Hossein Rezvan; Ali Khodadadi; Selman Ali
Abstract
Background: Leishmania is a pathogenic parasite which infects mononuclear cells in vertebrate hosts. Different strategies have been taken to develop immunity against Leishmania . DCs loaded with immunogenic antigen have resulted in different levels of Th1-type immune response and cytotoxic T lymphocytes ...
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Background: Leishmania is a pathogenic parasite which infects mononuclear cells in vertebrate hosts. Different strategies have been taken to develop immunity against Leishmania . DCs loaded with immunogenic antigen have resulted in different levels of Th1-type immune response and cytotoxic T lymphocytes (CTL) activity. Objective: To evaluate the potency of DCs primed with soluble Leishmania mexicana antigens (SLA) in developing CTL activity. Methods: DCs were loaded with SLA and injected to Balb/c mice. After two weeks the mice were sacrificed and their splenocytes were used as effector cells in a standard 4-hour cytotoxicity assay against DCs transfected with pcDNA3 containing L. mexicana gp63 gene. Results: Immunization of Balb/c mice with DCs loaded with SLA resulted in high levels of CTL activity against DCs transfected with pcDNA3 containing L. mexicana gp63 gene. Conclusions: The results indicate a high potency for DCs primed with Leishmania antigens in inducing CTL activity, which can be used for developing an immunogenic vaccine against Leishmania.
Seyed Mahmoud Sadjjadi; Sadreddin Mohseni Ardehali; Reza Adibmanesh; Ezzatollah Basiri
Abstract
Background: Different methods have been used for characterization of Leishmania promastigotes. Monoclonal antibodies are useful in characterization of Leishmania spp . both amastogotes and promastigotes. Objective: Comparing the characterization of Leishmania spp. promastigotes with immunoperoxidase ...
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Background: Different methods have been used for characterization of Leishmania promastigotes. Monoclonal antibodies are useful in characterization of Leishmania spp . both amastogotes and promastigotes. Objective: Comparing the characterization of Leishmania spp. promastigotes with immunoperoxidase test (Avidin-Biotin) techniques and an indirect immunofluorescent assay (IFA). Methods: Application of specific monoclonal antibodies for characterization of different Leishmania species. Immunoperoxidase tests (Avidin-Biotin) and indirect immunofluorescent assay (IFA) were employed for characterization of different Leishmnia promastigotes from culture. Five monoclonal antibodies including LXXVIII-2E5- A8 (D2) specific for L. donovani:L. infantum , IS2-2B4 (A11) specific for L. tropica, XCIV-H2- AB (T10) specific for L. tropica, XLVI-5B8- B3 (T1) specific for L. major, and T7 reactive to both L. major and L. tropica as well as an anti GP63 mAb were used. Results: The best result was obtained with the dilution of 1:50 for both mAb and conjugate. One hundred percent sensitivity and specificity was achieved for characterization of Leishmania promastogotes with both methods. Conclusion: As immunoperoxidase method needs less equipments compared to IFA technique, it would be a preferred method for characterization of promastigotes.
Ezzatallah Basiri; Sadreddin Mohseni Ardehali; Mehrnoosh Doroudchi; Fereidoun Mahboodi; Arsalan Kharazmi; Abbas Ghaderi
Abstract
Background : Production of monoclonal antibodies to Leishmania antigens assists the identification and characterization of these organisms. Objective: Production of monoclonal antibodies against epitopes on the gp63. Methods: Two murine monoclonal antibodies to gp63 were produced and characterized. The ...
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Background : Production of monoclonal antibodies to Leishmania antigens assists the identification and characterization of these organisms. Objective: Production of monoclonal antibodies against epitopes on the gp63. Methods: Two murine monoclonal antibodies to gp63 were produced and characterized. The reactions of both antibodies with soluble leishmanial antigens, purified gp63 and truncated recombinant gp63 molecules were studied by an ELISA assay. These two antibodies reacted with the crude soluble antigens prepared from 4 reference strains of Leishmania, 10 isolates from the patients, purified gp63 and recombinant gp63 molecules. However, no reaction with several non-leishmanial antigens was observed. Reaction of both antibodies with the intact recombinant gp63 and truncated molecules were compared. Results: The results indicated that the two antibodies specifically recognize two different epitopes on the gp63 molecule. Conclusion: Possible applications of such antibodies in searching for immunogenic epitopes are discussed.
Seyed Mahmoud Sadjjadi; Sadreddin Mohseni Ardehali; Reza Adibmanesh; Ezzatollah Basiri; Rahmatollah Salmanpour
Abstract
Background: Monoclonal antibodies have been employed extensively for the identification of Leishmania species, development of diagnostic tests and in the characterization of defined leishmanial antigens. Objectives: Identification and characterization of Leishmania spp. directly from cutaneous ...
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Background: Monoclonal antibodies have been employed extensively for the identification of Leishmania species, development of diagnostic tests and in the characterization of defined leishmanial antigens. Objectives: Identification and characterization of Leishmania spp. directly from cutaneous lesions of infected individuals. Methods: An immunoperoxidase test (Avidin-Biotin technique) using monoclonal antibodies was used for this purpose. One hundred and fifty individuals referring to Dermatology Clinic or Parasitology and Mycology Department of Shiraz University of Medical Sciences were chosen of whom a total of 28 individuals whose smears showed a large number of amastigotes after staining with Giemsa were included in this study. Five monoclonal antibodies designated: D2 (against L. donovani), A11 and T10 (against L. tropica), T1 (against L. major) and T7 (against L. tropica and L. major) were used. Amastigotes were identified by Labeled Avidin Biotin (LAB) method. Results: LAB method for identification of amastigotes in impression smears of patient lesions showed that 20 out of 28 cases (71%) were positive. Among these 12 (60%) and 7(35%) were identified as L. tropica and L. major respectively. Conclusion: The results showed that immunoperoxidase is suitable for in situ identification and characterization of Leishmania spp. at the species level.
