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1.
Chinese Journal of Infectious Diseases ; (12): 538-544, 2022.
Article in Chinese | WPRIM | ID: wpr-956451

ABSTRACT

Objective:To explore the dynamic changes of inflammatory cytokines and T lymphocyte activation in the peripheral blood of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients during anti-retroviral therapy (ART).Methods:Two hundred and six HIV/AIDS patients with ART at clinic of the Department of Infectious Diseases in Second Xiangya Hospital, Central-South University between January 2017 and December 2019 were selected as HIV infection group. They were followed up regularly and the blood samples before treatment and at month 6, month 12, month 24 of treatment were collected. Meanwhile, 52 healthy cases were enrolled in the healthy control group and their blood samples were collected. Enzyme-linked immunosorbent assay was used to detect the plasma concentrations of interleukin (IL)-6, hypersensitive C-reactive protein (hsCRP) and tumor necrosis factor (TNF)-α. Flow cytometry was used to detect the CD3 + CD4 + T lymphocytes count and the percentage of CD4 + CD38 + T lymphocytes and CD8 + CD38 + T lymphocytes in the peripheral blood mononuclear cells. Plasma HIV RNA viral load was determined using a quantitative real-time polymerase chain reaction technique. Statistical methods used paired t test and Pearson correlation analysis. Results:The concentrations of IL-6, hsCRP and TNF-α in HIV infection group were (13.42±2.35) pg/mL, (4 012.46±1 012.35) μg/L and (51.78±11.32) μg/L, respectively, which were higher than those in healthy control group ((6.14±0.78) pg/mL, (707.21±305.76) μg/L and (19.01±6.48) μg/L, respectively). The differences were all statistically significant ( t=12.56, 16.79 and 13.45, respectively, all P<0.001). They decreased gradually after initiation of ART in HIV infection group, and returned to normal levels at month 24 of ART. CD3 + CD4 + T cells count was (256.00±65.32)/μL and HIV RNA viral load was (4.467±4.244) lg copies/mL before ART in HIV infection group, which were negatively correlated ( r=-0.625, P=0.041). The percentages of CD8 + CD38 + T lymphocytes before treatment and at month 12 or month 24 of treatment in HIV infection group were higher than those in healthy control group. The differences were all statistically significant ( t=3.85, 6.84 and 2.57, respectively, all P<0.050). The percentage of CD8 + CD38 + T lymphocytes was positively correlated with HIV RNA viral load before ART ( r=0.768, P=0.026). The percentages of CD4 + CD38 + T lymphocytes before treatment and at month 12 or month 24 of treatment in the HIV infection group were lower than those in the healthy control group, and the differences were all statistically significant ( t=6.80, 1.10, and 2.11, respectively, all P<0.050). Conclusions:HIV infection could not only cause insufficiency in immune system, but also abnormal activation of immune system, which could get better gradually with ART.

2.
Chinese Journal of Infectious Diseases ; (12): 271-274, 2019.
Article in Chinese | WPRIM | ID: wpr-754660

ABSTRACT

Objective To analyze the impact of secondary infection on prognosis of liver failure. Methods A total of 384 hospitalized patients who were diagnosed with liver failure from January 2015 to Decembet 2017 in the Department of Infectious Diseases of the Second Xiangya Hospital of Central South University were retrospectively analyzed.The patients were divided into infected group and non-infected group according to whether they were complicated with infection during hospitalization .The cause of liver failure, the area and source of infection were recorded.The infected group was divided into bacterial group and fungal group.The liver and kidney function , international normalized ratio ( INR).The model for end-stage liver disease ( MELD ) score, hospitalization days , medical expenditure , and mortality were calculated and evaluated.T test was used for normally distributed continuous variables , and chi-square test was used for classified variables.Results A total of 384 hospitalized patients with liver failure were enrolled , including 321 males and 63 females with age of (45.5 ±13.4) years.There were 240 patients (62.5%, infected group) who had secondary infection during the whole course , and 144 patients (37.5%, non-infected group ) were not infected.Among the 384 patients, 328 patients (85.4%) were infected with hepatitis B virus, 8(2.1%) with hepatitis C virus, and 10(2.6%) with alcoholic hepatitis.As for the clinical types of liver failure , 187 patients (48.7%) were diagnosed with acute-on-chronic (subacute) liver failure and 158 (41.1%) with chronic liver failure.Among the 240 patients in the infected group, 122 patients (50.8%) had abdominal infection, 84 (35%) had pulmonary infection, 8(3.3%) had urinary tract infection, 13(5.4%) had biliary tract infection , and 11 ( 4.6%) had bloodstream infection.The levels of total bilirubin , creatinine, MELD scores, hospitalization days and medical expenditure in the infected group and non -infected group were statistically significant (all P<0.01) after 30 days in hospital.In the infected group, 362 various samples from 240 patients were submitted for bacterial culture , among which 87 samples were positive, including Candida in 15 samples, Aspergillus in 8 samples, Acinetobacter baumannii in 13 samples, Staphylococcus in 10 samples, Escherichia coli in 11 samples, Klebsiella pneumoniae in 14 samples, Bacillus faecalis in 4 samples, Bacillus pallid in 4 samples, Stenotrophomonas maltophilia in 4 samples and Aeromonas hydrophila in 4 samples.Among the 240 patients in the infected group , 182 patients were diagnosed with bacterial infection and 58 with fungal infection. There were significant differences in total bilirubin , serum creatinine, INR, MELD scores and mortality rate between the two groups ( all P<0.05).Conclusions The rate of secondary infection in patients with liver failure is not related with age.The development of secondary infection , especially fungal infection , worsens the prognosis of patients with liver failure.

3.
Journal of Central South University(Medical Sciences) ; (12): 537-544, 2013.
Article in Chinese | WPRIM | ID: wpr-814846

ABSTRACT

OBJECTIVE@#To observe the dynamic changes of 3 types of viral reservoir cells (NK cells, T lymphocytes and monocytes), and its relationship with treatment effect in Chinese HIV-1 infected patients receiving highly active antiretroviral treatment (HAART) for 2 years.@*METHODS@#A total of 40 chronic HIV-1-infected adults who initiated HAART were enrolled in this study and followed up for 2 years. Peripheral whole blood was obtained from each patient at baseline (0 month), 6, 12, 18 and 24 months. Real-time fluorescent quantitative PCR was used to detect the HIV-1 RNA in the plasma and HIV-1 DNA in NK cells, T lymphocytes and monocytes. All the data were statistically analyzed.@*RESULTS@#CD4 count increased with the decrease of the viral load during HAART. After HAART initiation, HIV-1 DNA showed a significant decrease in NK cells, T lymphocytes and monocytes. The HIV-1 DNA from T lymphocytes, NK cells and monocytes correlated positively with the HIV- 1 RNA (P<0.05) while NK cells and T lymphocytes correlated negatively with CD4+ T cell count. However we did not find significant correlation between CD4+ T cell count and HIV-1 DNA in monocytes at the baseline of HAART.@*CONCLUSION@#This study found that NK cell was an important HIV cellular reservoir besides T lymphocytes and monocytes. T lymphocytes may be the main long lasting HIV reservoir. HIV-1 proviral DNA may play an important role in the efficacy of treatment and monitoring the disease progression.


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Antiretroviral Therapy, Highly Active , DNA, Viral , HIV Infections , Drug Therapy , Virology , HIV-1 , Genetics , Killer Cells, Natural , Virology , Monocytes , Virology , RNA, Viral , Blood , T-Lymphocytes , Virology , Viral Load
4.
Journal of Chinese Physician ; (12): 308-312,317, 2012.
Article in Chinese | WPRIM | ID: wpr-597889

ABSTRACT

Objective The objective of this study was to investigate the course of certain common gamma cytokines ( IL-2 and IL-7 ) and their role on the control of the viral infection in a short term antiviral therapy.Methods A total of 35 adults with chronic HIV infection,responding to combined antiretroviral therapy (cART) guideline criteria were enrolled in this one year follow-up study.After signing an informed consent,20 ml blood were collected from each patient at base line,week 0,week 24 and week 48.1 ml serum collected from each patient was kept at -80 * C until use.Serum concentration of IL-2 and IL-7 was determined using ELISA kit from ebioscience Beijing.CD4 and CD8 cells were counted and quantified using flux cytometry,and serum HIV RNA was quantified using real time PCR.Results All patients had a mean baseline IL-2 level [ (9.67 ± 2.6 ) pg/ml ]lower than the controls [ ( 27.36 ± 5.05 ) pg/ml ].After treatment for 48 weeks,IL-2 increased[ ( 19.8 ± 3.3 ) pg/ml ].However,the mean baseline 1L-7 [ ( 81.74± 20.47 ) pg/ml ]in patients was higher than controls [ ( 2.06 ± 1.52 ) pg/ml ].After treatment for 48 weeks,IL-7 decreased [ (8.36 ± 2.16)pg/ml ].IL-2 showed a significant increase and positive correlation with CD4 cells after HAART initiation (0week:R =0.21,P =0.063,24week:R =0.24,P =0.033,48week:R =0.19,P =0.103; IL-7 showed a significant decrease after HAART initiation but it did not show correlation with CD4 cells.We noted there was a negative correlation between IL-2 and CD4 count in HAART baseline (R =0.28,P =0.012 ),but no correlation between IL-7 and CD4 count from 6 month after HAART.IL-2 showed negative correlation with HIV RNA ( R =- 0.17,P =0.032),but IL-7 showed a relationship with the HIV RNA Conclusions The increase of IL-2 coupled with the decrease of IL-7 revealed a partial restoration of immune response during HAART.However,the absence of relationship with HIV RNA suggested that these cytokines might not be directly involved in the reduction of viral load.

5.
Chinese Journal of Microbiology and Immunology ; (12): 512-516, 2011.
Article in Chinese | WPRIM | ID: wpr-415663

ABSTRACT

Objective To observe the Th17, IL-17 and Tr cells equilibrium state as well as their changes of HIV infected or AIDS suffered patients in one-year HAART treatment. Methods Select 33 HIV/AIDS patients received HAART treatment while 33 healthy volunteers as controls. Flow cytometry was used to analyze Th17 and Tr cells in venous blood at the time of pre-therapy, 6th, 12th month when IL-17 levels in serum are tested by ELISA. Results The ratio of Th17 cells in CD4 cells in HIV/AIDS patients and volunteers were (1.20±0.37)%, (2.50±1.03)%, (3.70±1.56)%, (4.70±1.43)%, respectively; The ratio of Tr cells were (9.16±3.33)%, (7.19±2.91)%, (5.53±1.88)%, (4.43±0.97)%, respectively; The levels of IL-17 in serum were (5.3±2.5) pg/ml, (7.7±2.4) pg/ml, (10.4±3.1) pg/ml, (17.7±6.6) pg/ml respectively. The Th17 cells' level was positively correlative with the amount of CD4 cells, negatively correlate with the count of viral load. However, the Tr cells level is positively correlative with the count of viral load, negatively relate to the quantity of CD4 cells. Conclusion HIV could make IL-17, Th17 cells and Tr cells lost their balance, but the immune equilibrium state may gradually recover after HAART treatment. Which indicates the IL-17, Th17 cells and Treg cells may play an important role in the pathogenesis of AIDS, and they are likely to be the effective indexes to observe the progress of AIDS and the treatment effect of highly active antiretroviral therapy(HAART).

6.
Journal of Central South University(Medical Sciences) ; (12): 1037-1045, 2011.
Article in Chinese | WPRIM | ID: wpr-669505

ABSTRACT

Although highly active antiretroviral therapy (HAART) can effectively reduce the HIV replication,complete recovery of CD4+ T cells does not always occur,even among patients with high virological control.Current researches on γ-chain cytokines have understood the biology and their crucial roles in initiating,maintaining,and regulating the immunologic homeostasis and the inflammatory processes.Due to the multiple functions such as the regulatory and effector cellular function in healthy and disease state,these molecules,their receptors,and their signal transduction pathways are promising candidates for therapeutic interference.The common γ-chain cytokines IL-2,IL-7,IL-15,and IL-21 are primary regulators of T cell homeostasis and thus have been considered prime immunotherapeutic candidates,both for increasing T cell levels/function and augmenting vaccine-elicited viral-specific T cell responses in immunocompromised AIDS patients.The objective of this review is to update the role of the common γ-chain cytokines IL-2,IL-7,IL-15,and IL-21 in HIV AIDS pathogenesis.

7.
Journal of Central South University(Medical Sciences) ; (12): 1059-1064, 2011.
Article in Chinese | WPRIM | ID: wpr-669503

ABSTRACT

To investigate the dynamics of interleukin-21 (IL-21) cytokine in the Chinese HIV patients undergoing highly active antiretroviral therapy (HAAPT).Methods A total of 25 adults with chronic HIV infections,responding to combined highly active antiretroviral therapy (HAART) guideline criteria were enrolled for a 1-year follow-up.After signing an informed consent,20 mL blood was collected from each patient at the base line,6 month and 12 month,respectively.CD4 and CD8 cell count was quantified by flux cytometry,serum HIV RNA quantified by real time PCR and IL-21 concentrations by ELISA.Results IL-21 levels increased gradually during the follow-up but did not reach the healthy levels.IL-21 correlated positively with the CD4 cells but not with CD8 T cells.HIV RNA correlated negatively with CD4 cell count but did not show any relationship with the CD8 cells.Conclusion IL-21 has potential role in the immunopathogenesis of HIV,and might be an important factor in immune construction during HAART.

8.
Chinese Journal of Microbiology and Immunology ; (12): 62-68, 2011.
Article in Chinese | WPRIM | ID: wpr-382664

ABSTRACT

Objective To investigate the immunological pathogenesis of immune reconstitution inflammatory syndrome (IRIS) during highly active antiretroviral therapy( HAART), in this prospective cohort study we analyzed the lymphocyte subsets, lymphocyte activation, changes in regulatory T cells, and levels of Th1 and Th2 cytokines in both IRIS and non-IRIS groups. Methods Two hundred and thirty-eight AIDS patients received HAART and participated prospective research cohort for 24 weeks follow-up. Forty-seven IRIS cases and 191 non-IRIS cases were enrolled in the IRIS group or non-IRIS group respectively. Blood samples were collected in both groups at pre- and post-HAART 12 weeks, 24 weeks. Using flow cytometer to detect the immunophenotypes of lymphocyte subsets (CD4 + CD45RA+ CD62L+, CD8+ CD45RA+ CD62L+naive T cells; CD4+ CD45RO+, CD8+ CD45RO+ memory T cells), activated T lymphocytes (CD4+CD38 +, CD8 + CD38 + cells), and regulatory T cell ( CD4 + CD25 + Foxp3 + ). Blood samples collected at pre-and post-HAART4 weeks, 12 weeks, 24 weeks and used ELISA to detect IL-2, IFN-γ, IL-4, IL-10and IL-7 cytokine serum levels. Results The percentages of CD4 + and CD8 + naive T cells and mlemory T cells exhibited no significant differences at the baseline, 12 weeks, 24 weeks of HAART initiation between both groups, but CD4 + and CD8 + memory T cells were demonstrated a trend towards to increase while compared to baseline during HAART. The percentages of CD4 + and CD8 + activated T cells are significantly higher at the baseline while compared to normal control and demonstrated a downward trend, but between both groups showed no significant difference. The percentages of CD4 + regulatory T cell was lower in IRIS group than non-IRIS group at the baseline, 12 weeks, 24 weeks and the onset of IRIS. Th1 cytokines, IL-2 and IFN-γshowed an upward trend during HAART at the levels of IRIS group had significantly increased at 4 weeks and the onset of IRIS. Th2 cytokines, IL-4 and IL-10 showed a downward trend during HAART,and the levels of IL-10 in IRIS group had significantly decreased at 4 weeks and the onset of IRIS. IL-7 was higher than normal control at the baseline in two groups and showed a downward trend during HAART. The level of IL-7 was higher than non-IRIS group at all follow-up points. Conclusion Memory T cells appear rapid increase in the early stage of HAART and may play a significant role in the inflammatory response of IRIS. CD4 + and CD8 + naive T cells, memory T cells and activated T cells showed no significant difference between IRIS and non-IRIS group within 24 weeks after HAART started. There was a significant reduction in the frequency of regulatory T cells in IRIS group without obvious upward trend during HAART, suggesting that the immune suppression function of regulatory T cells in IRIS was impaired. IL-2 and IFN-γ significantly increased while IL-10 significantly decreased at 4 weeks post-HAART initiation and onset of IRIS in IRISgroup than non-IRIS group, suggested that IRIS was related to cytokines environment disorder. That is, a significant increase in inflammatory cytokines, while the relative lack of non-inflammatory cytokines. The level of IL-7 decreased gradually after HAART started, and it was higher in IRIS group when compared to non-IRIS group in the first 24 weeks after HAART started. Also IL-7 may play a role in the pathogenesis of IRIS.

9.
Journal of Chinese Physician ; (12): 1158-1161, 2010.
Article in Chinese | WPRIM | ID: wpr-386542

ABSTRACT

Objective To determine the incidence, clinical manifestation and part of lymphokines which represent the balance of Th1 and Th2 in the role of the immunologic mechanisms for IRIS(immune restoration inflammatory syndromes)in patients initiating HAART(Highly Active Antiretroviral Therapy).Methods A prospective study of all patients initiating HAART was performed. A period of six months tracking initiating HAART was performed. The incidence of IRIS, time of occurrence and clinical disease spectrum were recorded. The main T lymphokines including IL-2, INF-γ, IL-4, IL-10 which on behalf of the balance of Th1 and Th2 were detected. To explore the immunopathologic mechanisms for IRIS, the levels of T lymphokines at pre-HAART, initiating HAART for 1 month, 3months and 6 months were compared in IRIS group and non-IRIS group, healthy group. Results A total of 212 patients were enrolled in this study. 59 patients were diagnosed as IRIS at a median of 21 days after HAART initiation (QR 19 days).The main disease spectrum included tuberculosis, herpes virus infections, pneumocystis jirovecii pneumonia. No matter in the IRIS group or non-IRIS group, the main lymphokines baseline of IL-2, INF-γ reduced and IL-4, IL-10 increased before HAART compared to healthy group (P < 0. 05), which had the tendency to restore balance relations initiating HAART. The lymphokines levels had significant difference between baseline and 6 months initiating HAART (P < 0. 05). The changed levels of lymphokines between IRIS group and non-IRIS group before HAART had significant difference compared to healthy group. IL-2, INF-γ increased level[(11.68 ± 2. 89) pg/ml vs (8.52 ±2.26) pg/ml; (22. 19 ± 6. 22) pg/ml vs (18.34 ±5. 35) pg/ml] and IL-10 decreased level [(19. 21 ± 4. 03) pg/ml vs (23. 19 ± 5.92) pg/ml] had significant difference between IRIS group and non-IRIS group initiating HAART I month(P <0. 05). Conclusions The incidence of IRIS during 6 months initiating HAART in HIV/AIDS was 27. 8%, IRIS usually occurred in 1 month initiating HAART. The most common disease spectrum was infectious disease, including tuberculosis and herpes virus infection. Lymphokine of Th1 and Th2 existed unbalance in IRIS group and non-IRIS group before HAART. The unbalance tendency in IRIS group was more obvious. All lymphokines had the trend to recover balance. IL-2, INF-γ significantly increased and IL-10 significantly decreased, which might involve the occurrence of the IRIS.

10.
Journal of Central South University(Medical Sciences) ; (12): 731-737, 2009.
Article in Chinese | WPRIM | ID: wpr-814281

ABSTRACT

OBJECTIVE@#To evaluate the long-term efficacy and safety of nevirapine (NVP)-based regimens for HIV-infected Chinese patients in routine clinical practice.@*METHODS@#From October 2002 to May 2004, 57 HIV-1-infected patients commenced highly active antiretroviral therapy (HAART), and were followed to December 2008. They originally received 2 nucleoside reverse transcriptase inhibitors (NRTIs) and nevirapine. HIV RNA levels, T lymphocyte subsets and safety were assessed. Blood routine test and main laboratory parameter changes were traced. If apparent side effects or virological failure appeared we would, if necessary, terminate the therapy or change the regimen.@*RESULTS@#Of the 57 subjects, 34 were followed-up for more than 4 years. After 5-6 years, 63.3% of the subjects (19/30) had HIV RNA levels<50 copies/microL, and the median increase in CD4(+) cell count from the baseline was 329 cells/microL. The mean decrease in CD8(+) cell count was 128 cells/microL. At the same time, the CD4(+) CD45RA+CD62L cell count and CD4(+)CD45RO(+) cell gradually increased, and the counts of CD8(+)CD38(+) cell declined gradually. These changes are apparent 2 years after HAART. The increase rate slowed down after 2 years. But they did not recover completely as well as healthy people at year 6. About 56% (32/57) of HIV-infected patients developed various drug-related side effects. The most common was gastrointestinal side effect, followed nervous disorder, baldness, and rashes, mostly happened in 6 months. Gamma-GT increased occurred in 29.8% of patients (17/57), and serum cholesterol and triglyceride elevated in 26.3% of the patients (15/57). Six patients developed lipodystrophy, mainly in female patients, and 25 patients showed abnormal blood picture and liver function, renal function changes and amylase elevation. Grade 3-4 adverse events occurred in 3 cases (2 peripheral neuropathy, and 1 suspected lactic acidosis). One subject experienced grade 3 rashes.@*CONCLUSION@#Antiretroviral therapy with NVP-based regimens is safe and effective by suppressing HIV viremia and producing continued CD4 cell increases in subjects with HIV or AIDS for 6 years.


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Anti-HIV Agents , Antiretroviral Therapy, Highly Active , Methods , CD4 Lymphocyte Count , China , Didanosine , Follow-Up Studies , HIV Infections , Drug Therapy , Virology , HIV-1 , Nevirapine , Reverse Transcriptase Inhibitors , Stavudine , Treatment Outcome , Viral Load
11.
Chinese Journal of Microbiology and Immunology ; (12): 1025-1030, 2009.
Article in Chinese | WPRIM | ID: wpr-380248

ABSTRACT

Objective To explore ability of the vpr gene of human immunodeficiency virus type 1 ( HIV-1 vpr) to induce cell G_2 arrest and apoptosis, and the influence when it mutated, the relationship between Vpr-induced G_2 arrest and apoptosis inductions. Methods Fourteen mutant vpr fragments selected from Chinese patients with HIV. Both eukaryotic expression vector pcDNA3.1( + ) and PCR products purified, double-cut by Hind Ⅲ and BamH Ⅰ and the cut products legated and transformed into competent cells JM109. The 14 reconstructed plasmids electronically transfected into Jurkat-cells, and established cells with pcDNA3. 1-vpr , pcDNA3. 1-vpr-Fs and pcDNA3. 1 blank cells, and without pcDNA3. 1 cell. Cells were harvested after 24 h. mRNA expression was detected by RT-PCR, the DNA content and percentage of apoptosis were monitored by flow cytometry. Results Transfected with 14 mutant HIV-1 Vpr protein, cells display different G_2 percentage and apoptosis ratio. HIV-1 vpr induce cell cycle G_2 arrest and apoptosis, wherase Vpr Fs with a C-terminal end truncation, vector pcDNA3.1( + ) and the blank cells can not. The G_2 percentage and apoptosis ratio reduced when transfected with vpr expressing mutating of 70V, 85P, 86G, 94G compared to the wild type. Subtype AE has a weaker potential to induce cell cycle G_2 arrest and apoptosis. Preliminary, we find that the higher G_2 percentage followed the higher ratio of apoptosis. Conclusion HIV-1 vpr can induce cell cycle G_2 arrest and apoptosis, wherase Vpr Fs with a C-terminal end truncation can not. We firstly found that mutated sites of 70V, 85P, 86G, 94G may reduce the ability of Vpr to induce cell cycle G_2 arrest and apoptosis, subtype AE of vpr in Chinese HIV-1 patients has a weaker potential to induce cell cycle G_2 arrest and apoptosis. Analysis of various mutations in the vpr gene revealed that the extent of Vpr-induced G_2 arrest correlated with the levels of apoptosis. And investigate the pathegenesis of HIV vpr. This can also make a good foundation for further study on gene therapy.

12.
Chinese Journal of Microbiology and Immunology ; (12): 545-549, 2008.
Article in Chinese | WPRIM | ID: wpr-382015

ABSTRACT

Objective To establish cell strain expressing the genes of HIV vpr and mutant HIV vpr-FS, and to explore cell apoptosis ability by HIV Vpr and Vpr-FS. Methods The recombinant plasmids were constructed by cloning HIV vpr and HIV vpr-FS genes into the eukaryotic expression vector pcDNA3.1respectively. To determine the primary structures of HIV vpr and HIV vpr-FS, plasmids were cleaved by restriction enzymes. After the plasmids were transfected into HeLa cells by liposome, the HeLa cells were selected with G418 selective medium, mRNA expression of HIV vpr or HIV vpr-FS of transfected cells was detected by RT-PCR, and Vpr and Vpr-FS protein expression were detected by Western blot assay respectively. The DNA content and the percentage of apoptosis in HeLa HIV vpr cell, HeLa HIV vpr-FS cell and HeLa pcDNA3.1 cell were monitored by flow cytometry and the DNA fragmentation was analyzed by agarose gel electrophoresis. Results BamH Ⅰ and Hind Ⅲ cleavaged products of pcDNA3.1-vpr and pcDNA3.1-vpr-Fincluded 342 bp length fragments suggesting that the length of DNA sequence containing HIV vpr (HIV vpr-FS) within pcDNA3.1 was the same as theoretical length. The HeLa cells transfected by pcDNA3.1-vpr or pcDNA3, l-vpr-FS and selected with G418 could express HIV vpr or HIV vpr-FS by RT-PCR, and express HIV Vpr or HIV Vpr-FS protein by Western blot. The results of flow cytometry and DNA fragmentation showed that there was significant different in the number of apoptotic cells between HeLa HIV vpr cell and HeLa HIV vpr-FS cell, but the difference between HeLa HIV vpr-FS cell and control group was not obvious. Conclusion Recombinant plasmids pcDNA3.1-vpr and pcDNA3. 1-vpr-FS were constructed successfully, and the cell strain expressing HIV Vpr and HIV Vpr-FS proteins was established. The HIV Vpr could induce host cell apoptosis, while the mutant of Vpr did not or weakened this ability. This study provides foundation for further study on HIV vpr gene.

13.
Journal of Chinese Physician ; (12)2000.
Article in Chinese | WPRIM | ID: wpr-526086

ABSTRACT

Objective To observe the changes of CD_(28) and soluble CD_(27)(sCD_(27)) among HIV-1-infected Chinese patients receiving highly active antiretroviral therapy(HAART) and to explore their clinical significance.Methods The HIV-RNA viral load(VL) and cell counts of CD_3~+CD_4~+,CD_3~+CD_8~+,CD_4~+CD_(45)RA~+CD_(62)L~+,CD_4~+CD_(28)~+,CD_8~+CD_(28)~+ and the content of sCD_(27) were determined in 28 cases of HIV-1-infected patients at the baseline and 12,24 and 48 weeks respectively after the treatment with three drug regimens based on two nucleoside reverse transcriptase inhibitors(NRTIs) and an Non-NRTI(NNRTI) for 1 year.Results The average of HIV-RNA VL before the treatment was 5.25 logs,and decreased to 3.30,1.92 and 1.74 logs after the treatment for 12,24 and 48 weeks respectively.The cell counts of CD_3~+CD_4~+,CD_3~+CD_8~+,CD_4~+CD_(45)RA~+62L~+,CD_4~+CD_(28)~+,CD_8~+CD_(28)~+ before the treatment were(236?85),(917?345),(58?15),(175?63) and(285?87) cells/?l,and changed into(327?112),(803?343),(114?25),(286?132) and(387?201) cells/?l after the treatment for 48 weeks respectively.The content of sCD_(27) decreased from(475?123) U/ml to(174?77) U/ml after the treatment for 48 weeks.The HIV RNA VL had significantly negative correlation with cell counts of CD_4~+T,CD_4~+CD_(28)~+ and CD_8~+CD_(28)~+,and there was positive correlation between HIV RNA VL and the level of sCD_(27).Conclusion The immune reconstitution and activation depend on the efficacy of antiretroviral therapy.The HIV RNA VL and cell counts of CD_4~+T are the representative parameters of immunologic and viral responses of HAART.The cell counts of CD_4~+CD_(28)~+ and CD_8~+CD_(28)~+ and their percentages show concomitant changes with viral load,and play an effective role in the progression and prognosis of AIDS.The level of sCD_(27) is positively correlated with HIV RNA VL and negatively with counts of CD_4~+T cell.The detection of sCD_(27) by enzyme linked immunosorbent assay(ELISA),which is much more convenient and cheaper than RT-PCR and flow cytometry,may serve as an important parameter to judge the pathogenetic condition of HIV-1-infected patients and to examine the therapeutic effects of HAART in the medical resource-limited area.

14.
Journal of Chinese Physician ; (12)2000.
Article in Chinese | WPRIM | ID: wpr-521678

ABSTRACT

Objective To obserre antiviral effect, immune rebuilding efficacy and side effects of didonosine, stavudine and nevirapine combination therapy for human immunodeficiency virus (HIV) type 1 infection. Methods Three medicines were administered for 8 HIV-1 infected cases. Plasma HIV RNA load, and the levels of CD 4 + T cell and CD 8 + T cell were detected before starting treatment and 4,12 and 24 weeks after starting treatment. Side effects and changes in laboratory's examinations were observed during the trial. Results After 12 and 24 week treatment, the plasma HIV-1 level reduced 2 28 logs and 2 63 logs, from a mean baseline of (283,125?187,217) copies/ml to (1,501?930) copies/ml and (669?477) copies/ml, the CD 4 + T cell counts increased from a mean baseline of (337?221) cells/ml to (393?301) cells/ml and (414?284) cells/ml, and CD 8 + T cell counts decreased from a mean baseline of (918?371) cells/ml to (823?315) cells/ml (812?305) cells/ml, respectively. Part of cases occurred mild or medium rash, nausea, headache, fatique, abdomina1 pain and diarrhea, and had blood transaminase increased, and leucocyte and hemoglobin decreased. Conclusions Didonosine, stavudine and nevirapine showed high anti-HIV and immune rebuilding effects in highly active antiretroviral therapy (HAART). The medicine regimen was well tolerated in a six-month clinical trial.

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