Cytotoxic T lymphocytes and viral evolution in primary HIV-1 infection

Efforts to develop immune-based therapies for HIV infection have been impeded by incomplete definition of the immunological correlates of protection. Despite many precedents demonstrating that CD8(+) cytotoxic T lymphocytes are key mediators of protective anti-viral immunity in non-human animal models, direct evidence that these effector cells control viral replication in HIV-1 infection has remained elusive. The first part of this paper describes a detailed immunological and genetic study founded on evolutionary considerations. Following infection with HIV-1, virus variants which escaped recognition by autologous cytotoxic T lymphocytes were shown to possess a selection advantage within the host environment. Cytotoxic T lymphocytes therefore exert anti-viral pressure in vivo. This observation provides compelling evidence that cytotoxic T lymphocytes comprise a significant element of anti-retroviral immunity. Subsequently, the quantification of peripheral cytotoxic T lymphocyte frequencies utilizing peptide-(human leucocyte antigen class I) tetrameric complexes is described. Five patients with qualitatively similar immunodominant cytotoxic T lymphocyte responses during symptomatic primary HIV-1 infection were studied longitudinally. Expansions of virus-specific CD8(+) lymphocytes comprising up to 2% of the total CD8(+) T cell population were observed in the acute phase of infection. Antigenic load was identified as an important determinant of circulating HIV-1-specific CD8(+) lymphocyte levels; however, significant numbers of such cells were also found to persist following prolonged therapeutic suppression of plasma viraemia. In addition, an analysis of antigenic sequence variation with time in this case series suggests that the early administration of combination anti-retroviral therapy may limit HIV-1 mutational escape from host cytolytic specificities. The implications of these preliminary data are discussed. The data presented suggest that vaccination protocols should aim to elicit vigorous cytotoxic T lymphocyte responses to HIV-1. Attempts to stimulate polyvalent responses to mutationally intolerant epitopes are likely to be most effective. Optimal management of HIV-1 infection requires an understanding of dynamic host-virus interactions, and may involve strategies designed to enhance cytotoxic T lymphocyte activity following periods of anti-retroviral drug therapy.     

Effects of organic and inorganic selenium supplementation on selenoenzyme activity in blood lymphocytes, granulocytes, platelets and erythrocytes

The blood selenium (Se) concentration in the U.K. population has declined by approx. 50% between 1974 and 1991, reflecting a large decrease in dietary Se supply, with intakes only half the reference nutrient intake of 1 microg/kg body weight. Tissue levels of Se are readily influenced by dietary intake. Therefore selenoprotein activity may be sub-optimal due to low Se status, and thus compromise normal cell function. To examine the effects of changing Se intake on selenoproteins, we have determined the relative effectiveness of organic selenomethionine and inorganic sodium selenite (50 microg of Se daily for 28 days) in modulating glutathione peroxidase activities in blood cells from 45 healthy men and women, from a U.K. population. Transient and acute changes in lymphocyte, granulocyte and platelet phospholipid-hydroperoxide glutathione peroxidase (GPx4) activity occurred by day 7 or 14 of sodium selenite treatment and by day 7 in lymphocytes from selenomethionine-treated subjects compared with controls taking a placebo. In contrast, GPx4 activity in granulocytes and platelets in the selenomethionine group increased gradually over the 28 days. Cytosolic glutathione peroxidase (GPx1) activity in these blood cells from both treatment groups increased gradually over the 28 days. For each cellular selenoenzyme activity a significant inter-individual difference (P<0.001) in the extent of the response to Se supplementation was observed, but this was not related to blood Se concentrations either before or after treatments. Significant inverse correlations were evident between baseline enzyme activities and percentage change in activity after 28 days of supplementation [e.g. lymphocyte GPx4, r=-0.695 (P<0.001)], indicating that pre-treatment activity may be sub-optimal as a result of poor Se status. The different and contrasting effects that Se supplementation had on blood selenoenzyme activities may be indicative of a difference in metabolic need for Se regulated at the level of Se-dependent cell function.     

HIV-1 proprotein processing as a target for gene therapy

The central role of endoconvertases and HIV-1 protease (HIV-1 PR) in the processing of HIV proproteins makes the design of specific inhibitors important in anti-HIV gene therapy. Accordingly, we tested native alpha(1) antitrypsin (alpha(1)AT) delivered by a recombinant simian virus-40-based vector, SV(AT), as an inhibitor of HIV-1 proprotein maturation. Cell lines and primary human lymphocytes were transduced with SV(AT) without selection and detectable toxicity. Expression of alpha(1)AT was confirmed by Northern blotting, immunoprecipitation and immunostaining. SV(AT)-transduced cells showed no evidence of HIV-1-related cytopathic effects when challenged with high doses of HIV-1(NL4-3). As measured by HIV-1 p24 assay, SV(AT)-transduced cells were protected from HIV-1(NL4-3) at challenge dose of 40 000 TCID(50) (MOI = 0.04). In addition, peripheral blood lymphocytes treated with SV(AT) were protected from HIV doses challenge up to 40 000 TCID(50) (MOI = 0.04). By Western blot analyses, the delivered alpha(1)AT inhibited cellular processing of gp160 to gp120 and decreased HIV-1 virion gp120. SV(AT) inhibited processing of p55(Gag) as well. Furthermore, high levels of uncleaved p55(Gag) protein were detected in HIV virus particles recovered from SV(AT)-transduced cells lines and primary lymphocytes. Thus, delivering alpha(1)AT using SV(AT) to human lymphocytes strongly inhibits replication of HIV-1, most likely by inhibiting the activities both of the cellular serine proteases involved in processing gp160 and of the aspartyl protease, HIV-1 PR, which cleaves p55(Gag). alpha(1)AT delivered by SV(AT) may represent a novel and effective strategy for gene therapy to interfere with HIV replication, by blocking a stage in the virus replicative cycle that has until now been inaccessible to gene therapeutic intervention.     

Prolonged, low dose alpha-tocopherol therapy counteracts intercellular cell adhesion molecule-1 activation

BACKGROUND: Up-regulation of ICAM-1 at the vascular endothelial level is one of the most important promoters in the slow progression of a healthy vessel to an atherosclerotic one. The current study aimed to evaluate whether low dose of the antioxidant alpha-tocopherol affects the circulating soluble (s) ICAM-1 in healthy subjects. METHODS: Either alpha-tocopherol E (50 I.U./day) or placebo was randomly, double-blindly given to 39 healthy male volunteers (mean age 41.6+/-5.9 years) over a period of 20 weeks. RESULTS: At the baseline, sICAM-1 levels were inversely correlated with alpha-tocopherol concentrations (r=-0.525, p<0.0001). Twenty weeks of alpha-tocopherol supplementation (n=20 subjects) significantly decreased the circulating sICAM-1 levels (from 149.2+/-18.4 to 131.5+/-17.2 microg l(-1), p<0.004) while it increased the alpha-tocopherol concentrations (from 25.8+/-5.0 to 31.2+/-5.7 micromol l(-1), p<0.003). No significant changes in plasma sICAM-1 and alpha-tocopherol levels were observed in placebo-treated subjects (n=19). In actively treated subjects, changes in circulating sICAM-1 were inversely correlated with changes in alpha-tocopherol concentrations (r=-0.597, p=0.005). CONCLUSIONS: Plasma sICAM-1 concentrations are stable in healthy subjects over a period of 20 weeks while they significantly decreased with low dose of alpha-tocopherol. Thus, antioxidant vitamins are likely to counteract with endothelial changes that could potentially trigger the atherogenetic process.     

The effectiveness of anti-retroviral drug therapy for HIV-1 is associated with HIV-1 proviral DNA levels and viral selection

The effect of combination anti-retroviral therapy regimens on HIV-1 proviral DNA levels in peripheral blood mononuclear cells was examined in 12 HIV-1-positive patients, using endpoint dilution polymerase chain reaction and serial cloning, and sequencing of the gag region of HIV-1. The major clone was defined as the most numerous of 10 analysed clones, and observation periods ranged from 8 months to 32 months (mean 19.7 +/- 10.2 months). In five patients (one with primary-stage HIV-1 infection) receiving three anti-retroviral drugs, HIV-1 RNA reduced to undetectable levels (i.e. < or = 100 copies/ml). HIV-1 proviral DNA and the number of major clones reduced in four of these patients. HIV-1 RNA levels reduced, but remained detectable, in five other patients. In the two remaining patients (both receiving two rather than three anti-retroviral drugs), HIV-1 RNA levels increased. These results suggest that the population of major clones may be affected when HIV-1 RNA levels reduce following combination regimens of anti-retroviral therapy.     

Adipose tissue fatty acid composition, serum lipids, and serum alpha-tocopherol in continuous ambulatory peritoneal dialysis patients living on the island of Crete.

OBJECTIVE: A pilot study to explore diet-related atherogenic patterns in continuous ambulatory peritoneal dialysis (CAPD) patients on the island of Crete. Cretans are well known for their high olive consumption and low atherosclerotic heart disease mortality, in general. DESIGN: Case-control study. SETTING: This was a hospital-based study initiated in 1991. Catchment area was the island of Crete, Greece. PARTICIPANTS: Seventeen patients admitted for treatment to the General State Hospital of Rethimnon. Controls were selected from the general population of the island and consisted of a random sample of 27 subjects from a total of 168 healthy subjects who visited the Preventive Medicine Clinic of the University Hospital for routine check-up. The control group was age- and sex-matched with patients. Both CAPD patients and controls had been residents of Crete for at least the last 10 years. MAIN OUTCOME MEASURES: Adipose tissue was aspirated, and a 12-hour fasting blood sample was collected for determination of serum lipid parameters and serum alpha-tocopherol levels; dietary data from a 3-day recall were recorded. RESULTS: No significant differences between the two groups were observed with respect to the mean macronutrient intake. The mean levels of serum triglycerides (p = 0.016) and serum alpha-tocopherol (p = 0.001) were significantly higher in CAPD patients compared to controls. Mean levels of total serum cholesterol, high density lipoprotein (HDL) cholesterol, and low density lipoprotein (LDL) cholesterol were not significantly different. In CAPD patients the mean total percentage of monounsaturated fatty acids (MUFA) was significantly higher (p = 0.006) than in controls. The mean total percentage of saturated fatty acids (p = 0.004), along with the mean percentage of omega-6 (p = 0.002), the mean value of the ratio omega-6/omega-3 (p < 0.0001), and the percentage of linoleic acid (p = 0.001) were significantly lower in CAPD patients than in the controls. Among subjects with higher levels of MUFA in the adipose tissue, the CAPD patients were twice as likely to have high serum alpha-tocopherol (p < 0.001), and 2.6 times more likely to be in high risk of high total cholesterol (TC)/HDL (p = 0.08) compared to the controls. However, CAPD patients with high levels of MUFA in the adipose tissue (above the average of 65%) were unlikely [odds ratio (OR) = 0.001, p < 0.001] to be at risk of high TC/HDL (above the average of 4.1), and maybe unlikely (OR = 0.08, but p = 0.1) to have low serum alpha-tocopherol, when compared with the CAPD patients with low levels of MUFA. CAPD patients with high TC/HDL are 0.15 times less likely (p = 0.1) to have high levels of serum alpha-tocopherol compared to those with low TC/HDL. CONCLUSION: Cretan CAPD patients demonstrate an interesting profile consisting of unexpectedly positive aspects when atherogenesis-related factors such as those of adipose tissue fatty acid composition, serum lipids, and serum antioxidant alpha-tocopherol are considered.     

Exacerbation of microcytic anemia associated with cessation of anti-retroviral therapy in an HIV-1-infected patient with beta thalassemia

We report a patient with Japanese minor β thalassemia and HIV-1 infection.The patient showed prolonged anemia, which was originally attributed to chronic parvovirus B19 infection. Twelve years later, the patient presented with exacerbation of microcytic anemia following cessation of anti-retroviral therapy; the exacerbation resolved when anti-retroviral therapy was resumed. Sequencing of the β globin gene revealed heterozygosity for a four-nucleotides deletion at codon 41/42 and minor β thalassemia was confirmed.Because HIV-1-infected patients frequently show anemia due to nutritional deficiencies, opportunistic infections, AIDS-related malignancies, drug treatment and a direct effect of HIV-1 on the bone marrow, it is likely to overlook other causes of anemia.Thalassemia should be considered in the differential diagnosis of anemia even in HIV-1 infected patients, when microcytic anemia without iron deficiency is observed.Our case suggested that active HIV infection may have worsened β thalassemia, and early introduction of anti-retroviral therapy is beneficial for the recovery of anemia.     

Molecular-based laboratory testing and monitoring for human immunodeficiency virus infections.

Applications of laboratory testing for human immunodeficiency virus type 1 (HIV-1) infection have made significant impact on clinical care of HIV-infected patients globally. As these technologies continue to evolve and new technologies emerge, unique and highly sensitive nucleic acid-based testing methods will offer more and better means for us to guide physicians in anti-retroviral treatment strategies and clinical management of HIV infected patients. In this review we discuss a variety of current molecular-based methods that are available for HIV testing including diagnosis, monitoring disease progression, and detection of drug resistance to anti-retroviral therapy. Newer approaches that could be used in future HIV testing are also introduced.     

Comparison of traditional Chinese acupuncture, minimal acupuncture at non-acupoints and conventional treatment for chronic sinusitis

OBJECTIVES: To compare traditional Chinese acupuncture, minimal acupuncture at non-acupoints and conventional treatment for chronic sinusitis. DESIGN: A three-armed single blind randomised controlled study. SETTING: In an outpatient specialist clinic, we recruited 65 patients with symptoms of sinusitis >3 months and signs of sinusitis on computed tomography (CT). INTERVENTIONS: We randomised patients to one of three study arms: (1) 2-4 weeks of medication with antibiotics, corticosteroids, 0.9% sodium chloride solution, and local decongestants (n=21), (2) 10 treatments with traditional Chinese acupuncture (n=25), or (3) 10 treatments with minimal acupuncture at non-acupoints (n=19). OUTCOME MEASURES: Change in sinus soft tissue swelling on CT, symptoms of sinusitis, and health-related quality of life (HRQoL), using the two component summary scales of the Short Form 36 and a rating scale. RESULTS: In the conventional treatment group, sinus soft tissue swelling was reduced over 4 weeks (p=0.04), and HRQoL improved over 12 weeks (p=0.01-0.05). Pairwise comparisons of changes in total symptom score between the groups showed signs of a difference between conventional medication and sham over 4 weeks (p=0.06). CONCLUSION: Sinus soft tissue swelling was reduced in the conventional treatment group over 4 weeks, and HRQoL improved over 12 weeks. Only a non-significant difference in symptom score change over 4 and 12 weeks was shown between conventional medication and traditional Chinese acupuncture.     

Oxidative stress and antioxidant status in patients undergoing prolonged exposure to hyperbaric oxygen

OBJECTIVES: To evaluate the condition of oxidative stress in patients undergoing prolonged exposure to hyperbaric oxygen (HBO) and the possible modifications of the antioxidant defense systems in the absence of antioxidant supplementation. DESIGN AND METHODS: Twelve patients exposed to 15 HBO treatments for pathological conditions related to hypoxia were included in the study. Oxidative stress indices as well as plasma and erythrocyte antioxidant levels were measured in blood samples collected both at the 1st and 15th HBO session. RESULTS: The repeated exposures to HBO led to a significant accumulation of plasmatic reactive oxygen metabolites (ROM) and malondialdehyde (MDA). After 15 HBO sessions, no relevant differences were detected for reduced glutathione (GSH), alpha-tocopherol, and retinol plasma levels; however, a significant decrease in erythrocyte superoxide dismutase (SOD) and catalase (CAT) activity was observed when compared to the 1st HBO exposure; glutathione peroxidase (GPx) activity remained almost unchanged. CONCLUSIONS: In the absence of antioxidant supplementation, the prolonged HBO treatment leads to a condition of oxidative stress that seems to affect in particular the response of the enzymatic antioxidant defense system; the possible relationship between the chemical modifications of the enzymes caused by oxygen reactive species and the consequent inactivation of the proteins is under investigation.     

Acute parvovirus B19 infection during anti-retroviral therapy

Human parvovirus B19 (B19) has been described as a causative agent of chronic anemia in human immunodeficiency virus type-1 (HIV-1)-infected patients. We report an HIV-1 infected patient who had been receiving anti-retroviral therapy who showed sudden pancytopenia. Primary B19 infection was confirmed by the detection of plasma viremia and seroconversion. Although clearance required a prolonged period of time, the patient eventually cleared the B19 viral DNA from the plasma. More than likely, highly active anti-retroviral therapy (HAART), including a protease inhibitor, played a role in clearing the virus. Received: April 7, 2000 / Accepted: March 6, 2001     

原发性肝癌中高尔基体蛋白73表达与淋巴细胞亚群的相关性

目的:探讨原发性肝癌患者高尔基体蛋白73(GP73)的表达与外周血淋巴细胞亚群的相关性及其可能的临床意义.方法:收集2018年6月～2020年7月确诊为原发性肝癌患者129例及健康对照者36例,检测外周血血清GP73的表达量并根据临界值将原发性肝癌患者划分为阳性组和阴性组;流式细胞术检测外周血淋巴细胞亚群百分数.采用S...     

Human monoclonal antibodies to HIV-1 infection

HIV-1, which causes AIDS, has infected over 50 million and killed over 20 million individuals world wide since the beginning of the epidemic two decades ago. The introduction of highly active anti-retroviral therapy(HAART) has resulted in the control of the disease progression. Furthermore, supervised treatment interruption(STI) of HAART might be expected significantly to boost immune function and slow the progression of AIDS. On the other hand, several combinations of human monoclonal antibodies(hu-mo-Ab) against HIV-1 has been identified, and demonstrates the efficacy of the triple combination of hu-mo-Ab with neutralizing activity in macaque model. Therefore, a combination of chemotherapy and immunotherapy, such as hu-mo-Ab will be prove to be the most effective route for HIV-1 therapy.     

Subpopulation blood lymphocyte composition in blood of patients with chronic hepatitis C during therapy with interferon

The number of lymphocytes expressing CD3, CD4, CD8, CD16, CD25, and CD19 antigens was studied in patients with chronic hepatitis C before and after 12-week therapy with interferon-alpha (IFN-alpha). The percentage of cells expressing CD4+, CD16+, and CD25+ antigens decreased significantly in untreated patients, while the percentage of CD8+ and CD19+ lymphocytes was increased (p < 0.05) vs. the control. After 3-month therapy with IFN-alpha the counts of CD4+ and CD25+ increased significantly in patients with chronic hepatitis C (p < 0.05 and p < 0.01, respectively) in comparison with the initial values. The treatment led to a significant (p < 0.05) decrease in the number of CD8+ cells in the blood. The number of cells expressing CD19+ decreased, but remained high in comparison with the control. These results indicate that cellular immune response is inadequate in patients with chronic hepatitis C. Time course of subpopulation composition of T lymphocytes during effective treatment with IFN-alpha indicates an important role of T-cellular component of immunity in the antiviral defense mechanisms in chronic HCV infection.     

Flow cytometric characterization of cell populations in bronchoalveolar lavage and bronchial brushings from patients with chronic obstructive pulmonary disease

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a serious, chronic inflammatory disease of the airway associated with cigarette smoking. Leucocytes are involved in the inflammatory process in the airways in COPD. There is a need for accurate characterization of cellular populations in bronchoalveolar lavage (BAL) due to variation in the predominant cell types reported, which were investigated mostly with manual counting techniques. METHODS: Bronchial brushings and BAL were obtained from human subjects undergoing fiber optic bronchoscopy. Flow cytometry was applied to identify various cell types. Quenching of autofluorescence of BAL-derived alveolar macrophages was achieved with eta-octyl beta-D-galactopyranoside and crystal violet. Comparisons of cell counts obtained with flow cytometric and manual counting methods were performed. RESULTS: Correlation analysis showed that manual cell counting methods overestimated the percentage of macrophages when compared with flow cytometric methods (R2 = 0.54). There was also a small tendency by manual counting to underestimate the percentage of lymphocytes and neutrophils. Using flow cytometry, the percentage and absolute numbers of alveolar macrophages and lymphocytes in BAL were not significantly different between patients with COPD and control subjects. The percentage and absolute numbers of neutrophils were higher in BAL from patients with moderate to severe COPD. CONCLUSIONS: This novel flow cytometric assay for identification of various cell types from heterogenous samples of BAL and bronchial brushing will allow further investigation of cell characteristics, such as cytokine production and receptor expression, and an accurate evaluation of apoptosis for different cell types and provide a rationale for urgently required effective treatments for COPD.     

The side-effects of cardiopulmonary bypass on the lungs: changes in bronchoalveolar lavage fluids

Although technical refinements have improved the safety of cardiac operations, postoperative dysfunction of lung and other organs occurs frequently after cardiopulmonary bypass (CPB). The aim of the present study was to search the aetiopathogenesis of pulmonary complications due to CBP. Ten patients with stable coronary artery disease, undergoing coronary artery bypass grafting (CABG) surgery, were included in the study. Forty bronchoalveolar lavage (BAL) fluid samplings were performed in the 10 patients. Samples were obtained at the following time periods: (1) preoperatively; (2) at the end of the first hour after anaesthetic induction; (3) at the conclusion of 30 min of crossclamp on CPB; and (4) at the conclusion of 20 h after the end of CPB, postoperatively. Cell contents of bronchoalveolar lavage fluid, alveolar macrophage viability, eosinophil cationic protein (ECP) levels and myeloperoxidase (MPO) concentrations were analysed in each bronchoalveolar lavage fluids. While the percentage of preoperative macrophages was 85.90% and the percentage of preoperative neutrophils was 2.40%, they were 77.00% and 11.30% in the postoperative samples, respectively. Mean alveolar macrophage viability was 96.20% preoperatively and 90.40% in the postoperative period. Preoperative eosinophil cationic protein mean concentration was < 2 microg/l and mean response value (RV) was 28.80. Preoperative mean myeloperoxidase concentration was 7.66 ng/ml. Postoperative eosinophil cationic protein mean response value was 63.40 and mean myeloperoxidase concentration was 59.25 ng/ml. There were significant differences between third and final samples with regard to both neutrophil percentages (p = 0.028) and MPO levels (p = 0.005). While the preoperative mean PaO2 value was 89.39 mmHg and mean SaO2 value was 97.12%, they were calculated in the postoperative arterial blood specimens of patients, without inhaling O2, as 65.31 mmHg and 93.84%. These changes between blood gas analyses reflect the impairment of the lungs (p = 0.009 and p = 0.007, respectively). Neither alveolar macrophage viability nor ECP levels changed significantly between consecutive periods. However, when the results of the first and fourth samples were compared, we saw the cumulative effects of CPB, in that alveolar macrophages lost their viability and ECP mean RVs rose. These changes were statistically significant (p = 0.027 and p = 0.013, respectively). However, postoperative ECP levels were not like those found in a patient with asthma. Also, changes between alveolar macrophage percentages (p = 0.028), between neutrophil percentages (p = 0.036) and between MPO concentrations (p = 0.005) were statistically significant. Again, changes in neutrophil percentages between first and final samples correlated with changes in MPO levels between same periods (r = 0.657, p = 0.039).     

Increased DNA oxidative susceptibility without increased plasma LDL oxidizability in Type II diabetes: effects of alpha-tocopherol supplementation

In vivo supplementation studies of the antioxidant alpha-tocopherol in human Type II diabetes have used surrogate, rather than direct, markers of oxidative damage/antioxidant protection and have used higher doses of alpha-tocopherol than used in coronary secondary prevention trials. We tested the hypothesis that oral alpha-tocopherol in a dosage regimen used in secondary prevention trials would reduce directly observed oxidatively induced single-strand breaks in lymphocyte DNA in Type II diabetes. We studied 40 people with Type II diabetes and 30 controls in a randomized, double-blind, placebo-controlled trial of 400 i.u. of oral alpha-tocopherol daily for 8 weeks. Lymphocyte DNA single-strand breaks and low-density lipoprotein (LDL) particle size and oxidizability were measured at baseline, after 8 weeks, and after 4 weeks washout. Polymorphisms in the gene for the antioxidant enzyme paraoxonase-1 gene (position 192) were measured. The diabetics had increased DNA oxidative susceptibility (P=0.008), without increased LDL oxidative susceptibility. There was a direct relationship between DNA oxidative susceptibility and baseline plasma alpha-tocopherol in the diabetes group alone (r=0.421, r(2)=0.177 and P=0.023), but DNA and LDL oxidative susceptibility were not influenced by alpha-tocopherol supplementation in either group in this regimen. Paraoxonase-1 gene polymorphisms did not contribute to LDL or DNA oxidative susceptibility or response to alpha-tocopherol. Increased DNA oxidative susceptibility, therefore, can occur in Type II diabetes without increased LDL oxidative susceptibility, but alpha-tocopherol supplementation in this regimen has no influence on DNA or LDL oxidative susceptibility in Type II diabetes or controls. Polymorphisms in the paraoxonase gene (position 192) are not associated with differences in oxidative susceptibility or responses to alpha-tocopherol.     

Ruxolitinib and Tofacitinib Are Potent and Selective Inhibitors of HIV-1 Replication and Virus Reactivation In Vitro

The JAK-STAT pathway is activated in both macrophages and lymphocytes upon human immunodeficiency virus type 1 (HIV-1) infection and thus represents an attractive cellular target to achieve HIV suppression and reduced inflammation, which may impact virus sanctuaries. Ruxolitinib and tofacitinib are JAK1/2 inhibitors that are FDA approved for rheumatoid arthritis and myelofibrosis, respectively, but their therapeutic application for treatment of HIV infection was unexplored. Both drugs demonstrated submicromolar inhibition of infection with HIV-1, HIV-2, and a simian-human immunodeficiency virus, RT-SHIV, across primary human or rhesus macaque lymphocytes and macrophages, with no apparent significant cytotoxicity at 2 to 3 logs above the median effective antiviral concentration. Combination of tofacitinib and ruxolitinib increased the efficacy by 53- to 161-fold versus that observed for monotherapy, respectively, and each drug applied alone to primary human lymphocytes displayed similar efficacy against HIV-1 containing various polymerase substitutions. Both drugs inhibited virus replication in lymphocytes stimulated with phytohemagglutinin (PHA) plus interleukin-2 (IL-2), but not PHA alone, and inhibited reactivation of latent HIV-1 at low-micromolar concentrations across the J-Lat T cell latency model and in primary human central memory lymphocytes. Thus, targeted inhibition of JAK provided a selective, potent, and novel mechanism to inhibit HIV-1 replication in lymphocytes and macrophages, replication of drug-resistant HIV-1, and reactivation of latent HIV-1 and has the potential to reset the immunologic milieu in HIV-infected individuals.     

Effects of alpha-tocopherol on platelet membrane function in cystic fibrosis

Patients with cystic fibrosis and their parents were reported to have abnormal platelet aggregation responses to prostaglandin E1. To determine whether this is a property of the platelets, we studied the adenosine 3':5'-cyclic monophosphate (cAMP) response of washed platelets to prostaglandin E1. The cAMP response to prostaglandin E1 was the same in platelets from obligate heterozygotes for cystic fibrosis and from those of healthy controls. Patients with cystic fibrosis who had deficient vitamin E levels (plasma alpha-tocopherol, less than 500 micrograms/dl) had significantly (p less than 0.01) reduced platelet cAMP response to prostaglandin E1 compared with patients who had sufficient vitamin E, and supplementation with water-miscible vitamin E in these patients resulted in significant increases in plasma alpha-tocopherol levels (p less than 0.01) and in cAMP response to prostaglandin E1 (p less than 0.05). Plasma alpha-tocopherol levels correlated significantly with platelet cAMP response to prostaglandin E1 in patients with cystic fibrosis (r = 0.58, p less than 0.05). However, plasma alpha-tocopherol level was unrelated to the lymphocyte and granulocyte cAMP response to prostaglandin E1 or to the platelet cAMP response to alpha 2-adrenergic stimulation. Our data suggest that patients with cystic fibrosis have no inherited defect in platelet cAMP response to prostaglandin E1. In patients who have sufficient vitamin E, cAMP responses to prostaglandin E1 are normal in all the formed elements of the blood.