Establishment Of The Rhesus Monkey Immunosuppression Model Induced By Cyclophosphamide

Objective:Establishing animal disease models is used more and more widely and more focused on the rodents recently,but their physiology are very different with human.Establishing non-human primate rhesus monkey animal disease models has an unparalleled advantage.The biochemical metabolism,reproductive physiology and organizational structure of the rhesus monkeys are highly similar to humans,and there are 98%homology with the rhesus monkeys and human genetic material.And as a model for biomedical research it has a hundred years of history and the data is very rich,such as the model of Parkinson’s disease,tuberculosis,and the diseases of liver transplantation.Rhesus monkey has long been used as an ideal laboratory animal medicine and biology research in all areas of modeling.But there was no use of non-human primate to establish immunosuppression model.Therefore,this study intends to use to build immunosuppressed rhesus model and evaluate its biologically relevant indicators,for the future development of new human drugs,the research of inmmunosuppressive mechanism,viral infection model to provide a more ideal and reliable animal model and the improver evaluation system.Methods:Rhesus monkey is chosed for the model animal,immunosuppresssntsis cyclophosphamide,to 5mg/kg,15mg/kg,30mg/kg dose of three groups.Before administration,Rhesus monkey are viewed as the normal group,which are the self-controlled.Blood biochemical tests:total bilirubin,alkaline phosphatase,total protein,aspartate aminotransferase,alanine aminotransferase,albumin,urea nitrogen,to monitor the rhesus liver and renal function;blood test which red blood cells,white blood cells,lyrmphocyte absolute,neutrophilsabsolute,hemoglobin and plateletsmonitor the immune status of rhesus monkeys;lymphocyte proliferation is measured by MTT;flow cytometry tests CD3+T lymphocyte,CD3+CD4+T lymphocyte,CD3+CD8+T lymphocyte content percentage;real-time PCR is used to detect the mRNA expression level ofINF-α,IFN-γ,IL-2.Result:(1)The result of biochemical index:at the seventh day and the fifth day after administration,the index of 30mg/kg group were significantly higher than normal,after stopping administration,it did not resume to normal levelsimmediately.15mg/kg and 5mg/kg group were always within the normal range,at before and after administration;(2)the result blood detection index:at the seventh day and the fifth day after administration,the index of 330mg/kg and 15mg/kg group were significantly lower than before administration,after stopping administration the index of 30mg/kg group is no significant increase in the trend,rising trend 15mg/kg group appeared,two of 5mg/kg group are no significant changes in indicators,one rhesus monkey of this group appear significantly increased at the seventh day after administration,after withdrawal,it is recover;(3)lymphocyte inhibition rate:the rate of 3 30mg/kg and 15mg/kg group were significantly higher than before administration,after stopping administration the index of 30mg/kg group is no significant recover in the trend,dropping trend 15mg/kg group appeared,two of 5mg/kg group are no significant changes in indicators,one rhesusmonkey of this group appear significantly droppedat the seventh day after administration,after withdrawal,it is recover;(4)CD3+T lymphocyte,CD3+CD4+T lymphocyte,CD3+CDg+T lymphocyte’s percentage:at the seventh day and the fifth day after administration,the percentage of 3 30mg/kg and 15mg/kg group were significantly lower than before administration,after stopping administration the percentage of 30mg/kg group is no significant increase in the trend,rising trend 15mg/kg group appeared,two of 5mg/kg group are no significant changes in percentage,one rhesus monkey of this group appear significantly increased at the seventh day after administration,after withdrawal,it is recover;(5)TNF-a,IFN-y,mRNA IL-2 relative expression levels:at the seventh day and the fifth day after administration,the relative expression of 3 30mg/kg and 15mg/kg group were significantly lower than before administration,after stopping administration the relative expression of 30mg/kg group is no significant increase in the trend,rising trend 15mg/kg group appeared,two of 5mg/kg group are no significant changes in relative expression,one rhesus monkey of this group appear significantly increased at the seventh day after administration,after withdrawal,it is recover.From the changes in the detection of indicators it can be conclued,30mg/kg appeared immune suppression at the fifth day and the seventh day after administration,and liver and kidney function damage occurs,after administration they are still in immunosuppressed state,do not immediately restored to normal levels;15mg/kg appeared immune suppression at the fifth day and the seventh day after administration,liver and kidney dysfunction are normal,after stopping,their immune system begin recover;two of 5mg/kg group are no significant changes in all index and there are normal,one rhesus monkey of this group appear significantly change in all index and it exhibit immune enhancer.Conclusion:ti is successful that establishing rhesus monkey immunosuppressive model;the changes in the indicators of 15mg/kg group and the liver and kidney function are both similar with human in immune index,other groups were not.so 15mg/kg group is suit for establishing human-related immunosuppression model;it is successful that establishing a comprehensive evaluation method of immunosuppression,the blood chemistry,blood test,flow cytometry,lymphocyte proliferation test and fluorescence quantitative PCR techniques are all the relevant sensitive indicators to evaluate the functional status of immune suppression;we find that high-dose group and middle-dose group showed immunosuppression,while the low dose group may cause immune enhancer.