Prospective study of CD8+ lymphocyte activation in relation to viral load in HIV-infected patients with > or = 400 CD4+ lymphocytes per microliter.

To investigate the temporal relationship between CD8+ lymphocyte phenotypic alterations, the CD4+ T cell decline, and plasma HIV RNA levels during the natural history of HIV infection, 33 treatment-naive HIV-infected patients with > or =400 CD4+ cells/microl were studied prospectively for 3 years. During the study period, 20 patients remained untreated, and only 6 received more than 6 months of therapy. A significant relationship was found between changes in plasma HIV RNA and changes in the proportion of CD38+CD8+ cells. Conversely, the number of CD4+ T cells lost per year was strongly related to the increase in the proportion of CD28-CD8+ T cells. A strong relationship between mean yearly changes in CD4+ T cell numbers and changes in HIV RNA was also observed. CD4+ T cell changes were associated with changes in both viral load and CD8+ T cell activation. These results provide support for the use of both virologic and immunologic parameters for prognosis and management during HIV infection.

Human T lymphotropic virus type I (HTLV-I) infection in neurological patients in Salvador, Bahia, Brazil.

HTLV-I infection represents a major health concern in endemic areas throughout the world, such as Salvador, the main city of Bahia State, with socio-demographic characteristics similar to sub-Saharan African cities, located in the Northeast of Brazil. In order to provide an estimate of the frequency distribution, and range of neurological manifestations potentially related to HTLV-I infection in this city, we conducted a cross-sectional clinical-epidemiological study to determine the prevalence of this infection in patients with neurological diseases. Patients exhibiting vascular diseases, tumoral diseases or trauma were excluded. Over a period of 16 months, we studied 322 consecutive patients with chronic neurological diseases, who attended the neurological clinics of two major hospitals in Salvador. Overall, the prevalence of HTLV-I infection among the patients was 20.9% (67/320). However, the prevalence among the 104 patients with chronic myelopathy was 50.0% (52/104). It was observed that the major prevalence of HTLV-I was between the ages of 40 and 60 years with a female predominance. Our data indicate that, in Salvador city, HTLV-I is associated with chronic myelopathies or myeloneuropathies, which seem to be the only neurological diseases associated with HTLV-I.

AIDS related eye disease in Burundi, Africa.

AIMS: To determine the prevalence of ocular manifestations in AIDS patients hospitalised in Bujumbura, Burundi, according to their CD4+ lymphocyte count, serological status for CMV and VZV, and general health status. METHODS: Prospective study of 154 consecutive patients who underwent general and ophthalmological examinations, including dilated fundus examination. AIDS was diagnosed on the basis of Bangui criteria and HIV-1 seropositivity. CD4+ lymphocyte counts were determined by the Capcellia method. CMV and VZV antibodies were detected with ELISA methods. RESULTS: The mean age was 37 (SD 9) years and 65% of the patients were male. Active tuberculosis was the most frequent underlying disease (61%). Almost all the patients (99%) were seropositive for CMV and VZV. Among the 115 patients for whom CD4+ lymphocyte counts were available, 86 (75%) had more than 100 cells x 10(6)/l. Ocular involvement comprised 16 cases of microangiopathy, six of opalescence of the anterior chamber, five of retinal perivasculitis, two of zoster ophthalmicus, two of viral retinitis, and one of opalescence of the vitreous. CONCLUSION: In Africa, the prevalence of ocular involvement in HIV infection is far lower than in Europe and the United States, possibly because most African patients die before ocular opportunistic infections occur.

Detection and localization by in situ molecular biology techniques and immunohistochemistry of hepatitis C virus in livers of chronically infected patients.

Hepatitis C virus (HCV) detection in the livers of chronically infected patients remains a debatable issue. We used immunohistochemistry, in situ hybridization (ISH) alone or after microwave heating with FITC-labeled probes, RT-PCR with unlabeled primers followed by ISH (RT-PCR-ISH), and in situ RT-PCR with FITC-labeled primers (in situ RT-PCRd) to localize the virus in 38 liver biopsy specimens from 21 chronically infected HCV patients treated with interferon-alpha (IFN-alpha). Biopsies were taken at the beginning and end of IFN-alpha treatment and 1 year later. Results were compared with that of HCV-PCR in serum. RT-PCR-ISH and in situ RT-PCRd showed HCV signal in all liver biopsies even in responders with seronegative HCV PCR. This signal was intranuclear, diffuse, or peripheral, in hepatocytes, bile ductule cells, and lymphocytes. Cytoplasmic signals were occasionally observed. Whereas the percentage of labeled hepatocytes remained constant, the number of labeled lymphoid follicles decreased after INF-alpha therapy. Immunohistochemistry resulted in the same pattern of positivity but it was weaker and inconstant. This study indicates the persistency of HCV latency in IFN-alpha responders 1 year after IFN-alpha treatment cessation, a finding that certainly deserves confirmation.

[Prevention of viral hepatitis in travelers and expatriots in a tropical and subtropical environment]. / Prévention des hépatites virales chez les voyageurs et les expatriés en milieu tropical et subtropical.

Hepatitis A and B are hyperendemic in tropical and, to a lesser extent, subtropical countries. This high level of endemicity is in sharp contrast with the low frequency of these infections in the industrialized world. As a consequence, the incidences of hepatitis A and B are high among travellers to or foreigners living in tropical or subtropical countries. Therefore, these subjects should be vaccinated against hepatitis A and B. Furthermore, the usual preventive measures should be maintained. Risk of infection with the hepatitis C and E virus are much lower. Given the increasing number of travellers to tropical and subtropical countries, imported hepatitis is a public health problem for industrialized countries. Preventive measures must, then, be reinforced.

Complete nucleotide sequence of an African human T-lymphotropic virus type II subtype b isolate (HTLV-II-Gab): molecular and phylogenetic analysis.

We report the first complete nucleotide sequence of an African human T-cell lymphotropic virus type II. This new strain, called HTLV-II-Gab (Gab), was obtained from the uncultured peripheral blood mononuclear cells of a 44-year-old healthy Gabonese male who lived in a remote rural area, with neither history of blood transfusion nor sexual intercourse with non-Africans. Using nested PCR, 25 overlapping fragments, representing the entire proviral genome, were obtained, cloned and sequenced. The overall nucleotide sequence comparison with the four other available complete HTLV-II genomes indicated that Gab was more closely related to the HTLV-II subtype b prototypes (98.9, 99.3 and 98.2% nucleotide similarity with G12, NRA and GU respectively) than to the subtype a prototype (95.1% nucleotide similarity with Mo). Restriction profiles studies and phylogenetic analyses confirmed that Gab was a subtype b strain. However, this strain represents a newly described restriction fragment length polymorphism subtype, closely related to one of the rare partially sequenced African isolates originating from a pygmy living in Cameroon (PYGCAM). Nevertheless, the very low genetic divergence observed between this new African strain and the American strains raises several questions on the origins and level of genetic variability over time of this human retrovirus.

Nutritional status of HIV-seropositive subjects in an AIDS clinic in Paris.

OBJECTIVE: To measure the prevalence and analyse the characteristics of malnutrition among subjects attending an AIDS outpatient clinic and a day care center, to improve the nutritional management of HIV-infected subjects. DESIGN: Prospective cross-sectional study. SETTING: AIDS clinic in a University Hospital in Paris. SUBJECTS: 124 HIV-seropositive adults attending the clinic. MAIN OUTCOME MEASURES: Evaluation of nutritional status using anthropometry, impedancemetry, plasma albumin and pre-albumin assays. Degree of malnutrition, defined by the percentage of body weight loss (BWL), calculated by reference to the usual body weight. RESULTS: Among the 124 subjects recruited (M:F sex ratio: 3.3, mean age: 36.3 +/- 7.2 y), 77 (62.1%, 95%CI: 53.9-70.3) had normal nutrition status (BWL < or = 5%), 16 (12.9%, 95%CI: 7.0-18.2) moderate malnutrition (5% < BWL < or = 10%), 21 (16.9% 95%CI: 10.3-23.5) intermediate malnutrition (10% < BWL < or = 20%), and 10 (8.1%, 95%CI: 3.3-12.9) severe malnutrition (BWL > 20%). BWL was related to the CDC class (variance analysis, P < 9 x 10(-5)) and CD4 cell count (P < 3 x 10(-5)). Malnutrition was observed even among CDC class A subjects (14.9%). BWL was also related to the body mass index (P < 3 x 10(-6)), lean body mass (P < 3 x 10(-5)), body fat (P < 7 x 10(-6)), and as assessed by impedancemetry, body cell mass (P < 10(-5)) an the extra/intra cellular water ratio (P < 2 x 10(-4)). The decrease in lean body mass was related to the decrease in body cell mass. CONCLUSIONS: Given its high frequency, malnutrition should be prevented, detected, monitored and treated from the early stages of HIV infection among patients attending AIDS clinics in order to improve survival and quality of life.

Correlation of CD8 lymphocyte activation with cellular viremia and plasma HIV RNA levels in asymptomatic patients infected by human immunodeficiency virus type 1.

The relationship between CD8 lymphocyte phenotypic alterations and virological parameters was studied in 47 asymptomatic subjects with human immunodeficiency virus type 1 (HIV-1) infection and CD4 T cell counts above 400/microliters. CD8 subsets were examined by means of three-color flow cytometry, using an extensive panel of monoclonal antibody combinations. Virological parameters were measured by both end-point dilution culture of peripheral blood mononuclear cells (PBMCs) and plasma and branched-DNA (bDNA) signal amplification of plasma HIV RNA. Whereas HIV-infected patients had a near-normal CD4 cell count (mean, 782 cells/microliter), several subsets of activated CD8 cells were markedly expanded relative to values in 23 HIV-seronegative controls. The PBMC cultures were positive in 38 cases and plasma HIV RNA was detected in 31. The percentage of CD4 cells correlated negatively with both cellular viremia and plasma HIV RNA levels. Conversely, a positive correlation was observed between viral load and the percentage of CD8 cells. Among CD8 lymphocytes, the CD38+CD8 and HLA-DR+CD8 subsets correlated best with viral load. Three-color analysis showed that the subpopulations involved in this relationship were CD38+HLA-DR+, CD38+CD28-, HLA-DR+CD28+, HLA-DR+CD57-, CD38+CD57-, CD38+CD45RO+, and HLA-DR+CD45RO+. Our data provide the first evidence that viral load correlates with subsets of activated CD8 lymphocytes in asymptomatic HIV-infected subjects who have near-normal numbers of CD4 lymphocytes.

HTLV-I infection among prostitutes and pregnant women in Kinshasa, Zaïre: how important is high-risk sexual behavior?

High-risk sexual behavior as risk factor for human T-cell lymphotropic virus type I (HTLV-I) infection was assessed in cross-sectional studies with 1,183 prostitutes and 1,166 pregnant women in Kinshasa, Zaire. Eighty six (7.3%) prostitutes were positive for HTLV-I. The seroprevalence among prostitutes from the regions along the equator was 12.7%, whereas among prostitutes from the other regions it ranged between 0 and 4.3%. In the prostitutes from the high-prevalence regions, but not in the prostitutes from the low-prevalence regions, HTLV-I infection was associated with increasing age [odds ratio (OR) = 1.1 per year increment], active syphilis (OR = 2.3), and human immunodeficiency virus (HIV) infection (OR = 2.0). Forty three (3.7%) pregnant women were HTLV-I seropositive. Among the women from low-prevalence regions, there was no significant difference in HTLV-I seroprevalence between prostitutes (4.3%) and pregnant women (3.5%). In a group of 409 prostitutes who were observed for a mean duration of 23 months, the incidence of HTLV-I infection was 0.7 per 100 women-years, whereas the incidence of HIV infection was 9.8 per 100 women-years. We conclude that in Kinshasa prostitution per se was not associated with an increased risk of HTLV-I infection.

Comparative study of single and nested PCR for the detection of proviral HIV2 DNA.

We have tested seven pairs of primers for the detection of HIV2 DNA by single PCR in positive cultures from 21 infected patients. Four of these primer pairs were then used in a comparative study of single and nested PCR for the detection of HIV2 in fresh lymphocytes from 33 patients infected by the virus. HIV2 DNA was detected in 17 of the 33 patients (51.5%) by single PCR and 29/33 (88%) by nested PCR. All the patients negative in both nested and single PCR were asymptomatic and had CD4+ lymphocyte counts of at least 500 x 10(6)/l. This lack of PCR sensitivity for the detection of proviral HIV2 DNA in fresh lymphocytes cannot be totally attributed to genetic variability and may be related to a low viral load in asymptomatic HIV2-infected patients.

High level of hepatitis C endemicity in Gabon, equatorial Africa.

To assess the prevalence of hepatitis C virus (HCV) infection, a community-based study was performed in eastern Gabon on 1172 subjects over 5 years of age. The prevalence of antibodies to HCV (anti-HCV) detected using second-generation enzyme-linked immunosorbent assay (ELISA) and confirmed by an immunoblot assay (RIBA 2), was 6.5%. Anti-HCV prevalence increased with age but was related to neither sex nor ethnic group. Among 30 subjects with positive ELISA results, 14 had HCV viraemia as shown by the polymerase chain reaction (11/12 RIBA positive, 2/15 RIBA negative, 1/3 RIBA indeterminate). We conclude that HCV is highly endemic in western equatorial Africa and that a high proportion of the population may be viraemic.

Viral infections and chemical exposures as risk factors for hepatocellular carcinoma in Vietnam.

A case-control study investigating risk factors for hepatocellular carcinoma (HCC) was conducted in Hanoi, in the north of Vietnam, between 1989 and 1992. Male cases of HCC (152) diagnosed in 2 hospitals were included. Hospital controls (241) admitted mainly to abdominal surgery departments were frequency-matched to cases for sex, age, hospital and place of residence (Hanoi, province). Odds ratios adjusted for matching variables and other potential confounders were estimated using unconditional logistic regression, or exact non-parametric statistical inference when numbers were small. Positivity for hepatitis B surface antigen (HBsAg) was the main risk factor for HCC in this sample. Five subjects (3 cases, 2 controls) had been infected by hepatitis C virus (HCV), and none of them were carriers of HBsAg, giving an OR of 38 associated with HCV infection among HBsAG-negative subjects. Alcohol drinking was associated with HCC and interacted with HBsAg positivity. Agricultural use of organophosphorous pesticides (30 liters/year or more) and military service in the south of Vietnam for 10 years or more were also associated with an increased risk of HCC. This study confirms the major role played by HBV infection and its association with HCC in south-east Asia. It also suggests how other factors such as alcohol consumption or exposure to chemicals may interact with HBV infection.

Population-based study of serum hepatitis B virus DNA in Gabon.

The prevalence of serum hepatitis B virus (HBV) deoxyribonucleic acid (DNA) was measured in a Gabonese community at high risk for HBV infection. Among 698 subjects 5 to 24 years old, the prevalence of HBsAg was 11.1% vs. 57.9% for anti-HBs and 7.2% for anti-HBc alone. The prevalence of HBeAg among HBsAg-positive subjects was 26.5% vs. 59.5% for anti-HBe. The prevalence of HBV DNA tested by a hybridization spot test was 2.1% in the overall population and 18.7% among HBsAg-positive subjects. HBV DNA was found in 15 of 21 HBeAg-positive subjects but in none of the subjects positive for anti-HBe or negative for both HBeAg and anti-HBe. HBV DNA was not detected in any HBsAg negative subjects. The prevalence of HBV DNA decreased with age. This low prevalence of HBV DNA contrasts with the high level of endemicity in the study population.

Absence of a relationship between antibodies to hepatitis C virus and hepatocellular carcinoma in Mozambique.

To study the relationship between hepatocellular carcinoma (HCC) and hepatitis C virus (HCV), sera from 178 patients with HCC and 194 blood donors from Maputo, Mozambique were tested for antibodies to HCV (anti-HCV) using a second generation enzyme-linked immunosorbent assay and a confirmatory test with six synthetic peptides as reagents. The presence of hepatitis B surface antigen (HBsAg) was tested using an enzyme immunoassay. The prevalence of anti-HCV was higher in patients with HCC than in controls, but the difference was not significant after adjustment for age. Therefore, this difference reflected a difference in the age structure between the two groups. The prevalence of HBsAg was higher in patients with HCC than in controls. There was a negative association between anti-HCV and HBsAg in patients with HCC that was not significant after adjustment for age. These serologic results, which contrast with previous reports, show the need for further studies on the relationship between HCC and HCV using second generation serologic tests and molecular biology techniques.

Rapid determination of human cytomegalovirus susceptibility to ganciclovir directly from clinical specimen primocultures.

We were able to rapidly determine the susceptibility of human cytomegalovirus to ganciclovir using a late antigen synthesis reduction assay directly on primocultures of clinical specimens. This test was compared with a conventional susceptibility assay which was performed with cell-free virus obtained after cytomegalovirus isolation and in vitro passages. Both tests produced similar results. The rapid test, unlike conventional assays, is able to provide a result within 5 days after receipt of the specimen and could thus play a direct role in the therapeutic decision.

Anti-human immunodeficiency virus effects of zidovudine in combination with double-stranded RNA poly I poly C in T cells and monocytes-macrophages.

A comparison of the activity against human immunodeficiency virus 1 of zidovudine (AZT) and poly I poly C double-stranded RNA both alone and in combination in MT4 cells and primary monocyte/macrophage (M/M) cultures was made. The inhibition of the HIV-induced cytopathic effect or reverse transcriptase production by AZT in MT4 cells was not modified by the combination of the two agents. In contrast, AZT inhibition of reverse transcriptase production in the supernatant of M/M cultures was enhanced by the addition of poly I poly C. The inhibitory effect of the drug combination was more marked in M/M than in MT4 cells, indicating that the evaluation of compounds involving the induction of an antiviral state should be tested not only CD4+ T cells but also in monocyte-macrophages.