Presence of Immunoglobulins on the Surface of Lesion-Derived Amastigotes of Leishmania major As a Tool for Isolation of Amastigote Stage from BALB/c Mice

The onset of infections by Leishmania parasites mainly caused by amastigote growth inside the macrophages. One of the important properties of lesion-derived amastigote is thought to be the attachment of various host proteins including immunoglobulins on the surface of amastigote. In this study, the presence of immunoglobulins on the surface of lesion-derived amastigotes was detected by Western blotting using three different peroxidase conjugated anti-heavy chain antibodies and peroxidase conjugated anti-mouse IgG antibody. Then, a new technique was developed for isolation of lesion-derived amastigotes. This technique simply consists of a microbiological plate covered by rabbit anti-mouse immunoglobulins. Overnight incubation of the infected cell suspension isolated from mice lesion on such plate at 4°C, was followed by gentle washing and isolation of the amastigotes. The results showed that the surface of amastigote has covered with different amount of immunoglobulins such as IgG, IgM, and IgA detected by pixel analysis software. With this technique, which was comparable with other techniques, pure amastigote was isolated. This is a simple and reliable method for isolation of real amastigotes from lesion of the infected BALB/c mice