| Autoimmune diseases have caused a huge social and economic burden on our country and the families of patients due to high morbidity,high mortality.It affected the life quality of people because it had difficulty in treating and controlling.Therefore,it is very important to clarify the pathogenesis of the disease for the early diagnosis,early treatment,and prognostic evaluation.With the development of high-throughput sequencing technology and immunotherapy technology,the sequencing technology of immune repertoire has quickly attracted the attention of the public,and the information of immune repertoire is becoming more and more widely used in various fields of clinical testing.This technology is used to monitor the changes in the immune system to determine the health status of the human body.It is used to detect the clonal expansion of lymphocyte receptors for efficacy evaluation.The same receptor clones shared between different patients are used to better monitor and analyze the overall immune status of various patients under physiological and pathological conditions.This study used the adaptive immune repertoire sequencing technology to study the interaction process between autoimmune diseases and the immune system.It clearly and intuitively displayed the characteristics of the immune repertoire and tried to explore the immunological mechanism of disease occurrence and development through various visualization software and tools.The main content of this research included the following aspects:1.Lupus erythematosus is an autoimmune disease.It has a wide variety of complex clinical symptoms and involves multiple organs.Because the disease is related to T and B lymphocytes,current studies rarely observe the effects of T cells and B cells in patients at the same time.In this chapter,we used 7-chain immune repertoire kits to construct the TCR repertoire and BCR repertoire of healthy people and patients.We analyzed the immunological characteristics of the TCR repertoire and BCR repertoire in patients and healthy people and compared their difference.The results showed:(1)immune repertoire sequencing found that the expression ratio of BCR receptors in peripheral blood of SLE patients was higher than that of TCR receptors.In patients,Ig A and Ig G were mainly expressed,while Ig M was mainly expressed in healthy people.It was revealed that Ig A and Ig G might be pathogenic autoantibodies in autoimmune diseases,and Ig M was a protective antibody.(2)The diversity of receptors in the peripheral blood of SLE patients was better than that in the skin,while the diversity of immune repertoire in the skin and peripheral blood of DLE patients was similar to that of healthy people.This result revealed the difference in the immune system between healthy people and patients with lupus erythematosus.At the same time,the difference in the expression of the immune repertoire in the peripheral blood and skin of the two types of lupus was revealed.It was conducive to better distinguish the two kinds of lupus in the clinic.2.In this experiment,rats were used to simulate the pathogenesis of diabetic patients.After the model was successfully established,the treatment group was treated with 700mg/kg/d Astragalus polysaccharide(APS)for 2 weeks.The normal group and the model group were not treated,and the same volume of normal saline was used for intragastric administration.The results showed that astragalus polysaccharide treatment for 2 weeks reduced the body weight and blood lipids of the rats,and increased the level of insulin.In addition,astragalus polysaccharides reduced the inflammatory infiltration of liver and pancreas tissues.It was revealed that Astragalus polysaccharide improved liver damage,increased glycogen synthesis,protected islet cells,and increased insulin secretion,thereby inhibiting the occurrence and development of diabetes.3.The peripheral blood of rats in each group at different stages was collected.Total RNA was extracted.The T cell receptor alpha chain repertoire was amplified by using an automated library building system.A sequencing library was constructed and sequenced on the sequencer.Finally,biological and informatics analysis tools were used for gene comparison,splicing and the establishment of CDR3 repertoire.The immune repertoire analysis tools were used to analyze the obtained effective sequences.The results showed that:(1)before the 4th week,the diversity trend of the immune repertoire in the model group and the astragalus polysaccharide(APS)group was the same,and after the 4th week,the changing trend in the APS group began to gradually approach the normal group.(2)Through further analysis of the V/J/VJ combination,it was found that the expression of 3 V gene subfamilies increased and the expression of 3 V gene subfamilies decreased significantly after treatment.V gene diversity changed in the APS group before and after treatment.(3)The expression of TRAJ49,TRAJ43,TRAJ42,and TRAJ56 sequences increased in the normal group and the model group,and the expression of these 4 J genes decreased after treatment in the APS group,revealing that these sequences might be related to the pathogenicity of the disease and could be used as a potential therapeutic marker.(4)Through comparative analysis of different VJ combinations,we found that 4 pairs of VJ combinations were highly expressed.Among them,the V gene family was mainly expressed by TRAV14-1 and TRAV21-DV12,while the J gene family was primarily expressed by TRAJ42,TRAJ49,TRAJ56 and TRAJ56.These results revealed that astragalus polysaccharide might help increase the diversity of TCR in rats,thereby regulating the rat’s immune system to make the blood lipid production of type 2 diabetic rats lower and increase insulin secretion. |
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