OBJECTIVETo study the relationship between simian acquired immunodeficiency syndromn (SAIDS) and autoimmunity in simian immunodeficiency virus (SIV)-infected monkeys.

METHODSIndirect immunofluorescence assays were performed to detect plasma or serum autoantibodies in SIV-infected monkeys. The heart, liver, spleen, lung, kidney, and lymph node of BALB/c mice, a strain of endothelial cell ECV304, and granulocytes were used as target antigens. These results were compared with HE stained slides of SIV-infected monkeys.

RESULTSThe levels of various autoantibodies, including anti-lymphocyte autoantibodies, anti-endothelial cell autoantibodies, and anti-granulocyte antibodies, increased after SIV infection in monkeys. Moreover, pathological examinations showed injuries in the lymphoid tissue and vascular pathological changes in cerebral cortex, submucosa of gastrointestinal tract, interstitial capillaries of myocardium, nephron of the kidney, and sinusoid cell of liver.

CONCLUSIONThe increased autoantibodies and the pathological changes of tissues and organs confirm the existence of autoimmunity in SIV-infected monkeys.

Animals ; Autoantibodies ; blood ; Autoimmunity ; Endothelial Cells ; immunology ; Granulocytes ; immunology ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Simian Acquired Immunodeficiency Syndrome ; immunology ; pathology ; Simian Immunodeficiency Virus

OBJECTIVETo observe T lymphocyte subsets and indicators of changes in viral load in sub-acute period in Chinese rhesus monkey model of AIDS SIVmac239. To explore Virology related index variation in sub-acute period of the Chinese rhesus monkey model of AIDS.

METHODTo replicate Chinese rhesus monkey model of AIDS, healthy Chinese rhesus monkey was inoculated with SIVmac239 viral strain. To observe changes in T lymphocyte subsets indexes and viral load after infection with the simian immunodeficiency virus (SIV) in sub-acute period on an animal model. The clinical symptoms of the animal model was recorded simultaneously.

RESULTDuring the 10 weeks after SIV acute infection, body weight and BMI index were relatively stable, the difference was not significant at all time points. Twelve monkeys were tested SIV positive by real-time PCR after three days of infection. On the 7th day after infection, 15 monkeys were tested SIV positive. Viral load increased rapidly, but reached a peak on the 10th-14th day after infection, then showed a level of volatility decline. T lymphocyte subsets showed significant changes, among them, CD3% and CD3 counts fluctuated upward trend and reached to the highest level in two weeks after infection; of CD4% and CD4 count changes were not synchronized, CD4% declined trend while the CD4 count was an increasing trend after the infection; of CD8% and CD8 counts fluctuate upward trend, and reached to a highest level in two weeks after infection ;the ratio of CD4/CD8 and the counts of CD4CD28 T cells decreased significantly in two weeks after infection; the former followed by a slow decline, the latter followed by a rapid rise. Three mouths after the infection 3 monkeys showed significant clinical symptoms. One of the rhesus monkeys had symptoms of diarrhea and two of them had reduced food intake.

CONCLUSIONThis experiments established standardization of Chinese Rhesus monkeys used in the research of AIDS and provide a detailed contents in the changes of sub-acute phase.

Acquired Immunodeficiency Syndrome ; Acute Disease ; Animals ; Cell Count ; Disease Models, Animal ; Macaca mulatta ; Male ; Simian Acquired Immunodeficiency Syndrome ; immunology ; Simian Immunodeficiency Virus ; physiology ; T-Lymphocyte Subsets ; immunology ; Viral Load

OBJECTIVETo develop a model of SHIV-KB9/Chinese origin rhesus (Ch Rh) macaques for vaccine research and to compare the pathogenesis of SHIV-KB9 in Ch Rh macaques with that reported in Indian rhesus (Ind Rh) macaques.

METHODSSeven mamu-A*01 negative Ch Rh macaques were inoculated intravenously with 1-10000 MID50 of SHIV-KB9. The monkeys were monitored for viral load, CD4, CD8, SHIV-specific antibody and virus genetic variation. The results were compared with those previously observed in Ind Rh macaques.

RESULTSAs compared to that observed in Ind Rh macaques, SHIV-KB9 in Ch Rh macaques displayed three identical disease progression patterns. However, the primary pattern was not identical between the two subspecies. The level of plasma viremia differed in SHIV-KB9-infected Ch Rh macaques which exhibited different outcomes from those in Ind Rh macaques. Generally, the values of viral load and the maintenance of CD4+ T cells were associated with humoral responses. Otherwise, the viral genetic distances (divergence, diversity) were larger in animals (M419, M425) with their CD4+ T cells profoundly depleted.

CONCLUSIONThe model of SHIV-KB9/Ch Rh macaques displays a relatively slow progression to AIDS compared with Ind Rh macaques, which may more accurately reflect the potential of candidate vaccines in humans.

Animals ; CD4-CD8 Ratio ; China ; Disease Progression ; India ; Macaca mulatta ; Simian Acquired Immunodeficiency Syndrome ; pathology ; Species Specificity ; Viral Load

OBJECTIVETo establish a real-time RT-PCR based plasma virus quantification method and monitor the plasma viral load of SHIV-CN97001 during its in vivo passages in rhesus macaques.

METHODSViral RNA standards were prepared by in vitro transcription and one-tube real-time RT-PCR were established and optimized using TaqMan EZ RT-PCR CORE REAGENTS and TaqMan probes and primers directed to the 91 bases within the conserved gag region of SHIV. Plasma viral RNA of 126 plasma samples from rhesus macaques of different viral passages was quantified.

RESULTSThe PCR system was optimized by using serial dilution of standards, and the viral RNA load was detected. The lowest limit of the standard curve reached 2x10(-2) copies/ml. The correlation (r>0.99) and the repetition (CV=4.14 percent) also met the requirement. It was revealed that the viral RNA load of third passage was the highest. Generally, the viral load peaks (10(5)-10(6) copies/ml) appeared at the fourteenth day after the infection or inoculation.

CONCLUSIONThe method of one-tube real-time RT-PCR was established successfully, which may provide a sensitive way to qualify SHIV viral load. This will contribute to the establishment and application of SHIV/rhesus macaque models. It was also found that the replicative ability of SHIV-CN97001 was enhanced during the first 2 in vivo passages.

Animals ; HIV ; genetics ; isolation & purification ; HIV Infections ; virology ; Humans ; Macaca mulatta ; RNA, Viral ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Serial Passage ; Simian Acquired Immunodeficiency Syndrome ; virology ; Simian Immunodeficiency Virus ; genetics ; isolation & purification ; Viral Load

OBJECTIVETo observe the effect of Chinese herbal extract HuNan A-1 (HNA-1) on the thymic output function in Simian immunodeficiency virus (SIV) chronically infected rhesus macaques.

METHODSEight Chinese rhesus macaques had been infected by SIVmac239 for 16 to 21 months, and then they were randomly divided into the treatment group and the control group, 4 in each group. Monkeys in the treatment group were administered with HNA-1 by gastrogavage, once daily for 2 successive months, while those in the control group were administered with equal volume of normal saline by gastrogavage, once daily for 2 successive months. The general condition and body weight of monkeys were observed. Plasma viral loads were detected using real-time fluorescent quantitative PCR assay. CD4 percentages and counts, as well as naive CD subsets were detected using flow cytometry. T-cell receptor excision circles (TREC) were detected using real-time fluorescent quantitative PCR assay. The thymus tissue was pathologically observed using routine HE staining. The correlation between lesions of the thymus tissue, CD4 counts, naive CD counts, and TREC were analyzed.

RESULTSThere was no statistical difference in body weight, viral loads, absolute CD ratios between the two groups after treatment (P > 0.05). The altered TREC multiple showed an obvious decreasing tendency in the control group, while it showed an increasing tendency in the treatment group (P < 0.05). In both groups, destroyed structures of the thymus tissue could be seen, filled with pink unstructured material. Increased connective tissues, lowered connective cell density, and confused arrangement could also be seen in the two groups, with no obvious difference. TREC contents were positively correlated with naive CD4 counts after removing extremum (r = 0.926, P = 0.001). Naive CD4 counts were positively correlated with CD4 counts (r = 0.961, P = 0.005).

CONCLUSIONSTREC content determination, as a marker of newly thymic emigrants, could be taken as a testing method for evaluating the thymic output function. Besides, HNA-1 treatment increased the thymic output significantly in SIV chronically infected monkeys. Correlation existed among TREC contents, naive CD4 counts, and pathologies of thymus tissues, especially in late infection stage.

Animals ; CD4 Lymphocyte Count ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Macaca mulatta ; Plant Extracts ; pharmacology ; Random Allocation ; Simian Acquired Immunodeficiency Syndrome ; drug therapy ; Simian Immunodeficiency Virus ; Thymus Gland ; drug effects ; Viral Load

OBJECTIVETo investigate the biological and clinical features of Chinese rhesus monkeys after intravenous (IV) and intrarectal (IR) challenge with SIVmac239 in rhesus monkeys of Chinese origin, and compare the differences between the routes of infection.

METHODSRhesus monkeys of Chinese origin were inoculated with SIVmac239 either by IV (n = 19) or IR (n = 6) routes. Simian immunodeficiency virus (SIV)-specific antibody titer, CD4 + T cell counting, plasma SIV load, lymph node pathology, and clinical manifestations were compared between these two groups 232 or 168 days after challenging.

RESULTSAll SIVmac239-inoculated animals became seropositive for SIV-specific antibodies. SIV-specific IgM was detected in IV groups as from day 10 but was not detected in IR for all the time points. Although SIV-specific IgG was detected as from day 30 in both groups, the IgG titers were ten-fold higher in IV group than in IR group after day 168. CD4 + T-cell counting decreased progressively in IV group but remained stable in IR group over time. Plasma SIV RNA loads peaked in all animals between day 10 and day 14 (10(7) copies/ml), then declined to "setpoint" (10(3) - 10(6) copies/ml) about 2 months later. Most inoculated animals manifested lymphadenopathy. Two animals in IV group and one in IR group died of simian AIDS between day 150 and day 210, as evidenced by the autopsies showing the depletion of lymph tissues, Pneumocystis carinii pneumonia and other opportunity infections. Conclusion IV or IR inoculation of SIVmac239 in Chinese rhesus monkeys will result in chronic SIV infection with a similar clinical feature of natural HIV infection, which provides an excellent experimental animal model for AIDS.

Animals ; CD4-Positive T-Lymphocytes ; metabolism ; China ; Disease Models, Animal ; Female ; Macaca mulatta ; virology ; Male ; Rectum ; virology ; Simian Acquired Immunodeficiency Syndrome ; immunology ; virology ; Simian Immunodeficiency Virus ; immunology ; pathogenicity ; Veins ; virology

OBJECTIVETo explore the effectiveness of the traditional Chinese herbal medicine Lingzhi (Ganoderma lucidum) preparation in treating simian acquired immune deficiency syndrome (SAIDS).

METHODSFive female adult Chinese rhesus monkeys were inoculated rectally with SIVmac239, and were all diagnosed as SAIDS by laboratory and clinical examinations 17 months later. Of these 5 monkeys, 3 (#393, #374, and #381; treatment group) were orally administered with Ganoderma lucidum (2 spores powder capsules plus 2 spores oil capsules on a daily basis), and the remaining other two monkeys (#348 and #361) served as control and did not receive treatment.

RESULTSAnimal #393 (treatment group), #361 (control group) and #348 (control group) died of SAIDS (opportunity infection) 3.5 months, 6 months, and 11 months later, respectively. Two animals (#374 and #381) survived. The necropsy revealed depletion and/or exhaustion of their lymphoid tissue. In the monkey #374, the peripheral CD4(+) T lymphocyte increased by 30% in the 6(th) month compared with the baseline level and then fluctuated. The plasma viral load gradually fell and reached about 1 log(10) in the treatment group, but remained stable in the control group. As shown by pathological examinations, the lymph node and spleen of monkeys #374 (treatment group) and #381 (treatment group) showed rehabilitation and reconstruction in the lymphatic tissue, thymus, nerve tissue of gyrus hippocampi, pituitary gland, pineal body, thyroid gland, adrenal gland, and ovary. In the control group, however, animals experienced depletion of lymph nodes, atrophy of spleen, disappearance of thymus, and other disorders in endocrine organs.

CONCLUSIONGanoderma lucidum preparation may have certain protective effect on the immune system, nervous system, and endocrine system of monkeys with SAIDS.

Animals ; Disease Models, Animal ; Female ; Macaca mulatta ; Materia Medica ; therapeutic use ; Reishi ; Simian Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Treatment Outcome

In this study, five rhesus macaques were inoculated intravenously with SIVmac251 to establish a model of simian autoimmune deficiency syndrome (SAIDS). Peripheral blood samples were collected at different time points to monitor changes in the total T cell number and T lymphocyte subset. Plasma viral loads, cytokine expression levels and anti-SIV antibody levels were also assayed to acquire certain basic indexes to evaluate disease progression in the rhesus macaque SAIDS model. During the acute stage of infection, plasma viral loads reached a peak at week 1 post-inoculation and lasted for approximately 3 to 44 weeks. The CD3+ CD4+ T lymphocyte count in peripheral blood also transitorily decreased. During the same period, the level of interferon-gamma show an increasing trend, whereas IL-12 levels decreased; IL-2, IL-4, IL-10 and TNF-alpha were maintained at normal levels or could not be detected. During the asymptomatic and ARC phases, plasma viral loads persisted above 10(4) RNA copies/mL and either increased or declined during the later stages of disease; CD3+ CD4+ counts showed a steadily declining trend and the ratio of CD4 to CD8 decreased during late-stage disease. Moreover, antibodies against viral proteins were detected in the plasma and showed a significant increasing trend, while there were no apparently changes in the levels of IFN-gamma, IL-12, IL-2, IL-4, IL-10 and TNF-alpha. In conclusion, the characteristics of the SIV animal models in our study are similar to those of patients with AIDS. Therefore, the rhesus macaque SIVmac251 infection models can be applied for further studies into AIDS.
Animals ; Antibodies, Viral ; blood ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; virology ; Cytokines ; genetics ; immunology ; Disease Models, Animal ; HIV Infections ; genetics ; immunology ; virology ; HIV-1 ; physiology ; Humans ; Macaca mulatta ; Male ; Simian Acquired Immunodeficiency Syndrome ; genetics ; immunology ; virology ; Simian Immunodeficiency Virus ; physiology ; Viral Load