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Use of Phage Display technology in development of canine visceral leishmaniasis vaccine using synthetic peptide trapped in sphingomyelin/cholesterol liposomes.

Toledo-Machado CM, Bueno LL, Menezes-Souza D, Machado-de-Avila RA, Nguyen C, Granier C, Bartholomeu DC, Chávez-Olórtegui C, Fujiwara RT - Parasit Vectors (2015)

Bottom Line: Leishmania parasites can cause visceral or cutaneous disease and are found in subtropical and tropical regions of the Old and New World.Selected peptide was formulated with aluminum hydroxide and liposomes and immunization was performed in BALB/c mice.Our results demonstrate that immunization with Peptide 5 when formulated with aluminum hydroxide and liposomes is immunogenic and elicited significant protection associated with the induction of mixed Th1/Th2 immune response against L. infantum infection.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. cmonerat@yahoo.com.br.

ABSTRACT

Background: Leishmania parasites can cause visceral or cutaneous disease and are found in subtropical and tropical regions of the Old and New World. The pathology of the infection is determined by both host immune factors and species/strain differences of the parasite. Dogs represent the major reservoir of Leishmania infantum (syn. L. chagasi) and vaccines are considered the most cost-effective control tools for canine disease.

Methods: Selection of immunodominant peptides was performed by Phage Display to identify sequences recognized by L. infantum naturally infected animals. Sera from Leishmania infected animals were used in the biopanning to selection of specific peptides. Serum samples from T. cruzi infected and healthy animals were used as control. After selection, synthetic peptides were produced in membrane (spot-synthesis) in soluble form and blotting and ELISA were performed for validation of serum reactivity. Selected peptide was formulated with aluminum hydroxide and liposomes and immunization was performed in BALB/c mice. Protection was determined by qPCR after challenge infection with virulent L. infantum.

Results: We reported the selection of Peptide 5 through Phage Display technique and demonstrate its ability to promote a state of immunity against L. infantum infection in murine model after immunization using liposomes as vaccine carrier. Our results demonstrate that immunization with Peptide 5 when formulated with aluminum hydroxide and liposomes is immunogenic and elicited significant protection associated with the induction of mixed Th1/Th2 immune response against L. infantum infection.

Conclusion: Peptide 5 is a promising vaccine candidate and the findings obtained in the present study encourage canine trials to confirm the effectiveness of a vaccine against CVL.

No MeSH data available.


Related in: MedlinePlus

Parasite load after challenge infection. The number of parasites in the spleen was measured by Real-Time PCR technique and showed by number of parasites per 20 ng of DNA total. DNA was extracted from the spleen samples of mice immunized with vaccines containing Phosphate Buffered Saline (PBS = black circles - control group), Leishmania infantum antigen (LiAg = black squares) or Peptide 5 (Pep 5 = black triangles). Mean ± standard deviation (SD) in each group is shown. Statistically significant differences in the parasite load between experimental groups are showed in the graph.
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Fig5: Parasite load after challenge infection. The number of parasites in the spleen was measured by Real-Time PCR technique and showed by number of parasites per 20 ng of DNA total. DNA was extracted from the spleen samples of mice immunized with vaccines containing Phosphate Buffered Saline (PBS = black circles - control group), Leishmania infantum antigen (LiAg = black squares) or Peptide 5 (Pep 5 = black triangles). Mean ± standard deviation (SD) in each group is shown. Statistically significant differences in the parasite load between experimental groups are showed in the graph.

Mentions: Parasite quantification was performed using DNA extracted from the spleen samples of mice from Peptide 5, LiAg and control groups. Animals immunized with Peptide 5 presented significant reduction of parasite burden (up to 98%), when compared to control and LiAg groups (Figure 5). Peptide 5 demonstrated an excellent effectiveness as vaccine antigen.Figure 5


Use of Phage Display technology in development of canine visceral leishmaniasis vaccine using synthetic peptide trapped in sphingomyelin/cholesterol liposomes.

Toledo-Machado CM, Bueno LL, Menezes-Souza D, Machado-de-Avila RA, Nguyen C, Granier C, Bartholomeu DC, Chávez-Olórtegui C, Fujiwara RT - Parasit Vectors (2015)

Parasite load after challenge infection. The number of parasites in the spleen was measured by Real-Time PCR technique and showed by number of parasites per 20 ng of DNA total. DNA was extracted from the spleen samples of mice immunized with vaccines containing Phosphate Buffered Saline (PBS = black circles - control group), Leishmania infantum antigen (LiAg = black squares) or Peptide 5 (Pep 5 = black triangles). Mean ± standard deviation (SD) in each group is shown. Statistically significant differences in the parasite load between experimental groups are showed in the graph.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4352561&req=5

Fig5: Parasite load after challenge infection. The number of parasites in the spleen was measured by Real-Time PCR technique and showed by number of parasites per 20 ng of DNA total. DNA was extracted from the spleen samples of mice immunized with vaccines containing Phosphate Buffered Saline (PBS = black circles - control group), Leishmania infantum antigen (LiAg = black squares) or Peptide 5 (Pep 5 = black triangles). Mean ± standard deviation (SD) in each group is shown. Statistically significant differences in the parasite load between experimental groups are showed in the graph.
Mentions: Parasite quantification was performed using DNA extracted from the spleen samples of mice from Peptide 5, LiAg and control groups. Animals immunized with Peptide 5 presented significant reduction of parasite burden (up to 98%), when compared to control and LiAg groups (Figure 5). Peptide 5 demonstrated an excellent effectiveness as vaccine antigen.Figure 5

Bottom Line: Leishmania parasites can cause visceral or cutaneous disease and are found in subtropical and tropical regions of the Old and New World.Selected peptide was formulated with aluminum hydroxide and liposomes and immunization was performed in BALB/c mice.Our results demonstrate that immunization with Peptide 5 when formulated with aluminum hydroxide and liposomes is immunogenic and elicited significant protection associated with the induction of mixed Th1/Th2 immune response against L. infantum infection.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Parasitologia, ICB, Universidade Federal de Minas Gerais, CP: 486 - CEP: 31.270-901, Belo Horizonte, Minas Gerais, Brazil. cmonerat@yahoo.com.br.

ABSTRACT

Background: Leishmania parasites can cause visceral or cutaneous disease and are found in subtropical and tropical regions of the Old and New World. The pathology of the infection is determined by both host immune factors and species/strain differences of the parasite. Dogs represent the major reservoir of Leishmania infantum (syn. L. chagasi) and vaccines are considered the most cost-effective control tools for canine disease.

Methods: Selection of immunodominant peptides was performed by Phage Display to identify sequences recognized by L. infantum naturally infected animals. Sera from Leishmania infected animals were used in the biopanning to selection of specific peptides. Serum samples from T. cruzi infected and healthy animals were used as control. After selection, synthetic peptides were produced in membrane (spot-synthesis) in soluble form and blotting and ELISA were performed for validation of serum reactivity. Selected peptide was formulated with aluminum hydroxide and liposomes and immunization was performed in BALB/c mice. Protection was determined by qPCR after challenge infection with virulent L. infantum.

Results: We reported the selection of Peptide 5 through Phage Display technique and demonstrate its ability to promote a state of immunity against L. infantum infection in murine model after immunization using liposomes as vaccine carrier. Our results demonstrate that immunization with Peptide 5 when formulated with aluminum hydroxide and liposomes is immunogenic and elicited significant protection associated with the induction of mixed Th1/Th2 immune response against L. infantum infection.

Conclusion: Peptide 5 is a promising vaccine candidate and the findings obtained in the present study encourage canine trials to confirm the effectiveness of a vaccine against CVL.

No MeSH data available.


Related in: MedlinePlus