Seropositive Intravenous Drug Users Research Articles

Lymphocyte subsets and natural killer cell cytotoxicity in intravenous drug users with HIV-1 infection among Thai population

Intravenous drug users (IVDUs) are among the high-risk groups who are most vulnerable to HIV infection. Several illicit drugs alter host immune function with increased incidence of infections including that of HIV. Many studies of the immune response of NK cells in HIV-1 seronegative IVDUs and HIV-1 seropositive IVDUs have been published from the Western countries and yet no data is available from Thailand. To determine natural killer cell cytotoxicity and lymphocyte subsets in Thai HIV-1 infected intravenous drug users. The NK cell cytotoxic function was determined using our well-established EGFP-K562 flow cytometric assay in 30 IVDUs with HIV-1 infection (IVH) comparing with those from the same number of non-infected IVDUs (IVX), HIV-1 seropositive individuals (HIV-1+ve) and healthy controls. The percentage and the absolute number of NK cells, helper CD4+ T cells and cytotoxic CD8+ T cells were also investigated. Among the study groups, IVH showed not only the lowest percentage of lytic activity by NK cells, but also a decline in the percentage and absolute count of NK cells. A decline in helper CD4+ T cells and an increase of cytotoxic CD8+ T cells of IVH group when compared to those of other 3 groups were also demonstrated. The failure of innate immune NK cell function and their number in IVH may support the involvement of additional components of the immune system in the control of HIV-1 disease.

Identification of HIV-1 specific T lymphocyte responses in highly exposed persistently seronegative Chinese

Studies of highly exposed persistently seronegative (HEPS) individuals may provide valuable information on mechanisms of protection and on vaccine design. Cellular immune responses play a critical role in containing human immunodeficiency virus. However, the cellular immune responses in HEPS individuals have not been thoroughly assessed at the entire viral genome level. Ten HEPS Chinese with a history of frequent penetrative vaginal intercourse (mean frequency, at least once a week), with some unprotected sexual contact occurring in the weeks or days immediately before enrollment, 25 HIV-1 seropositive individuals, 10 HIV-1-seronegative healthy individuals with low-risk sexual behavior and no history suggestive of exposure to HIV-1 infection were enrolled. HIV-1-specific T cell responses were comprehensively analyzed by an interferon-gamma Elispot assay against 770 overlapping peptides spanning all HIV-1 proteins. HIV-1-specific T-cell responses of interferon-gamma secretion were identified in 3 (30%) out of 10 HEPS individuals; the specific cytotoxic T lymphocytes were targeted at Pol (2/10), Env (2/10), and Tat (1/10). HIV-1-specific T-cell responses of interferon-gamma secretion were identified in 20 (80%) out of 25 seropositive intravenous drug users (IDUs), revealing that all HIV-1 proteins and protein subunits could serve as targets for HIV-1-specific CD8(+) T cell responses with 85% recognizing Gag, 80% recognizing Nef, 75% recognizing Pol, 60% recognizing Env, 55% recognizing Vpu, 45% recognizing Vpr, 20% recognizing Vif, 20% recognizing Tat and 15% recognizing Rev in these seropositive individuals. None of the seronegative healthy individuals gave the positive T-cell responses. About 30% of HEPS Chinese mounted HIV-1 specific T cell immune responses. Cell-mediated immunity against HIV-1 may be developed through non-productive infections.

Molecular and phylogenetic analysis of HIV-1 variants circulating among injecting drug users in Mashhad-Iran

Genetic and phylogenetic information on the HIV-1 epidemic in Middle-East Countries, and in particular in Iran, are extremely limited. By March 2004, the Iranian Ministry of Health officially reported a cumulative number of 6'532 HIV positive individuals and 214 AIDS cases in the Iranian HIV-1 epidemic. The intra-venous drug users (IDUs) represent the group at highest risk for HIV-1 infection in Iran, accounting for almost 63% of all HIV-infected population. In this regards, a molecular phylogenetic study has been performed on a sentinel cohort of HIV-1 seropositive IDUs enrolled at the end of 2005 at the University of Mashhad, the largest city North East of Tehran. The study has been performed on both gag and env subgenomic regions amplified by Polymerase Chain Reaction (PCR) from peripheral blood mononuclear cells (PBMCs) and characterized by direct DNA sequence analysis. The results reported here show that the HIV-1 subtype A is circulating in this IDUs sentinel cohort. Moreover, the single phylogenetic cluster as well as the intra-group low nucleotide divergence is indicative of a recent outbreak. Unexpectedly, the Iranian samples appear to be phylogenetically derived from African Sub-Saharan subtype A viruses, raising stirring speculations on HIV-1 introduction into the IDUs epidemic in Mashhad. This sentinel study could represent the starting point for a wider molecular survey of the HIV-1 epidemics in Iran to evaluate in detail the distribution of genetic subtypes and possible natural drug-resistant variants, which are extremely helpful information to design diagnostic and therapeutic strategies.

Characterization of a virtually full-length human immunodeficiency virus type 1 genome of a prevalent intersubtype (C/B') recombinant strain in China.

A molecular epidemiology study was conducted among more than 100 human immunodeficiency virus type 1 (HIV-1) subtype C seropositive intravenous drug users (IDUs) from China. Genotyping based on the envelope C2V3 coding region revealed the highest homology of the most prevalent virus strains circulating throughout China to subtype C sequences of Indian origin. Based on these results, a virtually full-length genome representing the most prevalent class of clade C strains circulating throughout China was directly amplified from peripheral blood mononuclear cells of a selected HIV-infected IDU and subcloned. Sequence analysis identified a mosaic structure, suggesting extensive intersubtype recombination events between genomes of the prevalent clade C and (B')-subtype Thai virus strains of that geographic region. Recombinant Identification Program analysis and phylogenetic bootstrapping suggested that there were 10 breakpoints (i) in the gag-pol coding region, (ii) in vpr and at the 3' end of the vpu gene, and (iii) in the nef open reading frame. (B')-sequences therefore include (i) several insertions in the gag-pol coding region; (ii) 3'-vpr, the complete vpu gene, and the first exons of tat and rev; and (iii) the 5' half of the nef gene. Breakpoints located in the vpr/vpu coding region as well as in the nef gene of 97cn54 were found at almost identical positions of all subtype C strains isolated from IDUs living in different areas of China, suggesting a common ancestor for the C/B' recombinant strains. More than 50% of well-defined subtype B-derived cytotoxic T-lymphocyte epitopes within Gag and Pol and 10% of the known epitopes in Env were found to exactly match sequences within in this clade C/B' chimeric reference strain. These results may substantially facilitate a biological comparison of clade C-derived reference strains as well as the generation of useful reagents supporting vaccine-related efforts in China.

The effect of saliva specimen collection, handling and storage protocols on hepatitis C virus (HCV) RNA detection by PCR.

Commercial assays can now be adapted to detect salivary anti-hepatitis C virus (HCV) antibodies, increasing the potential of saliva as a non-invasive diagnostic specimen suited to surveillance and epidemiological studies. However, current diagnostic algorithms involve confirmation of HCV infection by RT-PCR. Manipulation and storage conditions of serum can influence the stability of viral RNA. This study examined whether varying specimen collection, handling and storage protocols also affected subsequent HCV RNA detection by RT-PCR applied to saliva specimens. Whole unstimulated saliva, together with saliva samples collected in two commercially available devices (Salivette and Omnisal) were obtained from 50 HCV seropositive intravenous drug users. The specimens were subjected to a number of handling and storage conditions, including heat treatment and prolonged storage, then examined for HCV RNA by RT-PCR using primers derived from the 5' non-coding region (5'NCR). HCV RNA was detected in saliva samples from 25 (50%) of the patients. No single collection device or handling procedure identified all the subjects with HCV RNA positive saliva though whole saliva yielded the greatest number of positive results. Collection and processing of saliva specimens for RT-PCR analysis is complex. At present, detection of salivary HCV RNA by PCR is not sufficiently sensitive for use as a diagnostic tool in epidemiological studies.

Mutational analysis of the CCR5 and CXCR4 genes (HIV-1 co-receptors) in resistance to HIV-1 infection and AIDS development among intravenous drug users.

We analysed a group of Spanish intravenous drug users and controls to determine the role of mutations at the chemokine receptor-5/HIV-1 cofactor (CCR5), previously implicated in resistance to HIV-1 infection, and CXCR4 genes in susceptibility to HIV-1 infection. The complete coding sequence of both genes was amplified by the polymerase chain reaction from genomic DNA of 50 seropositive slow progressors and 10 long-term non-progressors, and analysed by the single-strand conformation polymorphism technique in a search for mutations. No mutation in CXCR4 was found, and delta ccr5 was the only mutation identified at the CCR5 gene. We genotyped (delta ccr5 allele) 150 HIV-1+ intravenous drug users and 250 healthy controls from the same population (Asturias, Northern Spain). Patients were divided into rapid progressors, presenting an event indicating progression to the acquired immunodeficiency syndrome (AIDS) in the 2 years after infection (100 patients), and slow progressors, remaining asymptomatic for 2-10 years (50 patients). The frequencies of the delta ccr5 allele were 0.105 and 0.040 in controls and HIV-1+ patients, respectively. Eighteen per cent of the controls (45/250) and 8% (12/150) of the patients carried the delta ccr-5 allele (P=0.013). The frequency of delta ccr5 carriers among rapid and slow disease progressors was 3 and 15%, respectively. A highly significant difference was found between rapid progressors and controls (P=0.0014). No patient (0/150) was delta ccr5 homozygous compared with 1% among controls. Thus, the delta ccr5 allele (the only CCR5 mutation found in our HIV-1 patients) was rare among seropositive intravenous drug users, suggesting that the absence of this mutation confers an advantage to the virus when infecting cells in vivo. In addition, patients carrying the delta ccr5 allele tend to show a slow progression towards HIV-1-related disease, remaining asymptomatic for longer periods of time.

Changes in Membrane Enzymes and Glycosphingolipids in Lymphocytes from HIV-1-Infected and Noninfected Intravenous Drug Users

The amounts of cell-surface glycosphingolipids and plasma membrane enzymes produced on the peripheral blood mononuclear cells (PBMNCs) isolated from 101 intravenous drug users (IDUs), of whom 91 were HIV-1 seropositive, were examined. Seronegative IDUs and age-matched healthy donors served as controls. The numbers of circulating CD3+, CD4+, and CD8+ T lymphocytes decreased during the advanced stages of the infection. There were also fewer CD4+ T-helper cells in HIV-1--seronegative IDU drug addicts. PBMNCs from HIV-1--seropositive subjects had abnormal surface enzyme kinetics. The phospholipase C had two pH optima, whereas the enzyme on normal cells has only one. The specific activity in cells from AIDS subjects was 4 times lower than that in normal PBMNCs. 5'-Nucleotidase showed a similar trend, whereas neutral endopeptidase activity did not correlate with the amounts of surface common acute lymphoblastic leukemia antigen (CALLA). These enzyme activities were decreased in HIV-seronegative IDUs. The subcellular distribution of enzymes and the profile of surface glycosphingolipids were also markedly changed, indicating the profound alterations in the membranes of PBMNCs from HIV-1--seropositive IDUs. These data suggest that intravenous drug use compromises the biochemical and structural integrity of the membrane surface of PBMNCs even before the onset of HIV.

Prognostic value of adenosine deaminase compared to other markers for progression to acquired immunodeficiency syndrome among intravenous drug users

This study was designed to determine the prognostic value of erythrocyte adenosine deaminase (ADA) as a possible indicator of progression to AIDS, and compare this with other known cellular and serological markers. At the end of a 3-year study, a cohort of 114 human immunodeficiency virus-1 (HIV-1) seropositive intravenous drug users (IVDUs) from the five different Center for Disease Control (CDC) groups was examined in order to estimate the prognostic relevance with respect to the progression to acquired immunodeficiency syndrome (AIDS) of each of the following markers at baseline value: number and percentage of CD4+ T cells, number of CD8+ T cells, CD4+/CD8+ ratio, IgA and beta 2 microglobulin and ADA levels, and the presence of HIV antigens. Moreover, 57 IVDUs belonging to II and III CDC groups were analyzed in a follow-up study at 6-month intervals, in order to evaluate and compare the behavior of each marker over time. The prognostic significance of each marker was assessed by computing the survival distribution and the Cox analysis in a multivariate model providing the set of markers with greatest predictive value. The levels of ADA and the CD4+/CD8+ ratio showed a linear association with disease staging, whereas beta 2 microglobulin and CD4+/CD8+ ratio were the best predictors for AIDS progression. A highly significant increase in ADA and beta 2 microglobulin was observed during follow-up. The results obtained among HIV-positive IVDUs clearly indicate that the erythrocyte ADA may be considered a reliable marker of the development of HIV infection from the intermediate stages of the disease onwards.(ABSTRACT TRUNCATED AT 250 WORDS)

Oral manifestations of HIV infection in homosexual men and intravenous drug users: Study design and relationship of epidemiologic clinical, and immunologic parameters to oral lesions

This article describes the baseline findings from a study designed to compare the oral manifestations of HIV infection in homosexual men and intravenous drug users. Both seropositive and seronegative persons were studied. A standard examination instrument was developed to record indexes of oral disease as well as to record the presence of oral lesions. The two groups differed in terms of education, race, socioeconomic status, employment status, housing, and smoking experience. The prevalence and type of oral lesions differed in the two seropositive groups. In seropositive homosexual men, white lesions on the tongue (28.4%) predominated; whereas for the seropositive intravenous drug users, oral candidiasis (43.0%) and gingival marginal erythema (33.3%) were most often detected. We also observed that seronegative intravenous drug users displayed a greater number of oral lesions than seronegative homosexual men. For seropositive homosexual men, lesion presence was significantly associated with decreased levels of CD4; positive associations were seen with current smoking, antiviral drug use, and antibiotic use, and a negative association was observed with current employment. In contrast, only exposure to antiviral drugs was significantly correlated with lesion presence for seropositive intravenous drug users. This baseline analysis from our longitudinal study suggests clear differences in oral manifestations of HIV infection between seropositive homosexual men and intravenous drug users and between seronegative homosexual men and intravenous drug users. Among other parameters, it is apparent that lifestyle, access to health care, and the condition of the oral cavity before infection influence the development of oral lesions in persons with HIV infection.

Cognitive behavior in asymptomatic (CDC stage II and III) HIV--seropositive intravenous drug users (IVDUs).

To evaluate cognitive impairment in the early stages of HIV infection in intravenous drug users (IVDUs) we have studied 39 consecutive HIV-infected subjects (CDC stage II-III) whose only known risk factor for the infection was intravenous heroin addiction. The control group was represented by 30 seronegative IVDUs. All subjects were tested with an extensive neuropsychological battery assessing general intellectual abilities and single cognitive functions. The patients differed from controls only for tests of attention and visual-motor abilities: 20% of asymptomatic seronegative and PGL patients showed alterations in two or more cognitive tests, as opposed to 3% of controls (p < 0.001). Our findings suggest that cognitive deficits seem to be present in a substantial percentage of IVDUs with asymptomatic HIV infection. Cognitive damage at this stage seems to selectively involve attention and visual-motor abilities, sparing general intellectual performances.

T-Lymphocyte Subsets in Intravenous Drug Users With HIV-1 Infection

<h3>To the Editor.</h3> —Recently, Margolick et al¹reported on changes in CD4 and CD8 T-lymphocyte subsets in intravenous drug users (IVDUs). In their words, "The principal finding was the slow rate of decline in CD4 lymphocyte counts in HIV-1 [human immunodeficiency virus type 1] seropositive IVDUs over a 2.5-year period of observation." (Actual observation period was 18 months.) The authors assert that this finding contradicts "the common perception that HIV-1 infection in IVDUs leads to rapid decline in CD4 lymphocytes." Howev Follow-up, Acquired Immunodeficiency Syndrome (AIDS) Incidence, and Rates of Change in CD4⁺Lymphocyte Numbers in the San Giovanni Hospital Subcohort of the MPH Study, by CD4⁺Lymphocyte Level at Baseline* Baseline CD4 Level, Cells/μL Developed AIDS No. (%) Incidence† Median Follow-up, Semesters 1.16 Changes/6 mo Median Mean &lt;150 4(44) 34.36 -42.92 10.90 150-299 20 4(20) 2.08 2.42 6.25 23.80 26 6(23) 6.37 -15.31 450-599 45 6(13) 3.19 3.66 -29.79

Changes in T-Lymphocyte Subsets in Intravenous Drug Users With HIV-1 Infection

To evaluate changes in T-cell subsets in prevalent human immunodeficiency virus type 1 (HIV-1) seronegative and seropositive intravenous drug users (IVDUs) and in HIV-1 seropositive IVDUs with known time of seroconversion. Cohort study with a median 18-month follow-up. Community-based clinic established to study the natural history of HIV infection in IVDUs. Eight hundred fifty-nine self-referred IVDUs aged 18 through 49 years who injected drugs within the last 10 years and who did not have an AIDS (acquired immunodeficiency syndrome)--defining illness; 152 were seronegative for HIV-1, 621 were seropositive, and 86 seroconverted during the study. Proportions and absolute numbers of lymphocytes and CD3, CD4, and CD8 T cells as determined at 6-month intervals by flow cytometry and complete blood cell counts with automated differential. Median numbers of CD4 lymphocytes at enrollment were 1061/microL (1.06 x 10(9)/L) for seronegative IVDUs, 508/microL for seropositive IVDUs, and 733/microL for those who seroconverted (enrolled a median of 4.5 months after seroconversion); the corresponding figures for CD8 lymphocytes were 628, 894, and 889/microL, respectively. Median rates of decline in absolute numbers and percentages of CD4 lymphocytes per 6 months were 7.6/microL (0.0%) for seropositive IVDUs and 55.1/microL (1.9%) for IVDUs who seroconverted (median follow-up after seroconversion was 12 months). Multivariate regression analysis that incorporated the within-individual correlation of the CD4 lymphocyte counts showed no significant change in these cells over time and no change due to use of drugs. Our data suggest that progression of HIV-1 infection in IVDUs, as reflected in decline of CD4 cell counts, is no more rapid than that reported for other risk groups.

Prevalence of Tuberculin Positivity and Skin Test Anergy in HIV-1—Seropositive and —Seronegative Intravenous Drug Users

--To identify differences in purified protein derivative (PPD) tuberculin positivity and skin test anergy rates by human immunodeficiency virus (HIV) serostatus, CD4+ lymphocyte count, and other risk factors in intravenous drug users (IVDUs); and to evaluate the appropriateness of the Centers for Disease Control (CDC)--recommended definition for a positive PPD tuberculin skin test result in HIV-1-seropositive patients. --Nested case-control and cross-sectional analyses. --Community-based cohort of IVDUs. --Two hundred sixty HIV-1-seropositive and -seronegative IVDUs, drawn from an unselected cohort, were skin-tested for sensitivity to PPD tuberculin, mumps, and Candida antigens using the Mantoux method. --Positivity to PPD tuberculin, skin test anergy. --Even using the CDC definition of an induration 5 mm or greater in diameter in HIV-1 seropositives, this group was substantially less likely to be PPD tuberculin positive than HIV-1 seronegatives (13.8% vs 25.2%; P = .02). In the HIV-1 seropositives the relative odds of being PPD positive varied depending on whether 10 mm or greater (odds ratio [OR], 0.3; 95% confidence interval [CI], 0.2 to 0.7), 5 mm or greater (OR, 0.5; 95% CI, 0.2 to 0.9), or 2 mm or greater (OR, 0.7; 95% CI, 0.4 to 1.3) was used to define a positive test result. The mean diameter induration in the HIV-1-seropositive group was 2.6 mm vs 5.4 mm in the seronegative group (P = .005). Skin test anergy (to mumps and Candida) appeared to explain the differential. Anergy was substantially higher in the HIV-1 seropositive group and increased as the CD4+ lymphocyte count fell (chi 2 for linear trend, 24.5; P less than .0001). An inverse linear trend for PPD positivity and CD4+ lymphocyte count was also observed (chi 2 for trend, 6.1; P = .01). In multivariate analyses, being 35 years of age or older and being HIV-1 seronegative were significantly associated with PPD positivity, while history of previous police arrest was of borderline significance. Only HIV-1 seropositivity was significantly associated with anergy. --These findings show that CDC-recommended definition of an induration 5 mm or greater in diameter for PPD tuberculin positivity in HIV-1 seropositives significantly underestimates the "true" infection rate (using the PPD positivity rate in HIV-1 seronegatives as the criterion standard). A definition of 2 mm or greater would appear to be a better cutoff for reducing misclassification in HIV-1 seropositives. This study also confirms that delayed-type hypersensitivity is seriously depressed in HIV-1 seropositive IVDUs and that anergy testing is mandatory to properly assess a negative PPD test result.

Utilization of health services in a cohort of intravenous drug users with known HIV-1 serostatus.

Intravenous drug users (IVDUs) with human immunodeficiency virus (HIV) infection and AIDS often have no health insurance or rely on public programs to finance their health care. We examined the independent contributions of HIV serostatus, clinical symptoms, CD4 cell counts, and health insurance to utilization of health services among 1881 intravenous drug users in Baltimore, Maryland. Participants in an ongoing natural history study of HIV were informed of HIV serostatus and seropositives were informed of CD4 cell counts; 6 months later, participants were administered a questionnaire concerning self-reported use of health services, insurance coverage, and HIV-related symptoms. Of 1881 participants, 67% had health insurance (including Medicaid), 48% had at least one outpatient visit, and 12% had at least one inpatient visit within the prior 6 months. The proportion of the study population that was HIV-1 seropositive was 32%. In multivariate analysis, the single most important predictor of health care utilization was the presence of two or more HIV-related clinical symptoms. HIV positive serostatus alone or known low CD4 counts were not significantly associated with use of health care services. These data suggest that HIV seropositive IVDUs are not receiving recommended preventive care. Additional efforts will be needed to ensure that HIV-seropositive drug users participate in currently recommended protocols for early treatment of asymptomatic HIV-1 infection.

Low Prevalence of Sexual Transmission of Hepatitis C Virus in Sex Partners of Seropositive Intravenous Drug Users

Low Prevalence of Sexual Transmission of Hepatitis C Virus in Sex Partners of Seropositive Intravenous Drug Users

HTLV-I/II seropositivity and death from AIDS among HIV-1 seropositive intravenous drug users

In a prospective study of street-recruited intravenous drug users (IVDUs) in Miami, Florida, USA, 107 (46·5%) of 230 IVDUs were infected with HIV-1. Of these HIV-1 seropositive individuals, 23 were also infected with human T-lymphotropic virus type I or II (HTLV-I/II). To test the hypothesis that HTLV-I/II infection has an effect on the survival of HIV-1 seropositive IVDUs, various baseline clinical and laboratory HIV-1 related indices in HTLV-I/II positive and negative groups were compared. Life table analysis and Cox's proportional hazards model were used to estimate the potential effect of HTLV-I/II infection on the survival patterns of people infected with HIV-1. IVDUs infected with both viruses were three times more likely to die from AIDS during follow-up than were those infected with HIV-1 only. This finding suggests that HTLV-I/II seropositivity may adversely affect the clinical outcome of HIV-1 seropositive patients.

Risk of AIDS in HIV seroconverters: A comparison between intravenous drug users and homosexual males

A multicentre cohort study was conducted in Italy to estimate the risk of developing AIDS in 261 intravenous drug users and 89 homosexual males for whom the seroconversion period was known. Four years after HIV seroconversion, AIDS incidence, estimated by Kaplan-Meier survival technique, was 13.8% for intravenous drug users and 16.2% for homosexual males; the difference was not statistically significant. These findings suggest that four years after seroconversion the risk of developing AIDS in HIV seropositive intravenous drug users is no higher than that of subjects who acquired HIV infection through sexual contact.