Prophylactic efficacy of reaferon in viral hepatitis A and acute respiratory infections in children

Reaferon, the analog of human alpha 2-interferon obtained by gene engineering techniques, was studied with a view to its use for the prevention of hepatitis A. The study involved children of preschool age in Tashkent. In a strictly controlled trial children aged 2-6 years received the preparation orally in a dose of 1 X 10(6) I. U. or the diluent alone used as placebo. The preparation was administered to 1,100 children and the placebo to 1,078 children. The preparation and placebo were administered twice a week for two months. On the whole, during that period hepatitis A morbidity in both test and control groups of children was the same (5.1% and 4.9% respectively), but among children of nursery age receiving Reaferon the incidence of hepatitis A and acute respiratory viral infections was lower than among those receiving the placebo, though this difference was statistically significant only for cases of acute respiratory infections.     

Influenza virus infection induces functional alterations in peripheral blood lymphocytes

This report describes alterations in functional responses to lectin-induced stimulation of peripheral blood lymphocytes and in the natural killer cell (NKC) activity, of college students, obtained during an outbreak of influenza A/Philippines/2/82(H3N2) virus infection. These results are compared with similar observations in college students with an acute, febrile, noninfluenzal respiratory illness that occurred during the same outbreak. The lymphopenia typical of influenza during acute illness was shown to be due to a reduction in both T and B cells without alteration in the CD4:CD8 ratio. In addition, phytohemagglutinin and concanavalin A responses were reduced and NKC activity was increased, while pokeweed mitogen reactivity was unaltered at the time of admission to the study. Patients with noninfluenzal illness showed early polymorphonuclear leukocytosis and a similar lymphopenia. Lymphocyte functions were virtually unchanged during acute illness in noninfluenza patients. The relatively specific alterations in lymphocyte responses to lectin-induced stimulation in influenza patients may indicate that the peripheral T cells are incapable of activation via the CD3 or CD2 activation pathways. In addition, increased NKC activity in the periphery may be reflective of increased NKC activity in the lung. Influenza-infected individuals with higher NKC activity at the time of admission to the study also took longer to recover. Finally, the early lymphopenia and the later neutropenia in the influenza-infected patient may represent migration of these cells from the circulation to the infected respiratory tract as a consequence of infection.     

Functional properties of lymphocyte subpopulations in hepatitis B virus infection. I. Suppressor cell control of T lymphocyte responsiveness

Hepatocellular injury in hepatitis B virus infection may be produced by an autoaggressive hepatocytotoxic immune response. To test the hypothesis that acquired suppressor cell defects may participate in such a response, we assessed the functional integrity of 2 suppressor cell populations in patients with type B viral hepatitis. Spontaneous suppression of the 1-way mixed lymphocyte response by radiation-resistant, adherent peripheral blood mononuclear cells decreases during the acute phase of disease, returns towards normal with clinical recovery, but remains depressed in patients with chronic hepatitis. The degree of spontaneous suppressor cell dysfunction correlates inversely with at least 1 biochemical parameter of hepatocellular injury (SGPT). The functional integrity of this suppressor cell fluctuates during chronic hepatitis and may reflect currently undefined biologic variables in this disease. Mitogen-induced suppression on lymphocyte activation by radiation resistant, nonadherent suppressor cells is also depressed in acute and chronic hepatitis, but it does not correlate with biochemical evidence of hepatocellular injury on an individual-patient basis. Documentation of these generalized defects of nonspecific suppressor cell function establishes a basis for the possible existence of specific anomalies of immuno-regulation that may permit the expression of normally suppressed auoaggressive hepatocytotoxic immune mechanisms in viral hepatitis.     

Plasma transforming growth factor beta1, metalloproteinase-1 and tissue inhibitor of metalloproteinases-1 in acute viral hepatitis type B

AIM: Antiproliferative, pro-apoptotic and immunosuppressive activity effects suggest crucial role of transforming growth factor (TGF)-beta1, metalloproteinase (MMP)-1 and its tissue inhibitor (TIMP)-1 in the pathogenesis of acute liver injury that in some patients precede development of chronic liver diseases and fibrogenesis. The aim of this study was to evaluate effect of acute HBV infection on plasma TGF-beta1, MMP-1 and TIMP-1 levels. METHODS: TGF-beta1, MMP-1 and TIMP-1 plasma concentrations were measured with an enzyme immunoassay in 39 patients with acute viral hepatitis type B. Baseline measurement was performed within the first week of jaundice and then weekly up to the fourth week of the disease. Results were compared to baseline and normal values and to liver function tests. RESULTS: Plasma concentrations of TGF-beta1, TIMP-1 and MMP-1 were significantly elevated in the first week of acute viral B hepatitis in comparison to normal. Analysis of individual values demonstrated significant positive correlation between plasma concentrations of TGF-beta1 and TIMP-1. There was no correlation between MMP-1 and TGF-beta1 or TIMP-1. Significant correlation was demonstrated between both TGF-beta1 and ALT or AST as well as between TIMP-1 and ALT, AST or bilirubin. Elevated baseline levels of both TGF-beta1 and TIMP-1 decreased gradually in consecutive weeks of the disease. TGF-beta1 but not TIMP-1 plasma concentrations were significantly lower in 3rd and 4th week than baseline values. MMP-1 concentration remained on baseline level in the 2nd week of the disease. However in the 3rd week its values increased suddenly but the significant difference in comparison to baseline was observed only in 4th week. CONCLUSIONS: These results indicate important role of TGF-beta1, TIMP-1 and MMP-1 in acute viral hepatitis, that seems to be connected first of all with hepatocytes damage. Their role in extracellular matrix metabolism during acute liver injury needs further evaluation.     

Functional and metabolic activity of peripheral blood neutrophils in acute viral hepatitis B and C

The functional metabolic activity of peripheral blood neutrophils in acute virus hepatitis B (VHB) and/or virus hepatitis C (VHC) was evaluated. 48 patients were examined; of these, VHB was diagnosed in 28 patients and VHC was diagnosed in 9 patients and the mixed form of virus hepatitis (VHB + VHC), in 11 patients. Determination of adhesive capacity of neutrophils, production of superoxidase anion in the nitro blue tetrazolium (NBT) test, activity of myeloperoxidase (MPO) and acidic phosphatase (AP), the amount of cation proteins (CP) was made. Most pronounced functional dysbalance of neutrophil leukocytes and considerable changes in biochemical characteristics of the activity of the infectious process in patients with the mixed form of virus hepatitis were established. These data demonstrated that in acute virus hepatitis B and C at the peak of the disease such characteristics of the functional activity of neutrophils as results of the NBT test, the activity of MPO and AP, as well as the amount of CP, were highly informative.     

Virus-specific CD8+ lymphocytes share the same effector-memory phenotype but exhibit functional differences in acute hepatitis B and C

Hepatitis B and hepatitis C viruses (HBV and HCV) are both noncytopathic and can cause acute and chronic infections of the liver. Although they share tropism for the same organ, development of chronic hepatitis is much more frequent following HCV infection, suggesting different mechanisms of viral persistence. In this study, we show that circulating HBV- and HCV-specific tetramer-positive CD8 cells during the acute phase of hepatitis B and C belong almost entirely to an effector-memory subset (CCR7(-) CD45RA(-)). Despite this phenotypic similarity, HBV- and HCV-specific CD8 cells show striking functional differences. HBV-specific tetramer-positive CD8 cells express high perforin content ex vivo, expand vigorously, and display efficient cytotoxic activity and gamma interferon (IFN-gamma) production upon peptide stimulation. A comparable degree of functional efficiency is maintained after the resolution of hepatitis B. In contrast, HCV-specific CD8 cells in the acute phase of hepatitis C express significantly lower levels of perforin molecules ex vivo and show depressed CD8 function in terms of proliferation, lytic activity, and IFN-gamma production, irrespective of the final outcome of the disease. This defect is transient, because HCV-specific CD8 cells can progressively improve their function in patients with self-limited hepatitis C, while the CD8 function remains persistently depressed in subjects with a chronic evolution.     

Evaluation of free and IgG-bound components of e antigen (HBeAg) in acute virus B hepatitis

Soluble (free) and insoluble (IgG-bound) hepatitis B e antigen (HBeAg) activities in 1,33 M ammonium sulfate solution was evaluated serially in sera from 12 patients with acute type B hepatitis followed up to 60 days. In three cases preclinical sera were available. In patients who showed clinical and biochemical resolution of hepatitis within 60 days, HBeAg free was the prevalent form of circulating HBeAg during the incubation period of disease, then gradually shifting to the IgG-bound form at the onset of jaundice. Finally HBeAg-bound was no longer demonstrable, followed by detection of anti-HBe antibody. On the contrary, in all patients with unresolved hepatitis both HBeAg-free and HBeAg-bound were detected with prevalence of HBeAg-free ratio. These data suggest a striking correlation between synthesis of HBeAg and HBV replication and may indicate that HBeAg-free and IgG-bound ratio reflects the stage and prognosis of acute HBV infection.     

Biological activity of human alpha-interferons studied using specific antisera

Preparation of highly active rabbit antisera (AS) to human recombinant alpha 2-interferon and their use for studying biological properties of natural and plasmid alpha-interferons are described. By exhaustion of AS by alpha 3-interferon there were prepared practically monospecific AS not reacting with antigenic determinants of alpha 3-interferon. It was found that alpha 3-interferon represented a significant portion of human lymphoblastoid interferons and was included in PH-labile alpha-interferon from serum of patients with Kaposi carcinoma. AS to alpha 2-interferon completely neutralized antiviral and antiproliferative activity of the homologous subtype alpha-interferon and stimulation of cytotoxicity of human natural killer cells induced by it. It neutralized also the same effects of the heterologous subtypes (alpha 3 and alpha F/D) and leukocytic interferon, but the neutralization level was lower. The results of the study confirmed the polyfunctional nature of the interferon molecule.     

Viral hepatitis with clinical and epidemiological manifestations similar to hepatitis A but of a different etiology

A case of viral hepatitis in man, appearing as the result of infection caused by pooled concentrated suspensions of fecal samples collected from patients having had repeated hepatitis infection during the period of 1-2 years, is described. Though this infection was similar to hepatitis A in many clinical and epidemiological signs, the possibility of its etiologic relationship with hepatitis virus A was positively excluded; there was also no evidence of the participation of hepatitis virus B in the process. Immunoelectron microscopy of excretions collected at the acute stage of the disease revealed the presence of spherical viral particles 27-30 nm in diameter. Antibodies capable of reacting with these particles were detected in the sera of patients having had 2 kinds of hepatitis and in the sera of patients having the 1 kind of hepatitis in the focus of infection where repeated cases of hepatitis had been observed. No such antibodies were found in the sera of patients with hepatitis A alone and in the set of standard sera specific to viruses causing hepatitis A and hepatitis non A, non B. The authors believe that 2 kinds of hepatitis with the fecal-oral mechanism of transmission exist and propose to name their causative agents hepatitis viruses A, type 1 and type 2.     

Indices of immunity and allergy in patients with viral hepatitis treated with prodigiozan

The immune and allergic status of patients with acute viral hepatitides A and B after treatment with prodigiosan, a bacterial lipopolysaccharide preparation, was studied. The drug produced as stimulating effect on cell-mediated and humoral immunity. The development of delayed specific drug sensitization was registered in two-thirds of the patients, which correlated with the regression of clinicobiochemical manifestations of hepatitis.     

Functional properties of lymphocyte subpopulations in hepatitis B virus infection. II. Cytotoxic effector cell killing of targets that naturally express hepatitis B surface antigen and liver-specific lipoprotein

Cytotoxic effector cell responsiveness to host and/or virus-determined hepatocyte surface membrane antigens has been postulated as an important pathogenetic determinant of hepatocellular injury in hepatitis B virus infection. Assuming that such effector cell populations would be detectable in peripheral blood, the present study was designed to examine 2 questions: first, whether target cells that normally express liver-specific protein (LSP) and hepatitis B surface antigen (HBsAg) are selectively destroyed by peripheral blood effector cells from patients with viral hepatitis; second, whether cytotoxic effector cell function emerges coincident with the development of defective suppressor cell activity in the same patients. No evidence of increased HBsAg or LSP specific cytotoxic effector cell activity was found in the peripheral blood natural killer (NK) or T killer cell populations of patients with acute or chronic viral hepatitis.     

Anti-pre-S2 antibody response in subjects vaccinated against hepatitis B and in naturally immunized subjects

Serum samples from individuals immunized with a pepsinized or non-pepsinized vaccine and from patients who had recovered from acute hepatitis B or who developed a chronic form of the disease, were analysed for the presence of antibody against the pre-S2 epitope of the hepatitis B virus.Anti-pre-S2 antibody was absent in all but one individual immunized with the pepsinized vaccine. Thirty-eight percent of the subjects who responded by anti-HBs production to the non-pepsinized preparation showed anti-pre-S2 antibody one year after complete vaccination. Among subjects who did not produce anti-HBs after immunization with this vaccine, 1 single individual produced anti-pre-S2 antibody. Anti-preS2 antibody was detectable after one year in 38% of the patients who recovered from acute hepatitis B, but in none of those with chronic hepatitis B. The kinetics of anti-pre-S2 antibody response to a booster injection was also analysed 1 month and 1 year after the 3rd injection and 1 month after the 4th injection of the non-pepsinized vaccine.     

Plasmapheresis in the treatment of hepatic coma complicating viral hepatitis]

Therapeutic plasmaphereses using CS 3000 Fenwal Cell Separator were performed in 4 women and 2 men, aged between 17 and 44 years, with hepatic coma complicating acute viral hepatitis type B. One to four plasma exchanges per patient were performed, usually at the volume of 3000 ml per procedure. Two patients at II and IVa period of the coma, according to Aboun classification, survived. Four patients at II, III and two at III/IV period of the coma died. The authors suggest that in some cases exchange of large volumes of plasma in the treatment of hepatic coma complicating acute viral hepatitis may be a lifesaving procedure.     

Epidemic focus of non-A, non-B viral hepatitis in a plasmapheresis unit]

Non-A, non-B hepatitis has been diagnosed in 12 blood donors in a plasmapheresis unit. The course of the disease has been symptomatic, accompanied by jaundice, fatigue, and nausea in 8 cases, and subclinical in the remaining 4 patients. Nine patients were followed-up to 2 years and only 2 patients liver biochemical tests were normalized permanently. The biopsies performed, a year after the acute phase of hepatitis period revealed chronic active disease in patients, chronic persistent hepatitis in 2 patients, acute hepatitis in one, and normal liver in one patient. Repeated liver biopsies, performed one year later, have basically shown similar lesions except one patient in whom chronic active hepatitis progressed to incipient liver cirrhosis. No symptoms of the disease have been usually noted in patients with chronic form of the disease, and liver function tests have occasionally been normal.     

Suppressor cell regulation of the spontaneously induced in vitro secondary antibody response.

The cytotoxic effect of peripheral blood lymphocytes from patients with chronic active hepatitis (CAH) on Chang liver cells was studied using a chromium-51 release assay. Cytotoxic effector cell activity was unaffected or slightly enhanced after the removal of glass-adherent mononuclear cells, indicating that the major effector cell is not a classical monocyte. Lymphocytes from 12 of 18 patients with CAH were cytotoxic, and the cytotoxic cells were contained within T-cell-depleted fractions. Further studies revealed that these effector cells were K cells, not B cells. K cells were identified as Fs- and complement receptor-bearing cells without detectable surface immunoglobulin. Serial studies in CAH showed continuous cytotoxicity in contrast to transient cytotoxicity in acute viral hepatitis, suggesting that the cytotoxic activity of lymphocytes is responsible for persistence of the disease.     

Recombinant human alfa2-interferon (reaferon) in ointment form in the treatment of various diseases of the skin and mucosa

Clinical trials of the gene engineered human alpha 2-interferon as ointment were performed. The ointment was used in the treatment of patients with relapsing herpes of various localization, relapsing aphthous stomatitis and pemphigus vulgaris. It was shown that the ointment was harmless and its therapeutic effect was favourable. The preparation is recommended for introduction into the practice of public health.     

Cellular immune response to hepatitis B virus-encoded antigens in acute and chronic hepatitis B virus infection

The proliferative response of PBMC to hepatitis B virus (HBV) envelope, core, and e Ag was analyzed prospectively in 21 patients with acute self-limited HBV infection and compared with the response of patients with chronic HBV infection and different levels of HBV replication (i.e., hepatitis e Ag (HBeAg)- or anti-HBe-positive) and liver damage (i.e., chronic active hepatitis or chronic asymptomatic carriers). Our results indicate that: 1) HBV-infected subjects who develop a self-limited acute hepatitis show a vigorous PBMC response to hepatitis B core Ag and HBeAg, as expression of T cell activation; 2) appearance of a detectable lymphocyte response to HBV nucleocapsid Ag is temporally associated with the clearance of HBV envelope Ag; 3) in patients with chronic HBV infection the level of T cell responsiveness to hepatitis B core Ag and to HBeAg is significantly lower than that observed during acute infection; 4) T cell sensitization to HBV envelope Ag in acute and chronic HBV infection is usually undetectable and when measurable is expressed transiently and at low levels. These results may reflect immune events of pathogenetic relevance with respect to evolution of disease and viral clearance.     

Alpha1 Na,K-ATPase and Na,K,2Cl-cotransporte/D3mit3 loci interact to increase susceptibility to salt-sensitive hypertension in Dahl S(HSD) rats

BACKGROUND: Essential (multigenic) hypertension is a complex multifactorial disease whose genetic etiology has not been unraveled on a major locus-effect investigative paradigm. As with other complex genetic diseases, applying an interacting loci paradigm could be critical in the elucidation of genetic determinants. Having defined the alpha1 Na,K-ATPase (alpha1NK) as a hypertension susceptibility gene in Dahl salt-sensitive (Dahl S) rats, we determined whether alphaINK interacts with another renal epithelial Na transporter to increase susceptibility to salt-sensitive hypertension. We focused on alpha1NK and Na,K,2Cl-cotransporter (NKC) as an a priori candidate interacting gene pair because they comprise a functionally linked Na transport system in renal thick ascending limb of Henle (TALH) epithelial cells and exhibit altered function in prehypertensive Dahl S rats in contrast to Dahl salt-resistant normotensive (Dahl R) rats. MATERIAL AND METHOD: Cosegregation analysis of alphaNK and NKC loci was done in a (Dahl S x Dahl R) F2 cohort characterized for blood pressure by radiotelemetry using the D2mghII microsatellite marker in the alpha1NK gene and the D3mit3 microsatellite marker close to the NKC gene (NKC/D3mit3 locus). Single locus and digenic analyses were performed to establish the individual and interactive genetic contribution to salt-sensitive hypertension. Molecular analysis was then done to support the NKC gene as the likely candidate gene interacting with alpha1NK in Dahl salt-sensitive hypertension pathogenesis. RESULTS: Compared with respective single locus analysis, digenic analysis of 96 F2 (Dahl S x Dahl R) hybrid male rats revealed cosegregation of alpha1NK and NKC/D3mit3 loci as interacting pair with salt-sensitive hypertension with markedly increased significance for systolic (one-way ANOVA p = 10(-6)), diastolic (p = 10(-5)), and mean arterial (p = 10(-6)) blood pressures. Concordantly, two-way ANOVA detected interaction between alpha1NK and NKC loci in determining the levels of systolic (p = 0.004), diastolic (p = 0.008), and mean arterial (p = 0.006) pressures. To unravel potential NKC molecular dysfunction(s) involved in hypertension pathogenesis, we investigated putative differences between Dahl S and Dahl R rats in nucleotide sequence and isoform gene expression of the renal-specific Na,K,2Cl-cotransporter. Molecular analysis revealed an inversion of alternatively spliced NKC-isoform ratios (4B:4A:4F) between Dahl S and Dahl R prehypertensive kidneys supported by four mutations in intron-3 immediately upstream to alternatively spliced exons 4B, 4A, and 4F. No nucleotide changes were detected within the aminoacid encoding exons of NKC. CONCLUSIONS: Altogether, these current data and previous characterization of the role of the Q276L alpha1NK molecular variant in Dahl S hypertension provide cumulative compelling evidence that alpha1NK and NKC/D3mit3 loci interact to increase susceptibility to hypertension in Dahl S rats and that NKC is the likely candidate gene that interacts with alpha 1NK. More importantly, the data substantiate gene interaction as an operative mechanism in multigenic hypertension.     

Intrinsic functional dysregulation of CD4 T cells occurs rapidly following persistent viral infection

Effective T-cell responses are critical to eradicate acute viral infections and prevent viral persistence. Emerging evidence indicates that robust, early CD4 T-cell responses are important in effectively sustaining CD8 T-cell activity. Herein, we illustrate that virus-specific CD4 T cells are functionally inactivated early during the transition into viral persistence and fail to produce effector cytokines (i.e., interleukin-2 and tumor necrosis factor alpha), thereby compromising an efficient and effective antiviral immune response. Mechanistically, the inactivation occurs at the cellular level and is not an active process maintained by regulatory T cells or antigen-presenting cells. Importantly, a small subpopulation of cells is able to resist inactivation and persist into the chronic phase of infection. However, the virus-specific CD4 T-cell population ultimately undergoes a second round of inactivation, and the cells that had retained functional capacity fail to respond to rechallenge in an acute time frame. Based on these results we propose a biological mechanism whereby early CD4 T-cell inactivation leads to a subsequent inability to sustain cytotoxic T-lymphocyte function, which in turn facilitates viral persistence. Moreover, these studies are likely relevant to chronic/persistent infections of humans (e.g., human immunodeficiency virus, hepatitis C virus, and hepatitis B virus) by providing evidence that a reservoir of virus-specific CD4 T cells can remain functional during chronic infection and represent a potential therapeutic target to stimulate the immune response and establish control of infection.     

Effect of tactivin in vitro on lymphocytes from patients with acute hepatitis B

In vitro experiments have revealed that Tactivin increases the number of lymphocytes and their functional activity in patients with acute virus hepatitis B. Under the action of Tactivin the increase of theophylline-resistant and theophylline-sensitive T-lymphocytes occurs in patients with acute virus hepatitis B in a moderate or severe form. In a mild form of acute virus hepatitis B and in healthy persons Tactivin produces no in vitro changes in T-lymphocyte subpopulations.