ABSTRACT
BACKGROUND: Most persons who are infected with human immunodeficiency virus type 1 (HIV-1) are also infected with herpes simplex virus type 2 (HSV-2), which is frequently reactivated and is associated with increased plasma and genital levels of HIV-1. Therapy to suppress HSV-2 reduces the frequency of reactivation of HSV-2 as well as HIV-1 levels, suggesting that suppression of HSV-2 may reduce the risk of transmission of HIV-1. METHODS: We conducted a randomized, placebo-controlled trial of suppressive therapy for HSV-2 (acyclovir at a dose of 400 mg orally twice daily) in couples in which only one of the partners was seropositive for HIV-1 (CD4 count, > or = 250 cells per cubic millimeter) and that partner was also infected with HSV-2 and was not taking antiretroviral therapy at the time of enrollment. The primary end point was transmission of HIV-1 to the partner who was not initially infected with HIV-1; linkage of transmissions was assessed by means of genetic sequencing of viruses. RESULTS: A total of 3408 couples were enrolled at 14 sites in Africa. Of the partners who were infected with HIV-1, 68% were women, and the baseline median CD4 count was 462 cells per cubic millimeter. Of 132 HIV-1 seroconversions that occurred after randomization (an incidence of 2.7 per 100 person-years), 84 were linked within couples by viral sequencing: 41 in the acyclovir group and 43 in the placebo group (hazard ratio with acyclovir, 0.92, 95% confidence interval [CI], 0.60 to 1.41; P=0.69). Suppression with acyclovir reduced the mean plasma concentration of HIV-1 by 0.25 log(10) copies per milliliter (95% CI, 0.22 to 0.29; P<0.001) and the occurrence of HSV-2-positive genital ulcers by 73% (risk ratio, 0.27; 95% CI, 0.20 to 0.36; P<0.001). A total of 92% of the partners infected with HIV-1 and 84% of the partners not infected with HIV-1 remained in the study for 24 months. The level of adherence to the dispensed study drug was 96%. No serious adverse events related to acyclovir were observed. CONCLUSIONS: Daily acyclovir therapy did not reduce the risk of transmission of HIV-1, despite a reduction in plasma HIV-1 RNA of 0.25 log(10) copies per milliliter and a 73% reduction in the occurrence of genital ulcers due to HSV-2. (ClinicalTrials.gov number, NCT00194519.)
Subject(s)
AIDS-Related Opportunistic Infections/drug therapy , Acyclovir/therapeutic use , Antiviral Agents/therapeutic use , HIV Infections/transmission , HIV-1 , Herpes Genitalis/drug therapy , Herpesvirus 2, Human , Acyclovir/adverse effects , Adolescent , Adult , Antiviral Agents/adverse effects , CD4 Lymphocyte Count , Female , Follow-Up Studies , HIV Infections/complications , HIV-1/genetics , HIV-1/isolation & purification , Herpes Genitalis/complications , Humans , Intention to Treat Analysis , Kaplan-Meier Estimate , Male , Patient Compliance , Pregnancy , RNA, Viral/blood , Unsafe Sex/statistics & numerical data , Young AdultABSTRACT
Peroxidase, H2O2, and a halide form a powerful antimicrobial system in phagocytes and tissue fluids, and certain microorganisms can serve as the source of H2O2 for this system. H2O2-generating Lactobacillus acidophilus (LB+) is present in the vagina of most normal women and peroxidase has been detected in vaginal fluid. LB+ at high concentration is viricidal to HIV-1, and, at levels where LB+ is ineffective alone, the addition of peroxidase (myeloperoxidase, eosinophil peroxidase) and a halide (chloride, iodide, bromide, thiocyanate) restore viricidal activity. LB+ can be replaced by H2O2, but not by non-H2O2-producing LB, and viricidal activity is inhibited by azide and catalase. The survival of HIV in the female genital tract and thus the likelihood of transmission may be influenced by the activity of the LB(+)-peroxidase-halide system in the vagina.
Subject(s)
HIV Infections/transmission , HIV-1/physiology , Hydrogen Peroxide/metabolism , Lactobacillus/physiology , Sexually Transmitted Diseases, Viral/transmission , Vagina/microbiology , Azides/pharmacology , Catalase/pharmacology , Chlorides/pharmacology , Female , HIV Infections/prevention & control , HIV-1/drug effects , Humans , Leukocytes/enzymology , Male , Peroxidase/blood , Peroxidase/pharmacology , Sexually Transmitted Diseases, Viral/prevention & controlABSTRACT
After observations that Macaca nemestrina were exceptionally susceptible to simian immunodeficiency virus and human immunodeficiency virus type-2 (HIV-2), studies of HIV-1 replication were initiated. Several strains of HIV-1, including a recent patient isolate, replicated in vitro in peripheral blood mononuclear cells (PBMCs) and in CD4-positive M. nemestrina lymphocytes in a CD4-dependent fashion. Eight animals were subsequently inoculated with either cell-associated or cell-free suspensions of HIV-1. All animals had HIV-1 isolated by cocultivation, had HIV-1 DNA in their PBMCs as shown by polymerase chain reaction, and experienced sustained seroconversion to a broad spectrum of HIV-1 proteins. Macaca nemestrina is an animal model of HIV-1 infections that provides opportunities for evaluating the pathogenesis of acute HIV-1 replication and candidate vaccines and therapies.
Subject(s)
Genes, gag , HIV Infections/physiopathology , HIV-1/physiology , Macaca nemestrina/microbiology , Virus Replication , Animals , Base Sequence , CD4 Antigens/physiology , Cysteine/metabolism , Databases, Factual , HIV Seropositivity , HIV-1/isolation & purification , HIV-1/pathogenicity , Humans , Lymphocytes/immunology , Lymphocytes/physiology , Methionine/metabolism , Molecular Sequence Data , Oligodeoxyribonucleotides , Oligonucleotide Probes , Viral Proteins/biosynthesis , Viral Proteins/isolation & purificationABSTRACT
Detection of human immunodeficiency virus-type 1 (HIV-1) on only one or a few occasions in infants born to infected mothers has been interpreted to indicate that infection may be transient rather than persistent. Forty-two cases of suspected transient HIV-1 viremia among 1562 perinatally exposed seroreverting infants and one mother were reanalyzed. HIV-1 env sequences were not found in specimens from 20; in specimens from 6, somatic genetic analysis revealed that specimens were mistakenly attributed to an infant; and in specimens from 17, phylogenetic analysis failed to demonstrate the expected linkage between the infant's and the mother's virus. These findings argue that transient HIV-1 infection, if it exists, will only rarely be satisfactorily documented.
Subject(s)
HIV Infections/virology , HIV-1/genetics , HIV-1/isolation & purification , Specimen Handling , DNA, Viral/analysis , DNA, Viral/genetics , Diagnostic Errors , Equipment Contamination , Female , Genes, env , HIV Infections/immunology , HIV Infections/transmission , Humans , Infant , Infant, Newborn , Infectious Disease Transmission, Vertical , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , RNA, Viral/analysis , T-Lymphocytes, Cytotoxic/immunology , Viremia/virologyABSTRACT
Myeloperoxidase (MPO), H2O2, and chloride form an antimicrobial system in neutrophilic polymorphonuclear leukocytes (PMN) effective against a variety of microorganisms. Normal human PMN, when stimulated with phorbol myristate acetate or opsonized zymosan, are viricidal to HIV-1. The viricidal effect was lost when chloride was replaced by sulfate and was inhibited by the peroxidase inhibitor azide and by catalase, but not by heated catalase or superoxide dismutase, implicating H2O2. Stimulated PMN from patients with chronic granulomatous disease (CGD) were not viricidal to HIV unless H2O2 or glucose oxidase (which generates H2O2) was added, and the viricidal activity of H2O2-supplemented CGD PMN was inhibited by azide, implicating endogenous MPO. Stimulated PMN from patients with hereditary MPO deficiency had decreased viricidal activity unless MPO was added, and the viricidal activity of MPO-supplemented, MPO-deficient PMN was inhibited by catalase, implicating endogenous H2O2. The data suggest that when PMN are stimulated, MPO released by degranulation reacts with H2O2 formed by the respiratory burst to oxidize chloride to a product (presumably hypochlorous acid) that is toxic to HIV-1. Our findings raise the possibility that this viricidal effect of stimulated PMN may influence the host defense against HIV-1.
Subject(s)
HIV-1/immunology , Neutrophils/immunology , Peroxidase/metabolism , Cells, Cultured , Chlorides/pharmacology , Humans , Hydrogen Peroxide/pharmacology , Neutrophils/enzymologyABSTRACT
Bacterial spinae from marine bacterium D71 are multi-subunit structures of a single protein. This protein, called spinin, is homogeneous by immunodiffusion and immunoelectrophoresis, amino acid composition, polyacrylamide gel electrophoresis with a number of buffer systems, sedimentation velocity and diffusion boundary analysis. Sedimentation equilibrium gives Mr = 19,000, while phosphate polyacryl-amide gel electrophoresis in presence of dodecyl sulfate gives Mr = 32,000. The lower Mr estimate for spinin is supported by sedimentation equilibrium in 6 M guanidine . HCl, and covalent cross-linking with dimethyl suberimidate or glutaraldehyde. The higher Mr value probably arises from an anomalous spinin-dodecyl sulfate interaction. Isoelectric focusing in polyacrylamide gel gives pI = 3.45; however, the focusing pattern also contains three distinct bands that may arise from hydrolysis of the spinin protomer during anodic migration. This study presents the first extensive physicochemical characterization of spinin and provides the basis for investigating the subunit assembly of spinae.
Subject(s)
Bacteria/ultrastructure , Bacterial Proteins , Amino Acids/analysis , Bacterial Proteins/isolation & purification , Chemical Phenomena , Chemistry , Electrophoresis, Polyacrylamide Gel , Immunodiffusion , Immunoelectrophoresis , Isoelectric Focusing , Macromolecular Substances , Molecular Weight , Ultracentrifugation , Water MicrobiologyABSTRACT
OBJECTIVE: Causes of indeterminate results of Western blot testing (IWB) for human immunodeficiency virus (HIV) type 1 include seroconversion, HIV-2 cross-reactivity, and autoimmune disease, but most IWB results remain unexplained. This case-control study assessed risk factors for IWB results, including early HIV infection, other retroviral infection, autoantibodies, and other medical conditions. DESIGN: Prospective study to determine HIV seroconversion rate, with a case-control design to assess other risk factors for IWB. Cases (persons with one or more repeatedly reactive HIV-1 enzyme immunoassay with IWB), their current sexual partners, and controls (persons with negative enzyme immunoassay and Western blot results) were recruited from blood banks, health department and prenatal clinics, and private providers in Washington and Oregon. RESULTS: Of 244 cases enrolled, 206 were followed up for 6 months or longer, and six (3.0%; 95% confidence interval [CI], 0.7% to 5.3%) with recent HIV risk behaviors seroconverted. The Western blot banding patterns differed among groups; cases usually had p17 or p24 bands, while controls and cases' sexual partners usually had polymerase bands. Conditional logistic regression indicated that independent risk factors for IWB among male cases and controls were a tetanus booster in the past 2 years (odds ratio, 3.2; 95% CI, 1.2 to 8.6) and sexual contact with a prostitute (odds ratio, 3.0; 95% CI, 1.0 to 9.5). Independent risk factors for women were parity (odds ratio, 1.2; 95% CI, 1.02 to 1.4) and autoantibodies, either rheumatoid factor or antinuclear antibodies (odds ratio, 2.3; 95% CI, 1.03 to 5.6). No cross-reactivity was detected with HIV-2, human T-lymphotrophic virus type 1, feline immunodeficiency or feline leukemia, or bovine immunodeficiency viruses. CONCLUSIONS: Evaluation of persons with reactive HIV-1 enzyme immunoassays and IWB should include an assessment of HIV risk and other possible risk factors, such as alloimmunization (ie, parity or recent immunization) or autoantibodies (ie, antinuclear antibodies and rheumatoid factor). The relationship of IWB among men who reported sex with prostitutes is intriguing and warrants further study.
Subject(s)
HIV Seropositivity/diagnosis , Adolescent , Adult , Aged , Autoantibodies/blood , Blotting, Western , Case-Control Studies , Cross Reactions , Deltaretrovirus Antibodies/blood , Female , HIV Seropositivity/epidemiology , HIV Seropositivity/immunology , Humans , Immunoenzyme Techniques , Logistic Models , Male , Middle Aged , Odds Ratio , Pregnancy , Pregnancy Complications, Infectious/diagnosis , Prospective Studies , Risk FactorsABSTRACT
OBJECTIVE: To develop a model to predict transmission of HIV-1 from men to women. DESIGN: HIV-1 in seminal plasma, and endocervical CCR5 receptors were correlated with epidemiological studies of HIV-1 transmission to develop a probabilistic model. SETTINGS: Semen samples were collected from patient subjects in Seattle Washington, Chapel Hill, North Carolina, and St. Gallen, Switzerland. Endocervical biopsy specimens were obtained from women in Chicago, Illinois. PARTICIPANTS: Eighty-six men (not receiving antiretroviral therapy) in whom CD4 cell count and semen volume were available, and 24 women in whom the number of endocervical CCR5 receptors were determined. MAIN OUTCOME MEASURES: Prediction of transmission of HIV-1 from men to women per episode of vaginal intercourse based on the absolute burden of HIV (volume x HIV RNA copies/ml seminal plasma). RESULTS: The model suggests efficient heterosexual transmission of HIV-1 when semen viral burden is high. When semen contains 100 000 copies of non-syncytium-inducing (NSI) HIV RNA the probability of HIV-1 transmission is 1 per 100 episodes of intercourse; conversely, with 1000 copies NSI HIV RNA in semen, transmission probability is 3 per 10 000 episodes of intercourse. CONCLUSIONS: This model links biological and epidemiological data related to heterosexual HIV-1 transmission. The model can be used to estimate transmission of HIV from men with high semen viral burden from inflammation, or reduced burden after antiretroviral therapy. The results offer a biological explanation for the magnitude of the HIV epidemic in places where earlier studies have shown men have high semen viral burden, such as in sub-Saharan Africa. The model can be used to develop and test HIV-1 prevention strategies.
Subject(s)
Cervix Uteri/metabolism , Disease Transmission, Infectious , HIV Infections/transmission , HIV-1 , Models, Biological , Models, Statistical , Receptors, CCR5/metabolism , Semen/virology , Viral Load , Female , HIV Infections/epidemiology , Humans , Male , United States/epidemiologyABSTRACT
OBJECTIVES: To evaluate the effect of the menstrual cycle in HIV-positive women on plasma and genital cytokine levels, interrelationships between vaginal and plasma cytokines, CD4 and CD8 T cell fluctuations, and genital and plasma viral loads. METHODS: Plasma and cervicovaginal lavage specimens were collected from 55 HIV-positive women with CD4 cell counts < 350 cells/microl during phases of the menstrual cycle. Samples were assayed for IL-1beta, IL-6, IL-4, IL-8, IL-10, TGFbeta, TNFalpha, INFgamma, MIP1alpha, MIP1beta, RANTES, and TNFR-II using enzyme-linked immunosorbent assays. CD4 and CD8 T cell expression was evaluated by flow cytometry. Repeated measures regression models were used to assess the effect of the menstrual cycle on cytokines and viral load. Multivariate repeated regression models were used to assess the correlation among selected cytokines and between selected cytokines and HIV viral load. RESULTS: Vaginal IL-1beta, IL-4, IL-6, IL-8, IL-10, MIP1beta, RANTES, TGFbeta, and TNFR-II were significantly elevated during menses but were not altered during other phases. Plasma cytokine levels were not altered during the menstrual cycle. A positive Candida culture increased vaginal IL-8 during menses, whereas vaginal discharge was associated with a reduction in vaginal IL-4, IL-10, and RANTES. CD4 and CD8 cell numbers did not vary with the menstrual cycle. Vaginal cytokine levels correlated only with vaginal viral load, in a sampling method-dependent manner. CONCLUSION: We provide evidence of elevated vaginal cytokine levels during menses, which appear to regulate vaginal and not plasma HIV shedding, suggesting that a menstrual cycle pattern exists for cytokine production in HIV-positive women impacting vaginal shedding of HIV.
Subject(s)
Cytokines/metabolism , HIV Infections/immunology , HIV-1/physiology , Menstrual Cycle/immunology , Vagina/virology , Adolescent , Adult , Cytokines/blood , Female , HIV Infections/virology , HIV-1/isolation & purification , Humans , Middle Aged , RNA, Viral/blood , T-Lymphocytes/immunology , Vagina/immunology , Viral Load , Virus Shedding/physiologyABSTRACT
OBJECTIVE: To determine whether combination antiretroviral therapy is associated with reduced detection of HIV-1 RNA and DNA in the anorectal mucosa of men who have sex with men (MSM). DESIGN: Cross-sectional study of 233 MSM recruited from community and clinic sites in Seattle, Washington between July 1996 and December 1997. METHODS: HIV-1 RNA and HIV-1 DNA were detected in anorectal swab specimens by polymerase chain reaction amplification assays. RESULTS: HIV-1 RNA was detected significantly less often in anorectal specimens from users of combination antiretroviral therapies, whether a protease inhibitor was received (15/89; 17%) or not (16/53; 30%), than in men not receiving therapy (43/88; 49%) (P < 0.001, P = 0.03, respectively). In contrast, HIV-1 DNA was detected only slightly less frequently in anorectal specimens obtained from men receiving protease inhibitors (35/81; 43%) or reverse transcriptase inhibitors alone (22/48; 46%) than in specimens from men not receiving therapy (45/78; 58%) (P = 0.07, P = 0.20, respectively). Among men with < 50 copies HIV-1 RNA/ml plasma, detection of HIV-1 RNA in anorectal specimens was rare (1/54; 2%) but detection of HIV-1 DNA was common (14/50; 28%). CONCLUSIONS: Combination antiretroviral therapy is associated with reductions in HIV-1 RNA, but HIV-1 DNA remains detectable in the anorectal canal of almost half of MSM receiving such therapy. Condom use during anal intercourse should be encouraged, regardless of plasma viral load response to potent antiretroviral therapy.
Subject(s)
Anti-HIV Agents/therapeutic use , DNA, Viral/analysis , HIV Infections/virology , HIV-1/isolation & purification , Intestinal Mucosa/virology , RNA, Viral/analysis , Rectum/virology , Adult , Aged , Cross-Sectional Studies , Drug Therapy, Combination , HIV Infections/drug therapy , HIV Protease Inhibitors/therapeutic use , HIV-1/genetics , Homosexuality , Humans , Male , Middle Aged , Polymerase Chain Reaction , Proviruses , Reverse Transcriptase Inhibitors/therapeutic use , Viral LoadABSTRACT
OBJECTIVES: To assess the specific contributions of assay variation and biological variation to the total variation of plasma HIV-1 RNA measured by the Roche Monitor assay and the extent to which batch assays reduced both assay variability and total variability compared with real-time determinations. DESIGN: A retrospective analysis of data obtained from three trials conducted by the Adult and Pediatric AIDS Clinical Trials Groups (ATCG), the Women and Infants Transmission Study (WITS) and the NIAID-sponsored Virology Quality Assurance Program. METHODS: Within-subject variation was assessed from stored, serially collected plasma samples from 663 subjects enrolled in the ACTG and WITS studies. Interassay and intra-assay variation were estimated from two of the clinical trials and 22 laboratories that participated in a quality assurance program and were used to estimate the effect of real-time testing on total variation. RESULTS: The total variation (standard deviation) from a random effects model was 0.26 log10 RNA copies/ml. The estimated interassay variation was 0.08 log10 and intra-assay variation was 0.12 log10 RNA copies/ml. Biological variation accounted for 56-80% of total variation. The effect of real-time testing compared with batch testing was minimal. CONCLUSION: Our estimates of total within-subject HIV-1 RNA variation support the current recommendation to obtain at least two specimens, preferably obtained less than 2 weeks apart, for viral RNA measurement before starting therapy. The major contribution of biological variation to the total variation supports the use of real-time HIV-1 RNA assays, provided that consistent specimen collection procedures are followed and acceptable assay proficiency is maintained.
Subject(s)
HIV Infections/virology , HIV-1/isolation & purification , RNA, Viral/blood , Adult , Anti-HIV Agents/therapeutic use , Clinical Trials as Topic , Confidence Intervals , Female , HIV Infections/drug therapy , HIV-1/genetics , Humans , Male , Retrospective StudiesABSTRACT
OBJECTIVE: To assess the variation in HIV-1 over the menstrual cycle, including RNA levels in the female genital tract, plasma HIV-1-RNA levels, CD4 cell counts, and culturable virus. DESIGN: A prospective analysis of 55 HIV-1-infected women. METHODS: Blood and genital tract specimens were collected weekly over 8 weeks, spanning two complete menstrual cycles. Applying repeated-measures models that used menses as the reference level, the variation in viral RNA levels was compared in endocervical canal fluid and cells (collected by Sno-strips and cytobrush, respectively) and ectocervicovaginal lavage (CVL) fluid. Repeated-measures models were also used to assess the variation in plasma CD4 cell counts and viral load. RESULTS: Shedding patterns differed among the three sampling methods, independent of genital tract co-infections. Genital tract HIV-1-RNA levels from CVL fluid and endocervical canal cytobrush specimens were highest during menses and lowest immediately thereafter (P = 0.001 and P = 0.04). The HIV-1-RNA level in endocervical canal fluid was highest in the week preceding menses (P = 0.003). The menstrual cycle had no effect on blood levels of RNA (P = 0.62), culturable virus (P = 0.34), or CD4 cell counts (P = 0.55). HIV-1-RNA levels were higher in endocervical canal fluid than in peripheral blood plasma during the late luteal phase (P = 0.03). CONCLUSION: HIV-1-RNA levels vary with the menstrual cycle in the female genital tract but not the blood compartment. HIV-1-RNA levels are higher in endocervical canal fluid than in blood plasma. These findings may have important implications for sex-specific pathogenesis, heterosexual transmission, and contraceptive hormone interventions in HIV-1-infected women.
Subject(s)
Genitalia, Female/virology , HIV Infections/virology , HIV-1/isolation & purification , Menstrual Cycle , Viremia , Adult , CD4 Lymphocyte Count , Female , HIV Infections/immunology , HIV-1/immunology , Humans , Luteal Phase , Prospective Studies , RNA, Viral/analysis , Therapeutic Irrigation , Viral LoadABSTRACT
OBJECTIVE: To determine the safety of the zidovudine (ZDV) regimen utilized in the Pediatric AIDS Clinical Trial Group (ACTG) 076 study. DESIGN: ACTG 076 was a randomized, double-blind, placebo-controlled trial which demonstrated that a ZDV regimen could prevent mother-to-child HIV-1 transmission. Infants were followed through 18 months of age and women were followed through 6 months postpartum. METHODS: Maternal complications, pregnancy outcomes, growth and development of the uninfected infants, and HIV-1 disease progression in the women were monitored prospectively. RESULTS: Maternal therapy was well tolerated. There was no serious pattern of adverse pregnancy outcomes associated with ZDV use. Amongst the ZDV-exposed infants, the only recognized toxicity was anemia within the first 6 weeks of life; the risk for anemia was not associated with premature delivery, duration of maternal treatment, degree of maternal immunosuppression, or maternal anemia. ZDV treatment was not associated with an increased incidence of newborn structural abnormalities. At 18 months of age, uninfected infants did not differ in growth parameters or immune function. No childhood neoplasias were reported in either group. In the women, at 6 months postpartum, there were no differences in clinical, immunologic, or virologic disease progression. CONCLUSION: There were no identified problems that would alter current recommendations for the routine use of ZDV for the prevention of mother-child HIV-1 transmission.
Subject(s)
Anti-HIV Agents/therapeutic use , HIV Infections/drug therapy , HIV Infections/transmission , HIV-1 , Infectious Disease Transmission, Vertical , Pregnancy Complications, Infectious/drug therapy , Zidovudine/therapeutic use , Anti-HIV Agents/adverse effects , Disease Progression , Double-Blind Method , Female , France , Humans , Infant , Infant, Newborn , Pregnancy , Pregnancy Complications/chemically induced , Pregnancy Outcome , United States , Zidovudine/adverse effectsABSTRACT
Fifty-two patients with acquired immunodeficiency syndrome were enrolled in this study to evaluate the relationship between cerebrospinal fluid (CSF) zidovudine concentrations and neurologic and human immunodeficiency virus (HIV) culture findings. Paired HIV-CSF culture and neurologic measurements were available in 30 and 45 patients, respectively. Twenty-nine patients were assessable for zidovudine CSF concentrations. Patients underwent lumbar puncture and neurologic testing before and after 8 weeks or more of oral zidovudine therapy (600 to 1500 mg/d). After 8 weeks of therapy, the frequency of HIV isolation from CSF cultures was unchanged. Significant neurologic improvement by examination was noted in 61.5% (32/52) of the patients. The median CSF zidovudine concentration among 29 patients was 0.047 mg/L (range, 0.015 to 0.198 mg/L). No correlation between CSF zidovudine concentration, cumulative dose, or HIV isolation from CSF and persistence or resolution of neurologic symptoms or signs was observed. The mechanisms by which zidovudine improves neurologic function are unclear and appear unrelated to direct clearance of virus from CSF.
Subject(s)
Acquired Immunodeficiency Syndrome/drug therapy , Zidovudine/therapeutic use , Acquired Immunodeficiency Syndrome/cerebrospinal fluid , Administration, Oral , Adult , Cerebrospinal Fluid/microbiology , Female , HIV/isolation & purification , Humans , Male , Middle Aged , Zidovudine/administration & dosage , Zidovudine/analysisABSTRACT
Peripheral blood mononuclear cells (PBMCs) from HIV-seronegative donors were infected in vitro with HIV-1. Infection was monitored by cytopathology, supernatant p24 antigen, and by immunocytochemical staining. After 14 days in culture, approximately 70-90% of the cells became infected with HIV, as indicated by cell fusion and immunostaining for virus. At this time, recombinant HuIFN-gamma was added to the cultures, followed by infection 24 h later with the intracellular protozoan parasites Toxoplasma gondii, Trypanosoma cruzi, or Leishmania chagasi. Percentages of intracellular parasites were determined at various points thereafter. Using a system capable of detecting both virus and parasite infection, we determined that (a) cells infected with HIV were capable of ingesting and/or being infected by each of these parasitic protozoa, (b) HIV-infected macrophages could be activated to inhibit the replication of all three parasites following treatment with IFN-gamma, and (c) cultures of HIV-infected macrophages could respond to IFN-gamma with increased oxidative burst activity. The degree of parasite infection or inhibition observed in infected cells was not significantly different from that observed in non-HIV-infected cells. From these observations, we concluded that HIV-1 infection does not render macrophages unresponsive to IFN-gamma activation for microbicidal activity.
Subject(s)
Eukaryota/growth & development , HIV Infections/parasitology , HIV-1/immunology , Interferon-gamma/pharmacology , Macrophage Activation , Macrophages/parasitology , Animals , Cells, Cultured , Eukaryota/drug effects , Eukaryota/ultrastructure , HIV Infections/immunology , HIV Infections/microbiology , HIV-1/drug effects , HIV-1/physiology , Humans , Leishmania/drug effects , Leishmania/growth & development , Leishmania/ultrastructure , Macrophages/immunology , Macrophages/microbiology , Respiratory Burst/drug effects , Toxoplasma/drug effects , Toxoplasma/growth & development , Toxoplasma/ultrastructure , Trypanosoma cruzi/drug effects , Trypanosoma cruzi/growth & development , Trypanosoma cruzi/ultrastructureABSTRACT
To characterize neurological and neuropsychological findings associated with human immunodeficiency virus type-I (HIV) infection, 77 seropositive homosexual or bisexual males with no or minor symptoms of HIV were compared prospectively to 44 HIV seronegative men by observers blinded to serological status of the subjects. Neurological symptoms and examination findings were not significantly different between seropositives and seronegatives except for cranial nerve findings, predominantly mild hearing impairment. Mean performance scores for a 15-test neuropsychological battery were within an unimpaired range for both groups, although for five tests, mean scores were significantly poorer in seropositives. After adjustment for vocabulary score, and demographic and psychosocial variables, the mean score of seropositives was significantly worse only for the Benton Visual Retention Test. Magnetic resonance (MR) images of brain were abnormal in 14 (27%) of 52 seropositives and one of 10 seronegatives (value was not significant). HIV was isolated from cerebrospinal fluid (CSF) in 31 (61%) of 51 seropositives. The only clinical or laboratory difference between CSF culture positives and negatives was a higher CSF immunoglobulin synthesis rate in the former subjects (medians of 10.3 versus 0.1 mg/day; p = 0.03). An additional 13 seropositive subjects had immunologic evidence of central nervous system HIV infection, defined by a serum-to-CSF HIV antibody ratio of less than 5.5. Intracranial abnormalities on MR imaging were associated with CSF immunologic responses to HIV. Nervous system involvement occurred in the vast majority of men with early HIV infection, but clinically significant impairment was uncommon.
Subject(s)
Central Nervous System Diseases/etiology , HIV Infections/complications , Adult , Bisexuality , Central Nervous System Diseases/epidemiology , HIV/isolation & purification , HIV Infections/epidemiology , HIV Infections/microbiology , Homosexuality , Humans , Leukocyte Count , Magnetic Resonance Imaging , Male , Neurologic Examination , Neuropsychological Tests , Substance Abuse, IntravenousABSTRACT
Eosinophils, when stimulated, release a variety of agents that can be toxic to ingested or extracellular targets. Among these systems is one that consists of eosinophil peroxidase (EPO), H2O2, and a halide. We report here that phorbol myristate acetate (PMA)-stimulated human eosinophils are virucidal to HIV-1 in a chloride-containing medium. When the eosinophil concentration is decreased to a level at which the virucidal effect is incomplete, the addition of bromide or iodide restored complete virucidal activity. The virucidal effect of eosinophils, PMA, and bromide under these conditions is inhibited by the peroxidase inhibitor azide and catalase, but not heated catalase or superoxide dismutase, implicating the EPO-H2O2-halide system. Purified EPO when combined with H2O2 in a chloride-containing medium is virucidal to HIV-1. When the EPO concentration is suboptimal, virucidal activity is increased by bromide, iodide, and, in this instance, thiocyanate and the virucidal activity of the bromide-supplemented system is inhibited by azide and catalase. Our findings, together with the demonstration that eosinophils express CD4 on their surface and, under some circumstances, can be productively infected with HIV-1, raise the possibility that biological oxidants formed by eosinophils can influence the pathogenesis of HIV-1 infection by their toxicity to eosinophil-associated or extracellular virus.
Subject(s)
Eosinophils/immunology , HIV-1/immunology , Bromides/pharmacology , Cells, Cultured , Eosinophil Peroxidase , Humans , Hydrogen Peroxide , Iodides/pharmacology , Peroxidases/metabolism , Tetradecanoylphorbol Acetate/pharmacologyABSTRACT
Previous use of the HIV-1 protease inhibitor saquinavir resulted in the infrequent appearance of mutations in the HIV-1 protease gene associated with resistance. We have examined the ability of saquinavir to select for resistance mutations. In multiple selections of HIV-1 in cell culture with saquinavir, similar patterns of mutations were reproducibly observed and the number of mutations increased with increasing selective pressure. In a small number of subjects who showed an antiviral response when saquinavir was added to their therapeutic regimen, similar mutations were detected in viral genomic RNA in vivo after 30 to 40 weeks of therapy. These results indicate that saquinavir can select for resistance mutations and suggest that the infrequent appearance of these mutations in vivo is the result of low drug exposure. These results also predict that the use of higher levels of saquinavir will lead to an even greater frequency of resistance mutations in patients who fail therapy.
Subject(s)
Anti-HIV Agents/pharmacology , HIV Protease Inhibitors/pharmacology , HIV Protease/genetics , Mutation , Saquinavir/pharmacology , Cell Line , Drug Resistance, Microbial/genetics , HIV Protease/drug effects , Humans , Reproducibility of ResultsABSTRACT
To determine safety and efficacy of tumor necrosis factor (TNF) and interferon-gamma (IFN gamma) in the treatment of patients with acquired immunodeficiency syndrome (AIDS)-related complex, a randomized, double-blind study was conducted. Twenty-five patients with AIDS-related complex and CD4 lymphocytes less than or equal to 500 x 10(6)/L attended an AIDS Clinical Trials Unit of a tertiary referral center. Patients were administered tumor necrosis factor (TNF) (10 micrograms/m2) or IFN gamma (10 micrograms/m2), or both intramuscularly three times weekly for 16 weeks. Side effects from all three preparations included fever, constitutional symptoms, and local reactions. No significant hematologic, hepatic, renal, or coagulation abnormalities were observed. CD4 lymphocyte counts, beta 2-microglobulin, p24 antigen levels, and anti-p24 antibody did not change significantly during therapy. Similarly, no significant change was noted in rates of HIV isolation from peripheral blood mononuclear cells or plasma. TNF and IFN gamma were tolerable after premedication with acetaminophen; however, no significant change in markers of human immunodeficiency virus infection was demonstrated. These cytokines alone do not appear to be of benefit, nor do they appear to hasten the progression of HIV infection.
Subject(s)
AIDS-Related Complex/drug therapy , Interferon-gamma/therapeutic use , Tumor Necrosis Factor-alpha/therapeutic use , AIDS-Related Complex/physiopathology , Adult , Biomarkers , Double-Blind Method , Drug Evaluation , Drug Therapy, Combination , Female , Follow-Up Studies , Humans , Injections, Intramuscular , Interferon-gamma/administration & dosage , Interferon-gamma/adverse effects , Male , Middle Aged , Tumor Necrosis Factor-alpha/administration & dosage , Tumor Necrosis Factor-alpha/adverse effectsABSTRACT
HIV-1 infection represents a dynamic interaction between the rapid turnover of virus, CD4+ cell proliferation and clearance. HIV-1 disease progression is assessed, in part, by the inverse relationship between virus burden and CD4+ cell count. However, there is enormous individual subject variability between virus burden in the peripheral blood and CD4+ cell count with subsequent disease progression, suggesting that there must be virologic and immunologic modifiers of the inverse relationship between virus load and CD4+ cell count. To investigate these modifiers, we have used a Cartesian coordinate plot analysis to describe the inverse relationship between viral burden and the peripheral blood CD4+ cell count. Subjects from several clinical studies with CD4+ cell counts ranging from < 50 to > 600 cells/microL and varying viral burdens were studied. The analysis described the effect of various virologic and immunologic modifiers on this inverse relationship, for example, viral resistance, viral phenotype and the effect of very low CD4+ cell counts, and specifically addressed individual subject variation in assessing the association between the viral and immunologic parameters that define disease progression and response to antiretroviral therapy. As such, the Cartesian coordinate plot analysis method provides one approach to investigating the individual subject response to antiretroviral therapy.