• Applied Microscopy
• /
• v.37 no.1
• /
• pp.43-52
• /
• 2007

In order to observe the localization of excretory, purified and infected antigenic protein in the tissue of Trichinella spiralis larvae, immunogoldlabeling methodology using IgG and protein A-gold complex was implemented. T. spiralis larvae obtained from rat muscle were initially cultured in medium, and secreted excretory antigen was collected for 1 or 3 days. Purified antigenic protein was obtained from homogenized T. spiralis larvae. Rabbits were then immunized with 1 or 3 days secreted excretory protein and purified 45 kDa protein, and IgG was purified from collected serum. Serum, against infected antigen, collected from rat on 1 and 4 weeks after infection with T. spiralis larvae, and IgG was purified from collected serum. T. spiralis larvae were embedded in Lowicryl HM20 medium. Then they were finally treated with immunized IgG and protein A-gold complex (particle size; 15 nm) and observed under electron microscope. In T. spiralis larvae tissue, the tissue antigen reacted with rabbit IgC antigen Day 1 secreted excretory protein, infected antigenic protein and purified 45 kDa protein. But different distribution pattern of labeled gold particles were observed. When Day 1 secreted excretoy protein was used, gold particle labeling was observed specifically on the cuticle, basal layer, esophagus interstitial matrix (EIM) and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. In a separate group of tissue, the antigen reacted with rabbit IgG against Day 3 secreted excretory protein. Labeled gold particles were specifically distributed on the surface layer of cuticle, EIM and ${\alpha}_0$ granules of stichocyte of the worm. In case of using infected antigenic protein, gold particle labeling was specifically distributed on the cuticle and EIM of the worm. When purifed 45 kDa protein was used gold particle labeling was specifically distributed on the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. Therefore, excretory antigens appeared to originate from the cuticle and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte for the first day but the cuticle layer associated with globular proteins and ${\alpha}_0$ granules of stichocyte after 3 days and infected antigens appeared to originate from the cuticle for 1 and 4 weeks after infection. These results suggest that excretory and infection specific antigens are secreted into the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte and 45 kDa protein may be contained these specific antigens.