Whole blood as an alternative to peripheral blood mononuclear cell for detection of total HIV-1 DNA.

BACKGROUND: A more time saving, convenient, reproducible, and scalable method is needed to assess total HIV-1 DNA levels. METHODS: Frozen whole blood and peripheral blood mononuclear cell (PBMC) samples both 200 µl at the same point were used to detect total HIV-1 DNA. Automatic extraction of total HIV-1 DNA was used to ensure the consistency of sample extraction efficiency. The detection reagent was HIV-1 DNA quantitative detection kit and real-time quantitative PCR was utilized. RESULTS: Of the 44 included patients, 42 were male and 2 were female, with a median age of 33 years. Thirty-three cases were collected after receiving antiviral treatment, with a median duration of treatment of 3 months, and the other 11 cases were collected before antiviral treatment. The median viral load was 1.83 log10 copies/mL, the median CD4 and CD8 count were 94 and 680 cells/µL, and the median CD4/CD8 ratio was 0.18. The results of the two samples were 3.02 ± 0.39 log10 copies/106 PBMCs in PBMC samples and 3.05 ± 0.40 log10 copies/106 PBMCs in whole blood samples. The detection results of the two methods were highly correlated and consistent by using paired t test (P = 0.370), pearson correlation (r = 0.887, P < 0.0001) and intra-group correlation coefficient (ICC = 0.887, P < 0.0001) and bland-altman [4.55% points were outside the 95% limits of agreement (- 0.340 ~ 0.390)]. CONCLUSIONS: The results of the whole blood sample test for total HIV-1 DNA are consistent with those of PBMC samples. In a clinical setting it is recommended to use whole blood samples directly for the evaluation of the HIV reservoir.

[Clinical and Laboratory Characteristics of Disseminated Non-tuberculous Mycobacterial Disease].

Objective To explore the clinical and laboratory characteristics and the prognosis of disseminated non-tuberculous mycobacteria(NTM)diseases in human immunodeficiency virus(HIV)negative patients. Methods Cases of disseminated NTM disease were retrospectively collected in Peking Union Medical College Hospital from January 2012 to October 2018.Clinical manifestations,laboratory findings,treatment,and prognosis of these cases were retrieved from the electronic medical record system. Results Among the 23 HIV negative patients with disseminated NTM disease,21 had underlying diseases,with rheumatoid immune disease(n=7)as the most common one.The main clinical manifestation was fever(n=23).Laboratory tests showed anemia [hemoglobin(85.78±25.47)g/L],hypoalbuminemia [albumin 29(27-32)g/L],elevated erythrocyte sedimentation rate [(85.73±43.78)mm/h] and hypersensitive C-reactive protein [(112.00±70.90)mg/L],and reduction of lymphocyte count [0.69(0.29-2.10)×10 9/L].Lymphocyte subset analysis indicated reduction in CD4 + T cells [213(113-775)/µl],CD8 + T cells [267(99-457)/µl],B cells [39(4-165)/µl],and NK cells [88(32-279)/µl] and elevation of human leukocyte antigen-D related(HLA-DR),and CD38 expression in CD8 + T cells [HLA-DR +CD8 +/CD8 +,60(40-68)%;CD38 +CD8 +/CD8 +,81(65-90)%].The most common species of NTM was Mycobacterium intracellular(n=6).Lymphocyte,CD8 + T cell,B cell,and NK cell counts were significantly lower in dead patients than surviving patients(P =0.045,P=0.045,P=0.032,and P=0.010,respectively). Conclusions Disseminated NTM disease in HIV negative patients is mainly manifested as fever,anemia,hypoalbuminemia,and elevated inflammatory indicators.It is more likely to occur in immunocompromised patients.Patients with decreased lymphocytes,CD8 + T cells,B cells and NK cells tend to have a poor prognosis.

[The influence of long-term nucleotide reverse transcriptase inhibitors on lipids metabolism in HIV/AIDS patients].

OBJECTIVE: To evaluate the influence of long-term nucleotide reverse transcriptase inhibitors (NRTIs) on lipids metabolism in HIV/AIDS patients and correlating clinical factors. METHODS: A total of 118 HIV/AIDS patients were divided into 3 groups: untreated group (40 patients), highly active antiretroviral therapy (HAART) for 1 - 2 years group (37 patients) and HAART over 5 years group (41 patients), with 20 healthy individuals as the control group. Clinical lipodystrophy (LD) was defined as concordance between patient's report of change and physical examination. Fat mass (FM) was measured by dual-energy X-ray absorptiometry (DXA). RESULTS: There was no significant difference in the incidence of LD between HAART for 1 - 2 years group and HAART over 5 years group (51.2% vs 40.5%, P = 0.345). The prevalence of LD was 2.4 folds with stavudine (d4T) treatment compared with zidovudine (AZT)-containing regimens (61.6% vs 23.5%, P = 0.001). Based on DXA measurements, FM of total body and limbs were significantly lower in the HAART over 5 years group than that in the control group, the untreated group and the HAART for 1 - 2 years group (P < 0.05). Trunk FM was significantly lower in the HAART over 5 years group than the untreated group and the HAART for 1 - 2 years group (P < 0.05). FM of total body and trunk were significantly lower in patients without LD in the HAART over 5 years group than patients without LD in the HAART for 1 - 2 years group (P < 0.05). FM was correlated positively with body weight and BMI. Limbs FM was correlated negatively with peripheral blood triglyceride concentration. CONCLUSIONS: HIV/AIDS patients with NRTIs therapy have high prevalence of LD, which mainly occurs 1 - 2 years after therapy, and increases with d4T treatment compared with AZT-containing regimens. There was no significant difference in the incidence of LD between the HAART for 1 - 2 years group and the HAART over 5 years group. FM was significantly decreased after long-term HAART in the patients with or without LD. DXA can evaluate LD objectively and guide further clinical treatment.

[Influence of long-term highly active antiretroviral therapy on bone mineral density in HIV/AIDS patients].

OBJECTIVE: To evaluate the influence of highly active antiretroviral therapy (HAART) on bone mineral density (BMD) in HIV/AIDS patients and correlating clinical factors. METHODS: 149 HIV patients were divided into 3 groups:untreated group with 41 patients, HAART for 1-2 years group with 60 patients, HAART over 5 years group with 48 patients; 20 healthy individuals included as a control group. BMD-T score and BMD-Z score were measured by dual-energy X-ray absorptiometry (DXA). RESULTS: BMD-Z score of right hip was significantly lower in HAART over 5 years group (0.16 ± 0.82) than untreated group (0.61 ± 1.09) (P = 0.039). BMD-Z score of right femoral neck was significantly lower in HAART over 5 years group (-0.002 ± 0.87) than untreated group (0.55 ± 1.08) (P = 0.012). BMD-Z score of HAART for 1-2 years group was not significantly decreased. BMD-Z score of right hip and right femoral neck were correlated negatively with HAART duration. The incidence of osteopenia/osteoporosis in HAART for 1 - 2 years group (31.7%) and HAART over 5 years group (31.3%) were significantly higher than untreated group (12.2%) (P < 0.05). Body weight was revealed as a risk factor of osteopenia/osteoporosis. CONCLUSION: BMD of right hip and right femur neck were significantly lower in HAART over 5 years group. The incidence of osteopenia/osteoporosis were significantly higher in patients receiving HAART. BMD were correlated negatively with HAART duration. Patients in long-term HAART combined with risk factors such as old age or lower body weight should be checked by DXA regularly.

[An analysis of clinical characteristics of forty-six AIDS phobia patients].

OBJECTIVE: To summarize the clinical characteristics of AIDS phobia patients and establish the preliminary clinical diagnostic criteria. METHODS: The clinical information of 46 AIDS phobia patients was collected and summarized. General demographic data, clinical manifestations and laboratory results were analyzed. RESULTS: The clinical characteristics of AIDS phobia patients include: (1) With or without high-risk behavior of HIV-1 infection; (2) Patients repeatedly demanded HIV/AIDS related laboratory tests, suspected or believed in HIV-1 infection with daily life affected; (3) The main complaints were non-specific including influenza-like symptoms (headache, sore throat and so on), fasciculation, formication, arthrodynia, fatigue and complaint of fever with normal body temperature; physical examination did not reveal any positive physical sign except white coated tongue; (4) Symptoms mainly appeared 0-3 months after the high-risk behavior while HIV-1 antibody kept negative; (5) T lymphocyte subsets test was carried out in 23 patients and showed 19 (82.6%) with CD(4)(+) T lymphocyte count > 500/µl, the remaining 4 were 300 - 500/µl, with the lowest count of 307/µl. Few patients had inversed CD(4)(+)/CD(8)(+) ratio but without excessive CD(8)(+)T lymphocyte activation. CONCLUSIONS: AIDS phobia is a complicated physical and mental disease, whose diagnosis and treatment still need further investigation.

[Association between the change regularity of peripheral blood mononuclear cell mitochondrial deoxyribonucleic acid content and human immunodeficiency virus-related lipodystrophy].

OBJECTIVE: To investigate the change regularity of peripheral blood mononuclear cell (PBMC) mtDNA (mitochondrial deoxyribonucleic acid) content and its association with HIV-LD (human immunodeficiency virus-related lipodystrophy) in HAART (highly active antiretroviral therapy). METHODS: At baseline, Months 6 and 24 of therapy, the cryopreserved PBMC were collected from 33 patients on a regular follow-up at our clinic. Among them, 17 had HIV-LD. Then total DNA was extracted and mtDNA content quantified by real-time PCR (polymerase chain reaction). RESULTS: The HIV/AIDS patients had a lower content of PBMC mtDNA (2(-ΔΔCt)) than the healthy controls at baseline (9.578 vs 17.195, P < 0.01). The mtDNA content was lower in the HIV-LD group than that in the no LD (NLD) group at each time point of therapy (13.619 vs 5.775, 6.360 vs 1.387, 7.170 vs 1.266, all P < 0.05). In the HIV-LD group, the half- and 2-year PBMC mtDNA content was markedly lower than those at baseline (both P < 0.05). And the change of mtDNA content (within half a year) was earlier than the onset of clinical HIV-LD at one year later. In the NLD group, the PBMC mtDNA content have an insignificant change after therapy. The mtDNA content decreased significantly in stavudine (d4T)-containing regimen group after treatment (P < 0.01), but showed no significant change in zidovudine (AZT)-containing regimen group after therapy. CONCLUSION: The decreased content of PBMC mtDNA after HIV infection and during HAART therapy is associated with HIV-LD. Nucleoside reverse transcriptase inhibitor, especially d4T, plays an important role in the progression of HIV-LD.

[Safety study of 52-week highly active antiretroviral therapy in 198 HIV/AIDS Chinese patients].

OBJECTIVE: To evaluate the safety profiles of three nevirapine-based therapies for antiretroviral-naive Chinese adults infected with HIV-1 (human immunodeficiency virus-1). METHODS: For this prospective multicentric randomized trial, a total of 198 antiretroviral-naive HIV-1 positive patients were recruited from 13 research centers in China. They were randomly assigned to receive three NVP-based antiretroviral therapies for 52 weeks: Group A, AZT (zidovudine) + DDI (didanosine) + NVP (nevirapine); Group B, D4T (stavudine) + 3TC (lamivudine) + NVP; Group C, AZT + 3TC + NVP. Their clinical events and laboratory examinations were monitored at baseline and the end of weeks 4, 8, 12, 24, 36 & 52 post-HAART (highly active antiretroviral therapy) to evaluate the occurrence of adverse events (AEs). The chi-square or Fisher's exact test was employed to compare the rates of AEs among three treatment groups. Multivariate logistic regression analyses were used to identify the factors associated with hepatotoxicity. For all tests, P < 0.05 was considered as statistically significant. RESULTS: During the 52-week HAART, 968 cases of AEs occurred in 188 patients (95.0%). Only 37.4% experienced grade 3/4 AE. And 37 patients withdrew because of HAART-related AEs (18.7%). The common AEs were hepatotoxicity, bone morrow suppression, gastrointestinal disorders, rash and hyperlipidemia, etc. Most instances of AEs occurred during the early 12 weeks. The total count of AEs for each group had no statistic significant difference (P = 0.403). Bone marrow suppression was more strongly associated with an AZT-containing HAART and it was especially prone to gastrointestinal disorders when combined with DDI. The introduction of D4T or DDI led more frequently to peripheral neuropathy and hyperlipidemia. Logistic regression analysis indicated that presence of hepatotoxicity was associated with a higher baseline level of CD4 (CD4 count > 250/µl) (OR = 2.08, 95%CI: 1.114 - 3.882, P = 0.021). CONCLUSION: The common reasons of discontinuing HAART are hepatotoxicity, gastrointestinal disorders, bone marrow suppression and rash. The occurrence of AEs should be vigorously monitored especially during the early 3 months of HAART. The HIV/AIDS patients with a CD4 count of > 250/µl shall avoid any NVP-containing regimen.

[The impact of highly active antiretroviral therapy on bone mineral density in human immunodeficiency virus infected patients].

OBJECTIVE: To evaluate the influence of highly active antiretroviral therapy (HAART)on bone mineral density(BMD) of human immunodeficiency virus (HIV) infected patients and correlating clinical factors. METHODS: The clinical data from 2007 to 2008 were analyzed, including 50 patients treated with HAART (named treated group), 12 HIV-infected antiretroviral-naive patients (named untreated group) and 20 healthy people (named control group). Lumbar, femoral neck, femur, femoral greater trochanter and whole body BMD were measured by dual energy X-ray absorptiometry. The data were respectively analyzed. RESULTS: There were 19(38.0%) patients with osteopenia and 1 (2.0%) patient with osteoporosis in the treated group. There were 6 (50.0%) patients with osteopenia and 2 (16.7%) patient with osteoporosis in the untreated group. There were 5 (25.0%) patients with osteopenia, no one with osteoporosis in the control group. The prevalence of osteopenia/osteoporosis was statistically higher in the untreated group than that in the control group (P=0.02). The BMD of femur, femoral neck and greater trochanter [(0.97±0.14), (0.91±0.13), (0.76±0.12) g/cm2] in the HIV-infected group (including the treated and untreated group) were significantly lower than that in the control group [(1.04±0.12), (0.98±0.14), (0.84 ± 0.11) g/cm2, P<0.05]. There were no significantly differences in the BMD between the untreated group and the treated group. In the treated group, osteopenia/osteoporosis correlated with body weight less than 60 kg (r=0.074, P=0.004) and the viral load before HAART (r=5.103, P=0.021). CONCLUSIONS: The prevalence of osteopenia and osteoporosis in antiretroviral-naive HIV-infected patients is higher. The BMD of HIV-infected patients are reduced compared with the healthy people. The BMD is similar among HIV-infected patients irrespective of antiretroviral treatment. Body weight less than 60 kg and the viral load before HAART are the risk factors of osteopenia/osteoporosis for the HIV-infected antiretroviral patients.

[Change of plasma TH1/TH2 cytokines levels in patients with hemorrhagic fever with renal syndrome].

OBJECTIVE: To observe changes in T cell subsets and TH1/TH2 secreted cytokines in the plasma of patients with hemorrhagic fever with renal syndrome (HFRS). METHODS: Totally 22 patients with HFRS (9 mild cases and 13 moderate cases) were enrolled. Blood samples were taken 1, 4, and 12 weeks after presentation. T cell subsets were tested by flow cytometry (FCM), and the expression of cytokines in plasma were analysed with enzyme-linked immunosorbent assay (ELISA). Another 16 healthy blood donors were enrolled as the control group. RESULTS: CD3 + CD8 + T lymphocytes increased at week 1 and 4 (P < 0.01), which was more significant in mild cases than in moderate cases (P < 0.05). The change of CD3 + CD4 + T lymphocytes during the disease course were not significantly different from that in control group (P > 0.05). One week after presentation, TH1 [interleukin (IL)-2 and interferon-gamma (IFN-gamma)] and TH2 (IL-6, IL-10) cytokine productions were significantly higher in HFRS patients than in the control group (P < 0.01); IL-2 and IL-10 remained high levels during the whole observation period, and were still significantly higher than in the control group (P < 0.01). At week 4, the plasma IL-5 level was significantly higher in HFRS patients than in the control group (P < 0.01), and were still significantly higher than in the control group at week 12 (P < 0.01). At week 1 and 4, the plasma INF-gamma levels were significantly higher in moderate patients than in mild patients (P < 0.05); at week 12, the plasma IL-10 level was significantly higher in moderate patients than in mild patients(P < 0.05). CONCLUSIONS: CD3 + CD4 + T lymphocytes remarkably increases at the early stage of disease in patients with mild HFRS. The early cell mediated immune response is helpful for disease control. The cytokines INF-gamma and IL-10 increase more obviously in moderate patients, indicating that cytokines also are key pathogenic factors of HRFS.

[A study of HIV-related lipodystrophy syndrome in 55 HIV-infected Chinese adult patients].

OBJECTIVE: To study the prevalence, clinical characteristics and risk factors of HIV-related lipodystrophy syndrome (HIV-LD) in our cohort of HIV-1 infected Chinese adults. METHODS: In a cross-sectional study, 55 HIV-infected patients were recruited from the HIV clinic of Peking Union Medical College Hospital; most of them were undergoing the first-class highly active antiretroviral therapy (HAART) of today in China. Lipoatrophy or lipohypertrophy was defined if there was concordance between the report of fat change and clinical examination of the participants. Whole body dual-energy X-ray absorptiometry (DEXA) scanning was performed. RESULTS: Prevalence of clinical body fat redistribution in the present study was 47.3%. Comparing with non-LD patients, HIV-LD patients had elder age and longer exposure to HAART (P < 0.05). HAART exposure and stavudine (d4T) usage were two independent risk factors for HIV-LD. CONCLUSIONS: HIV-related fat redistribution does exist in Chinese HIV population. Peripheral lipoatrophy occurs commonly in HIV-infected adults but is not associated with increased trunk fat. HAART exposure and especially d4T usage are independent risk factors for HIV-LD.

[Adipokines and highly active antiretroviral therapy related lipodystrophy: clinical study of 52 cases].

OBJECTIVE: To investigate the prevalence of glucose and lipid abnormalities in AIDS patients treated with highly active antiretroviral therapy (HAART) and difference thereof between the HIV-lipodystrophy (LD) and non-HIV-LD groups, and to compare the plasma levels of adiponectin (APN) and leptin (LEP) and their relationship to metabolic disturbance and fat redistribution in these 2 groups. METHODS: Fifty-two HIV-infected patients were divided into HIV-LD group and non-HIV-LD group according to the patients' reports and doctors' evaluation. Body composition was assessed by whole body dual-energy X-ray absorptiometry. Plasma samples were analyzed for cholesterol, triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), insulin, APN, and LEP. The prevalence of dyslipidemia and hyperinsulinemia, the difference of adipocytokine levels, and the relationship of adiponectin, leptin with lipids, insulin as well as fat mass in different body regions were analyzed between the groups. RESULTS: The prevalence rates of hypercholesterolaemia, hypertriglyceridaemia, and low HDL-C level were 17.3%, 50.0%, and 17.3% respectively. The rate of hyperinsulinemia and any kind of dyslipidemia were 25.0% and 59.6%. Compared with non-HIV-LD patients, HIV-LD patients had higher TG level, and lower HDL-C and APN levels. In the HIV-LD group, the APN level was correlated positively with limb/total body fat, but negatively with trunk/total body fat, and was an independent predictor of HDL-C and insulin level. However, LEP was positively correlated with the levels of total body fat, limb fat, and trunk fat in both groups. CONCLUSION: The prevalence rates of dyslipidemia and insulin resistance are high in Chinese HIV/AIDS patients receiving HAART, especially in the HIV-LD group. The APN concentration in the HIV-LD patients is closely related to fat redistribution and independently predicts the levels of HDL-C and insulin. LEP can serve as a biomarker of total body fat mass.

[The reconstitutional profiles of peripheral blood natural killer cell, gammadelta T lymphocyte and CD(4)(+)T cell in human immunodeficiency virus infected patients during one-year antiretroviral therapy].

OBJECTIVE: To investigate the different reconstitutional profiles for acquired (CD(4)(+)T cell) and innate (NK cell, gammadelta T lymphocyte) immunity after highly active antiretroviral therapy (HAART). METHODS: The CD(3)(+)CD(4)(+), CD(3)(+)CD(4)(-)CD(8)(-), CD(3)(-)CD(16)/CD(56)(+), CD(4)(+)CD(45)RA(+)CD(62)L(+) and CD(4)(+)CD(45)RA(-) subsets were measured by flow cytometry. The dynamic changes of these subsets after HAART initiation were assessed in 59 patients who were followed for 12 months in regular 3-month visits. RESULTS: At baseline the cell counts of CD(4)(+)T cells including its naïve and memory subsets, NK cell and gammadelta T cells in HIV/AIDS patients were all significantly lower than those of healthy individuals. There was a decrease of 2.33 lg copies/ml in HIV-1 RNA from baseline noted 1 month after initiation of treatment which was sustained through 12 months. CD(4)(+)T cell count showed a bi-phase increase during treatment. The first rapid increase was mainly memory CD(4)(+)T cells and this followed by the second slow but steady increase of naïve CD(4)(+)T cells. Increases in NK cell and gammadelta T cell were noted at 3 months of HAART and this restoration were different quantitatively when compared with the one in CD(4)(+)T cells. CONCLUSION: HAART could induce a different quantitative restorational patterns in peripheral CD(4)(+)T cells, NK cells and gammadelta T cells.

[The dynamics of T lymphocyte subsets in hemorrhagic fever with renal syndrome].

OBJECTIVES: To investigate the T cell subsets changes in hemorrhagic fever with renal syndrome (HFRS) patients. METHODS: 22 HFRS patients who were diagnosed in Qin Huang Dao Third Hospital from April 2005 to July 2005 were enrolled in this study and divided into two groups according to clinical manifestations. T cell subsets of the 22 patients were monitored at week 1, 4 and 12. Another 56 subjects were enrolled as healthy controls. RESULTS: B cell count was normal during the 12 weeks in all the subjects. NK cell decreased significantly at week 1, and recovered at week 4 rapidly. CD(4)(+)T cell count was normal throughout the course of the disease, but the percentage of memory phenotype increased at week 1 and 4, reaching(64.1 +/- 17.5)% and (59.9 +/- 10.1)%, but recovered at week 12. CD(4)(+)CD(28)(+)T cells were normal throughout the entire study. CD(8)(+)T cell count increased dramatically at week 1 and 4, but finally recovered at week 12. The count of CD(8)(+)CD(28)(-)T cells increased significantly at week 1 in low-grade goup, but in median-grade group, this increase lagged to week 4 and was not as significant as in low-grade group. The percentage of CD(38)(+) or HLA-DR(+) subsets of CD(8)(+)T cell increased at week 1, 4. CONCLUSION: The results confirmed the relationship between HFRS progression and cellular immunity. It revealed that, at the early stage of HFRS, rapid and effective cytotoxicity T lymphocyte response may contribute to clear Hantavirus away and improve HFRS symptom.

[Change of plasma pro-inflammatory cytokines levels in patients with hemorrhagic fever with renal syndrome].

OBJECTIVE: To observe the changes of the plasma pro-inflammatory cytokines levels in patients with hemorrhagic fever with renal syndrome (HFRS). METHODS: Enzyme-linked immunosorbent assay (ELISA) was performed to detect the plasma pro-inflammatory cytokines levels of 22 HFRS patients (9 mild cases and 13 moderate cases) 1, 4, and 12 weeks after they were diagnosed. Sixteen healthy blood donors were recruited as control group. RESULTS: The levels of interleukin (IL)-1beta, IL-6, IL-10, tumor necrosis factor (TNF)-alpha, and IL-8 in HFRS patients were significantly higher than those in control group 1 week after they were diagnosed (all P < 0.01). The levels of IL-6 and TNF-alpha in HFRS patients returned to the normal levels four weeks after the diagnosis, while those of IL-1beta, IL-8, and IL-10 remained significantly higher than those in control group 12 weeks after the diagnosis (all P < 0.01). The IL-8 and IL-10 levels in mild HFRS patients were significantly higher than those in moderate HFRS patients at the same period (all P < 0.05). CONCLUSION: Abnormal expressions and secretion of pro-inflammatory cytokines occurs during the disease course of HFRS.

Clinical outcomes and immune reconstitution in 103 advanced AIDS patients undergoing 12-month highly active antiretroviral therapy.

BACKGROUND: Highly active antiretroviral therapy (HAART) produces profound suppression of HIV replication, substantial increase in CD4(+) T cells, and partial reconstitution of the immune system. However, the numbers of subjects were small in previous Chinese studies. This study evaluated the efficacy and side effects of HAART in Chinese advanced AIDS patients. METHODS: One hundred and three antiretroviral drug naive AIDS patients were enrolled in this study and were divided into two groups by their baseline CD4(+) count: < 100 cells/microl or > or = 100 cells/microl. Clinical, virological and immunological outcomes were monitored at baseline and at 1, 3, 6, 9 and 12 months during the course of treatment with HAART. RESULTS: One patient died and another was lost from the follow-up. For the remaining 101 HIV/AIDS patients at the 12th month during the HAART, the plasma viral load (VL) was reduced to (3.2 +/- 0.7) lg copies/ml, the CD4(+) count increased to (168 +/- 51) cells/microl [among which the naive phenotype (CD45RA(+)CD62L(+)) increased to (49 +/- 27) cells/microl and the memory phenotype (CD45RA(-)) increased to (119 +/- 55) cells/microl], and the percentage of CD4(+)CD28(+) cells increased. At the same time, there was a significant reduction of CD8(+) T cell activation. In the 69 patients with the baseline CD4(+) count < 100 cells/microl, 37 had a VL < 50 copies/ml; while in the 34 patients with the baseline CD4(+) count > or = 100 cells/microl, 25 had a VL < 50 copies/ml, the difference between the two groups was statistically significant. The CD4(+) T cell count showed a two-phase increase during HAART and a significant positive correlation was shown between the change of CD4(+) count and plasma VL. Over 12 months of HAART, 10 patients had gastrointestinal side effects, 13 peripheral neuritis, 7 hepatic lesions, 8 hematological side effects, 8 skin rashes, 10 lipodystrophy and 1 renal calculus. CONCLUSIONS: Immune reconstitution as well as the significantly improved clinical outcomes is observed in Chinese advanced AIDS patients after HAART. Side effects are common during HAART and require clinical attention.

Association between Nef-specific CD8 T-cell responses and disease progression in HIV-1 subtype B infection.

BACKGROUND: The correlation between HIV-1 Nef-specific CD8 T-cell responses and markers of HIV-1 disease progression still remains unclear. This study analysed and compared the role of HIV-1 Nef-specific CD8 T-cell responses in patients with different disease status. METHODS: Two groups of patients with HIV-1 subtype B infection were selected according to CD4 count and clinical manifestations: long-term nonprogressors (LTNPs, n = 20) and advanced progressors (APs, CD4 count < 500 cells/microl, n = 34). Nef-specific CD8 T-cell responses were studied by interferon-gamma ELISpot assay against 3 pools of HIV-Nef peptides. RESULTS: Nef-specific CD8 T-cell responses did not correlate with viral load or CD4 count in all patients and no significant differences were found in the magnitude of Nef-specific CD8 T-cell responses between groups LTNPs and APs (670 SFC/10(6) peripheral blood mononuclear cells vs 1107 SFC/10(6) peripheral blood mononuclear cells, P = 0.255). Further comparisons showed that there were also no significant correlations observed in group LTNPs, but Nef-specific CD8 T cells correlated negatively with viral load (r = -0.397, P = 0.020) and positively with CD4 count (r = 0.364, P = 0.034) in group APs. CONCLUSION: These data suggest that different correlation patterns between Nef-specific CD8 T-cell responses and disease progression exist in LTNPs and APs. Although a negative association was observed with concurrent plasma HIV RNA in APs, Nef-specific CD8 T-cell responses might fail to play a protective role in different stages of HIV-1 infection.

[The clinical outcome and immune reconstitution in 45 advanced AIDS patients undergoing highly active antiretroviral therapy for 12 months].

OBJECTIVE: To investigate the efficacy and side effects of highly active antiretroviral therapy (HAART) in Chinese AIDS patients. METHODS: 45 antiretroviral drug-naive AIDS patients were enrolled and divided into two groups by their baseline CD(4) count < 100/microl or > or = 100/microl. Clinical, virological and immunological outcomes as well as side effects were followed at baseline and at the end of month 1, 3, 6, 9, 12 after receiving HAART. RESULTS: Among the 45 HIV/AIDS patients included, by the end of 12 months of HAART, the plasma viral load (VL) got a mean reduction by 2.8 lg copies/ml, CD(4) count had a mean gain of 187/microl, among which the naive phenotype increased by 68/microl and the memory phenotype by 119/microl. The CD(4)(+)CD(28)(+) T cell percentage went up from (62.5 +/- 25.8)% to (82.6 +/- 15.6)% (P < 0.001); and there was a significant reduction of CD(8)(+) T-cell activation. In the 31 patients with their baseline CD(4) count < 100/microl, 11 had a VL < 50 copies/ml, and 14 had fluctuations in their VL; while in 14 patients with their baseline CD(4) count > or = 100/microl, 10 had a VL < 50 copies/ml and 2 had fluctuations in their VL, respectively, with statistic significance between the two groups. CD(4) count showed a bi-phase increase during HAART and there was significant positive correlation between the change of CD(4) count and plasma VL. Throughout the 12 months of HAART, 39 patients had gastrointestinal side effects, 15 peripheral neuritis, 3 hepatic lesions, 4 hematological side effects and 1 renal calculus. 9 patients had adjustment of their initial therapy because of side effects. CONCLUSIONS: Immune reconstitution as well as significant therapeutic effect was observed in advanced Chinese AIDS patients after HAART. Side effects were common during HAART, so close clinical attention is needed.

[The correlation of HIV-1 viral load to expression of CD8+ T lymphocyte activators CD38 and HLA-DR].

OBJECTIVE: To investigate the correlation of CD38 and HLA-DR abnormal activating expression on CD8+ T with plasma viral load (VL) and evaluate the possibility of the economical CD38 and HLA-DR test to substitute VL assay in HIV/AIDS patients. METHODS: A multi-point correlation study of the percentage of CD38 and HLA-DR on CD8+ T by flow cytometry with plasma VL by bDNA was performed in 103 HIV/AIDS patients during a 12-month highly active anti-retroviral therapy (HAART). The cutoff values of CD38 and HLA-DR were evaluated with ROC area, sensitivity and specificity for predictive VL < 50 copies/ml, < 500 copies/ml, > 1000 copies/ml and > 10,000 copies/ml respectively. RESULTS: The level of CD38 and HLA-DR on CD8+ T in 103 patients decreased gradually with the reduction of VL during a 12-month HAART. The correlation of CD38 and HLA-DR with VL in the year of HAART was 0.424, 0.376, 0.335, 0.326, 0.297, 0.285 and 0.377, 0.318, 0.333, 0.312, 0.361, 0.358 with significant P value. Moreover, the overall correlation of CD38 and HLA-DR with VL were 0.483 (P < 0.001) and 0.477 (P < 0.001). Depending on optimal ROC, sensitivity and specificity for the substitute method, the cutoffs percentage of CD38 were < 68.5% and < 72.5% for predictive VL < 50 copies/ml and < 500 copies/ml as well as > 39.5% and > 46.5% of HLA-DR cutoff to predict VL > 1000 copies/ml and > 10,000 copies/ml. CONCLUSION: The detection of CD38 and HLA-DR percentage expression on CD8+ T can be available for prediction about HIV VL assay as a substitute method to survey the disease progression and HAART outcome in some resource-limited areas of China.

[Characteristics of immunophenotypic alterations in 263 HIV/AIDS patients].

OBJECTIVE: To explore the characteristics of immunophenotypic alterations of HIV-infected persons/AIDS patients--people living with AIDS (PLWA). METHODS: The clinical data and anti-coagulated blood samples of 263 treatment naive PLWA and 56 healthy controls were collected. Flow cytometry was used to determine the sets of peripheral lymphocytes: B cell, NK cell, CD4(+) T cell including the functional subset (CD28(+)CD4(+) cell), naïve subset (CD4(+)CD45RA(+)CD62L(+) cell), and memory subset (CD4(+)CD45RA(-)cell) of CD4(+) T cell, CD8(+) T cell including the activated subset (CD8(+)CD38(+) cell). Branch DNA (bDNA) assay was used to detect the plasma viral load. RESULTS: The mean CD4(+) T cell count, naïve CD4(+) T cell percentage, and CD28 expression rate in CD4(+) T cells of the PLWA were 205 (348, 63) x 10(6) cells/L, 18.5 (32.0, 6.5)%, and 86.1 (94.0, 68.3)% respectively, all significantly lower than those of the healthy controls [787 (1058, 615) x 10(6) cells/L, 35.4 (45.5, 30.0)%, and 95.7 (97.6, 91.0)% respectively, all P < 0.01]. The percentage of CD38 expression in CD8(+) T cells of the PLWA was 84.3 (92.7, 69.0)%, significantly higher than that of the controls [42.6 (50.6, 36.1)%, P < 0.01]. In the PLWA the CD4(+) T cell count was positively correlated with its CD28 expression (r = 0.480, P < 0.01), and the percentage of CD38 expression in CD8(+) T cells was positively correlated with eh plasma viral load (r = 0.331, P < 0.01). The PLWA were divided into 3 groups according to the CD4(+) T cell count: Group A with the he CD4(+) T cell count < 200 x 10(6) cells/L, Group B with the CD4(+) T cell count of 200 - 350 x 10(6) cells/L, and Group C with the CD4(+) T cell count > 350 x 10(6) cells/L. In comparison with Groups B and C the plasma viral load, activated CD8(+) T cell subset proportion, and percentage of memory CD4(+) T cells of Group A were all significantly higher, and the naive CD4(+) T cell percentage and CD28 expression rate were both significantly lower (all P < 0.01). There were no significant differences in the percentage of memory CD4(+) T cells, CD28 expression, and CD8(+) T cell activated subset proportion between Groups B and C. CONCLUSION: The major immunophenotypic alternations in the PLWA in China include significantly lower counts of CD4(+) T cells and their naive subsets, marked down-regulation of CD28 expression and extremely activated CD8(+) T cells. Distinct features of the immunophenotypic alteration may exist in different disease stages. The CD4(+) T cell count < 200 x 10(6) cells/L may predict more severe immunodeficiency.

[Dynamic changes of T lymphocyte subsets in the long-term follow-up of severe acute respiratory syndrome patients].

OBJECTIVE: To study the dynamic changes of T lymphocyte subsets in severe acute respiratory syndrome (SARS) patients. METHODS: Sequential anti-coagulated blood samples were collected from 62 seropositive SARS patients during the first week, the second week, the first month, the 2nd-3rd month, and 1 year after disease onset. T-lymphocyte subsets including CD4 + T cells, CD8 + T cells, and naive and memory CD4 + T cells were detected by flow cytometry. Samples from 56 healthy blood donors were also detected as healthy controls. RESULTS: A rapid restoration in T-lymphocyte subsets was observed during a short period after infection. During the 2-3 months after disease onset, CD8 + T cell count returned to the normal level. At the same time point, CD4 + T cell and naive CD4 + T cell counts increased to (534 +/- 197)/mm3 and (175 +/- 75)/mm3 respectively. Total lymphocytes, T lymphocyte, CD4 + T and naive CD4 + T cell were still significantly lower than the normal levels even one year after disease onset. CONCLUSION: The periphery T lymphocyte subsets in SARS infection shows transient decrease and then rapid recovery.