Studies On Trichinellla Spiralis Muscle Larve ES Antigen-induced Apoptosis In Mice Thymus Lymphocytes

Trichinosis is a kind of serious zoonosis in the world which is caused by Trichinella spiralis. T. spiralis muscle larvae Excretory-Secretory antigen produced during the growth stage is a good antigen for detection and immunity of Trichinosis. The recent study indicated that apoptosis induced by parasitic infection plays an important role in mechanism of disease. Apoptosis is needed in parasites and related with growth, development, infection and turnover of parasites. There is rare information of Trichinosis and apoptosis internal and external. In order to study the relationship between apoptosis and infection of T. spiralis, mice thymus lymphocytes cultured in vitro was used as experiment materials in this study induced by T. spiralis muscle larvae ES antigen with some items, such as the activity of cells, cell cycle and morphology and biochemistry of apoptosis, detected. The results showed as follows:1. The IC50 of ES antigen on mice thymus lymphocytes was detected by trypan blue method and MTT method respectively. That is, 24.96±0.46μg/ml of 6h, 17.36±0.68μg/ml of 12h, 11.09±0.17μg/ml of 24h, 5.35±0.86μg/ml of 48h. The results showed that ES antigen is harmful to mice thymus lymphocytes, and the sensitivity increases along with time going.2. AO/EB double fluorescence coloration was used to detect apoptosis induced by T. spiralis muscle larvae ES antigen in mice thymus lymphocytes from the morphological aspect. It revealed that ES antigen induced lymphocytes death mainly by the way of apoptosis.3. The effectof ES antigen on DNA damage in mice thymus lymphocytes was detected by the method of DNA Ladder analysis. The result exhibited obvious ladder-shaped straps and showed that ES antigen caused apoptosis of lymphocytes by DNA damage, which exhibited time-dependent effect.4. Flow cytometry assay was used to detect cell cycle and DNA contents of mice thymus lymphocytes, and the result indicated that ES antigen induced apoptosis of mice thymus lymphocytes.The phenomenon of T. spiralis muscle larvae ES antigen induced mice thymus lymphocytes in vitro was detected in this study from morphology and biochemistry, and the results showed that ES antigen induced apoptosis of mice thymus lymphocytes with time-dependent effect, thereby, indicated ES antigen can cause immune suppression of host. This finding will play an important roll in pathogenesis, prevention and therapy of infection of T. spiralis afterward.