Behnam Mohammadi-Ghalehbin; Gholamreza Hatam; Bahador Sarkari; Mehdi Mohebali; Zabih Zarei; Shahab Bohlooli
Volume 14, Issue 4 , December 2017, , Pages 293-305
Abstract
Background: Canine visceral leishmaniasis (CVL) caused by Leishmania infantum is endemic in the northwest and south of Iran. An appropriate vaccine can help to prevent and control visceral leishmaniasis in both humans and animals. Few studies have confirmed that the fucose-mannose ligand (FML) antigen ...
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Background: Canine visceral leishmaniasis (CVL) caused by Leishmania infantum is endemic in the northwest and south of Iran. An appropriate vaccine can help to prevent and control visceral leishmaniasis in both humans and animals. Few studies have confirmed that the fucose-mannose ligand (FML) antigen of Leishmania donovani produced protective immunity in dogs against CVL. Objective: To evaluate the immune responses of vaccinated dogs against FML antigen of L. infantum. Methods: We isolated the FML antigen from native L. infantum and vaccinated the dogs with FML-saponin in an endemic area of VL in Iran to evaluate the immune responses of vaccinated dogs against this antigen. Results: Our results indicated a significant increase in the expression of IFN-γ, IL-10 and IL-13, but not IL-12A, gene transcripts in PBMCs of FML-saponin vaccinated dogs in comparison with controls. Our findings showed a significant difference in the ratio of IFN-γ/IL-10 mRNA expression in FML-saponin vaccinated dogs in comparison with two control groups. Moreover, a significant level of anti-FML antibodies was detected in serum of vaccinated dogs. Conclusion: These findings showed that FML-saponin stimulates both Th1 and Th2 immune responses with predominant Th1 and strong humoral immune responses to produce protective immunity against CVL.
Amir Savardashtaki; Bahador Sarkari; Farzane Arianfar; Zohreh Mostafavi-Pour
Volume 14, Issue 2 , June 2017, , Pages 111-122
Abstract
Background: Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory. Objective: The current study aimed at sub-cloning a gene, encoding the B8/1 subunit ...
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Background: Cystic echinococcosis (CE), as a chronic parasitic disease, is a major health problem in many countries. The performance of the currently available serodiagnostic tests for the diagnosis of CE is unsatisfactory. Objective: The current study aimed at sub-cloning a gene, encoding the B8/1 subunit of antigen B (AgB) from Echinococcus granulosus, using gene optimization for the immunodiagnosis of human CE. Methods: The coding sequence for AgB8/1 subunit of Echinococcus granulosus was selected from GenBank and was gene-optimized. The sequence was synthesized and inserted into pGEX-4T-1 vector. Purification was performed with GST tag affinity column. Diagnostic performance of the produced recombinant antigen, native antigen B and a commercial ELISA kit were further evaluated in an ELISA system, using a panel of sera from CE patients and controls. Results: SDS-PAGE demonstrated that the protein of interest had a high expression level and purity after GST tag affinity purification. Western blotting verified the immunoreactivity of the produced recombinant antigen with the sera of CE patients. In an ELISA system, the sensitivity and specificity (for human CE diagnosis) of the recombinant antigen, native antigen B and commercial kit were respectively 93% and 92%, 87% and 90% and 97% and 95%. Conclusion: The produced recombinant antigen showed a high diagnostic value which can be recommended for serodiagnosis of CE in Iran and other CE-endemic areas. Utilizing the combination of other subunits of AgB8 would improve the performance value of the introduced ELISA system.
Hamid Reza Rahimi; Tahereh Mohammadzadeh; Seyed Mahmoud Sadjjadi; Bahador Sarkari; Farzaneh Zahabiun
Volume 14, Issue 2 , June 2017, , Pages 123-133
Abstract
Background: Echinococcosis is a zoonotic parasitic disease caused by the larval stage of Echinococcus granulosus. Several native and recombinant antigens, derived from different stages of E. granulosus life cycle, have been used for vaccine trials. In vitro reared adult worms are good candidates for ...
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Background: Echinococcosis is a zoonotic parasitic disease caused by the larval stage of Echinococcus granulosus. Several native and recombinant antigens, derived from different stages of E. granulosus life cycle, have been used for vaccine trials. In vitro reared adult worms are good candidates for vaccination as they do not produce fertile egg/s and do not have any risk of contamination for researchers. Objective: To evaluate different antigens derived from in vitro reared E. granulosus adult worms for the immunization of BALB/c mice against secondary hydatidosis. Methods: Viable protoscoleces (PCSs) of sheep hydatid cyst were cultivated in S.10E.H media. Excretory secretory (E/S) and crude antigens were prepared from reared adult worms. A total of fifty BALB/c mice, each 8-weeks-old, were divided into 5 groups of 10 mice. Three groups were subcutaneously immunized with crude, E/S and immunodominant antigens on days 1 and 28. The fourth group received only PBS and the fifth group had no injection. Three weeks following the second immunization, all groups were challenged, intraperitoneal, with viable PSCs. After the autopsy of the mice and opening their abdominal wall, cysts were counted and measured followed by histopathological observations. Results: The highest protective immunity (98.7%) against hydatidosis was induced by crude antigen, followed by E/S and immunodominant antigens. Conclusion: Antigens (crude antigens in particular) derived from in vitro reared E. granulosus adult worms, and their different protein components are suitable candidates for the vaccination of intermediate hosts against hydatidosis.
