| Background:Echinococcus granulosus is a chronic infectious disease caused by larvae infected with Echinococcus granulosus.The disease is infected all over the world,causing heavy burdens and economic losses to agriculture and animal husbandry.China is also one of the high incidence areas of the disease.The main cause of human infection with echinococcosis is caused by unfortunate eating of its eggs.Ingested eggs will gradually hatch in the stomach and duodenum of the human body,releasing hexacoccidia,invading the intestinal mucosa,and developing into echinococcus through the blood circulation and parasitizing the liver,causing liver hydatidosis;Hexagrams can pass through the liver,reach the lungs,brain,and bone marrow to cause lesions in these parts.Echinococcus granulosus can survive in humans for years to decades.At present,the treatment of fine-grained echinococcal disease mainly includes surgical resection,benzimidazole(BZ)drug treatment,subcutaneous puncture,and observation(monitoring of inactive cysts).At present,the preferred treatment method is still surgical resection,but when multiple cysts appear in multiple organs or surgical contraindications,in order to reduce recurrence,drugs still need to be treated accordingly.To date,the preferred drugs for the treatment of Echinococcus granulosus are the benzimidazoles,including albendazole and mebendazole.However,benzimidazole drugs can cause more side effects when treating echinococcus disease,such as obvious gastrointestinal reactions and low absorption efficiency.In addition,BZ as an antiparasitic drug,the more obvious deficiency is that some patients will still relapse after treatment.Therefore,we need to develop new drugs to treat Echinococcus granulosus.Objective:To study the therapeutic effect and immune mechanism of IL-28B on Echinococcus granulosus infection model mice.Methods:Intraperitoneal injection of protococcidiosis in female BALB/c mice to establish an Echinococcus granulosus infection model,the mice were randomly divided into the following 5 groups after 18 weeks of infection:Untreated group:Infected with Echinococcus granulosus protococcus,after 18weeks of modeling,the drug was started,and only PBS was given for intervention.rAd-EGFP group:Infected with Echinococcus granulosus protococcus,after 18weeks of modeling,the drug was given,and rAd-EGFP 100?l(10~8PFU/ml)was injected intramuscularly.The drug was given once a week.ABZ group:Albendazole,infected with Echinococcus granulosus protococcus,after 18 weeks of modeling,the drug was started,and albendazole was given by gavage,100 mg/kg/day.rAd-mIL-28B group:Infected with Echinococcus granulosus protococcus,after18 weeks of modeling,began to administer,intramuscular injection of rAd-mIL-28B100?l(10~8PFU/ml),once a week.ABZ+rAd-mIL-28B group:ABZ combined with rAd-mIL-28B treatment group,infected with Echinococcus granulosus protococcus,after 18 weeks of modeling,began to administer,albendazole intragastric,100mg/kg/day At the same time,rAd-mIL-28B100?l(10~8PFU/ml)was injected intramuscularly at the same time,and the drug was given once a week.After 6 weeks of treatment,the mice were sacrificed,the number of cysts of Echinococcus granulosus was counted,the cyst weight was weighed,and the inhibition rate of the cyst growth of Echinococcus granulosus was calculated by the drug;the liver and spleen were weighed to calculate the spleen index,Liver index;HE staining of cysts and liver to detect pathological changes of cysts and liver tissues in mice;observation of ultrastructure of cyst wall of mouse hydatid cysts under scanning electron microscope;detection of spleen and liver by flow cytometry T lymphocyte subsets;ELISA detection of serum IL-10,TGF-?,IFN-?expression levels and IL-10,TGF-?expression levels in peritoneal fluid;protein chip detection of serum Th1,Th2and Th17 cytokines level.Result:ABZ+rAd-mIL-28B can significantly reduce the weight of cysts of Echinococcus granulosus,and the combined treatment shows a superimposed therapeutic effect.Compared with the untreated group,the ABZ+rAd-mIL-28B group(cyst wet weight 5.8±1.4g,cyst growth inhibition rate 55.7%)significantly reduced cyst wet weight(p<0.05);meanwhile,the inhibition rate of this group was significantly higher than rAd-EGFP(12.6±4.0g,3.8%),ABZ alone treatment group(7.1±1.7g,45.8%)(p<0.05).H&E staining results showed that the horny layer of the cysts in the Untreated group had a uniform texture and a complete structure;the horny layer of the mouse cysts in the rAd-mIL-28B group and ABZ+rAd-mIL-28B group became significantly incomplete and the number of adventitia appeared Infiltration of many lymphocytes.The ultrastructure of the cysts was observed with an electron microscope.The surface of the germinal layer of the cyst in the Untreated group was smooth and the structure was complete;the germinal layer of the cystic cyst wall after treatment in the rAd-mIL-28B group and ABZ+rAd-mIL-28B group lost its With complete cell structure,the cells show a certain degree of rupture,shrinkage,and shedding.Flow cytometry results showed that ABZ group(5.64%±1.66%),rAd-mIL-28B group(4.78%±0.77%),ABZ+rAd-mIL-28B group(4.53%±0.54%)and Untreated group((8.85%±3.67%))The percentage of Foxp3+Treg cells in the spleen of mice decreased significantly(p<0.05).ELISA results showed that rAd-EGFP(5261.22pg/ml±7106.11pg/ml),rAd-mIL-28B group(6927.72pg/ml±7034.37pg/ml)and ABZ+rAd-mIL-28B group(2786.49pg/ml±2789.42pg/ml)Serum IL-10 expression was significantly higher than that in the untreated group(646.63pg/ml±559.4pg/ml)(p<0.05);rAd-mIL-28B group(393.09pg/ml)ml±234.98pg/ml),ABZ+rAd-mIL-28 group(450.34pg/ml±383.41pg/ml)compared with Untreated group(870.08pg/ml±228.67 pg/ml),the expression of TGF-?in serum has Significantly decreased(p<0.05);the expression of IFN-?in serum rAd-mIL-28B group(628.87pg/ml±467.16pg/ml),ABZ+ABZ+rAd-mIL-28B group(783.89pg/ml±645.42pg/ml).Compared with the untreated group(352.90pg/ml±569.90pg/ml),the expression of IFN-?was significantly increased(p<0.05);the ABZ group(59.72pg/ml±36.84pg/ml),ABZ+rAd-mIL-28 group(59.39pg/ml±45.44pg/ml))IL-10 expression in peritoneal fluid was significantly decreased(p<0.05)compared with Untreated group(165.11pg/ml±41.58 pg/ml);compared with Untreated group(122.14pg/ml±81.09 pg/ml)compared with rAd-mIL-28B group(628.87pg/ml±467.16pg/ml),ABZ+rAd-mIL-28 group(999.76pg/m±l587.60pg/ml)the expression of IFN-?in peritoneal fluid increased significantly(p<0.05).Protein chip results showed that compared with normal mice,the expression of Th1 cytokine IFN-?in the rAd-mIL-28B group was significantly increased(p<0.05),and the expression of Th17 cytokines IL-17,IL-17F,IL-23,etc.Significantly increased(p<0.05),and the expression of Th2 cytokine IL-4 decreasedConclusion:After rAd-mIL-28B treatment,the percentage of Foxp3+T cells in mice infected with Echinococcus granulosus decreased,and the expression level of Th1type cytokines increased,indicating that rAd-mIL-28B can reduce Tregs,own-regulate immune suppression,and enhance Th1 type cellular immunity.Answer.So as to achieve the effect of treating Echinococcus granulosus. |
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