| Objective:Many experiments have testified that neonates are on the physiological low immune status, especially pre-mature infants,which have much lower immune functions inducing to high incidents of infection diseases and high mortality. Moreover,many allergic diseases and auto-immune diseases are also associated with the imbalance of tolerance reaction and immune response during the new-born period Therefore,the set up of normal immune system for the new-born neonates can not only decrease neonate mortality,but also avoid some diseases,such as bronchial asthma,eczema,food allergy,which occur later on. Every kind of neonate immune cells is on the different stage of maturity,taking variant times to reach maturity.It is very complicated for setting up the mature immune system,and it is influenced by many factors,such as different stage of gestation,stimulation of antigen, nutrition and immune ingredients in mother milk,diseases,drugs and so on.Most other experiments observed the T and B lymphocytes,helper cells and cytokines functions after different antigen stimulation.My experiment uses flow cytometry to measure the quantities of neonatal rat spleen CD3,CD20 and CD25 cells,measure the swallow rate of macrophages,measurement of PFC and neonatal rat intestinal sIgA by using fluoroimmunoassay,in order to fully explain the whole process of setting up normal neonatal rat immune system.By using cell culture,analysis the interaction of relocation of bacterial in MLN and CD25/CD3,and their influence by antibiotics,explain bacterial antigen and antibiotics influence on neonatal rat immune system set up,therefore provide evidence for clinic protection,improve neonates normal immune system set up and decrease the rate of infection and mortality.Methods:1.animal groups:160 SD rats,male and female mixed,multi-colons,including 40 new-born rats,30 each neonatal 3day,7 days,14 days and 30 days rats。10 new-born rats were randomly chosen from 40 new-born rats,as antibiotics treatment observing group；Each different age rats were divided into 3 groups。2.Sampling and experiment methods 2.1The quantities of CD3,CD20,CD25:rats of observing group were killed at 0,3,7,14,30 days.The rats of treating group were injected with penicillin (1500u/g /day)into intyaperitoneal once per day from 0 days to 6 days after being born and were killed at 7 days.Each rat's spleen was sampled and made into mononuclear cell's supernatant.Use flow cytometry to measure the quantities of CD3,CD20 and CD25 cells.2.2Hemolytic plaque test:SRBC intraperitoneal inoculation.The animals were killed after 4 days treatment.Each rat's spleen was sampled and was made into spleen cell's supernatant.and then added to Bengal gelatine with SRBC.They were poured into a plate.Measure PFC after hybridization and count the number of PFC per 106 spleen cells.2.3The percentage of phagocytosis:Intraperitoneal injection with the starch-gravy was done to the rats.Then CRBC was injected after 3 days.Liquid in abdominal cavitywas sampled and was made into thin slices stained with Wright,measure the macrophage which swallowed CRBC per 100 macrophages.2.4sIgA in the intestine:the substance in the typhl was sampled after the rats were killed.Measure sIgA by using fluoroimmunoassay.2.5the rate of bacteria translocation:The MIN was sampled after the rats were killed,and was weighted and diluted.Then thedilutions were plated to culture bacterial.The rat was thought to occur bacteria translocation if the sample grew bacteria.The rate of bacteria translocation=the number of the rats occurred bacteria translocation per group/10×100%.3Statistic analysis:all data was analysed by using SAS software,the data was used mean±stand error,p<0.05 was decided as significant.Results:1 The description of immune function's setting up in the neonatal rats.1.1 cellular immune function according to quantities of CD3,CD25:The quantities of CD3 was 22.0±2.6 on new-born rats,increasing to 45.16±3.0 after being born 30 days.The quantities of CD25 was 0.5±0.6on...
|
PDF Full Text Request