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Protective immunity against Trichinella spiralis infection induced by a multi-epitope vaccine in a murine model.

Gu Y, Wei J, Yang J, Huang J, Yang X, Zhu X - PLoS ONE (2013)

Bottom Line: To increase more effective protection, the epitope 8F7 that was found to induce the highest protection in this study was combined with two other previously identified epitopes (YX1 from Ts-Pmy and M7 from Ts-87) to formulate a multi-epitope vaccine.This protection is significantly higher than that induced by individual-epitope peptides and is associated with high levels of subclasses IgG and IgG1.These results showed that a multi-epitope vaccine induced better protective immunity than an individual epitope and provided a feasible approach for developing a safer and more effective vaccine against trichinellosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology, School of Basic Medical Sciences, Capital Medical University, Beijing, PR China.

ABSTRACT
Trichinellosis is one of the most important food-borne parasitic zoonoses throughout the world. Because infected pigs are the major source of human infections, and China is becoming the largest international producer of pork, the development of a transmission-blocking vaccine to prevent swine from being infected is urgently needed for trichinellosis control in China. Our previous studies have demonstrated that specific Trichinella spiralis paramyosin (Ts-Pmy) and Ts-87 antigen could provide protective immunity against T. spiralis infection in immunized mice. Certain protective epitopes of Ts-Pmy and Ts-87 antigen have been identified. To identify more Ts-Pmy protective epitopes, a new monoclonal antibody, termed 8F12, was produced against the N-terminus of Ts-Pmy. This antibody elicited significant protective immunity in mice against T. spiralis infection by passive transfer and was subsequently used to screen a random phage display peptide library to identify recognized epitopes. Seven distinct positive phage clones were identified and their displayed peptides were sequenced. Synthesized epitope peptides conjugated to keyhole limpet hemocyanin were used to immunize mice, four of which exhibited larval reduction (from 18.7% to 26.3%, respectively) in vaccinated mice in comparison to the KLH control. To increase more effective protection, the epitope 8F7 that was found to induce the highest protection in this study was combined with two other previously identified epitopes (YX1 from Ts-Pmy and M7 from Ts-87) to formulate a multi-epitope vaccine. Mice immunized with this multi-epitope vaccine experienced a 35.0% reduction in muscle larvae burden after being challenged with T. spiralis larvae. This protection is significantly higher than that induced by individual-epitope peptides and is associated with high levels of subclasses IgG and IgG1. These results showed that a multi-epitope vaccine induced better protective immunity than an individual epitope and provided a feasible approach for developing a safer and more effective vaccine against trichinellosis.

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Specific binding of positive phage clones to mAb 8F12 by Western blot (A) and immunoprecipitation (B) analyses.(A). Seven positive phage clones were transferred to a PVDF membrane and a wild-type M13 phage was used as a negative control. The mAb 8F12 recognized a single band with a MW of approximately 60 kDa (displayed peptide co-expressed with phage coat protein pIII) in all seven positive clones, but not in the M13 control. M:Marker; Lanes 1-7: seven positive phage clones; Lane 8: Wild-type M13 phage. (B). Immunoprecipitation of positive phage 8F7 with mAb 8F12 at non-denatured and denatured conditions. M:Marker; Lane 1: mAb 8F12 pulled phage 8F7 in non-denaturing lysis buffer; Lane 2: mAb 8F12 pulled wild-type M13 phage in non-denaturing lysis buffer; Lane 3: mAb 8F12 pulled phage 8F7 in denaturing lysis buffer; Lane 4: mAb 8F12 pulled wild-type M13 phage in denaturing lysis buffer; Lanes 5: phage 8F7 only; Lane 6: mAb 8F12 pulled rTs-Pmy-N.
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pone-0077238-g003: Specific binding of positive phage clones to mAb 8F12 by Western blot (A) and immunoprecipitation (B) analyses.(A). Seven positive phage clones were transferred to a PVDF membrane and a wild-type M13 phage was used as a negative control. The mAb 8F12 recognized a single band with a MW of approximately 60 kDa (displayed peptide co-expressed with phage coat protein pIII) in all seven positive clones, but not in the M13 control. M:Marker; Lanes 1-7: seven positive phage clones; Lane 8: Wild-type M13 phage. (B). Immunoprecipitation of positive phage 8F7 with mAb 8F12 at non-denatured and denatured conditions. M:Marker; Lane 1: mAb 8F12 pulled phage 8F7 in non-denaturing lysis buffer; Lane 2: mAb 8F12 pulled wild-type M13 phage in non-denaturing lysis buffer; Lane 3: mAb 8F12 pulled phage 8F7 in denaturing lysis buffer; Lane 4: mAb 8F12 pulled wild-type M13 phage in denaturing lysis buffer; Lanes 5: phage 8F7 only; Lane 6: mAb 8F12 pulled rTs-Pmy-N.

Mentions: Ten positive phage clones were obtained after three rounds of biopanning with mAb 8F12. ELISA results showed that each positive phage clone was able to specifically bind to mAb 8F12, but not to the BSA negative control (Figure 2). Western blotting showed that a single band with a MW of approximately 60 kDa (displayed peptide co-expressed with phage coat protein pIII) was recognized in all seven sequenced phage clones by mAb 8F12 but not in the M13 plain phage (Figure 3A). Positive phage clone 8F7 (as a representative) could be recognized and pulled down by mAb 8F12 at both non-denatured and denatured conditions through immunoprecipitation (Figure 3B).


Protective immunity against Trichinella spiralis infection induced by a multi-epitope vaccine in a murine model.

Gu Y, Wei J, Yang J, Huang J, Yang X, Zhu X - PLoS ONE (2013)

Specific binding of positive phage clones to mAb 8F12 by Western blot (A) and immunoprecipitation (B) analyses.(A). Seven positive phage clones were transferred to a PVDF membrane and a wild-type M13 phage was used as a negative control. The mAb 8F12 recognized a single band with a MW of approximately 60 kDa (displayed peptide co-expressed with phage coat protein pIII) in all seven positive clones, but not in the M13 control. M:Marker; Lanes 1-7: seven positive phage clones; Lane 8: Wild-type M13 phage. (B). Immunoprecipitation of positive phage 8F7 with mAb 8F12 at non-denatured and denatured conditions. M:Marker; Lane 1: mAb 8F12 pulled phage 8F7 in non-denaturing lysis buffer; Lane 2: mAb 8F12 pulled wild-type M13 phage in non-denaturing lysis buffer; Lane 3: mAb 8F12 pulled phage 8F7 in denaturing lysis buffer; Lane 4: mAb 8F12 pulled wild-type M13 phage in denaturing lysis buffer; Lanes 5: phage 8F7 only; Lane 6: mAb 8F12 pulled rTs-Pmy-N.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3795051&req=5

pone-0077238-g003: Specific binding of positive phage clones to mAb 8F12 by Western blot (A) and immunoprecipitation (B) analyses.(A). Seven positive phage clones were transferred to a PVDF membrane and a wild-type M13 phage was used as a negative control. The mAb 8F12 recognized a single band with a MW of approximately 60 kDa (displayed peptide co-expressed with phage coat protein pIII) in all seven positive clones, but not in the M13 control. M:Marker; Lanes 1-7: seven positive phage clones; Lane 8: Wild-type M13 phage. (B). Immunoprecipitation of positive phage 8F7 with mAb 8F12 at non-denatured and denatured conditions. M:Marker; Lane 1: mAb 8F12 pulled phage 8F7 in non-denaturing lysis buffer; Lane 2: mAb 8F12 pulled wild-type M13 phage in non-denaturing lysis buffer; Lane 3: mAb 8F12 pulled phage 8F7 in denaturing lysis buffer; Lane 4: mAb 8F12 pulled wild-type M13 phage in denaturing lysis buffer; Lanes 5: phage 8F7 only; Lane 6: mAb 8F12 pulled rTs-Pmy-N.
Mentions: Ten positive phage clones were obtained after three rounds of biopanning with mAb 8F12. ELISA results showed that each positive phage clone was able to specifically bind to mAb 8F12, but not to the BSA negative control (Figure 2). Western blotting showed that a single band with a MW of approximately 60 kDa (displayed peptide co-expressed with phage coat protein pIII) was recognized in all seven sequenced phage clones by mAb 8F12 but not in the M13 plain phage (Figure 3A). Positive phage clone 8F7 (as a representative) could be recognized and pulled down by mAb 8F12 at both non-denatured and denatured conditions through immunoprecipitation (Figure 3B).

Bottom Line: To increase more effective protection, the epitope 8F7 that was found to induce the highest protection in this study was combined with two other previously identified epitopes (YX1 from Ts-Pmy and M7 from Ts-87) to formulate a multi-epitope vaccine.This protection is significantly higher than that induced by individual-epitope peptides and is associated with high levels of subclasses IgG and IgG1.These results showed that a multi-epitope vaccine induced better protective immunity than an individual epitope and provided a feasible approach for developing a safer and more effective vaccine against trichinellosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology, School of Basic Medical Sciences, Capital Medical University, Beijing, PR China.

ABSTRACT
Trichinellosis is one of the most important food-borne parasitic zoonoses throughout the world. Because infected pigs are the major source of human infections, and China is becoming the largest international producer of pork, the development of a transmission-blocking vaccine to prevent swine from being infected is urgently needed for trichinellosis control in China. Our previous studies have demonstrated that specific Trichinella spiralis paramyosin (Ts-Pmy) and Ts-87 antigen could provide protective immunity against T. spiralis infection in immunized mice. Certain protective epitopes of Ts-Pmy and Ts-87 antigen have been identified. To identify more Ts-Pmy protective epitopes, a new monoclonal antibody, termed 8F12, was produced against the N-terminus of Ts-Pmy. This antibody elicited significant protective immunity in mice against T. spiralis infection by passive transfer and was subsequently used to screen a random phage display peptide library to identify recognized epitopes. Seven distinct positive phage clones were identified and their displayed peptides were sequenced. Synthesized epitope peptides conjugated to keyhole limpet hemocyanin were used to immunize mice, four of which exhibited larval reduction (from 18.7% to 26.3%, respectively) in vaccinated mice in comparison to the KLH control. To increase more effective protection, the epitope 8F7 that was found to induce the highest protection in this study was combined with two other previously identified epitopes (YX1 from Ts-Pmy and M7 from Ts-87) to formulate a multi-epitope vaccine. Mice immunized with this multi-epitope vaccine experienced a 35.0% reduction in muscle larvae burden after being challenged with T. spiralis larvae. This protection is significantly higher than that induced by individual-epitope peptides and is associated with high levels of subclasses IgG and IgG1. These results showed that a multi-epitope vaccine induced better protective immunity than an individual epitope and provided a feasible approach for developing a safer and more effective vaccine against trichinellosis.

Show MeSH
Related in: MedlinePlus