteins might be the early specific diagnostic antigens for trichinellosis.

Identification of stage-specifically expressed genes of Trichinella spiralis by suppression subtractive hybridization

Parasitology ◽  
2007 ◽  
Vol 134 (10) ◽  
pp. 1443-1455 ◽  
Author(s):  
M. Y. LIU ◽  
X. L. WANG ◽  
B. Q. FU ◽  
C. Y. LI ◽  
X. P. WU ◽  
...  
Keyword(s):  
Suppression Subtractive Hybridization ◽  
Serine Proteases ◽  
Zinc Finger Protein ◽  
Trichinella Spiralis ◽  
Gene Families ◽  
Subtractive Hybridization ◽  
Cdna Clones ◽  
Dnase Ii ◽  
Finger Protein ◽  
Old Adult

SUMMARYNewborn larvae (NBL) and adult (Ad) stage-specifically expressed genes or members of gene families of Trichinella spiralis were identified by suppression subtractive hybridization (SSH)†. Six cDNA clones were identified as NBL stage-specific, including 1 member of the T. spiralis gene family encoding glutamic acid-rich proteins, 2 clones encoding novel serine proteases, 2 closely related clones encoding proteins that are members of a deoxyribonuclease II (DNase II)-like family and 1 clone with no similarity to known genes. Four stage-specific clones encoding homologues of retinoid X receptor, caveolin, C2H2 type zinc finger protein and a putative protein with no homology to known sequences were obtained from 3-day-old adult worms. One gene specifically up-regulated in the 5-day-old adult worms encoding a putative cuticle collagen was also identified.

scholarly journals Characterisation of a Plancitoxin-1-Like DNase II Gene in Trichinella spiralis

2014 ◽  
Vol 8 (8) ◽  
pp. e3097 ◽  
Author(s):  
Chengshui Liao ◽  
Mingyuan Liu ◽  
Xue Bai ◽  
Pan Liu ◽  
Xuelin Wang ◽  
...  
Keyword(s):  
Trichinella Spiralis ◽  
Dnase Ii

scholarly journals Advances in the sequencing of the genome of the adenophorean nematode Trichinella spiralis

Parasitology ◽  
2008 ◽  
Vol 135 (8) ◽  
pp. 869-880 ◽  
Author(s):  
M. Mitreva ◽  
D. P. Jasmer
Keyword(s):  
Genome Sequence ◽  
Evolutionary Biology ◽  
Trichinella Spiralis ◽  
Control Strategies ◽  
Cell Interaction ◽  
Parasitic Nematodes ◽  
Molecular Characteristics ◽  
Dnase Ii ◽  
Interspecific Comparisons ◽  
Insight Into

SUMMARYThe adenophorean nematodes are evolutionarily distant from other species in the phylum Nematoda. Interspecific comparisons of predicted proteins have supported such an ancient divergence. Accordingly, Trichinella spiralis represents a basal nematode representative for genome sequencing focused on gaining a deeper insight into the evolutionary biology of nematodes. In addition, molecular characteristics that are conserved across the phylum could be of great value for control strategies with broad application. In this review, we describe and summarize progress that has been made on the sequencing and analysis of the T. spiralis genome. The genome sequence was used in preliminary analyses for the investigation of specific questions relating to the biology of T. spiralis and, more generally, to parasitic nematodes. For instance, we evaluated an unusually large DNase II-like protein family, predicted proteins of prospective interest in the parasite-host muscle cell interaction, anthelmintic targets and prospective intestinal genes, the encoded proteins (potentially) linked to immunological control against other nematodes. The results are discussed in relation to characteristics that are broadly conserved among evolutionary distant nematodes. The results lead to expectations that this genome sequence will contribute to advances in research on T. spiralis and other parasitic nematodes.

Molecular cloning and characterization of a serine proteinase inhibitor from Trichinella spiralis

Parasitology ◽  
2001 ◽  
Vol 123 (1) ◽  
pp. 77-83 ◽  
Author(s):  
I. NAGANO ◽  
Z. WU ◽  
T. NAKADA ◽  
A. MATSUO ◽  
Y. TAKAHASHI
Keyword(s):  
Recombinant Protein ◽  
Proteinase Inhibitors ◽  
Trichinella Spiralis ◽  
Serine Proteinase ◽  
Predicted Amino Acid Sequence ◽  
Serpin Gene ◽  
Specific Expression ◽  
Crude Extracts ◽  
Muscle Larvae

We produced a recombinant protein from a cDNA library from muscle larvae of Trichinella spiralis which had proteinase inhibitory activity. The predicted amino acid sequence of the clone had an identity of only 30% to the serine proteinase inhibitors (serpins) from Caenorhabditis elegans or Brugia malayi. At the putative reactive region, however, the identity was about 50%. The recombinant protein expressed in Escherichia coli inhibited 82% of the activity of the serine proteinase (trypsin). Stage-specific expression of this protein was suggested from the following experiments. Antibody against the recombinant protein could stain proteins migrating at about 42 kDa (which is the expected size from the sequence) in crude extracts from newborn larvae and 18-day post-infection (p.i.) muscle larvae, but it failed to stain any proteins in crude extracts from 30-day p.i. muscle larvae. Production of mRNA transcript for the serpin gene was restricted largely to the newborn larvae and to 18-day p.i. muscle larvae. The antibody reacted with the stichocytes of the larvae at 18 days p.i., but did not react with the muscle larvae at 24 days and 30 days p.i.

The protease phenotype and crystalline nature of the granules of intraepithelial mast cells in Trichinella spiralis-infected mice

1995 ◽  
Vol 53 ◽  
pp. 818-819
Author(s):  
D.S. Friend ◽  
N. Ghildyal ◽  
M.F. Gurish ◽  
K.F. Austen ◽  
R.L. Stevens
Keyword(s):  
Small Intestine ◽  
Mast Cells ◽  
Epithelial Cells ◽  
Lamina Propria ◽  
Trichinella Spiralis ◽  
Intestinal Epithelial ◽  
Carboxypeptidase A ◽  
C3h Mice ◽  
Granule Morphology ◽  
Crystalline Nature

Trichinella spiralis induces a profound mastocytosis and eosinophilia in the small intestine of the infected mouse. Mouse mast cells (MC) store in their granules various combinations of at least five chymotryptic chymases [designated mouse MC protease (mMCP) 1 to 5], two tryptic proteases designated mMCP-6 and mMCP-7 and an exopeptidase, carboxypeptidase A (mMC-CPA). Using antipeptide, protease -specific antibodies to these MC granule proteases, immunohistochemistry was done to determine the distribution, number and protease phenotype of the MCs in the small intestine and spleen 10 to >60 days after Trichinella infection of BALB/c and C3H mice. TEM was performed to evaluate the granule morphology of the MCs between intestinal epithelial cells and in the lamina propria (mucosal MCs) and in the submucosa, muscle and serosa of the intestine (submucosal MCs).As noted in the table below, the number of submucosal MCs remained constant throughout the study. In contrast, on day 14, the number of MCs in the mucosa increased ~25 fold. Increased numbers of MCs were observed between epithelial cells in the mucosal crypts, in the lamina propria and to a lesser extent, between epithelial cells of the intestinal villi.

Effectiveness of zona pellucida protein ZPB as an immunocontraceptive antigen

Reproduction ◽  
2000 ◽  
pp. 19-32 ◽  
Author(s):  
ML Martinez ◽  
JD Harris
Keyword(s):  
Menstrual Cycle ◽  
Recombinant Protein ◽  
Antibody Titre ◽  
Zona Pellucida ◽  
Protein A ◽  
Papio Cynocephalus ◽  
Cynomolgus Monkeys ◽  
Menstrual Cycles ◽  
Normal Menstrual Cycle

Immunization of female mammals with native zona pellucida (ZP) proteins is known to cause infertility. Since each human ZP protein is now available as a purified recombinant protein, is it possible to compare the immunocontraceptive potential of each ZP protein. A breeding study was conducted in cynomolgus monkeys (Macaca fasicularis) after immunization with recombinant human ZP (rhZP) proteins (ZPA, ZPB, ZPC) separately and in combinations. This study demonstrated that immunization with recombinant human ZPB (rhZPB) protein caused cynomolgus monkeys to become infertile for 9-35 months. A second study was conducted in baboons (Papio cynocephalus), which yielded a similar result. The baboons immunized with rhZPB became infertile for 9 to > 20 months. During the time of maximum antibody titre, some animals experienced disruption of the menstrual cycle, but eventually all of the animals resumed normal menstrual cycles. Control animals and animals immunized with other rhZP proteins all became pregnant before any of the rhZPB-treated animals. This is the first study in which a recombinant ZP protein has consistently induced infertility in a primate without permanent disruption of the normal menstrual cycle.

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