The Treatment Of The Chinese Rhesus Monkey AIDS Model By Bushenjiedu Formulation And Its Effect On Innate Immune Mechanism Of Gastrointestinal Tract Mucosa

Objectives:To apply the treatment of bushenjiedu formulation(formulation of kidney-notification and detoxification)to the Chinese Rhesus Monkey AIDS model and to observe its efficacy;to study the innate immune mechanism of gastrointestinal tract mucosa of the Chinese Rhesus Monkey AIDS model after the treatment of bushenjiedu formulation;and to explore the balance between immune activation and immune tolerance in AIDS treatment.Methods:(1)Use the SIVmac239 injection to establish the Chinese Rhesus Monkey AIDS model.Observe the healthy group,the model group in the acute phase,and the model group in the chronic phase over the time of infection.The observation includes the clinical symptom scores;the routine blood test;the T lymphocyte subsets in the lymph node and the blood;the changes of innate immune factors in the blood plasma;the SIV viral load in the blood plasma;and the pathological state of the gastrointestinal tract mucosa and the lymph node.(2)Apply bushenjiedu formulation to the treatment of Chinese Rhesus Monkey AIDS model.To make an objective evaluation of the efficacy,we compared the treatment group with the model group on the 30th day,the 50th day,the 80th day and the 100th day of the treatment.The comparisons include the routine blood test,the T lymphocyte subsets in the lymph node and the blood,the SIV viral load in the blood plasma,and the pathological state of the gastrointestinal tract mucosa and the lymph node.(3)To study innate immune regulatory mechanism of AIDS and the balance mechanism between immune activation and immune tolerance,we adopted several methods including Western Blotting and Immunohistochemistry to test the changes of innate immune factors in the blood plasma and the gastrointestinal tract mucosa(such as MBL,a-defensins,SIgA,IL2,IL15),the changes of the protein receptors of the signaling pathway in the innate immune system(such as CCR5,MAVS,NF-?B,TLR4,TLR7),and the changes of the CD56,CD103,CDllc,CD80,and CD86.Resulis:(1)The Chinese Rhesus Monkey AIDS model is successfully established:The monkeys have been in the middle and late course of the disease after infection of one to two years.The values of the hematologic fluctuations are within the normal range.There are significant differences of the logarithmic values of the viral load in the blood plasma among AIDS model group,treatment group,and healthy group with model group and treatment group very high level of viral load and healthy group no viral load.When comparing the T lymphocyte subsets' indictors in the blood of the AIDS model group and the treatment group with[the T lymphocyte subsets'indictors in the blood of]the SIV acute phase group and the healthy group,it is found that the differences among the counts of CD3,CD4,CD8,and CD4/CD8 have statistical significance.Pathological manifestations of lymph node,ileum and stomach tissue in Monkey AIDS model is similar to that of human AIDS patients.(2)The changes of the innate immune factors in the blood plasma of the Chinese Rhesus Monkey AIDS model:in the SIV acute infection phase of the Chinese Rhesus Monkey,the MBL concentration declines over time of the infection and returns to a certain level when entering the AIDS phase.In the SIV acute infection phase of the Chinese Rhesus Monkey,the?-defensins concentration in the blood plasma increases and returns to a relatively low level when entering the AIDS phase.(3)The efficacy of bushenjiedu formulation on treatment of Chinese Rhesus Monkey AIDS model:? The application of bushenjiedu formulation to treatment of the Chinese Rhesus Monkey AIDS model can significantly increase the counts of CD3,CD4%,CD4,CD8%,CD8,and can maintain the level of CD4/CD8.? When comparing the treatment group with the healthy group it is found that bushenjiedu formulation can significantly increase the counts of CD4%and CD4/CD8,and these increased counts are close to the normal range.? When comparing the treatment group with the model group,it is found that the application of bushenjiedu formulation on AIDS Chinese Rhesus Monkey can significantly increase the counts of CD4 and CD8,and sustain the level of CD4/CD8.? When comparing the treatment group with the model group,it is found that bushenjiedu formulation has a certain[level of]efficacy to remove the SIV virus in the blood plasma.After 30 days of treatment,two monkeys(No.147 and No.357)have no detected SIV virus in their blood plasma after repeated detections.The decline of the virus of the other treatment group reaches 1.85 log10,and the average decline reaches 0,95 log10 after 50 days of treatment.? When comparing pathological manifestations of the treatment group with that of the model group,it is found that bushenjiedu formulation can improve the inflammatory cell infiltration on the propria of the gastric mucosa;decrease the collagen deposition on the lymphoid follicles and submucosa;prevent inflammatory cell infiltration on the intestinal mucosa and the formation of Peyer' s patches;improve the dissolution and the failure of the lymphoid follicles in the lymph node;stimulate the lymphoid hyperplasia;and improve blood circulation.? When comparing the immunohistochemistry of the lymph node of the treatment group with that of the model group,it is found that bushenjiedu formulation can increase the in-situ CD4 and CD8 lymphocytes generated by the lymph node.? The treatment group shows better improvement of clinical symptoms than model group.No severe clinical symptoms and side effects were detected in the treatment group,while one of the monkeys in the model group died of systemic failure of AIDS.(4)The regulatory mechanism of the innate immune system of Chinese Rhesus Monkey AIDS model after the treatment of bushenjiedu formulation:? The MBL in the blood plasma of the Chinese Rhesus Monkey AIDS model is below the healthy level and gradually decreases along with the disease progression,which difference has a significance(P<0.01).After the treatment of bushenjiedu formulation,the level of MBL in the blood plasma gradually increases and reaches a healthy level,and is significantly higher than the model group,which difference has a significance(P<0.01).Comparing with the model group,the Chinese Rhesus Monkey AIDS treatment group has a significantly-raised level of MBL in the gastric mucosa after the treatment of bushenjiedu formulation,which difference has a significance(P<0.01).? The a-defensins in the blood plasma of the Chinese Rhesus Monkey AIDS model is below the healthy level,which difference has a significance(P<0.01).After the treatment of bushenjiedu formulation,the level of MBL in the blood plasma[of the Chinese Rhesus Monkey AIDS model]gradually increases and passes the healthy level along with the treatment,and is significantly higher than the level of the model group,which difference has a significance(P<0.01).Compared with the model group,the Chinese Rhesus Monkey AIDS treatment group has a significantly-raised level of a-defensins in the gastric mucosa after the treatment of bushenjiedu formulation,which difference has a significance(P<0.01).? The IL2 in the blood plasma of the Chinese Rhesus Monkey AIDS model decreases along with the disease progression,which difference has a significance(P<0.01).After the treatment of bushenjiedu formulation,the level of IL2 in the blood plasma gradually increases and passes the healthy level,and is a significantly higher than the model group,which difference has a significance P<0.01.Compared with the model group,the Chinese Rhesus Monkey AIDS treatment,group has a significantly-raised level of IL2 in the intestinal mucosa after the treatment of bushenjiedu formulation,which difference has a significance(P<0.05).? After the treatment of bushenjiedu formulation,the level of IL15 in the blood plasma gradually increases and passes the healthy level,and is significantly higher than the model group,which difference has a significance(P<0.01).Compared with the model group,the Chinese Rhesus Monkey AIDS treatment group has a raised level of IL15 in the intestinal mucosa and was raised most a significantly on the 50th day of the treatment,which difference has a significance(P<0.01).? The SIgA in the blood plasma of the Chinese Rhesus Monkey AIDS model decreases along with the disease progression,which difference has a significance P<0.01.After the treatment of bushenjiedu formulation,the level of SIgA in the blood plasma of the Chinese Rhesus Monkey AIDS treatment group gradually increases and passes the healthy level over time of the treatment,and is significantly higher than that level of the model group,which difference has a significance(P = 0000).Compared with the model group,the Chinese Rhesus Monkey AIDS treatment group has a significantly-raised level of SIgA in the gastric mucosa after the treatment of bushenjiedu formulation,which difference has a significance(P<0.05).? Compared with the model group,the Chinese Rhesus Monkey AIDS treatment group can decrease the protein expression of CCR5 in the intestinal mucosa,NF ? B(p65)in the intestinal mucosa,NF ? B(p65)in the gastric mucosa,and TLR4 in the intestinal mucosa,after the treatment of bushenjiedu formulation,all of which differences have a significance(P<0.01).Compared with the model group,decreased level of protein expression of CCR5 in the gastric mucosa,TLR7 in the intestinal mucosa,and TLR4 in the gastric mucosa was found in the Chinese Rhesus Monkey AIDS treatment group,after the treatment of bushenjiedu formulation,[all of]which differences have significance(P<0.05).? Compared with the model group,increased level of protein expression of MAVS in the intestinal and gastric mucosa was found in the Chinese Rhesus Monkey AIDS treatment group after the treatment of bushenjiedu formulation,which difference has a significance(P<0.01).? Compared with the model group,increased level of protein expression of CD56 in the intestinal mucosa,CD56 in the gastric mucosa,CDllc in the intestinal mucosa,CDllc in the gastric mucosa,and CD103 in the gastric mucosa was found in the Chinese Rhesus Monkey AIDS treatment group,after the treatment of bushenjiedu formulation,all of which differences have significance(P<0.01).? Compared with the model group,decreased level of protein expression of CD80 in the intestinal mucosa,CD80 in the gastric mucosa,and CD86 in the gastric mucosa was found in the Chinese Rhesus Monkey AIDS treatment group after the treatment of bushenjiedu formulation,all of which differences have significance(P<0.01).Conclusion:(1)After being infected with SIV,the Chinese Rhesus Monkeys develop the illness into AIDS in one to two years.The clinical symptoms,the T lymphocyte subsets in the blood,the pathological states of the lymph node,and the gastrointestinal tissues are all different from those of the model group in the acute phase but all similar to those of human AIDS patients.(2)The application of bushenjiedu formulation to treatment of Chinese Rhesus Monkey AIDS model can improve clinical symptoms,increase the volume of CD4T and CD8T lymphocyte subsets in the peripheral blood,particularly the increase of CD8T lymphocyte subsets.Besides,the bushenjiedu formulation can inhibit SIV replication.(3)The application of bushenjiedu formulation to the treatment of Chinese Rhesus Monkey AIDS model stimulates the generation and the strengthening of the function of NK cells and dendritic cells(DC)' in the immune system.The combination of dendritic cells and T-cells largely generates multiple types of innate immune factors(MBL,?-definsin,IL2,IL15,and SIgA),activates the proliferation of the T-cells,induces the generation of CTL,and inhibits the replication of the SIV virus.The use of bushenjiedu formulation in the treatment of the Chinese Rhesus Monkey AIDS model initiates the signaling pathway of MAVS,decreases the expression of CD80 and CD86,reduces the combination of the virus and the damages of CD 4+T lymphocyte,increases the level of CD 4 + T lymphocyte in the blood and the lymph node,decreases the expression of CD80 and CD86,forms the immune tolerance,and prevents the injuries caused by the excessive immune activation.Therefore,the use of bushenjiedu formulation in the treatment of the Chinese Rhesus Monkey AIDS model has achieved a balance between immune activation and immune tolerance and has deferred disease progression.