Long-term analysis of the resistance development in HIV-1 positive patients treated with protease and reverse transcriptase inhibitors: correlation of the genotype and disease progression.

In this study, 27 HIV-1-positive patients on long-term highly active antiretroviral therapy (HAART) in the Czech Republic were followed for a period of up to 7 years. Variability of the HIV-1 protease (PR) sequence common in the Czech Republic was observed. Under the pressure of inhibitors of protease (PRIs) and reverse transcriptase (RTIs) mutations in PR were detected. Development of resistance to PRIs was followed by a decrease in CD4 count and increase in viral load. The dynamics of viral load closely corresponded to the accumulation of specific primary mutations in PR and RT. Out of 27 patients 18 developed resistance to PRIs and the prolonged therapy led to the accumulation of a higher number of amino acid changes associated with the resistance and, consequently, cross-resistance to several PRIs was observed. These multi-resistant variants of HIV-1 with mutations in PR could not be inhibited sufficiently with PRIs that are currently available in clinical practice. Efficient yet temporary suppression of viral replication was achieved by a lopinavir (LPV) treatment.

Genetic subtypes of HIV type 1 viruses circulating in the Czech Republic.

We analyzed the genetic diversity of HIV-1 strains circulating in the Czech Republic. Phylogenetic analysis of the env and gag gene sequence fragments from 39 isolates revealed that the majority of these strains (32 of 39, 82%) were of subtype B; other genetic subtypes identified were A, C, F, and recombinant circulating form CRF01_AE. The isolates that did not cluster with subtype B originated almost exclusively from a heterosexual route of transmission. The molecular epidemiological data are suggestive of multiple entry of HIV-1 infection into the Czech Republic and show that the genetic pattern of the HIV-1 strains circulating in this country corresponds to that found in other European countries.

Seroprevalence of HHV 8 antibodies among the general population and HIV positive persons in the Czech Republic.

BACKGROUND: The seroprevalence rates of herpesvirus 8 (HHV 8) antibodies were determined for the general Czech population and HIV-positive individuals. OBJECTIVES: Six hundred and sixty six serum samples from the general Czech population and 129 serum samples from HIV-positive persons were tested for the presence of antibodies to the HHV 8 lytic and latent antigens. STUDY DESIGN: HHV 8 antibodies were detected by the indirect immunofluorescence test. RESULTS: In the general Czech population, only 2.4 and 0.3% of the serum samples tested positive for antibodies against the lytic and latent HHV 8 antigens, respectively. As many as 34.9 and 10.9% HIV positive individuals had antibodies to the HHV 8 antigens, respectively. Only three of them have developed Kaposi's sarcoma (KS) to date. At the time of KS diagnosis, the three patients had antibodies to both HHV 8 antigens. HIV-positive homo/bisexuals were at significantly higher risk of acquiring HHV 8 infection compared with HIV-positive heterosexuals. The increase in HHV 8 seroprevalence was associated with progression of the HIV infection from stage A to stage B. No correlation was found between the HHV 8 seroprevalence and CD 4+T-lymphocytes counts or the HIV viral load. CONCLUSIONS: Among the general Czech population, the HHV 8 seroprevalence is as low as in the West European countries. The mean HHV 8 seroprevalence rate in HIV-positive individuals was 34.9% and was comparable with those reported in other low seroprevalence countries.

Human immunodeficiency virus 1 strains resistant to nucleoside inhibitors of reverse transcriptase in isolates from the Czech Republic as monitored by line probe assay and nucleotide sequencing.

The genetic resistance to nucleoside inhibitors of the reverse transcriptase (RT) of human immunodeficiency virus I (HIV-1) isolates in the Czech Republic was examined by a line probe assay (LiPA) and nucleotide sequencing. The results of LiPA analysis of 294 blood specimens obtained from 156 patients revealed a high incidence of mutations in the RT gene related to resistance to various drugs (67.3%) in various combinations. Mutations in RT gene (M41L, K70R and T215Y/F) conferring the resistance to zidovudine (ZDV) were most frequent (62.6%), that (M184V) responsible for the resistance to lamivudine (3TC) was less frequent (33.7%), while those linked to the resistance to dideoxyinosine (ddl) and dideoxyinosine together with dideoxycytidine (ddl/ddC) were rather rare (6.5% and 5.1%, respectively). LiPA gave a high rate of uninterpretable results due to codon hybridization failure, especially in HIV-1 isolates of non-B subtype. Thirty-two specimens were analyzed also by direct sequencing of a part of RT gene. The results obtained by LiPA and the sequencing were highly concordant for codons successfully analyzed by both methods, but the sequencing provided information also about the codons that could not be analyzed by LiPA. A high prevalence of resistant strains in the Czech Republic and their heterogeneity justifies a regular HIV-1 resistance testing. LiPA turned out as a fast, powerful and most reliable tool for such a purpose. However, due to an increasing diversity of HIV-1 strains circulating in the Czech Republic, LiPA cannot replace the nucleotide sequence analysis.

Clinical importance of assessment of anti-HCV IgM antibodies in chronic hepatitis C.

In the majority of patients with acute hepatitis C the anti-HCV IgM antibodies in serum were present, however, some patients with chronic hepatitis C were positive for anti-HCV IgM too. The aim of this study was to assess the presence of anti-c22 IgM in patients with chronic hepatitis C and to determine whether the positivity for anti-c22 IgM has an impact on the histological finding in the liver. A total of 88 patients were examined (44 women, 44 men), mean age 48 years. The first group comprised 24 patients positive for both anti-HCV IgG and anti-c22 IgM, the second group 38 patients positive for anti-HCV IgG only, and the third group 26 patients negative for both anti-HCV IgG and anti-c22 IgM. Of 62 anti-HCV-IgG-positive subjects 24 (39%) were positive also for anti-c22 IgM. Of 24 patients who received a blood transfusion 9 (37.5%) were positive for anti-c22 IgM. The mean serum alanine aminotransferase (ALT) activity was significantly higher in subjects with anti-c22 IgM than that in subjects without them (p = 0.006), however, the difference in aspartate aminotransferase (AST) was not significant (p = 0.09). Histological examination was performed in 46 patients. Two-thirds of the patients with anti-c22 IgM had either cirrhosis or chronic active hepatitis (CAH) while only one third of the anti-HCV-positive patients without anti-c22 IgM had CAH or cirrhosis. The results showed that approximately 40% of the patients with CAH and cirrhosis had anti-c22 IgM, a significantly higher serum ALT activity and more serious histological finding in the liver than anti-HCV-positive patients without anti-c22 IgM.

[Indicators of progression of HIV infection and their application to therapy]. / Ukazatele progrese infekce HIV a jejich vyuzití v terapii.

The authors evaluated in a group of 217 HIV positive patients the mutual relationship of the number of CD4+T lymphocytes and the level of the viral load of HIV RNA. Using correlation analysis evidence was provided of a not very marked negative correlation of the two indicators. As it was assumed that the relationship of the two parameters is influenced by the applied therapeutic procedures which reduce in particular the viral load, the two parameters were evaluated in relation to treatment. The closest relationship of the two investigated parameters was found in the group treated by monotherapy with zidovudine, followed by the group treated with a combination of two preparations (two nucleoside inhibitors of reverse transcriptase). In the group treated by three preparations (two nucleoside inhibitors of reverse transcriptase and a protease inhibitor) the relationship of the two parameters was least close. Investigation of the level of the viral load of HIV RNA and number of CD4+T lymphocytes is of major importance for the introduction of antiretrovirus treatment and selection of a suitable combination of antiretrovirus preparations. It makes it also possible to follow up the effectiveness of this treatment.

[Clinical significance of IgM anti-HCV determination in chronic hepatitis C]. / Klinický význam stanovení IgM anti-HCV u chronické hepatitídy C.

BACKGROUND: In the majority of patients with acute viral hepatitis C the early antibody IgM anti-HCV in serum is positive. However, a substantial portion of the patients with chronic hepatitis C has also positive IgM anti-HCV as a sign of the continuing replication of the virus. The objective of the work was to assess the presence of IgM anti-HCV in patients with confirmed chronic hepatitis C, in subjects with HBsAg negative chronic hepatitis and in excluded blood donors. Moreover, the authors assessed the relationship between IgM anti-HCV positivity and the activity of serum transaminases and whether the presence of IgM anti-HCV has an impact on the histological finding in the liver. METHODS AND RESULTS: 88 patients were examined (44 women and 44 men), average age 48 years. In 47 subjects histological examinations of the liver were made. IgG anti-HCV were assessed by the Monolisa anti-HCV Sanofi Pasteur test and IgM anti-c22 by an Abbott kit IgM HCV EIA: With regard to the results of the serological examination the patients were divided into three groups which were mutually compared. Group 1 comprised 24 patients with a positive IgG and IgM anti-HCV, group 2 38 patients with a positive IgG anti-HCV only and group 3 26 patients with a negative IgG and IgM anti-HCV. Of 88 examined patients 62 had positive IgG anti-HCV (70%). Of 62 IgG anti-HCV positive subjects 24 (39%) had positive IgM anti-c22. A total of 24 patients had blood transfusions (39%) but only 9 of them had positive IgM anti-c22 (37.5%). The mean ALT serum activity was significantly higher in subjects with positive IgM than in those without IgM (p = 0.006), however, for AST the difference was not significant (p = 0.09). Comparison of patients with a confirmed histological finding in the liver revealed that two-thirds of patients with a positive IgM anti-c22 either suffered from cirrhosis or chronic active hepatitis, while anti-HCV positive patients without IgM anti-c22 had cirrhosis or chronic active hepatitis only in one third of the cases. CONCLUSIONS: The results suggest that in chronic hepatitis C some 40% of the patients have positive IgM anti-c22; these subjects have a significantly higher serum ALT activity and a more advanced histological finding in the liver than subjects without IgM anti-c22. Assessment of IgM anti-c22 is important not only for diagnosis but also for treatment of chronic HCV infection with antiviral drugs.

[A relapsing and protracted form of viral hepatitis A: comparison of adults and children]. / Relapsující a protrahovaná forma virové hepatitidy A: porovnání dospelých s detmi.

In recent years in children and adults with acute viral hepatitis A relapses and a protracted course of the disease were described. The authors followed up 37 patients with viral hepatitis A (20 children and 17 adults) and compared the clinical course of the disease, the period of hospitalization, persistence of IgM anti-HAV antibody in serum, the incidence of relapses and protracted forms of the disease. In adults the mean hospitalization period was longer (28 days as compared with 19), the average serum bilirubin value was higher (94 mumol/l as compared with 51 mumol/l), there were more cases with obvious jaundice (59% as compared with 30%) and the early serum antibody IgM anti-HAV persisted longer (19 weeks as compared with 14 weeks). Relapses of the disease were equally frequent (12% vs. 10%), however adults had more often a protracted course of hepatitis (23.5% vs. 10%). The observed differences were not statistically significant. Almost half the cases of hepatitis affected several members of the family. The results suggest that viral hepatitis A in adult age has a more severe course than in children. The authors recommend in cases with an elevated transaminase serum activity more frequent check-up examinations to avoid missing of a relapse, and to examine repeatedly IgM anti-HAV as in protracted forms of hepatitis IgM anti-HAV may persist even when the transaminase activity is normal.

Infection-associated hemophagocytic syndrome complicated by infectious lymphoproliferation: a case report.

The case of a 7-year-old boy with virus-associated hemophagocytic syndrome (VAHS) and serologically proven parvovirus B-19 infection is described. The patient with VAHS presented with fever, hepatosplenomegaly, pancytopenia, and hyperlipidemia type IV. After induction therapy with VP-16 and prednisone, partial remission was achieved. Despite maintenance therapy, reinductions, and the addition of cyclosporine A for 3 months, several relapses occurred. The therapy was stopped because of life-threatening complications (Klebsiella sepsis, neutropenic enterocolitis, and stercoral peritonitis). The complications were treated successfully. The patient status was stabilized after splenectomy. However, hepatomegaly progressed slowly and the hyperlipidemia endured. Ten months after the diagnosis leukocytosis with absolute T lymphocytosis appeared. Reactivation of VAHS was suspected and intravenous immunoglobin and then antilymphocyte immunoglobulin ALG therapy were started. The resultant decrease in leukocytosis was prompt, but lymphopenia did not occur. Virostatic treatment with foscarnet was introduced based on human herpesvirus-6 seroconversion. Twenty-six months after the diagnosis, the patient is well, without any sign of VAHS or lymphoproliferation.