SIVmac239 Inducing Immune-neuro-endocrine Deficiencies Syndrome In Rhesus Macaques And Protection Of Immune ?

Nowadays, the AIDS prevention and control situation remains tense all over the world and China. AIDS is an complex and multisystemic symdrome. Scholars from home and abroad have been paying more attention to the pathogenesis of immune-neuro-endocrine disorder. So it is an urgent issue for medical technician to find out the role which traditional Chinese medicine can play in AIDS prevention and control. In TCM, AIDS is characterized by infecting with immunotoxin evil and weak in five Zang. Evil is the direct pathogeny and main pathogenesis is primordial Qi is damaged and weak in five Zang. Main remedy are included in fortify the spleen and replenish Qi, clearing away the heat-evil and expelling superficial evils, activating blood circulation to dissipate blood stasis, supplementing Qi and nourishing Yin etc. Immune I, whose principle of treatment is benefiting vital energy and detoxication, is effective in AIDS in clinic.The aim of this experiment is to study the pathogenesis of AIDS neuroendocrine-immune system as well as to exam the function of Immune I,which is an effective prescription in clinical. This study took 16 male Chinese rhesus macaques as experimental subjects. According to their weight, the rhesus macaques were divided into two groups randomly, the normal group(normal, n=4) and the infection group(n=12). Model was built by using venous infection SIVmac239 in the infection group. The infections were confirmed by testing plasma SIV RNA with fluorescence quantitative RT-PCR two weeks later. According to CD3+CD4+ absolute counting, the infection group were dividend into two groups randomly 2 months later, the model group (model, n=6)and the treatment group (n=6). Two months after being infected, the treatment group were given drug treatments by pouring Immune I into stomach. The medical cares were taken once a day,6 times a week. The dose was 0.58g/kg, as many as 2.9 times of the usual adult dosage. A full treatment takes 3 months and the experiment takes two full treatments. There was one month of interval between two full treatments, and one month drug withdrawal after two full treatments. Euthanasia was performed in the end.The main outcome measures including:(1) General state of health, including weight, appetite, oral mucous membrane, coated tongue, excrement, temperature, biochemical indicators etc.(2) The proportion and absolute counting of peripheral blood CD3+CD4+and CD3+CD8 tested by FCM (flow cytometry).(3) The proportion of peripheral blood CD4+CD25+Foxp3+ tested by FCM.(4) The content of SIV RNA in plasma tested by fluorescence quantitative RT-PCR.(5) The damage level of lymphonodus is detected by inguinal lymph nodes or ALN biopsy respectively before infection, two month after infection and after a full treatment.(6) Pathological changes observed through HE stain in the immune system such as thymus, inguinal lymph nodes, ALN, mesenteric lymph node, spleen. New cells were revealed by immunohistochemistry Ki-67.(7) Pathological changes observed through HE stain in the nervous system such as medulla, pons, midbrain, cerebrum, hypothalamus, cerebellum, pineal body, ganglion of chest and waist. (8) Pathological changes observed through HE stain in the endocrine system and the reproductive system such as hypophysis, the adrenal gland, testicles, epididymis, the prostate and seminal vesicle.The results indicated that:(1) Since the Chinese rhesus macaques were infected with SIVmac239, the symptoms included weight loss, loss of appetite, ulcer and poor micro circulation in oral mucosa, diarrhea, cough, fever, biochemical indicators changing etc. After two full treatments of immune I, the rhesus monkeys began to gain weight. The occurrence of opportunistic infections decreased and oral mucosal micro circulation got improved. ALB, GGT, CHO and HDL raised after the rhesus macaques being infected by SIVmac239. After two months of immune I treatment, ALB and GGT obviously decreased, while it has no significant impact on CHO and HDL.(2) After rhesus macaques being infected with SIVmac239, the number of lymphocyte CD3+CD4+T and the percentage between CD3+CD4+ and CD3+CD8+ both decreased which were nearly the same as symptom of infected with HIV. While immune I can evidently restrain the decline of the percentage between CD3+CD4+ and CD3+CD8+ T lymphocyte.(3) 1-2 weeks after rhesus macaques being infected with SIVmac239, SIV RNA reached its peak (about 107copies/ml).4 weeks later, it was basically stable (104-106copies/ml). The process was similar to the copying of HIV in human bodies. However, the immune I had no significant impact on it.(4) According to the lymph node biopsy, the lymph node stayed original state at first. The size of lymph follicle was normal and no germinal center was observed. Two months after being infected, lymph follicle is proliferative and mildly degenerated (±?+) but not drained. Meanwhile, germinal center expanded, and cells'differentiation is active in light zone. Lymphocytes and reticulum cells were observed in medullary sinus. Five months after infection, lymph follicle proliferation was aggravated and deposition of immune complexes can be observed in germinal center which led to regression (±?++) of certain cells. Vascular endothelial cells puffed up and angiogenesis took place with lymphocytic infiltrating.(5) Ten months after infection, Lymph nodes of normal group are mild degenerated (2 cases±?+in 4cases) and not drained. Lymph nodes of all model group are proliferative (++?+++),3 cases degenerated(++?+++) and 3 cases drained(++?+++). germinal centers are atrophica and even disappeared. Paracortex is atrophica. Lymphocytic and macrophagic infiltration is in capillary and small vessels and resulting in microcirculation disturbance, germinal centers are atrophica and even disappeared. Cells in lymphatic tissue, cathepsin and disappeared. A great quantity of immune complexes generated in germinal center which led to much more severe cells regression. There were a great number of lymphocytes and reticulum cells in medullary sinus. Lymph nodes of Immune I group are proliferative (++?+++),3 cases degenerated(±?+) and not drained. There are no phenomenon of cells in lymphatic tissue, cathepsin and disappeared along sinus tract. Immune complexes generating in germinal center is less than model group.(6) Lymph node Ki-67 stain showed that there were only a few new cells in germinal center 5 months after infection with SIV. After the treatment of immune I, the germinal center contained much more Ki-67 positive cells and new cells.(7) Since the rhesus macaques were infected with SIVmac239, thymus of 3 cases in model group is obviously atrophica and 2 cases vestigial. The cortex as well as medulla of thymus got atrophied. Hassall's corpuscles got blastocolysis and non-keratinized. The symptoms also included enlargement of splenic corpuscles, hyperplasia and degeneration. Besides, it led to lymphocytes and reticulocyte in medullary sinus. The immune I can release the lesion of the thymus and the spleen in the form of decrease of cells degeneration caused by immune complex deposition.(8) Since the Chinese rhesus macaques were infected with SIVmac239, different levels of lesion occurred in medulla spinal cord, medulla, pons, midbrain, cerebrum, hypothalamus and cerebellum. Cells in nucleuses decreased and got pyknotic and then turn into akaryote. Acidophilic stain of cells aggravate, deterioration of cytoplasm started. Cell nucleus, nucleolus and it's corpuscles partly vanished. And part of chromatin turned into suspensoid. The branches from neurocytes got shorten and even disappeared. Infiltration of lymphocytes in vessel, nodule of glial cells and demyelination appear in severe cases. medulla oblongata?The lesion level of ganglions was related to chronic diarrhea. The immune I can release the ganglions lesion.(9) Since the rhesus macaques were infected with SIVmac239, hypophysis, the adrenal gland, thyroid, testicles, epididymis produced marked changes. Eosinophils and basicyte in hypophysis bdecreased sharply while chromophobe cells increased. In adrenal, cytoplasm in belt layers is rich in protain and reticular layers were neat. Also, most of the AMCCs were empty. There are only spermatogenous cells but no spermatocyte or spermatozoa within the testis seminiterous tubules. And also spermatozoa were barely in epididymis. The size of thyroid acinus varied and the colors of colloid were divers. With the treatment of immune I, cytoplasm in belt layers is rich in steroid and poor in protain. Chromaffin cells are plump. Spermatogenous cells, spermatocyte and a few spermatozoa can be observed in the testis seminiterous tubules. So the treatment of immune I can greatly ease the above-mentioned symptoms.Conclusions:After infected with SIVmac239, the rhesus macaques'symptoms included weight loss, loss of appetite, oral mucosa ulcer and poor micro circulation, diarrhea, pneumocystis carinii pneumonia etc. The ratios of CD3+CD4+T lymphocyte and CD3+CD4+/CD3+CD8+ gradually declined.1-2 weeks after rhesus macaques being infected, SIV RNA reached its peak. And then,4 weeks later, it was basically stable. After being infected, lymph follicle proliferation got ingravescent, deposition of immune complexes led to regression of a certain cells and lymph follicle drained in the end. The cortex as well as medulla of thymus got atrophied. Hassall's corpuscles got blastocolysis and non-keratinized. The symptoms also included enlargement of splenic corpuscles, hyperplasia and degeneration. Besides, Lymphocytes and reticulum cells were observed in medullary sinus. Different levels of lesion occurred in spinal cord, medulla, pons, midbrain, cerebrum, hypothalamus and cerebellum. Cells in nucleuses decreased and got pyknotic and then turn into akaryote. Acidophilic stain of cells aggravate, deterioration of cytoplasm started. Cell nucleus, nucleolus and its corpuscles partly vanished. And part of chromatin turned into suspensoid. The branches from neurocytes got shorten and even disappeared. Infiltration of lymphocytes in vessel, nodule of glial cells and demyelination appear in severe cases. The secretory function of pituitary body and adrenal gland as well as sperm-producing function of testis seminiterous tubules declined. Besides, there was hypothyroidism. All in all, neuroendocrine-immune system had been damaged.After two full treatments of immune I,the general appearance of the rhesus macaques got greatly improved. Besides, it can slightly raise the level of CD4+T lymphocytes and the ratio between CD3+CD4+ and CD3+CD8+. Immune I can ease the lesions of lymph gland, thymus gland and spleen, maybe resulting from adjusting autoimmunity caused by SIV. And it can protect the central nervous system and endocrine system.