Premature lesions of the carotid vessels in HIV-1-infected patients treated with protease inhibitors.

OBJECTIVES: To evaluate the presence of premature atherosclerotic lesions of epiaortic vessels in HIV-1-infected protease inhibitor-(PI) treated patients compared with PI-naive patients and healthy individuals. DESIGN: One-hundred and two HIV-1-positive patients, including 55 treated with PI for at least 12 months and 47 either naive or treated with PI-sparing regimens, were subjected to epiaortic vessel ultrasonography. These data were compared with those obtained from 104 healthy individuals. METHODS: Intima characteristics, pulsation and resistance indexes, and minimal, peak and mean speed were evaluated using a colour power doppler. Atherosclerotic plaques were described. Independent risk factors and values for glycaemia, cholesterolaemia and triglyceridaemia were considered. Statistical analysis included the chi-square test, Mantel-Haenszel test, odds ratio and logistic regression analysis. RESULTS: Of the PI-treated patients, 29 out of 55 (52.7%) presented acquired lesions of the vascular wall at ultrasonography, whereas similar lesions were found in seven out of 47 (14.9%) PI-naive patients. Of the 104 healthy individuals, seven cases (6.7%) of intimal medial thickness were noted. A slightly significant correlation was found between carotid lesions and age, male sex and hypercholesterolaemia, whereas cigarette smoking, hypertriglyceridaemia and Centers for Disease Control and Prevention stage significantly increased the risk of vascular lesions (P= 0.022, P= 0.017 and P= 0.079 respectively). However, the highest significance regarded use of PI (P= 0.011). These results were confirmed by logistic regression analysis. CONCLUSIONS: These data demonstrate a higher than expected prevalence of premature carotid lesions in the PI-treated compared with PI-naive patients. If confirmed, a periodic ultrasonographic study of the vascular wall should be included in the follow-up of HIV infected patients.

Epidemiological, clinical and therapeutic evaluation of the Italian cholera epidemic in 1994.

In the period between 18 October and 4 December 1994, 12 indigenous cases of cholera were registered in the southern Italian region of Puglia, 10 of them were diagnosed in our Departments of Infectious Diseases. All patients were infected by consumption of raw fish or mussels. The patients had an elevated mean age and most were affected with systemic pathologies. The clinical course was mild and rarely complicated, although frequently the characteristic riziform diarrhoea was absent. In all patients V. cholerae serotype Ogawa biotype El Tor, was isolated; one patient was co-infected by Salmonella typhi. All strains showed resistance to cotrimoxazole and tetracycline. Nine of ten patients were treated with oral ciprofloxacin at 1 g/day for 10 days resulting in disappearance of the symptoms within a median of 36 hours and negative fecal cultures within a median of 24 hours. Our data suggest that Italy is at high risk of infection imported from nearby nations. The resistance to commonly used antibiotics for treatment of cholera and the good response to ciprofloxacin suggest including fluoroquinolones among the drugs of first choice geographical areas involved in the circulation of resistant strains of V. cholerae O1.

Preliminary data on the establishment of permanent cell lines continuously producing HIV-1: factors affecting the transmissibility and adaptability of "wild" HIV-1 isolates.

This study examines some technical aspects of the transmission to and permanent adaptability in continuous cell lines of wild HIV-1 isolates. Three cell systems (the lymphocytic cell lines Molt-3 and H-9 and the monocytoid cell line U-937) and two transmission protocols (cell to cell and cell-free) were used. Two different replicative behaviours were observed among isolates efficiently transmitted: a) transmissibility but not adaptability (consisting in a limited length of viral replication); aa) transmissibility and adaptability (consisting in a stable and long term virus production). The second type of replication was confined to viruses from patients with severe immunodeficiency. Technical and viral factors can affect the rate of transmissibility and adaptability: the modality of infection (cell to cell transmission appeared to be the most efficient) and the tropism of the virus (some viruses could infect only one T cell line).

Development of antibody to hepatitis C virus (HCV) in acute and chronic non-A, non-B post-transfusion hepatitis.

The antibody to hepatitis C virus (anti-HCV) was measured by an immunoassay in 507 serum samples from 94 patients with acute and chronic post-transfusion non-A, non-B hepatitis (NANB) and in 436 healthy blood donors. Anti-HCV was found in 70.8 of patients with acute hepatitis, in 78.2 with chronic hepatitis, and in 1.4 of healthy blood donors. In acute hepatitis, anti-HCV appeared in the serum from 4 to 34 weeks after transfusion and from 1 to 30 weeks after the onset of the overt disease. Three patients with resolving hepatitis (21%) and 2 who developed chronic hepatitis (10%) lost anti-HCV during a mean follow-up period of 28 months. Among the 36 patients with chronic hepatitis, 2 (6%) lost anti-HCV after 12 months and 8 years respectively. These data indicate that in recent years HCV has been the major etiologic agent of acute and chronic transfusion-associated hepatitis (TAH) in our geographical area. The late appearance of anti-HCV from the onset of clinical and biochemical signs of acute hepatitis in more than 70% of patients limits the diagnostic utility of this assay for an earlier serological diagnosis of acute NANB hepatitis. Additional studies are required to determine the diagnostic significance of this antibody in chronic NANB hepatitis.

In vitro production of HIV-1 specific antibodies from "at risk" seronegative subjects.

We investigated a group of well characterized seronegative subjects "at risk" for HIV-1 infection including heterosexual partners of HIV-1 infected subjects and intravenous drug abusers. Pokeweed mitogen (PWM)-stimulated peripheral blood mononuclear cells (PBMCs) were studied for their ability to produce antibodies against HIV-1 structural proteins in vitro. Viral activity by means of HIV-1 isolation from PBMCs and presence of serum p24 antigen were also tested. In 7/42 cases (16.6%) HIV-1 immunoreactive specific antibodies were found, mostly directed against the envelope proteins (gp 120/160). Remarkably, none of these in vitro antibody producers yielded HIV-1 isolation in cell cultures or had detectable serum levels of p24 antigen.

HIV-1 isolation from small amounts of whole blood: a technical evaluation.

We have evaluated a simple and sensitive culture technique for isolation of Human Immunodeficiency Virus Type-1 (HIV-1) from small amounts of whole blood. Data shown in the paper demonstrate that: 1) cell cultures from small amounts of heparinized whole blood (HWB) allow a high isolation rate in infected subjects at all stages of diseases; 2) among asymptomatic subjects the HIV-1 isolation rate is increased in cell cultures from HWB, with respect to cell cultures from peripheral blood mononuclear cells; 3) cultural results from HWB are not influenced by the presence of detectable serum p24 antigen, but a good correlation was found with the titre of anti p24 antibodies in serum; 4) continuous cell lines (such as Molt-3 cells) instead of peripheral blood mononuclear cells can be used, obtaining good results, for HIV-1 isolation from HWB; 5) frozen samples of HWB can be used in cell cultures for HIV-1 isolation; 6) the type of anticoagulant (Heparin or EDTA) used for the collection of blood does not influence viral replication in cell cultures from whole blood; 7) viral isolation from HWB is highly sensitive; amounts so small as five microliters of whole blood are sufficient, in some cases, to obtain viral replication in cell cultures; 8) the minimal dose of HWB sufficient to infect cell cultures (HWB M.D.I.) varied among different patients. Although this work failed to establish a correlation between this parameter and the clinical and immunological status of patients, it is conceivable that HWB M.D.I. could give information about viral load in blood and have a prognostic significance; 9) the HWB M.D.I. rise in patients treated with Zidovudine, suggesting that this method could be employed in the virological evaluation of trials with antiretroviral drugs.

Lack of HBV and HDV replicative activity in HBsAg-positive intravenous drug addicts with immune deficiency due to HIV.

The effects of HIV infection on HBV and HDV replication and liver damage were evaluated by comparing the findings from 48 anti-HIV-positive HBsAg chronic carriers with those from 22 matched anti-HIV-negative subjects. The state of HBV/HDV infection was also related to the degree of immunodeficiency of the anti-HIV-positive patients. Most patients were intravenous drug addicts (IVDA) (84.2%); male homosexuals represented only a small proportion (7.1%). Serum HBV-DNA was detected more frequently in anti-HIV-negative than in anti-HIV-positive patients (50% vs. 35%) despite evidence of HDV replication in the anti-HIV-negative group (P = 0.02). Seroconversion from ongoing to inactive HBV infection occurred in 45% of anti-HIV-negative patients as well as in 23% of anti-HIV-positive patients (P = ns). The difference in severity of liver damage between the two groups was not statistically significant (P = 0.84). Furthermore, in the anti-HIV-positive subjects, HBV and/or HDV activity was detected in 63% of patients with mild immunodeficiency (CDC groups II and III with a total CD4 count greater than 400/mm3) and also in 75% of ARC-AIDS patients (CDC groups IV A-IV C) (P = ns). Severe hepatic disease occurred in subjects with CD4 counts above or below 400/mm3 (13 vs. 6, respectively). In conclusion, the data do not demonstrate that HBV or HDV infections are modified by HIV. The epidemiological background of the patients investigated and the extensive spread of hepatitis viruses in Italy before the appearance of HIV may account for the lack of relationship between HIV and HBV/HDV infections.

[Analysis of the replication kinetics in primary culture of isolates of HIV-1 from infected patients in various stages of the disease]. / Analisi della cinetica replicativa in coltura primaria di isolati di HIV-1 da soggetti infetti in vario stadio di malattia.

The authors have studied the replicative kinetics and the induction of cytopathic effects of HIV-1 in primary co-cultures from infected subjects at various stages of the disease. Cultures from subjects with ARC or AIDS yielded HIV-1 replication more precociously and at higher levels compared to those from asymptomatic subjects; cytopathic effect "in vitro" were observed more frequently and earlier in cell cultures from ARC/AIDS subjects. Presented data indicate that the clinical and immunological deterioration during HIV-1 infection is related to viral replicative activity and suggest that the study of HIV-1 replicative kinetics in primary co-cultures may be helpful in predicting who will progress to AIDS.

[Isolation of human immunodeficiency virus (HIV) from whole blood]. / L'isolamento del virus della immunodeficienza umana (HIV) da sangue intero.

The authors describe a simple and sensitive technique for HIV isolation from small amounts of heparinized whole blood. This method demonstrated a high efficiency in detecting HIV at all stages of disease and appeared more sensitive with respect to viral isolation from peripheral blood mononuclear cells. Although further studies are needed to better understand the biological significance of a positive cultural result obtained by this method, HIV isolation from whole blood can be routinely employed, especially when small amounts of blood are available.

Different biological properties of paired HIV-1 isolates from peripheral blood mononuclear cells and cerebrospinal fluid.

Seven paired HIV-1 isolates from peripheral blood mononuclear cells (PMCs) and cerebrospinal fluid (CSF) of infected subjects at various stages of the disease were studied for their capacity to replicate in continuous cell lines (Molt-3 and U-937 cells) and to induce cytopathic effects "in vitro". Obtained results indicate that paired HIV-1 isolates from PMCs and CSF of the same patient can differ in their replicative activity "in vitro", suggesting that, at least in some cases, CSF isolates may represent a distinct subtype of HIV-1.

HIV-1 variability and progression to AIDS: a longitudinal study.

HIV-1 replicative activity and its relation to the clinical and immunological evolution of infection was studied in a group of 150 HIV-1 seropositive Italian i.v. drug abusers over a 1 year period. HIV-1 was isolated from 90 (60%) subjects; two groups of isolates were distinguished, according to replicative activity "in vitro" and ability to induce cytopathic effects in cell cultures, and were termed "rapid-high" and "slow-low" viruses, in agreement with other workers. Rapid-high viruses were recovered more frequently from patients with ARC/AIDS, while slow-low viruses seemed related to the asymptomatic period of infection. The replicative properties of HIV-1 seem to affect strongly the course of disease. In fact, an important CD4 cell decline occurred in asymptomatic subjects with rapid-high virus infection; asymptomatic subjects with negative viral cultures or with slow-low viruses showed no such decline. Asymptomatic subjects with negative viral cultures had no signs of disease during the observation period, while 9% with slow-low virus and 45% with rapid-high virus progressed to AIDS.

HIV isolation from whole blood: a new approach to HIV detection.

The authors present a simple and sensitive culture technique for isolation of Human Immunodeficiency Virus (HIV) from small amounts of heparinized whole blood. This method appeared to be highly sensitive at all stages of disease. Among asymptomatics HIV was isolated from 41.3% of subjects by using peripheral blood mononuclear cells in a standard co-cultivation technique and in 89.6% of cases using whole blood. In the group of AIDS patients, in contrast, HIV isolation rate of the two techniques was the same (94.4%). The presence of detectable p24 Ag in plasma did not affect cultural results and no false positive was observed in the control group. Although further studies are needed to better understand the biological significance of a positive cultural result obtained by this method, HIV isolation from whole blood can be routinely employed, especially when small amounts of blood are available.

[Virological monitoring in subjects with HIV infection under treatment with zidovudine (AZT)]. / Monitoraggio virologico in soggetti con infezione da HIV in trattamento con zidovudina (AZT).

The authors report preliminary results of the virological evaluation (performed by serum p24 antigen detection and HIV isolation from peripheral blood lymphocytes) of Zidovudine therapy in 64 anti-HIV positive subjects. Data presented indicate that the administration of therapeutic doses of the drug can efficiently and precociously inhibit viral replication, suggesting that the clinical benefits of the therapy may be related to the anti-viral activity of the drug. However, the anti-viral efficacy of Zidovudine seems to be of limited length; in fact a reactivation of viral replication was observed in most patients during the study.

[Technical note on the isolation of HIV from peripheral lymphomonocytes: use of PHA]. / Note tecniche sull'isolamento dell'HIV da linfomonociti periferici: impiego della PHA.

As a part of ongoing study about the role that different technical factors may play in influencing the sensibility of HIV isolation procedures, the authors have evaluated the effects of PHA stimulation of infected cells on HIV replication in cell cultures. Data presented demonstrate that the use of PHA in cell cultures for HIV isolation causes a slower viral replication and, in some cases, inhibits HIV growth.

[Isolation of the human immunodeficiency virus (HIV) from the cerebro-spinal fluid]. / L'isolamento del virus della immunodeficienza umana (HIV) dal liquido cerebro-spinale.

The authors describe a simple and available technique for HIV isolation from cerebrospinal fluid and report preliminary results obtained. Exposed data indicate that neurological involvement occurs early in the natural history of HIV infection and that the virus may be recovered in CSF at all stages of the disease, regardless of immunological conditions of patients. Virological evaluation of CSF may be important in understanding pathogenetic aspects of HIV infection and in clinical management of infected patients.

[Correlation of viral replication activity, the clinical phase of the disease and immunologic deterioration in HIV infection]. / Correlazioni tra attività replicativa virale, stadio clinico di malattia e deterioramento immunologico nelle infezioni da HIV.

The authors have studied the relationship among HIV replicative activity, expressed by HIV isolation from peripheral blood lymphocytes and p24 antigenemia detection, and the evolution of infection, in terms of stage of disease and T4 cells absolute number, in a group of 120 infected subjects. Data presented indicate that the expression of viral gene products is related both to clinical and immunological deterioration and suggest that viral replication plays a central role in the progression of the disease.

[The discordance between the presence of identifiable p24 antigen in the blood and the isolation of HIV from the blood is not influenced by methodological factors]. / La discordanza tra presenza di antigenemia p24 dosabile e isolamento dell'HIV dal plasma non è influenzata da fattori di ordine metodologico.

To clarify the biological, clinical and prognostic relevance of HIV isolation from plasma and explain the relation to p24 antigenemia, we studied these two markers in 67 anti-HIV positive subjects in different stages of the disease. The results suggest that discordances between these two parameters are not dependent on methodological problems and may be attributed to yet unexplained biological phenomena. Some hypotheses to explain the observed discrepancies are suggested.