Normand Lapointe

Use of PGMY Primers in L1 Consensus PCR Improves Detection of Human Papillomavirus DNA in Genital Samples

ABSTRACT The novel PGMY L1 consensus primer pair is more sensitive than the MY09 and MY11 primer mix for detection and typing with PCR of human papillomavirus (HPV) DNA in genital specimens. We assessed the diagnostic yield of PGMY primers for the detection and typing of HPV by comparing the results obtained with PGMY09/PGMY11 and MY09/MY11/HMB01 on 299 genital samples. Amplicons generated with PGMY primers were typed with the line blot assay (PGMY-line blot), while HPV amplicons obtained with the degenerate primer pool MY09/MY11/HMB01 were detected with type-specific radiolabeled probes in a dot blot assay (standard consensus PCR test). Cervicovaginal lavage samples (N = 272) and cervical scrape samples (N = 27) were tested in parallel with both PCR tests. The PGMY-line blot test detected the presence of HPV DNA more frequently than the standard consensus PCR assay. The concordance for HPV typing between the two assays was 84.3% (214 of 255 samples), for a good kappa value of 0.69. Of the 177 samples containing HPV DNA by at least one method, 40 samples contained at least one HPV type detected only with PGMY-line blot, whereas positivity exclusively with the standard consensus PCR test was found for only 7 samples (P < 0.001). HPV types 45 and 52 were especially more frequently detected with PGMY than MY primers. However, most HPV types were better amplified with PGMY primers, including HPV-16. Samples with discordant results between the two PCR assays more frequently contained multiple HPV types. Studies using PGMY instead of MY primers have the potential to report higher detection rates of HPV infection not only for newer HPV types but also for well-known genital types.

Immunological and clinical differences between juvenile and adult onset of systemic lupus erythematosus

Introduction: Systemic lupus erythematosus (SLE) in children usually follows a more severe course than in adults, but sometimes in the previous studies reported there are many confounding factors Objective: To analyse the immunological and clinical characteristics of SLE juvenile onset and SLE adult onset. Methods: We studied 179 patients with SLE, 49 patients were aged 6 – 18 yrs at onset of disease. Anti-dsDNA antibodies were detected by radioimmunoassay and antibodies to extractable nuclear antigens (ENA): anti-nRNP, anti-Sm, anti-Ro/SS-A and anti-La/SS-B antibodies by ELISA, counterimmuno-electrophoresis and immunoblotting. Results: Juvenile-onset SLE shows a higher frequency of cutaneous vasculitis (44.8% vs 27.6%; P < 0.05), seizures (18.3% vs 7.6%; P < 0.05) nephropathy (67.3% vs 48.4%; P < 0.025), and discoid lupus erythematosus (26.5% vs 13.8%; P < 0.05). The incidence of articular manifestations is lower than in adults (85.7% vs 96.1%; P < 0.025). No significant differences were found between the two groups in relation with the prevalence of antinuclear antibodies. Conclusions: Juvenile-onset SLE has more frequent neurological and renal manifestations than adult-onset SLE, but immunological markers are similar in both groups. These features suggest the most severe clinical manifestations in the juvenile-onset SLE group are not related with the presence of studied antibodies by different methods.

Comparison between vaginal tampon and cervicovaginal lavage specimen collection for detection of human papillomavirus DNA by the polymerase chain reaction

The aim of the study was to compare the accuracy of self‐administered vaginal tampon (VT) specimens for the detection of human papillomaviruses (HPVs) with that of cervicovaginal lavage specimens (CVL). Two hundred seventy‐four paired VT and CVL specimens were collected prospectively from women at risk of sexually transmitted diseases. Specimens were treated and amplified with the polymerase chain reaction (PCR). Each woman served as her own control. One hundred and forty‐four of 272 (52.9%) CVLs and 159 of 271 (58.7%) VTs contained HPV DNA sequences (correlation of 88%). The sensitivity and specificity of vaginal tampons reached 93.9% (138/147) and 80.5% (99/123), respectively. HPV typing results were concordant for 99 negative paired samples and 114 paired samples positive for the same type(s) (correlation of 78.9%). It is concluded that these sampling methods collect cells from different areas of the genital epithelium, highlighting the importance of further assessment of the comparative predictive value of HPV detection in each sample. J Med Virol 51: 42–47, 1997.

A national review of vertical HIV transmission

Objectives:Prevention of vertical HIV transmission has evolved significantly in Canada over the last two decades. The aim of this analysis is to describe the surveillance programme used, rate of vertical HIV transmission and changing epidemiology of HIV-affected pregnancies in Canada. Design:National perinatal HIV surveillance programme. Methods:From 1990, annual retrospective data was collected on demographic and clinical characteristics of HIV-infected mothers and their infants referred to 22 participating sites across Canada either before/during pregnancy or within 3 months after delivery. Factors impacting HIV transmission and demographic features were explored. Results:Two thousand, six hundred and ninety-two mother–infant pairs were identified. The overall rate of vertical HIV transmission was 5.2%, declining to 2.9% since 1997. The rate of transmission for mothers who received HAART was 1%, and 0.4% if more than 4 weeks of HAART was given. Forty percent of women delivered by caesarean section, with no difference in transmission rate compared with vaginal delivery for women treated with HAART (1.4 vs. 0.6%, P = 0.129) but significant risk reduction for those who did not receive HAART (3.8 vs. 10.3%, P = 0.016). Black women were the largest group; proportions of black and aboriginal women increased significantly over time (P < 0.001 for both). Heterosexual contact was the most common risk category for maternal infection (65%), followed by injection drug use (IDU) (25%). Conclusion:Vertical HIV transmission in Canada has decreased dramatically for women treated with HAART therapy. All pregnant women should be evaluated for HIV infection and programmes expanded to reach vulnerable populations including aboriginal, immigrant and IDU women.

Sexuality in Montreal women living with HIV

The impact of learning a positive HIV test result on the sexuality of 161 women (47 injection drug users (IDU), 53 non-IDU women of Haitian or African origin (non-IDU-HA), and 61 non-IDU Caucasian women (non-IDU-C) was assessed using closed and open-ended questions. Self-reported CD4+ count correlated with any post-test (p = 0.001) and past month sexual activity (p = 0.007). After learning their HIV status, 110 women (68%) were sexually active, 48 (44%) of these within 1 month. After resuming sexual activity, 84% underwent a sexual adjustment period (median duration 8.5 months). IDU women were more likely to have frequent sex, be anorgasmic, and prefer sex less often. Consistent partner condom use was low in general (19% for IDU, 30% for non-IDU-HA, and 62% for non-IDU-C) and by partner type (new regular partner 58%, same regular partner 36%, casual partner 29%). Sexual satisfaction tended to decline post-test and then increase to higher than pre-test levels. Counselling focused on the safe and satisfying aspects of sex may assist women with HIV infection in sexual decision-making. Facilitating the access of IDU women with HIV infection to medically supervised drug provision and to detoxification and rehabilitation programmes can weaken the link between drug use and sex work.

HIV-associated Lipodystrophy Syndrome: A Review of Clinical Aspects

Approximately two years after the introduction of highly active antiretroviral therapy for the treatment of HIV infection, body shape changes and metabolic abnormalities were increasingly observed. Initially, these were ascribed to protease inhibitors, but it is now clear that nucleoside reverse transcriptase inhibitors also contribute to lipodystrophy syndrome. The syndrome groups together clinical conditions describing changes in body fat distribution that include lipoatrophy, lipoaccumulation or both. However, there does not appear to be a direct link between lipoatrophy and lipoaccumulation that would support a single mechanism for the redistribution of body fat. Currently, there is no clear definition of lipodystrophy, which explains the difficulty in determining its prevalence and etiology. There are no current guidelines for the treatment of fat distribution abnormalities that occur in the absence of other metabolic complications. The present article reviews the current state of knowledge of the definition, symptoms, risk factors, pathogenesis, diagnosis and treatment of the morphological changes associated with lipodystrophy syndrome.

Longitudinal Assessment of Changes in HIV-Specific Effector Activity in HIV-Infected Patients Starting Highly Active Antiretroviral Therapy in Primary Infection

Both the magnitude and breadth of HIV-specific immunity were evaluated longitudinally on samples collected from six subjects starting highly active antiretroviral therapy (HAART) preseroconversion (group 1), 11 recently infected subjects starting HAART postseroconversion (group 2), five subjects starting HAART in the second half of the first year of infection (group 3), and six persons starting treatment in the chronic phase of infection (group 4). HIV-specific immunity was measured by IFN-γ ELISPOT, detecting the frequency of cells responding to a panel of HLA-restricted HIV-1 peptides. Intracellular cytokine staining was used to detect the frequency of HIV-1 Gag p55-specific CD4+ and CD8+ T cells in a subset of participants. The magnitude and breadth of HIV-specific responses persisted in all group 1 subjects and in 5 of 11 (45%) group 2 subjects. Both of these parameters declined in 6 of 11 (55%) group 2 and in all group 3 and 4 individuals. All persons who maintained detectable numbers of HIV-1 Gag p55-specific CD4+ and CD8+ T cells after starting HAART preserved the intensity and breadth of their HIV-specific effector response. Our results show that HIV-specific immunity can be preserved even if HAART is initiated beyond the acute phase of infection.

Sexual behavior and pregnancy outcome in HIV-infected women

Sexual behavior and pregnancy outcome data for 392 HIV-infected women were analyzed. During the 6 months before study entry, 71.2% (279 of 392 women) were sexually active. In multivariate regression, women with baseline CD4+ > or = 200/microl were more likely than women with CD4+ < 200/microl to be sexually active (adjusted odds ratio [OR] = 1.75; 95% confidence interval [CI], 1.06-2.88; p = .03). Consistent condom use was reported with 58.4% (149 of 255) of steady male partners and 65.7% (23 of 35) of casual partners. Overall, 90.3% of 279 sexually active women were using contraception. Among women aged between 15 and 44 years (n = 320), the incidence of pregnancy in the year before HIV diagnosis was 27.5 per 100 person-years (PY) (95% CI, 22.1-33.9) compared with 8.3/100 PY (95% CI, 6.8-10.2) in the time since HIV diagnosis (p < .001). The incidence of therapeutic termination of pregnancies conceived in the 20 weeks before HIV diagnosis (10.6/100 PY) was more than triple that after diagnosis (3.1/100 PY; p = .001). After publication of results of zidovudine prophylaxis of mother-to-child transmission, pregnancy rates did not increase, but the incidence of therapeutic abortion dropped from 4.3/100 PY to 1.4/100 PY (p = .009). Knowledge of sexual behavior, including pregnancy frequency and outcome, can assist in tailoring counseling for HIV-infected women concerning sexual and reproductive health.

Romantic relationships and sexual activities of the first generation of youth living with HIV since birth

Abstract HIV-infected children, now maturing into adolescence and adulthood, must cope not only with adolescent developmental issues, but also with a chronic, socially stigmatised and sexually transmittable illness. Little research on this first generation of survivors has focused on romantic involvement and sexuality. This study, which employs a mixed-method embedded strategy (qualitative supported by quantitative), describes the perspectives of youth living with HIV since birth concerning: (1) romantic involvement and sexuality; and (2) risk management including the risk of HIV transmission and partner serostatus disclosure. Eighteen adolescents aged 13–22 from Montreal, Canada, participated in individual semi-structured interviews and completed self-report questionnaires. Most youths participated in non-penetrative sexual activities. Ten participants reported having had vaginal and three anal intercourses, at an average age of 14 for girls and 15 for boys. All sexually active youth reported having used a condom at least once. Of those who reported that their first sexual relationship was protected, over half had taken risks in subsequent relationships (e.g., unprotected sex, multiple partners, etc.). Interviews conducted with sexually inactive youths illustrate the interrelatedness of romantic involvement, sexual initiation and potential serostatus disclosure. Involvement in a sexual relationship would not be conceivable unless the partner was informed of their serostatus. For sexually active participants, risk management implies HIV transmission and partner disclosure. These youths have emotional issues regarding disclosure in romantic relationships and few risked potential rejection by disclosing. Condom use acts as a reminder of the infection and a barrier to intimacy. The narratives illustrate how risk perception changes and becomes relative with time and experience, especially when the viral load is undetectable and when past experience has convinced the adolescent that his/her partner might not become infected. Findings reinforce the need to prioritise sexual health issues for young people with perinatally acquired HIV.

Characterization of humoral and cell-mediated immune responses directed against hepatitis C virus F protein in subjects co-infected with hepatitis C virus and HIV-1.

Background:Hepatitis C virus (HCV) F protein is encoded in an alternate reading frame overlapping the core protein region. Its precise sequence, biological function and mode of expression are currently unclear. This study was conducted to examine the prevalence and characteristics of host humoral and cell-mediated immune responses directed against F protein in patients co-infected with HCV and HIV-1. Methods:Mutations were introduced to allow the expression of HCV-1a F protein in the absence of core. This recombinant and a truncated form lacking the first 11 amino acid residues shared with core were expressed in Escherichia coli, and their amino acid sequences were verified by mass spectrometry. Vaccinia-F protein recombinants were used to test F protein-specific cytotoxic T lymphocyte (CTL) activity. The binding of F protein-derived peptides to HLA-A*0201 was studied to identify putative CTL epitopes. Results:Sera from 23 of 39 patients infected with various HCV genotypes recognized the truncated form, including 13 of 25 subjects co-infected with HIV-1, indicative of antigenic crossreactivity and consistent with the conservation of F protein coding sequences between HCV genotypes. Crossreactive F protein-specific CTL precursors were detected in nine of 11 HCV-infected subjects, including seven of nine patients co-infected with HCV and HIV-1. Finally, three novel putative HLA-A*0201-restricted CTL epitopes were identified. Conclusion:These results indicate that patients co-infected with HCV and HIV-1 can mount immunoglobulin and CTL responses directed against HCV F protein that are fully comparable in scope and magnitude with those observed in individuals infected with HCV alone.