| Angiostrongylus cantonensis is an extracellular parasite with intermediate and terminal hosts.It can infect most mammals and cause serious infection to the human nervous system.Currently,more and more cases of A.cantonensis have been reported in the world.To establish a rapid,sensitive,simple and highly specific detection method is of great significance for the rapid screening of the disease.In this study,monoclonal antibody12D5 was applied to investigate the alleviating of eosinophilic meningitis caused by A.cantonensis.At the same time,macrophages would be activated into different phenotypic functions through different factor-mediated channels.Firstly,Recombinase Polymerase Amplification(RPA)and Lateral Flow Strip-Based Recombinase Polymerase Amplification(LF-RPA)detection methods for isothermal amplification of recombinase polymerase were established,and it was applied to screen the intermediate host,and analyze the infection status of A.Cantonensis with it in Shenzhen.The result showed that the established RPA method has the high sensitivity and maneuverability,and A.cantonensis positive brown cloud agate screw number 10 years ago showed a trend of increase from shenzhen nanshan district,with the trendance of decrease compared to other regions.Secondly,the mouse model of A.cantonensis infection with monoclonal antibody 12D5 was established.The effect of monoclonal antibody 12D5 on the pathological state of infected mice was investigated by means of mental behavior observation,pathological tissue section,tissue immunofluorescence and blood-brain barrier permeability test.The results showed that the pathological changes of the infected group was significantly reduced compared with that of the untreated group.In M1/M2 polarization ratio,it was found that the immune response of mice presented Th1 type in the early stage of A.cantonensis infection by tissue immunofluorescence assay,.with the extension of the infection period,the immune response changed from Th1 type to Th2 type,and the macrophages also changed from M1 to M2.Monoclonal Ab 12D5 may affect the immune response by inhibiting eosinophilic synthesis,or directly alter the M2 polarization pathway to down-regulate the inflammatory response.Finally,m RNA and miRNA profiles from spleen of mice infected with A.cantonensis after treatment of monoclonal antibody 12D5 was analyzed respectively.After RNA extraction,library construction and sequencing,the biological function analysis also were made,it was found that the miRNAs expression profile from spleens of infected mice injected with monoclonal antibody 12D5 was significantly different from that of mice not immunized with antibody.The main functions of the genes differentially expressed on m RNA focused on intercellular junctions,biological processes and biological regulatory processes,among which the Golgi membrane and chromosome functions related to m RNA enrichment was the highest.Differentially expressed m RNA pathways were mainly concentrated in apoptosis related pathways,C-type lectin receptor signaling pathway related pathways and connection signaling pathways.The differentially expressed miRNA genes are associated with three pathways: cell and glutamate synaptic function,cell cycle related pathways,interaction pathways in stimulating nervous tissues,and RAS signaling pathways.In conclusion,monoclonal antibody 12D5 significantly alleviates the pathological symptoms of eosinophilic meningitis in infected mice,and induces the transformation of the macrophage immune response from Th1 type to Th2 type,thereby downregulating the inflammatory response.Meanwhile,the m RNA and miRNA expression profiles of A.cantonensis infected mice were changed by monoclonal antibody 12D5. |
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