[Effect of anti-retroviral therapy on body composition changes: a literature review]. / Variazioni della composizione corporea associate alla terapia antiretrovirale: una rassegna della letteratura.

Protein-energetic malnutrition, characterized by both lean mass and fat depletion, was common in the pre-HAART era, and was associated with shortened survival and diminished quality of life. The pathogenesis of protein-energy malnutrition was multifactorial, and nutritional treatments were largely ineffective in the absence of disease stabilization. The introduction of HAART brought markedly improved outcomes, including a decrease in the incidence of malnutrition. However, other nutritional and metabolic alterations were noticed, and included changes in body shape, both lipoatrophy and lipohypertrophy, as well as changes in metabolism, notably hyperlipidemia and insulin resistance. These conditions, though sometimes occurring together, may occur independently, suggesting a complex, multifactorial cause. Several mechanisms have been hypothesized, including impairment to adipocyte differentiation and adipokine regulation, production of proinflammatory cytokines and mitochondrial toxicity. The role of the single drug class is still unclear, because both PI and NRTI have been associated with the syndrome, and the therapeutic protocols include both groups. Most of the medical therapies proposed for lipodystrophy are ineffective, and even if surgery remains an alternative, it is not associated with long lasting outcomes.

Inhibition of HIV-1 productive infection in hepatoblastoma HepG2 cells by recombinant tumor necrosis factor-alpha.

OBJECTIVE: To evaluate the role of liver cells in and the effect of tumor necrosis factor-alpha (TNF-alpha) on HIV-1 replication. METHODS: Human hepatoblastoma HepG2 cells were infected with various strains of HIV-1 and the effect of TNF-alpha treatment, either before or after infection, was monitored by p24 antigen assays. Northern blot analysis and gel retardation assays were performed to determine the expression of CD4 and HIV-1 trans-acting region (TAR)-binding proteins in these cells, respectively. RESULTS: HepG2 cells are CD4+ and support active HIV-1 replication, producing infectious virions, as measured by both p24 production and ability to infect T-cell lines with the virus produced by HepG2 cells. In contrast to the stimulatory effect of TNF-alpha on HIV-1 replication in T-cells and monocytes, up to 200 U/ml TNF-alpha treatment, at various times, either before or after HIV-1 infection, substantially inhibited p24 antigen production in HepG2 cells without causing any remarkable cytotoxicity. Gel-retardation assay revealed enhancement of a DNA-binding protein in TNF-alpha-treated HepG2 cells that binds to a specific sequence of the HIV-1 TAR, compared with the untreated control. CONCLUSIONS: These results indicate the importance of cellular factor(s) in HIV-1 infection and suggest that cytokines in different tissues can induce opposite effects. TAR-binding protein may act as an inhibitory factor for HIV-1 replication in the HepG2 cell line.

Sensitive ELISA for IgG and IgM anti-albumin autoantibodies not influenced by HBsAg-associated polyalbumin receptors.

An enzyme immunoassay for the detection of IgG and IgM anti-polymerized albumin autoantibodies (AAA) is described. It was found that polyalbumin receptors on HBsAg particles interfere in the detection of IgG AAA when polymerized human albumin (pHSA), but not polymerized bovine albumin (pBSA), is used as coating antigen. Polyalbumin receptors do not appear to interfere in the detection of IgM AAA, with either pHSA or pBSA as coating antigen. All normal sera showed evidence of AAA, of both IgG and IgM classes. Levels of IgG and IgM AAA in sera from most type A and type B acute hepatitis patients were above the range of normal controls. ELISA detection of AAA distinct from HBsAg reactivity can help in understanding the role of these autoantibodies in HBV infection.

Identification of a human immunodeficiency virus type 1 TAR binding protein in human hepatoblastoma HepG2 cells that trans-activates HIV-1 LTR-directed gene expression.

Recently, we have shown that the human immunodeficiency virus (HIV-1) long terminal repeat (LTR) directed chloramphenicol acetyltransferase (CAT) gene is efficiently expressed in human hepatoblastoma HepG2 cells and these cells can support productive HIV-1 replication. In this study we show that HepG2 cells contain a nuclear factor that binds to the HIV-1 trans-activating region (TAR), which we named HepG2-derived TAR binding protein (HTBP). Gel retardation assays using synthetic oligonucleotide probes carrying different mutations in the TAR region and competition DNA mobility-shift experiments using these oligonucleotides revealed the binding site encompassing between +7 and +13 nucleotides (5'-TCTGGTT-3') in the HIV-1 LTR. An in vivo CAT competition assay using -65HIV-1 LTR CAT as a reporter plasmid and various competitor plasmids containing these mutated oligonucleotides also demonstrated that HTBP can influence the HIV-1 LTR-directed CAT gene expression in HepG2 cells by interaction with a specific sequence in the TAR region.

Relation of changes in hepatitis B virus replication markers to the outcome of interferon treatment in patients with chronic hepatitis.

The relationship between the behavior of hepatitis B virus (HBV) replication markers and the response to treatment with recombinant alpha-2b interferon (IFN) was investigated in 11 patients with chronic hepatitis. At the end of 6 months of treatment, 4 patients showed a complete response to IFN: 2 more patients had seroconversion to HBeAb after 8 and 9 months of follow-up, respectively. The response to IFN was partial in the remaining patients. Pre-treatment levels of HBV DNA in patients showing complete response were lower than pre-treatment levels in patients with partial response: in addition, serum HBV DNA clearance during the treatment was associated with sustained remission more frequently than changes in the HBeAg/HBeAb system.

Quantitative viremia test for early prediction of a biochemical response in patients with chronic hepatitis C treated with interferon.

To evaluate the importance of the changes in viremia as an early predictor of the outcome of interferon (IFN) therapy, we assayed the levels of hepatitis C virus (HCV)-RNA in stored serum samples obtained from 34 patients with chronic hepatitis C who showed different biochemical responses to therapy. Serum samples obtained before the start of therapy and after 1 and 3 months were used, and viremia levels were determined by "branched DNA (bDNA)" technique. Viremia levels at 1 month of therapy were lower than pre-therapy levels in all 19 patients who had shown a persistent normalization of ALT during therapy (responder patients). The bDNA test was negative, i.e. the levels of viremia were below the sensitivity threshold of the method, in 12 (63.1%) patients at 1 month and in 13 (68.4%) at the 3rd month of therapy, whereas the bDNA test was negative in none of the 15 non-responder patients at the 1st month and in only 2 (13.3%) of them at the 3rd month of therapy. The bDNA test was superior to the ALT test both in predicting the non-response and the biochemical response to IFN after 1 month of therapy. The bDNA test results, instead, were not predictive of the duration of the response to IFN, either at the 1st or 3rd months of therapy. These results seem to indicate the usefulness of measuring the HCV-RNA levels at the beginning and after 1 month of IFN therapy in order to envisage or exclude a possible biochemical response early on in treatment.

Levels of anti-albumin autoantibodies in acute and chronic hepatitis B virus infection.

Levels of anti-albumin autoantibodies (AAA) of the IgG class were determined by ELISA in sera of patients with acute and chronic hepatitis B virus (HBV) infection. Mean AAA levels were higher than normal in both acute and chronic hepatitis patients. AAA levels were higher than the upper normal limits in practically all patients with acute self-resolving hepatitis, and decreased to normal levels when the patients recovered. Enhanced IgG AAA levels were observed in many patients with chronic hepatitis and serological markers of HBV replication. Elevated AAA levels were not associated with either more elevated transaminase levels or more severe histological forms of chronic hepatitis. The results of this study suggest that the interaction of albumin with HBV determinants is involved in AAA elevations, probably by mediating an increase in albumin immunogenicity. Moreover, the fall in AAA levels in the recovery phase of acute hepatitis, the coexistence of elevated levels with HBeAg and HBV-DNA, and the lack of correlation between AAA levels and different evolutive forms of chronic hepatitis, seem to exclude AAA from playing a relevant role, be it protective or damaging, during HBV infection.

[Role of hepatitis C virus in mixed cryoglobulinemia in patients with chronic hepatitis]. / Ruolo del virus dell'epatite C nella crioglobulinemia mista in pazienti con epatite cronica.

To evaluate the role played by hepatitis C virus (HCV) in mixed cryoglobulinemia in subjects with chronic hepatitis, we studied 72 consecutive patients: 43 had HCV-related chronic hepatitis, 19 HBV-related chronic hepatitis, and 10 chronic hepatitis of other etiology. We used second generation ELISA and RIBA to test for mixed cryoglobulinemia and anti-HCV antibodies in both serum and cryoprecipitates; HCV RNA were evaluated by "nested" PCR. Serum levels of rheumatoid factor and complement were also determined. The immunoglobulins in the cryoimmunoprecipitate were characterized by immunofixation electrophoresis. Cryoglobulinemia was present in 47% of the patients with chronic hepatitis C but in none of the sera of patients with HBV-related chronic hepatitis nor in those with chronic hepatitis of non-viral etiology. Type II mixed cryoglobulinemia was observed in 45% of the cases, and type III in 55%. HCV RNA and anti-HCV antibodies were present in all the cryoimmunoprecipitates. Ninety-five percent of the cryoglobulinemic patients had serum rheumatoid factor and 80% of them had low serum levels of C4. Our data indicate that mixed cryoglobulinemia is frequently associated with HCV-related chronic hepatitis, and that HCV and anti-HCV antibodies play an essential role in the development of mixed cryoglobulinemia.

[Diagnosis of HCV infection by 3rd-generation ELISA in HCV-RNA positive hypertransaminasemic patients with 2nd-generation ELISA negative results]. / Diagnosi di infezione da HCV con ELISA di 3 generazione in pazienti con ipertransaminasemia cronica HCV-RNA positivi ma negativi all'ELISA di 2 generazione.

To evaluate the improved sensitivity of 3rd-generation assays for the detection of ani-HCV antibodies in diagnosing cases of HCV infection, we have re-tested by 3rd-generation ELISA test (ELISA-3) serum samples from immunocompetent patients with chronic hypertransaminasemia who were HCV-RNA positive but tested negative with 2nd-generation ELISA (ELISA-2). Out of 21 HCV-RNA positive/ELISA-2 negative samples, 3 (14.3%) were ELISA-3 positive. Among the ELISA-3 reactive samples, two were indeterminate by RIBA-3 (one was reactive with c1 00 and the other with c22), and one was negative. These results demonstrate that even in the clinical setting ELISA-3 improves the diagnosis of HCV infection. The improvement seems to be related to a better reactivity of HCV peptides rather than to the inclusion of the new determinant NS5. However, the sensitivity of the tests for the detection of anti-HCV antibodies remains to be improved.

Biochemical response to interferon of chronic hepatitis in drug-addict patients infected with human immunodeficiency virus.

The effects of IFN treatment were retrospectively evaluated for 18 drug-addict patients with symptomatic HIV infection and chronic hepatitis C. Most of the patients were receiving concomitant treatment with zidovudine. Seven out of the 18 patients (39%) stopped IFN after less than three months, most of them for non-compliance. Among the 11 patients who completed a 6-12 month period of IFN treatment, 3 (27%) normalized and maintained normal ALT levels during therapy: for 2 of them the response was sustained after IFN discontinuation. The response to IFN therapy was neither correlated to the CD4+ levels nor to the clinical stage of the HIV infection. Instead, the response seemed to be influenced by pre-therapy ALT levels and liver histology. Tolerance to IFN treatment was good. These data show that IFN may be indicated in the therapy of chronic HCV infection for HIV-positive patients.

[Experience at a branch center of the National HIV Protease Inhibitor Registry]. / L'esperienza di un centro periferico sul registro nazionale degli inibitori delle proteasi di HIV.

In the present study we show our experience as one of the Centers (center 006) of the Italian Index of Protease Inhibitors (PI). In this multicentric, observational study we studied HIV-positive naive patients for antiretroviral therapy with protease inhibitors. From 8/1/97 to 31/1/98 in our department 64 patients were enrolled and followed up through an electronic medical sheet in order to evaluate efficacy, tolerability, toxicity and compliance of treatment with PI. Our results show a reduction in AIDS-related mortality and a decrease in AIDS defining diseases: in particular data show an increase in CNS pathologies in comparison with other opportunistic events. Moreover, in patients non complaints to therapy, the progression of the disease was more frequent. The side effects more frequently observed during PI treatment were hypertrigliceridaemia and lipodystrophy; gynaecomastia and hyperprolactinaemia were even present, perhaps as expression of direct metabolic action of HAART therapy.

Value of the test for serum HCV-RNA in clinical practice.

The test to identify genomic RNA is the only tool currently available to directly evidence the presence of the hepatitis C virus (HCV) in infected subjects. In this review we examined the most commonly used qualitative and quantitative methods to detect HCV-RNA in serum or plasma, focusing particular attention on commercially available methods. Furthermore, we report the clinical conditions in which the viremia test is useful or even mandatory for the diagnosis of HCV infection, and comment on the usefulness of the test for monitoring patients in the course of antiviral therapy.

Serodiagnosis of typhoid fever by enzyme-linked immunosorbent assay determination of anti-Salmonella typhi lipopolysaccharide antibodies.

Serum samples from 22 patients with proven typhoid fever, 60 febrile nontyphoidal patients, and 120 healthy subjects were tested for immunoglobulin A (IgA), IgG, and IgM anti-Salmonella typhi lipopolysaccharide antibodies by an enzyme-linked immunosorbent assay. The levels of all three classes of immunoglobulin anti-lipopolysaccharide were higher in typhoid patients than in controls; the test for IgM anti-lipopolysaccharide gave the best discrimination between typhoid and nontyphoidal sera. The absorbance values obtained with the enzyme-linked immunosorbent assay for IgM anti-lipopolysaccharide were highly correlated to the titers of anti-O agglutinins. However, the enzyme-linked immunosorbent assay was much more specific than the Widal test, and hence it could be a useful tool for the serological diagnosis of typhoid fever with a single blood sample.

Selectively increased adenosine deaminase levels in T cells during typhoid fever.

ADA and PNP levels were measured in T-enriched and T-depleted subpopulations of peripheral mononuclear cells from patients with typhoid fever. ADA activity was significantly increased in T-enriched, but normal in T-depleted preparations of typhoid patients. Elevated levels of ADA in T cells were observed from the initial phase up to 30 days from the onset of the disease. Unlike ADA changes, PNP modifications were transitory, and affected both T-enriched and T-depleted subpopulations. The usefulness of ADA determination in T cells to detect and follow up T-cell activation during typhoid fever is suggested.

Different levels of lymphocyte adenosine deaminase in active and inactive forms of chronic liver disease.

Adenosine deaminase (ADA) activity was measured in peripheral blood mononuclear cells (PBMC) from patients with chronic liver disease (CLD). Mean ADA levels were decreased in patients with untreated chronic active hepatitis (CAH). ADA levels in patients with CAH in remission under immunosuppressive treatment and in patients with inactive cirrhosis (IC) were similar to control values. ADA levels were not related to any of the biochemical parameters examined, or to the percentage of T cells evaluated on the same PBMC preparations. Our data suggest that biochemical and/or histological remission in CLD may be associated with normalization of ADA levels in PBMC.

[Adenosine deaminase activity in T-enriched and T-depleted preparations of human peripheral lymphocytes]. / Attività adenosina deaminasi in preparazioni T-arricchite e T-deplete di linfociti umani periferici.

Adenosine deaminase activity has been determined in T-enriched and T-depleted preparations of peripheral mononuclear cells from 6 normal subjects. T-enriched populations showed ADA levels lower than unseparated mononuclear cells (1.84 +/- 0.30 vs. 2.36 +/- 0.59 nmol NH3/min/10(6) cells). The highest values were found in T-depleted populations (2.46 +/- 0.39). Considering T cells percentages in our mononuclear cell preparations, T cells account for 64% of the biochemical ADA activity of peripheral blood mononuclear cells and the remaining 36% is due to non-T cells. The method for cell separation and enzymatic activity determination we used could be highly suitable for monitoring ADA levels in pathological conditions.