Enzymatic characteristics and preventive effect of leucine aminopeptidase against Echinococcus multilocularis

Wang, Lei; Wei, Wei; Zhou, Pei; Liu, Haisheng; Yang, Baoliang; Feng, Lin; Ge, Ri‐Li; Li, Runle; Tang, Feng

doi:10.1016/j.actatropica.2021.106066

Cited by 9 publications

(14 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…LAP is involved in the nutrient absorption of the parasite and its invasion in the host, and it can prevent not only the infection of E. multilocularis infection but also its metabolism and infiltration. The recombinant E. multilocularis LAP can significantly reduce the number and size of cysts in mice infected with E. multilocularis ( 9 ). GLUT is related to glucose absorption and energy supply in most parasites .…”

Section: Discussionmentioning

confidence: 99%

“…Because of this, the current cure rate of AE is only 30-40% per year ( 7 ). Recent studies have suggested that vaccination is an effective means for the control and prevention of E. multilocularis infection ( 8 ), and immunization with recombinant proteins of parasites, such as leucine aminopeptidase (LAP), EMY162, EM95 and Tetraspanin (TSP), could protect against E. multilocularis infection and infiltration in host livers ( 9 ). EMY162 and EM95 are secreted proteins expressed in all life stages of E. multilocularis ( 10 ); and TSP plays an important role in the growth and development of E. multilocularis ( 11 ).…”

Section: Introductionmentioning

confidence: 99%

“…This work provides a basis for the development of effective and safe vaccines targeting the epitopes of E. multilocularis ( 12 , 13 ).However, the use of a single epitope vaccine might not be adequate to prevent E. multilocularis infection because different types and amounts of proteins would be expressed at different developmental stages of E. multilocularis ( 14 ). In this study, a multi-epitope vaccine GILE was designed using bioinformatics and molecular biology techniques based on the dominant epitopes of E. multilocularis EMY162, LAP and GLUT1 (EmEMY162, EmLAP, and EmGLUT1) identified in our previous studies ( 9 , 10 , 15 ), and its immune protective effect against E. multilocularis infection was evaluated in mice.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice

Zhou

Zhou²,

Huayu³

et al. 2023

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

IntroductionThe objective of this study is to construct a multi-epitope vaccine GILE containing B-cell and T-cell epitopes against Echinococcus Multilocularis (E. multilocularis) infection based on the dominant epitopes of E. multilocularis EMY162, LAP, and GLUT1.MethodsThe structure and hydrophobicity of GILE were predicted by SWISSMODEL, pyMOL, SOPMA and VMD, and its sequence was optimized by Optimum™ Codon. The GILE gene was inserted into pCzn1 and transformed into Escherichia coli Arctic express competent cells. IPTG was added to induce the expression of recombinant proteins. High-purity GILE recombinant protein was obtained by Ni-NTA Resin. BALB/c mice were immunized with GILE mixed with Freund’s adjuvant, and the antibody levels and dynamic changes in the serum were detected by ELISA. Lymphocyte proliferation was detected by MTS. The levels of IFN-g and IL-4 were detected by ELISpot and flow cytometry (FCM). T cells were detected by FCM. The growth of hepatic cysts was evaluated by Ultrasound and their weights were measured to evaluate the immune protective effect of GILE.ResultsThe SWISS-MODEL analysis showed that the optimal model was EMY162 95-104―LAP464-479―LAP495-510―LAP396-410―LAP504-518―EMY162112-126. The SOPMA results showed that there were Alpha helix (14.88%), Extended strand (26.25%), Beta turn (3.73%) and Random coil (45.82%) in the secondary structure of GILE. The restriction enzyme digestion and sequencing results suggested that the plasmid pCzn1-GILE was successfully constructed. The SDSPAGE results indicated that the recombinant protein was 44.68 KD. The ELISA results indicated that mice immunized with GILE showed higher levels of serum antibodies compared to the PBS group. The FCM and ELISpot results indicated that mice immunized with GILE secreted more IFN-g and IL-4. Immunization with GILE also led to a significant decrease in the maximum diameter and weight of cysts and stimulated the production of CD4+ and CD8+ T Cell.DiscussionA multi-epitope vaccine GILE with good immunogenicity and antigenicity has been successfully constructed in this study, which may provide important theoretical and experimental bases for the prevention and treatment of E. multilocularis infection.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice

Zhou

Zhou²,

Huayu³

et al. 2023

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Vaccination with rEm-LAP significantly decreased both the number and size of cysts in an E . multilocularis metacestode-infected mouse model [24] . Our results suggested that the protection of rEg.P29 against CD4 + T cell-deficient mice with E .…”

Section: Discussionmentioning

confidence: 99%

Immunoprotective effect and mechanism of rEg.P29 against CD4<sup>+</sup> T cell-deficient mice with <italic>Echinococcus multilocularis</italic> infection

Li,

Zhu,

et al. 2023

ABBS

View full text Add to dashboard Cite

Alveolar echinococcosis (AE) is a zoonotic parasitic disease caused by infection with the larval stage of Echinococcus multilocularis and a major challenge to human public health. Vaccines are the most effective way to prevent and control infectious diseases. We previously revealed that the Echinocuccus granulosus recombinant protein P29 is a good vaccine candidate against E . granulosus . However, the protective and immunological mechanism of rEg.P29 against E . multilocularis remain unclear. In this study, CD4 + T cell-deficient mice are transferred with spleen CD4 + T cells isolated from wild-type mice and subjected to rEg.P29 immunization, and then these immunized mice are infected with E . multilocularis . The cyst inhibition rate is calculated by weighing the body and cyst weights. The level of antibody is detected by ELISA. Flow cytometry is used to detect the level of IFN-γ production by CD4 + T and CD8 + T cells. The cytokines in culture supernatant are detected by ELISA. The expressions of CD44 and CD62L on memory T cells are determined by flow cytometry. The results show the cyst inhibition rate is 41.52% after adoptive transfer of CD4 + T cells. Furthermore, the levels of IgG, IgM, IgA and IgE in serum are significantly increased compared with those in the PBS group. The IFN-γ-secretion by CD8 + T cells and the level of IFN-γ in culture supernatant are obviously increased; and the number of CD4 + T cells is increased, but the number of IFN-γ producing CD4 + T cells has no significant difference compared with PBS group. In addition, the number of CD44 + CD62L ‒ CD8 + memory T cells in the spleen is significantly increased, while the number of CD44 ‒ CD62L + CD8 + memory T cells is not significantly altered. Collectively, rEg.P29 can alleviate E . multilocularis infection by inducing humoral immune responses and CD8 + T cell responses.

show abstract

“…The cytokine profile based on the mRNA expression showed that immunization with Em CRT induced splenocytes to produce both Th1 (IFN-γ and IL-2) and Th2 cytokines (IL-4, IL-5 and IL-10). However, the IL-4 was mostly induced to the highest level, combining with the dominant IgG1 response over IgG2a, and it indicates that the immunization of Em CRT formulated with FA induced both Th1 and Th2 responses, with Th2 dominant [ 37 , 38 , 39 ]. In general, our data indicated that Em CRT is a highly immunogenic antigen and induces a strong antibody response and Th1/2 cytokine response in immunized mice that may contribute to the protective immunity against the E. multilocularis infection in this study.…”

Section: Discussionmentioning

confidence: 99%

Immunization with EmCRT-Induced Protective Immunity against Echinococcus multilocularis Infection in BALB/c Mice

et al. 2022

View full text Add to dashboard Cite

Alveolar echinococcosis (AE) is a severe parasitic zoonosis caused by the larval stage of Echinococcus multilocularis. The identification of the antigens eliciting acquired immunity during infection is important for vaccine development against Echinococcus infection. Here, we identified that E. multilocularis calreticulin (EmCRT), a ubiquitous protein with a Ca2+-binding ability, could be recognized by the sera of mice infected with E. multilocularis. The native EmCRT was expressed on the surface of E. multilocularis larvae as well as in the secreted products of metacestode vesicles and protoscoleces (PSCs). The coding DNA for EmCRT was cloned from the mRNA of the E. multilocularis metacestode vesicles and a recombinant EmCRT protein (rEmCRT) was expressed in E. coli. Mice immunized with soluble rEmCRT formulated with Freund’s adjuvant (FA) produced a 43.16% larval vesicle weight reduction against the challenge of E. multilocularis PSCs compared to those that received the PBS control associated with a high titer of IgG, IgG1 and IgG2a antibody responses as well as high levels of Th1 cytokines (IFN-γ and IL-2) and Th2 cytokines (IL-4, IL-5 and IL-10), produced by splenocytes. Our results suggest that EmCRT is an immunodominant protein secreted by E. multilocularis larvae and a vaccine candidate that induces partial protective immunity in vaccinated mice against Echinococcus infection.

show abstract

Enzymatic characteristics and preventive effect of leucine aminopeptidase against Echinococcus multilocularis

Cited by 9 publications

References 47 publications

A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice

A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice

Immunoprotective effect and mechanism of rEg.P29 against CD4<sup>+</sup> T cell-deficient mice with <italic>Echinococcus multilocularis</italic> infection

Immunization with EmCRT-Induced Protective Immunity against Echinococcus multilocularis Infection in BALB/c Mice

Contact Info

Product

Resources

About

Enzymatic characteristics and preventive effect of leucine aminopeptidase against Echinococcus multilocularis

Cited by 9 publications

References 47 publications

A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice

A multi-epitope vaccine GILE against Echinococcus Multilocularis infection in mice

Immunoprotective effect and mechanism of rEg.P29 against CD4&lt;sup&gt;+&lt;/sup&gt; T cell-deficient mice with &lt;italic&gt;Echinococcus multilocularis&lt;/italic&gt; infection

Immunization with EmCRT-Induced Protective Immunity against Echinococcus multilocularis Infection in BALB/c Mice

Contact Info

Product

Resources

About

Immunoprotective effect and mechanism of rEg.P29 against CD4<sup>+</sup> T cell-deficient mice with <italic>Echinococcus multilocularis</italic> infection