Identification and characterization of Leishmania amastigote and axenic form antigens

AA Abdelhaleem; EM Elamin; SM Bakheit; MM Mukhtar

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Identification and characterization of Leishmania amastigote and axenic form antigens

Author: Abdelhaleem, A.A ¹ ; Elamin, E.M ² ; Bakheit, S.M. ³ ; Mukhtar, M.M. ⁴
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1. Tropical and Endemic Diseases Research Unit, Medical Research Centre, Jazan University, P.O. Box 114, Jazan 42145, Saudi Arabia
2. Faculty of Medical Laboratory Sciences, Alzaeim Alazhari University, P.O. Box 1432, Khartoum Bahri 79371 Sudan
3. Institute of Endemic Diseases, University of Khartoum, P.O. Box 321, Khartoum 11111, Sudan
4. Bioscience Research Institute, Ibn Sina University, P.O. Box 10995, Khartoum 11111, Sudan
Publication Type:Journal Article
From:Tropical Biomedicine 2019;36(4):866-873
CountryMalaysia
Language:English
Abstract: This study was aimed to identify and characterize Leishmania amastigote, and axenic form antigens. Two in vitro techniques were used to change leishmania parasite isolates from promastigote form to amastigotes and amastigote like (axenic) forms. The main strategy relied upon in vitro infection of murine macrophages cell line J774 with leishmania promastigote, at 37°C with 5% CO2, while the second technique relied upon the culture of promastigote at 37°C with low pH (5.5), and 5-10% CO2. Proteins were extracted and fractionated utilizing 12% Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE). Antigens were recognized using both immune dot blot and western blot procedures. PCR was performed for recognition of leishmania parasites in infected J774 macrophages. L. major was quicker in infectivity of macrophages cell line than L. donovani. Shared proteins ranging from 26-116 kDa were identified by SDS PAGE in all stages. Immune Dot-blot method showed positive outcomes, while western blot identified an exceptional antigen band of 16 kDa in amastigote, this unique band could be of value in diagnosis and vaccination of leishmaniasis. PCR results confirmed presence of both isolates demonstrating that coinfection is conceivable, and no indications of hereditary recombination at kinetoplast DNA (kDNA) were identified in macrophages simultaneously infected by L. major and L. donovani.
Full text:8.2019my1072.pdf