Analysis of serum adenosine deaminase (ADA) and ADA1 and ADA2 isoenzyme activities in HIV positive and HIV-HBV co-infected patients

Objectives

To determine adenosine deaminase (ADA) activity as a possible diagnostic marker in HIV and HIV–HBV co-infected patients.

Design and methods

Blood samples were collected from 72 healthy, 33 HIV positive and 30 HIV–HBV co-infected subjects. Blood CD4+ cell count was recorded and serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total ADA, and ADA1 and ADA2 isoenzyme activities were determined.

Results

Serum ALT, AST, total ADA and ADA2 isoenzyme activities were significantly higher in HIV positive and HIV–HBV co-infected groups compare to the control (p < 0.05), whereas serum ALP showed no differences between groups. CD4+ cell counts markedly decreased in all patients and showed a significant inverse correlation with ADA activities (R² = 0.589, p < 0.001).

Conclusions

Serum ADA was significantly increased in HIV and HIV–HBV co-infections. Therefore, because of its low cost and simplicity to perform, ADA activity might be considered as a useful diagnostic tool among the other markers in these diseases.     

Significance of cerebrospinal fluid adenosine deaminase isoenzymes in tuberculous (TB) meningitis

Adenosine deaminase (ADA) exists as two isoenzymes, ADA(1) and ADA(2). It appears that the ADA(2) isoenzyme originates mainly from monocytes and macrophages. In tuberculous pleural effusions most of the ADA activity consists of ADA(2). The aim of this prospective study was to analyse ADA isoenzymes in the CSF of patients with meningitis to investigate whether the expected rise of the ADA(2) isoenzyme would occur in tuberculous meningitis. ADA isoenzyme analysis was performed on the CSF of 15 patients with tuberculous and 11 patients with bacterial meningitis by an automated kinetic enzyme coupled assay in the presence and absence of a specific ADA inhibitor. The ratio of ADA(2)/ADA(Total) was > 0.8 in 14/15 patients with tuberculous meningitis. In bacterial meningitis the ratio was > or =0.8 in 10/11 patients. The ADA(2) isoenzyme is the major contributor to increased ADA activity in the CSF of patients with tuberculous meningitis, probably reflecting the monocyte-macrophage origin of the ADA.     

Serum adenosine deaminase activity in women with hyperemesis gravidarum

BACKGROUND: Serum adenosine deaminase (ADA) activity increases in diseases where cellular immunity is stimulated. Since hyperemesis gravidarum is characterized by enhanced cell-mediated immunity, serum ADA activity may be altered. The present study evaluated the relation between serum ADA activity and changes in cell-mediated immunity as causes of changes in ADA activity in hyperemesis gravidarum. METHODS: Serum activities of total ADA and its isoenzymes, ADA1 and ADA2, were measured in women with hyperemesis gravidarum (n = 24) and normal pregnancies (n = 24). Peripheral blood lymphocyte and monocyte counts were also measured. RESULTS: In hyperemesis gravidarum, serum total ADA and ADA2 activities averaged 16.8 +/- 0.5 and 13.3 +/- 0.7 U/l, respectively, which were significantly higher than those in normal pregnancies (10.2 +/- 0.5 and 7.8 +/- 0.5 U/l, respectively) (p < 0.05). The mean values for ADA1 activity in women with hyperemesis gravidarum and normal pregnancies were similar. The increase in total ADA activity was accompanied by the increase in lymphocyte and monocyte counts. CONCLUSIONS: These results suggest that increased serum total ADA activity reflects increases in ADA2 activity, which may be at least in part attributed to enhanced cell-mediated immunity in hyperemesis gravidarum.     

Serum adenosine deaminase: isoenzymes and diagnostic application.

Human adenosine deaminase (ADA; EC 3.5.4.4) consists of three isoenzymes: ADA1, ADA1+CP, and ADA2. We developed an electrophoretic technique to distinguish between these three isoenzymes. The isoenzyme pattern was studied in tissue and cell homogenates, as well as in serum from normal subjects and from patients with increased serum ADA who had either hepatitis, infectious mononucleosis, tuberculosis, pneumonia, rheumatoid arthritis, or acute lymphoblastic leukemia (ALL). The highest ADA activity was found in lymphocytes and monocytes. ADA2 could be detected only in monocytes (18% of total ADA activity). It was also the predominant isoenzyme in the sera of controls and all disease groups, except for ALL--the only condition evaluated that is not of an inflammatory nature. We conclude that serum ADA reflects monocyte/macrophage activity or turnover in most diseases studied. The exception is ALL, where serum ADA most probably originates from lymphocyte precursors.     

Adenosine deaminase activity in the serum and malignant tumors of breast cancer: the assessment of isoenzyme ADA1 and ADA2 activities

OBJECTIVE: The potential relationship between adenosine deaminase activity and cancer progression was examined by investigating the activity of total ADA and its isoenzymes in serum and simultaneously in the cancerous tissue of each patient with breast cancer. METHODS: Total ADA and its isoenzymes were measured using the Giusti method. ADA2 activity was measured in the presence of a specific ADA1 inhibitor, EHNA. RESULTS: Our results indicated that ADA2 and total ADA activities were higher in serum and malignant tissues than those of corresponding controls (P < 0.05). Tumor ADA2 and total ADA activities were significantly (P < 0.05) correlated with lymph node involvement, histological grade and tumor size, whereas their levels in serum were significantly (P < 0.05) correlated with menopausal status and patient age. CONCLUSION: Although serum and tumor total ADA activity and its ADA2 isoenzyme were both found to be increased, distinct correlation patterns were observed with some of the prognostic factors. It can be speculated that increased ADA and isoenzyme activities in serum originated from sources other than the breast tumors.     

Pharmacokinetics and bioavailability of fluconazole in two groups of males with human immunodeficiency virus (HIV) infection compared with those in a group of males without HIV infection.

Fluconazole pharmacokinetics, including absolute bioavailability, were determined for one group of controls (n = 10) and two groups of people with human immunodeficiency virus (HIV) infection (those with CD4+ T-cell counts of less than [n = 4] or greater than [n = 9] 200 cells per mm3). Twenty subjects received four doses of fluconazole; three doses were oral (50, 100, and 400 mg), and one dose was intravenous (either 50, 100, or 400 mg). The other three subjects received one or two doses. The groups were comparable in terms of the weight, body mass index, and estimated creatinine clearance of the subjects, but the people with HIV infection were older. Pharmacokinetic parameters indicated linearity in all subjects; the area under the plasma concentration-time curve and the maximum concentration increased in proportion to the dose. The fraction of an oral dose of fluconazole absorbed approximated unity in all three groups of subjects. The mean (+/- standard deviation) plasma clearance of fluconazole was lowest in the group of subjects with low CD4+ T-cell counts; the value for this group was 0.74 +/- 0.19 liter/h, compared with 0.97 +/- 0.19 liter/h in the group with HIV infection and CD4+ T-cell counts of greater than 200 cells/mm3 and 1.18 +/- 0.23 liter/h in the group of control subjects (P < 0.05). The volume of distribution was lower in those with HIV infection (P = 0.04, corrected for weight). The half-life was longest in people with HIV infection and low CD4+ T-cell counts (P = 0.01). This study has shown that some differences do exist between the pharmacokinetics of fluconazole in people with HIV infection and those in noninfected controls.     

Immunological abnormalities in human immunodeficiency virus (HIV)-infected asymptomatic homosexual men. HIV affects the immune system before CD4+ T helper cell depletion occurs.

To investigate the effect of persistent HIV infection on the immune system, we studied leukocyte functions in 14 asymptomatic homosexual men (CDC group II/III) who were at least two years seropositive, but who still had normal numbers of circulating CD4+ T cells. Compared with age-matched heterosexual men and HIV-negative homosexual men, the CD4+ and CD8+ T cells from seropositive men showed decreased proliferation to anti-CD3 monoclonal antibody and decreased CD4+ T-helper activity on PWM-driven differentiation of normal donor B cells. Monocytes of HIV-infected homosexual men showed decreased accessory function on normal T cell proliferation induced by CD3 monoclonal antibody. The most striking defect in leukocyte functional activities was observed in the B cells of HIV-infected men. B cells of 13 out of 14 seropositive men failed to produce Ig in response to PWM in the presence of adequate allogeneic T-helper activity. These findings suggest that HIV induces severe immunological abnormalities in T cells, B cells, and antigen-presenting cells early in infection before CD4+ T cell numbers start to decline. Impaired immunological function in subclinically HIV-infected patients may have clinical implications for vaccination strategies, in particular the use of live vaccines in groups with a high prevalence of HIV seropositivity.     

Sex differences in adenosine deaminase activity of stroke patients.

BACKGROUND: Adenosine deaminase (ADA) catalyzes the irreversible hydrolytic deamination of adenosine to inosine. The purpose of this study was to determine the plasma activities of total adenosine deaminase (ADA T), and its isoenzymes, ADA1 and ADA2, and ADA1/ADA2 ratio of male and female ischemic stroke patients. METHODS: We determined activities of plasma ADA T, ADA1, ADA2 and ADA1/ADA2 ratio in 30 patients (15 men and 15 women) with acute ischemic stroke within 12 h of the onset of the attack, as well as in 30 control subjects (15 men and 15 women) of comparable age. RESULTS: There were significant differences between the ADA1 activity and ADA1/ADA2 ratio in male and female stroke patients (p<0.05). Compared with male stroke subjects, females had higher ADA1 activity and ADA1/ADA2 ratios. There were no significant differences between activities of ADA T and ADA2 in men and women of the stroke and control groups. In addition, the Canadian Neurological Scale in men was significantly higher than that of women in the stroke group (p<0.05). CONCLUSIONS: Our results suggest that the primary mechanism in men with ischemic stroke might involve the reduction of ADA1 activity. The reduction is probably an adaptation mechanism for induced increase in adenosine availability and protection of brain to ischemic injury.     

Quantitative gel-electrophoretic determination of serum amylase isoenzyme distributions.

I report a direct, sensitive, quantitative method for determining serum amylase isoenzyme activity with commercially available reagents. Day-to-day reproducibility (CV) was 3--4% for the isoenzymes in normal serum; within-run precision was 8, 3, and 2% for low, normal and high isoenzyme activities. Amylase isoenzymes, separated into the pancreatic and salivary types by electrophoresis in polyacrylamide gel, are then quantified by directly incubating the gels in soluble-starch solution, staining with iodine, and densitometry. The proportion of pancreatic isoenzyme (47 normal sera) was 43 +/- 8% (mean +/- SD). Isoenzyme activities as low as 2% of normal can be measured accurately in 10 micro L of serum. The reproducibility, precision, and sensitivity indicate that the method is applicable to differential diagnosis of hyperamylasemia or hypoamylasemia, and is suited for monitoring the subtle changes in serum amylase isoenzyme distribution that may accompany disease progression or therapy.     

Thermodynamic study of beta-N-acetylhexosaminidase enzyme heterogeneity in human seminal plasma

BACKGROUND: It has been suggested that the activity of beta-N-acetylhexosaminidase (Hex) in seminal plasma may be used as a biochemical marker of azoospermia. The purpose of our study was to evaluate this hypothesis using a thermodynamic procedure developed to determine total Hex activity and that of its isoenzymes in this biological fluid. METHODS: Using the substrate 3,3'-dichlorophenolsulphoftaleinil N-acetyl-beta-D-glucosaminide, a highly significant difference (p<0.001) is found between the activation energy of Hex A (41.5 kJ/mol) and of Hex B (72.3 kJ/mol), making it possible to determine the activity of these isoenzymes from the apparent activation energy of the total Hex in seminal plasma. RESULTS: A significant difference between the normozoospermic and azoospermic groups was only found for Hex A isoenzyme activity (p<0.05), although with considerable overlapping between the values of both groups. Significant partial correlations were found for the total Hex, Hex A and Hex B activities with the immobile spermatozoa count (p<0.01) and for total Hex and Hex B with the dead spermatozoa count (p<0.05). In turn, Hex A had a significant partial correlation with the live spermatozoa count (p<0.05); however, Hex activity in seminal plasma of acromosomal origin appears to be of little importance in quantitative terms. CONCLUSIONS: It was not possible to confirm that total Hex activity in seminal plasma, or even of its isoenzymes Hex A and Hex B, is a suitable biochemical marker of azoospermia (efficiency< or =67%). The thermodynamic procedure described may be a useful alternative for the study of the Hex enzyme heterogeneity in spermatozoa.     

Evaluation of cerebrospinal fluid adenosine deaminase activity in HIV-seropositive subjects and its association with lactate dehydrogenase and protein levels

The purpose of this study was to investigate the role of ADA as additional marker of HIV infection as well as its association with other biochemical markers. This study included 55 patients, 26 being diagnosed as HIV positive and 29 patients diagnosed as HIV negative. Glucose, total protein, lactate dehydrogenase, and adenosine deaminase (ADA) activity were measured on cerebrospinal fluid (CSF). ADA activity on CSF was statistically different in HIV-seropositive subjects compared with HIV-negative subjects. The sensitivity and specificity of ADA activity on CSF was 50 and 82.76%, respectively. ADA activity was positively correlated with lactate dehydrogenase and protein in patients with HIV positive and it was negatively correlated with glucose levels. ADA determination in CSF could add information about inflammatory processes in patients with HIV infection.     

Immunological abnormalities in asymptomatic homosexual men: correlation with antibody to HTLV-III and sequential changes over two years

A prospective study on 100 homosexual male volunteers was designed to examine immunological function in relation to sexual activity and infection with the human T cell lymphotropic virus Type III (HTLV-III). Complete data were available for 71 men. In a comparison with 100 age-matched heterosexual men, the study group of 100 men had a significantly higher mean serum IgG level (12.1 +/- SD 2.7 g/l vs. 10.9 +/- 2.4 g/l, p less than 0.01) and a significantly lower mean number of CD4 (T4) cells (845 +/- 310 X 10(-6)/l vs. 1128 +/- 375; p less than 0.01). For the study group, seropositivity for anti-HTLV-III was present initially in 22 per cent and was associated with a higher mean level of serum IgG and lower mean number of CD4 cells. Among seropositive homosexual men a low CD4/8 ratio was attributable to low numbers of CD4 cells in those without lymphadenopathy and to high numbers of CD8 cells in those with lymphadenopathy. For the seronegative homosexual men, a low CD4/8 ratio as a result of an increased CD8 cell count was present in 12 of 60, and was associated with numerous sexual partners and semen culture positive for cytomegalovirus. In two seropositive subjects a low CD4/8 ratio due to a decrease in the CD4 cell count was predictive of the development of AIDS by some two years. For the 71 men with complete data over two years, indices of cell-mediated immunity, including mean counts of CD4 cells, the CD4/8 ratio, and score for recall of cutaneous delayed type hypersensitivity increased during the first year but not during the second year in both seropositive and seronegative subjects. These increases occurred in association with changes in sexual practices and activity, but could not be attributed to any one particular factor.     

Functional aerobic impairment in adolescents seropositive for HIV: a quasiexperimental analysis

OBJECTIVE: To determine the degree to which cardiorespiratory insufficiency limits physical performance in adolescents seropositive for human immunodeficiency virus (HIV). DESIGN: Quasiexperimental, case series design. SETTING: Rehabilitation physiology laboratory. PARTICIPANTS: Seventeen adolescents (12 women, 5 men; age, 18 +/- 2 yr; weight, 74.7 +/- 19.3 kg; height, 170 +/- 9 cm) with HIV infection (viral load, 22,043 +/- 55,869 copies/mL; CD4 count, 499 +/- 210/mL) who were free of comorbid conditions limiting treadmill performance. MAIN OUTCOME MEASURES: Spirometric measurements of oxygen uptake and anaerobic threshold obtained from a peak exercise treadmill test using the modified Bruce protocol. RESULTS: Measured peak oxygen consumption (VO2) was 42% +/- 19% lower than expected (p < .025), suggesting a significant functional aerobic impairment (FAI) or peak VO2 less than 73% of expected values. Peak VO2 was only slightly higher (p < .05) than the oxygen uptake requirements for the most intense activities of daily living (ADL). Anaerobic threshold was only slightly higher (p < .05) than minimum ADL intensities. CONCLUSIONS: Cardiorespiratory insufficiency and FAI limited the ability to perform even low levels of physical activity in these adolescents with mild HIV seropositivity. Disability identified by quantification of FAI may affect implementation of the American with Disabilities Act and public health policy.     

The effect of Escherichia coli-derived lipopolysaccharides on plasma levels of malondialdehyde and 3-nitrotyrosine.

The aim of this study was to determine the effect of Escherichia coli (E. coli)-derived lipopolysaccharide on rat plasma low density lipoprotein (LDL), malondialdehyde and 3-nitrotyrosine levels (an indicator of protein nitration). Six hours after intraperitoneal administration of E.coli, plasma LDL was measured electrophoretically and malondialdehyde level was measured by spectrophotometric method. Plasma malondialdehyde was significantly (p<0.001) elevated in E. coli-injected rats (4.97 +/- 1.33; n=10) in comparison to control animals (1.83 +/- 0.5; n=10). In addition, plasma 3-nitrotyrosine level, determined by reverse-phase HPLC, was also increased in the infected group (2.84 +/- 1.17 to 0.22 +/- 0.13; n=10). This increase was statistically significant (p<0.001). An increased level of oxidation of lipids and 3-nitrotyrosine was observed as a result of free radical-mediated damage in plasma. In conclusion, asymptomatic infections may increase the risk of atherosclerosis by inducing free radical formation and a consequent increase in the oxidation of LDL.     

The safety profile and antiviral activity of the combination of stavudine, didanosine, and nelfinavir in patients with HIV infection

We assessed the safety profile, tolerability, and antiviral effect of 12 weeks of triple combination therapy with stavudine (d4T), didanosine (ddI), and nelfinavir in patients who had not previously received therapy with d4T, ddI, or a protease inhibitor. We also assessed the effect of the buffered tablet formulation of ddI on the pharmacokinetics of nelfinavir. The study had a single-arm, open-label design and enrolled patients aged > or =18 years who had HIV infection and > or =10,000 plasma HIV RNA copies/mL. Patients received the full recommended doses of oral d4T, ddI, and nelfinavir. Efficacy was assessed in terms of change from baseline in plasma HIV RNA and CD4+ cell counts, as well as in terms of the proportion of patients achieving HIV RNA levels <400 copies/mL. The first 10 patients enrolled in the study were included in a substudy to determine the effects of the buffered tablet formulation of ddI on the pharmacokinetic profile of nelfinavir. A dose of ddI was given 1 hour before nelfinavir, after which the maximum plasma concentration (Cmax), time to Cmax (Tmax), and area under the concentration-time curve (AUC) of nelfinavir were determined. A total of 22 patients entered the trial, of whom 1 (5%) had AIDS, 12 (55%) had symptomatic HIV infection, and 9 (41%) were asymptomatic. The median baseline CD4+ cell count was 315 cells/microL (range, 70-709 cells/microL), and the median plasma viral load was 4.8 log10 copies/mL (range, 4.0-5.6 log10 copies/mL). ddI had no clinically significant effects on the plasma pharmacokinetics of nelfinavir. At the end of 12 weeks of treatment, the mean (+/- SE) decrease in plasma viral load was 1.36+/-0.24 log10 copies/mL, and 8 of 16 patients (50%) achieved plasma HIV RNA levels <400 copies/mL; the mean (+/- SE) increase in CD4+ cell count was 111.4+/-31.7 cells/microL. Patients who were judged to be compliant with antiretroviral therapy (ie, who missed <7 days of all 3 study drugs during 12 weeks of treatment) experienced mean decreases in viral load exceeding 2.0 log10 copies/mL, and 6 of 7 patients achieved HIV RNA levels <400 copies/mL after 12 weeks of therapy. Although 95% of patients reported an adverse event of grade 1 or higher, only 1 patient experienced a grade 3 or 4 adverse event (maculopapular rash) related to nelfinavir. As reflected in the Cmax, Tmax, and AUC, administration of ddI 1 hour before nelfinavir did not influence the pharmacokinetic profile of the protease inhibitor. Triple drug therapy with d4T, ddI, and nelfinavir was well tolerated and associated with few clinically significant toxicities. This treatment resulted in substantial reductions in viral load and improvements in CD4+ cell count over 12 weeks.     

The role of adenosine in glycine-induced glomerular hyperfiltration in rats.

Ingestion of a high-protein diet or infusion of amino acids induces glomerular hyperfiltration and hyperemia. We have investigated the role of endogenous adenosine in glycine-induced hyperfiltration. Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were measured in conscious chronically instrumented rats. Glycine (3.7 mg/min, i.v.; n = 6) significantly increased GFR and ERPF from 0.92 +/- 0.07 to 1.13 +/- 0.08 and 3.28 +/- 0.24 to 3.69 +/- 0.19 ml/min.100 g, respectively. In the presence of adenosine deaminase (ADA, 2 U/kg.min, n = 6), glycine-induced glomerular hyperfiltration and hyperemia were blunted. The small changes in GFR (from 0.86 +/- 0.06 to 0.90 +/- 0.10 ml/min.100 g) and ERPF (from 3.60 +/- 0.57 to 3.83 +/- 0.53 ml/min x 100 g) were not statistically significant. Erythro-9-(2-hydroxy-3-nonyl) adenosine hydrochloride (100 micrograms/kg.min, n = 6), an ADA inhibitor, reversed the effect of ADA. Injection of 8-phenyltheophylline (10 mg/kg, n = 6), an adenosine A1 receptor antagonist that alone did not affect GFR, abolished the glycine-induced glomerular hyperfiltration (GFR from 1.02 +/- 0.08 to 0.93 +/- 0.08 ml/min.100 g, P > .05). 8-phenyltheophylline, which itself decreased ERPF, also significantly decreased the ERPF response to glycine (3.47 +/- 0.26 to 2.78 +/- 0.14 ml/min x 100 g). Thus, endogenous adenosine, acting at adenosine A1 receptors, plays an important role in the glomerular hyperfiltration and hyperemia induced by glycine.     

A radioimmunosorbent technique for the assay of B- and C-types of carbonic anhydrase in human tissues.

A radioimmunosorbent technique for the assay of the human carbonic anhydrase isoenzymes HCA B and HCA C in tissue fluids was developed. The sensitivity of the method was 0.2 ng/ml and the precision was 5% in duplicate determinations for both enzymes. The presence in a tissue of up to 20 times higher concentrations of one isoenzyme will not interfere with the assay of the other. Haemolysates contained (mean +/- SE, n = 11) 12.1 +/- 0.52 and 1.5 +/- 0.06 mg enzyme/g Hb, and serum 0.63 +/- 0.12 and 0.2 +/- 0.02 microgram/ml of HCA B and HCA C, respectively. Pilot experiments indicated that the isoenzymes can be determined also in tissues, i.e. urine, saliva and cerebrospinal fluid, where catalytic methods previously have indicated absence of or only weak carbonic anhydrase activity. N-terminals of both enzymes were not antigenic.     

Conversion of MM creatine kinase isoforms in human plasma by carboxypeptidase N

This study was undertaken to identify the carboxypeptidase(s) (CPase) in plasma mediating sequential conversion of the tissue isoform of the MM isoenzyme of creatine kinase (MM3 CK) to MM2 and MM1 isoforms and to elucidate relationships between CPase activity measured in plasma and observed rates of isoform conversion in vitro. Purified MM3 was incubated at 37 degrees C in plasma from normal subjects and patients with acute myocardial infarction. Isoforms were quantified by chromatofocusing. Preincubation with antiserum to CPase N prevented conversion of added MM3 to MM2 and MM1. Isoform conversion rates in the absence of antibody were proportional to plasma CPase N activity assayed spectrophotometrically by hydrolysis of furylacryloyl-L-alanyl-L-lysine substrate (r = 0.89, n = 8). Plasma CPase N activity varied by nearly 300% among individuals, but average activity was similar in samples from normal subjects (267 +/- 45 [SD] U/L, n = 18), those from outpatients with angina (289 +/- 43 U/L, n = 9), and those obtained at hospital admission from patients with acute infarction (Q wave: 279 +/- 70 U/L, n = 16; non-Q wave: 272 +/- 61 U/L, n = 14) or unstable angina (280 +/- 71 U/L, n = 11). In patients with Q wave infarction, CPase N activity increased by 43% +/- 25% between 48 hours and 72 hours (P less than 0.005 compared with admission) with a concomitant change in the rate of conversion of isoforms. Thus, the rate of conversion of isoforms in individual subjects can be estimated by assay of CPase N activity in plasma.(ABSTRACT TRUNCATED AT 250 WORDS)     

Adenosine and hemodialysis in humans.

BACKGROUND: Infections and hypotension are serious complications that develop during hemodialysis (HD) treatment. Adenosine (ADO), a strong hypotensive and immunosuppressive agent, may participate in these two HD complications, because high concentrations of ADO metabolites are found in dialyzed human plasma. ADO, which is released by endothelial cells, is quickly transformed into inosine (INO) by plasmatic ADO deaminase (ADA) and mononuclear cell ADO deaminase (MCADA). In plasma, the degradation of ADO into INO and its uptake by red blood cells (RBC) are both very rapid, resulting in the short half-life of ADO in blood. METHODS: Using liquid chromatography, we evaluated ADO and INO plasma concentrations before and after HD session. RESULTS: Before the HD session, ADO and INO plasma concentrations were higher in hemodialyzed patients than in controls and in peritoneally dialyzed patients. At the end of the HD session, ADO plasma concentration was increased. ADO plasma concentration for the undialyzed patients was in the same range as that of the controls. Before HD, ADA activity was higher in hemodialyzed patients (559 +/- 349 IU) than in controls (219 +/- 48 IU), and the activity rose during the session (665 +/- 135 IU). ADA activity in the undialyzed patients (222 +/- 80 IU) was in the same range as that of the controls (219 +/- 48 IU). Before the HD session, the MCADA activity (247 +/- 144 IU) was lower than in controls (624 +/- 99 IU). HD did not modify ADO RBC uptake. ADO inhibited mononuclear cell proliferation and interferon-gamma production in humans. Finally, as much as 50 microM INO does not inhibit ADO uptake by RBC and does not modify ADA and MCADA activities. CONCLUSIONS: These data indicate that chronic HD inhibited MCADA activity and increased ADO plasma concentration. Both high ADO plasma concentration and low MCADA activity may be involved in dialysis-induced immune system failure and thereby favor infectious diseases.     

CD4^＋T淋巴细胞在无症状人类免疫缺陷病毒感染者早期接受扶正排毒片治疗后的变化

背景:无症状人类免疫缺陷病毒(Human Immunodeficiency Virus,HIV)感染者不适宜抗病毒治疗,中医药早期干预对无症状带毒期HIV感染者会产生一定的治疗意义。目的:观察扶正排毒片早期干预HIV感染者CD4 T淋巴细胞水平及血浆病毒载体量变化。设计:观察对比试验。单位:河南中医学院艾滋病研究所。对象:于2005-11随机抽取HIV感染者较多的某乡69例有偿献血感染的无症状HIV感染者,男39例,女30例,年龄30～61岁,平均(43±8)岁,平均病程(13±3)年。纳入标准:①符合中华人民共和国国家艾滋病无症状HIV感染诊断标准(2005年修订版)。②年龄18～65岁。③200/mm3≤CD4 T淋巴细胞计数<600/mm3。④未使用抗病毒药物及其它药物治疗者。⑤签署知情同意书。排除标准:①严重肝肾功能不全,或合并心脑血管、肺和造血系统等严重原发性疾病,精神病患者。②原发性免疫缺陷患者,激素、化疗等引起的继发性免疫缺陷患者,血液病患者。③妊娠或哺乳期妇女。随机抽取的某个县的25例无症状HIV感染者为空白组,纳入标准、排除标准与治疗组相同。方法:治疗方案:治疗组感染者给予扶正排毒片(黄芪、生白术、防风、白花蛇舌草、甘草等11味,河南省中医研究院奥林特制药厂生产,产品批号:050601),每次5片,相当于生药6.6g,3次/d,温开水冲服。3个月为1疗程,共观察4个疗程。空白组不服药。主要观察指标:①检测治疗组感染者治疗前和每疗程结束后CD4 、CD8 及CD4 /CD8 值,空白组只检测6个月前后CD4 T淋巴细胞计数。②采用RT-PCR检测方法进行治疗组患者病毒载量检测。结果:治疗组感染者69例及空白组感染者25例均进入结果分析。①空白组感染者6个月前后CD4 较治疗前明显下降,差异有统计学意义(P<0.05)。②随机抽取23例治疗组感染者进行病毒载量检测,与疗前对比,疗后6个月有9例血浆病毒载量下降较多,疗后1年12例血浆病毒载量下降较多。疗前和疗后6个月及1年对比有效率均达91.30%。结论:扶正排毒片早期干预HIV感染者可提高其免疫功能,对部分病例具有降低病毒载量的作用。