| ObjectiveThe study was to evaluate the efficacy and safety of regimen combining Indinavir with Efavirenz in Chinese HIV/AIDS patients.Methods1. Subjects; Ten HIV - seropositive individuals confirmed by Western blot were recruited from Liaoning province. Their CD4 counts were < 350cells/l and HIV - 1 RNA >55000 copies/ml (RT -PCR assays) and without any other antiviral therapy within 6 months. No severe disfunction in liver and kidney was found .2. HA ART regimen; Indinavir, a protease inhibitor, should be taken orally l000mg every 8 hours. Efavirenz, a non - nucleoside reverse transcriptase inhibitor, should be taken orally 600mg daily. The duration was 6 months. No other antiviral medicine was involved during therapy and no drug and alcohol a-buse was permitted.3. Follow - up and monitoring: Viral load, CD4 true count and expression of HLA - DR and CD38 on CD4 and CDS T cells, drug resistance genes, clinical manifestations and adverse events were monitored before and 1,3,6 months after treatment. Routine test of blood and urine, function of liver and kidney as well as blood sugar, cholesterol and amylase were tested to monitor the side effect of HAART. The adherence to antiretroviral therapy was measured by the proportion of pills taken out of the total number of pills prescribed.4. Sample collection: The serum and plasma were stored in -80℃ within4 hours after obtaining blood samples. The analysis of T lymphocyte subsets wasperformed by flow cytometer within 6 hours after obtaining blood samples.5. HIV viral load assay: HIV - 1 viral load was measured in thawed plasma samples using a quantitative RT - PCR assay ( COB AS AMPLJCOR, Roche Company, Switzerland) according to the instructions provided with the HIV -1 RNA monitor Test Kit version 1.5. The detectable range was 400 - 7. 5 x 10s copies/ml.6. Detection of CD4 absolute counts: Pipette 20l of TriTEST CD4/CD8/ CD3 reagent and add 50l anticoagulated whole blood into the bottom of the TruCOUNT Tube using reverse pipetting and incubate for 15 minutes in dark at room temperature (20 - 25 ). Add 450 jxl IX FACS Lysing Solution to the tube, incubate for 15minutes in the dark at room temperature and analyze the samples on the flow cytometer using FACS MULTISET software to acquire CD3 + , CD8+ and CD4 +counts and corresponding ratios.7. Detection of HLA - DR and CD38 expression on CD4 and CD8 T cells Fluorirnetric analysis was performed using the monoclonal antibodies ( Becton Dickinson). A mixture of three antibodies consisting of CD4 - FITC or CD8-FITC, CD38 - PE, HLA - DR - Percp was added to 100l of undiluted blood. After a 30 - min incubation at room temperature, the erythrocytes were lysed u-sing FACS lysing solution, washed once with wash solution consisting of PBS, resuspended in 0.2ml of wash solution, and directly subjected to cytofluorimet-ric analysis performed by FACS Calibur ( Becton Dickinson) on a Macintosh Computer.8. Genotypic drug - resistance testing of HIV - 1: HIV - 1 RNA was extracted from 200 l of plasma by using QIAamp Viral RNA Mini Kit. ( QIA-GEN, Germany) , HIV -1 RNA pol region genes were reverse transcribed and amplified by nested PCR. The amplified fragments were directly sequenced u-sing DNA auto - sequencer ABI -377 (Applied Biosystems, USA). The results of amino acid sequence were analyzed by BioEdit software and compared with consensus sequence of subtype B, C, D, G, and B/C on Internet for determination the exist of drug resistance genes.9. Statistical Analysis: Parametric data for comparisons of means were analyzed using Paired - Samples t test with SPSS 11.0 software.Results1. AIDS -correlated clinical manifestations-. After treatment for 6 months, fatigue of 4 cases was relieved and weight of 6 cases increased averagely 6.5 kg. Pruritus of two cases were relieved and rash of one case disappeared. The platelets of one case increased from 71 x 109/L to 112 x 109/L after therapy.2. Viral load; The average level of viral load was 3. 58 x 10scopies...
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