The Molecular Mechanism Of Protein Regulation In Response To Babesia Infection In Mice

Babesiosis is a hemozoon disease caused by Babesia.It is reported that the cost cause by Babesiosis as much as hundreds of millions of dollars each year in the worldwide.Mammals are intermediate hosts of Babesia,and ticks are terminal hosts of Babesia.In recent years,more and more new cases of Babesiosis have been reported,which has posed a new threat to human health and livestock development.It is mainly parasitic in the red blood cells of the mammals and is mainly transmitted between mammals or humans and mammals by hard ticks.The host of Babesiosis may present with symptoms such as hyperthermia and anemia,which is cause by mutual antagonism between Babesia and the host.Part of the host can resist and eliminate Babesia through rapid activation of the immune system,although Babesia damage to the host seriously.But the host of this immune regulation mechanism is not clear.The mechanism of host autoimmune regulation is not clear at present.In this experiment,DIA quantitative technology and modified proteomic technology were used to study dynamic change of expression for spleen,heart and blood proteomics and phosphorylated modification proteomics.The key proteins were found in the development of disease,which can be used as diagnostic markers for disease diagnosis marker or target molecules so as to provide new clues and basis for the prevention detection and treatment research.1.It was found that the spleen structure was destroyed seriously at the peak of infection through the spleen histopathology of the mice infected with Babesia.In the later stage of infection,brown pigment granules and many worms were encapsulated in the spleen tissue.Ultramicroscopic pathology show that the number of phagosomes and lysosomes was the highest,mitochondria were swollen and cristae disappeared at the peak of infection.The phagosomes and lysosomes decreased obviously and the mitochondria recovered at late infection.At the early stage of infection,the proportion of CD4~+T lymphocytes was significantly decreased in the body,suggesting that the immunity of Babesia was mainly mediated by cellular immunity.At the same time,Treg cells significantly reduced after Babesia infection with 5 day,there is still no recovery until the infection of 19 day.It is suggest that the body immune response to Babel infection may be gradually enhanced.No effect to resist Babesia infection for the immune response of body,because of Th1cell-mediated immune responsed in the early stage of the infection mainly.2.Proteomics analysis for spleen tissue in different periods of mice infected with Babesia using DIA quantitative proteomic methods.The number of proteins was identified2250,2214,2270,2425 and 2241 at the five time points after the infection of Babesia spleens.1403 proteins of intersection were identified in the five periods.GO enrichment analysis results show that the difference of identified proteins are mainly involved in regulating cell process to stimulate the reaction process of the immune system and antioxidant activity,such as biological processes;KEGG pathway analysis found that these proteins are mainly involved in the metabolic pathway chemokine signaling pathway T/B cell receptor signaling pathway complement and blood coagulation cascade,etc.our analysis the role and molecular mechanism of these proteins in the host immune response against infection.3.Phosphorylated proteomics analysis for spleen tissue in different periods of mice infected with Babesia using DIA quantitative proteomic methods.2261 phosphopeptides were identified in five periods,involving 2470 phosphorylation sites and corresponding 1046proteins.KEGG analysis revealed that immune-related signaling pathways were enriched at different stages.Of which,CD45 plays an important role in immune regulation.The spleen of mice can remove Babesia and restore the original function quickly through spleen cells regulate the immune response by phosphorylating and dephosphorylating proteins.4.Proteomics analysis for heart tissue in different periods of mice infected with Babesia found that 1092 proteins of intersection were identified in the five periods.The change trend of proteins was identified by Cluster analysis.For 331 protein,GO and KEGG analysis showed that these pathways involved in biological process in a variety of stress,including complement and blood coagulation system oxidative phosphorylation protein synthesis metabolism related energy metabolism,these may be related to red blood cells rupture and hemolytic anemia,following the body was infected with Babesia.5.Phosphorylated proteomics analysis for heart tissue in different periods of mice infected with Babesia found that 1362,1370,1370,1370 and 1382 proteins were identified in five periods.GO analysis indicated that these proteins mainly involved in the binding activity,and participated in the material metabolism,biological regulation,catalytic activity,localization,the cell component organization and the biological synthesis process.KEGG pathway analysis revealed that the most of proteins involved in the metabolic pathway,followed by the insulin signaling pathway.Among the various signaling pathways,the Babesia infection had obviously influence on the energy metabolism related signaling pathway.6.Proteomics analysis for serum in different periods of mice infected with Babesia.304proteins were identified in the seven periods.The heat map results showed that most proteins of the infected mice were up-regulated during the infection and recovery period,including proteins involved in the inflammatory response,involving in the immune defense and neutralizing the virus.GO and KEGG analysis found that infection period and recovery of the differentially expressed proteins were enriched by hydrolysis enzyme activity、serine peptidase inhibitors activity、peptide enzyme inhibitors activity.The large number of infection and the recovery quantity changes of protein expression mainly involves the complement and blood coagulation cascade signaling pathways.This study presents the data how mammals regulate the immune response through protein changes the host immune,has important significance for response to Babesia infection.The results of this study indepth understand that the pathogenic mechanism of Babesiosis,as well as laid a foundation for the effective diagnosis,prevention and treatment of Babesiosis.