Analysis And Application Of SRSF3 Protein Phosphorylation Mediating Immune Response In 3D4/21 Cells Infected With Toxoplasma Gondii

Toxoplasma gondii is a eukaryotic organism that is widely parasitic in warm-blooded animals and can form nanoworm membranes in host cells to resist immune system clearance and help itself with nutrient uptake to establish long-term infection.Its infection can lead to Toxoplasma gondii ocular inflammation,encephalitis,and myocarditis in the host.Infection of Toxoplasma gondii in pregnant animals can lead to fetal malformations,miscarriage,and stillbirth.Due to the wide infectivity of Toxoplasma gondii and the serious damage to animals,the human toxoplasmosis and farmed animal toxoplasmosis caused by Toxoplasma gondii are the key control objects in my country.Toxoplasmosis purification in pig farms plays a key role in the growth and reproduction of pigs.Doing a good job in the prevention and control of toxoplasmosis in pigs can reduce the economic losses caused by toxoplasmosis in the farms,such as abortion and death,and can also cut off human consumption caused by toxoplasmosis.Route of infection in pork containing Toxoplasma gondii cysts.In addition,pigs are used as donor animals for organ transplantation,and the purification of pig toxoplasmosis can also greatly reduce the probability of human toxoplasmosis infection caused by organ transplantation.After Toxoplasma gondii infects host cells,it will cause an immune response of the host cells,which in turn stimulates the production of specific immune effector cells that target the foreign invading parasitic protozoa-Toxoplasma gondii,thus limiting the growth and reproduction of Toxoplasma gondii effect.Conversely,the invasion of Toxoplasma gondii will also bring about a series of changes in the physiological and biochemical state of host cells,such as changes in the level of phosphorylation modification of cells.Macrophages are classic immune effector cells.Selecting them as experimental cells can more intuitively reflect the changes in the levels of cytokines and protein phosphorylation after host cells are infected with Toxoplasma gondii,thereby further revealing the molecular biological mechanism of Toxoplasma gondii invading cells.In this experiment,LC-MS/MS technology was used to determine the phosphorylation level and proteomics of porcine alveolar macrophages 3D4/21 after Toxoplasma gondii infection.The results showed that,compared with the uninfected group,3D4/21 after infection with Toxoplasma had general phosphorylation modification differences in its intracellular proteins.The KEGG pathway points to the complement system signaling pathway,NF-κB signaling pathway and TNF signaling pathway.Results The protein SRSF3 with significant difference in phosphorylation modification and the protein NF-κB with significant difference in expression level were quantitatively detected.In order to further confirm that the differentially expressed protein phosphorylation of the Spliceosome pathway is significantly higher,the spliceosome protein SRSF3 was selected and the corresponding verification experiments were carried out;the differentially expressed protein NF-κB and its corresponding I-κB were also analyzed by Western blotting.The results were consistent with those reported in the omics report.To verify whether NF-κB is activated,immunofluorescence staining was performed on 3D4/21 cells infected with Toxoplasma RH,and the results showed that NF-κB was nuclear translocated in 3D4/21 cells infected with Toxoplasma gondii.It showed that the phosphorylation of spliceosome proteins was generally increased after 3D4/21 cells were infected with Toxoplasma gondii,and to a certain extent,it promoted the production of high levels of inflammatory mediators.