Sexual Transmission of Hepatitis Viruses: In order to determine the contribution of sexual transmission of the various hepatitis virus types, 191 heterosexual patients with sexually transmitted diseases (STD) and 390 well-matched blood donors were analysed for the prevalence of hepatitis A, B, C and D viruses in their serum. As compared to blood donors, STD patients had a significantly higher risk to acquire types A (47,0 vs. 20,8%), B (30,9 vs. 4,1%) and C (4,7 vs. 0,5%) hepatitis. An acute hepatitis A or B infection was not recognized on the basis of lgM-anti-HAV or lgM-anti-HBV tests. Probably due to its higher concentration in the serum, HBV is transmitted sexually more effectively than HCV. Whether the increased prevalence of HAV in STD patients is directly related to sexual transmission or other concurrent risk factors for this disease (e.g. travelling) is uncertain. The study clearly shows the need for active immunization against hepatitis A and B in STD patients and promiscuous heterosexual persons. Condoms may prevent spread of HCV similar to HIV.

Sulphoevernan is a sulphated alpha-1----3, 1----4 polyglucan (Mr 20,000) with a helical structure. This compound effectively inhibits both human immunodeficiency virus type 1 (HIV-1) and type 2 infection of cells in vitro at concentrations around 0.5 micrograms/ml. Moreover, the compound completely inhibits HIV-1-induced syncytium formation at a concentration of 1 microgram/ml. Competition experiments with 35S-labelled sulphoevernan revealed that the mannose-specific lectin from Narcissus pseudonarcissus prevented binding of sulphoevernan to HIV-1, whereas the antibody OKT4A did not reduce the amount of sulphoevernan bound to MT-2 cells. These data indicate that the non-cytotoxic polymer sulphoevernan binds to the virus rather than to the host cell. In vivo studies, using Rauscher leukaemia virus in NMRI mice, revealed that, at a daily dose of 20 mg/kg, the animals were protected against virus-induced increases in spleen weight. From these in vitro and in vivo data we conclude that sulphoevernan has potential in the treatment of acquired immunodeficiency syndrome.

Different groups of patients were analysed for antibody to hepatitis-C-virus (anti-HCV). A high prevalence was found in individuals with parenteral exposure (chronic Non-A, Non-B hepatitis, 77.5%, drug addicts 84.5%), while blood donors had a prevalence of 0.51%, this was significantly higher in patients with chronic type B hepatitis (30%), in homosexuals (22.5%) and in patients with different types of autoimmune hepatitis (57.2%). This indicates that differential diagnosis of chronic hepatitis and indications for alpha interferon therapy is not possible by simply anti-HCV testing. Further studies are required to establish the diagnostic value of the anti-HCV test.

The mRNAs of transiently expressed cytokine genes contain AUUUA-rich sequences in the 3' untranslated regions. In order to examine whether the AU-specific endoribonuclease V (EC 3.1.27.8) described previously by us transinactivates those mRNA species, we introduced a 51-nucleotide ATTTA sequence from tumor necrosis factor into the 3' untranslated region of beta-globin gene. Transcripts of that construct, synthesized in vitro, were prone to endoribonuclease V digestion at those AU-rich sequences. Stimulation of human macrophages with lipopolysaccharide resulted in a shift of the association state of the enzyme from the nuclear matrix-associated to the free form. This shift was strongly prevented by the hepatitis B surface antigen (HBsAg) and more weakly by hepatitis B nucleocapsid antigen and hepatitis B antigen of the X region. HBsAg and, to a lesser extent, hepatitis B nucleocapsid antigen and hepatitis B antigen of the X region inhibited the release of alpha interferon, tumor necrosis factor alpha, and granulocyte-macrophage colony stimulating factor, while it had no effect on interleukin-1 production from stimulated macrophages. Using the human hepatoma cell line PLC/PRF/5, we provide further experimental evidence that endoribonuclease V acts in trans as a posttranscriptional inactivator for nuclear matrix-associated cytokine transcripts. These results suggest that those cytokine transcripts which contain reiterated (overlapping) AUUUA sequences are degraded by nuclear matrix-associated endoribonuclease V. This degradation was comparably high in cells incubated with HBsAg or cells which produced this antigen.

Human PBMC from HIV-1-infected individuals produced ex vivo in response to vesicular stomatitis virus only low amounts of IFN-alpha. This impairment was significant as early as Walter Reed (WR) stage 2; at WR stage 4-5, the production was almost zero. At WR stage 2 of infection, IFN-alpha mRNA was exclusively found in association with polyribosomes, indicating that IFN-alpha gene was transcriptionally inactive under the experimental conditions used. A similar decrease of the level of transcripts as a function of the progression of the disease was also observed for the IFN-gamma mRNA. In contrast, TNF-alpha production was strongly enhanced in PBMC from HIV-1-infected individuals after stimulation with LPS compared to the TNF-alpha production of activated PBMC from healthy donors. Almost parallel with the increase of the level of the transcript for TNF-alpha, the level of TNF-beta increases as well. Data are presented which show that the increased TNF-alpha production is due to a longer half-life of TNF-alpha transcripts in PBMC from infected individuals. These results let us suggest that the up-regulation of TNF-alpha gene expression in PBMC from HIV-infected individuals is controlled predominantly on the posttranscriptional level, whereas transcriptional events regulate the level of IFN-alpha transcripts. This assumption is supported by run-on experiments which revealed that the extent of transcription of TNF-alpha gene is almost identical in nuclei from stimulated PBMC of noninfected and HIV-infected donors, whereas the transcription of IFN-alpha gene is strongly suppressed in nuclei from HIV-infected individuals at WR stages 3 and 6.

The production of interferon (IFN) from cultured peripheral blood mononuclear cells (PBMC) after virus or mitogen stimulation was evaluated in 141 patients positive for antibodies to human immunodeficiency virus type 1 (HIV-1). IFN-alpha production by PBMC of patients at Centers for Disease Control (CDC) stage II (Walter Reed [WR] 1) was comparable to that produced by PBMC of healthy controls. However, cells of patients at early CDC stage III (WR 2) produced significantly lower titers of IFN-alpha (P less than .001), and IFN-alpha was almost absent at CDC stage IV (WR 6) (P less than .001). IFN-gamma production was altered in patients at late CDC stage III (WR 4-5) and CDC IV (WR 6). A strong correlation between the disappearance of antibodies to core proteins and low IFN-alpha level was observed. IFN-alpha levels were significantly diminished in patients positive for HIV antigen. Reduced IFN-alpha production paralleled the HIV-1-related depletion of CD4+ lymphocytes and might serve as an additional parameter in defining the stage of HIV-1 infection.

For more than two years, all HIV-positive women were seen in the Department of Obstetrics and Gynaecology in cooperation with the Department of Internal Medicine of the University of Mainz, for medical care and therapy, if necessary. Of 40 HIV-infected female patients who were treated, 25 women were followed up every three to six months. Most of the latter live in rural areas or outside the cities in the State Rhineland Palatinate, FRG. 52% of them are or were former intravenous drug abusers, and 44% had been infected by sexual transmission. Only one-third use condoms during intercourse, another third reject this kind of protection. The rest are without cohabitation for various reasons. 23 pregnancies are known in this group and in 20% of the cases HIV infection were discovered during prenatal care. Gynaecological and obstetrical findings are reported. 80% of the patients have had a recurrent candida infection of the vagina, which was detected by culture. Immunologically, 75% of the patients belong to stage III (according to the CDC classification). Out of this group, more than 50% show less than 400 CD4 (+) cells/microliters. A positive result for HIV-Ag, anti-p41 and negative anti-p24 was seen in five cases. One of the four patients with AIDS died of a rapidly growing cancer of the cervix.

Six individuals with protracted acute type B hepatitis were treated with recombinant alpha-A-interferon for 12 weeks. Two females eliminated the HBV completely, while 4 males did not respond. Response was preceded by a flare-up of the liver disease. It appears that responses to interferon are not higher in protracted acute type B hepatitis than in progressed chronic active hepatitis B. This assumption has to be proven in larger studies on a statistical basis.

Hepatitis B virus and the human immunodeficiency virus are similarly transmitted. Individuals with preexisting HIV infection have a higher chance to become HBsAg carriers than do anti-HIV negative persons. Cytotoxic T cells with specificity for HBcAg, that are under the control of HBcAg-specific helper T cells, are responsible for liver injury. There is good evidence that HIV infection lowers inflammatory activity, is associated with milder liver histology, high levels of viral replication and low seroconversion rates. In addition interferon alpha therapy is less effective in anti-HIV positive subjects. The immune response against HBsAg is helper T-cell dependent and vaccination against hepatitis B is of low effectiveness. In addition, vaccination against hepatitis B may activate the HIV disease and is, therefore, presently not to be recommended.

Recombinant human α-interferon is now under intensive investigation as therapy for chronic Type B hepatitis. Recent reports have suggested that prolonged α-interferon therapy may induce autoimmune reactions. We have evaluated the problem of autoimmunity related to α-interferon therapy by testing for 15 different antibodies in the sera of 31 patients treated with α-interferon. No patient had autoantibodies before treatment; 27 (87%) of 31 patients developed at least one autoantibody. Eleven patients had antinuclear antibodies and 21 had smooth muscle antibodies, both of which usually developed during α-interferon therapy. In contrast, antibodies to endocrine organs such as thyroid microsomal, thyroglobulin and parietal cell antibodies arose in 12 patients, but usually several months after α-interferon treatment. The appearance of these autoantibodies did not correlate with disease activity or response to α-interferon. No patient developed autoantibodies specifically associated with autoimmune liver diseases such as liver kidney microsomal antibodies, autoantibodies to soluble liver antigen and the primary billiary cirrhosis-specific subtypes of antimitochondrial antibodies. These results suggest that prolonged α-interferon therapy can induce autoantibody production and, in susceptible patients, may lead to autoimmune disorders.

Six patients with chronic type B hepatitis and concurrent infection with the immunodeficiency virus were treated with 600 mg azidothymidine (AZT)/day and 3 X 10(6) units of interferon-alpha (IFN-alpha) every other day for a total of 4 months. None of the patients treated lost the hepatitis B virus (HBV). HBV-DNA concentrations were not significantly influenced by this treatment. Human immunodeficiency virus (HIV) infection was also not affected except for a transient rise in CD 4-positive cells in 2 individuals, who had initially low CD 4-positive cells. Treatment did not influence the presence of HIV-Ag in the serum. In conclusion, a combination therapy of IFN and AZT does not seem to be beneficial at the doses given and the time involved.

Twenty homosexual men [13 anti-human immunodeficiency virus (HIV)-positive, seven anti-HIV negative] without HBsAg, anti-HBs, and anti-HBc were vaccinated with three 20 micrograms doses of a recombinant hepatitis B vaccine. All anti-HIV-positive homosexuals were nonresponders independent of the initial number of CD4-positive cells. Among seven anti-HIV-negative individuals, five responded. After three doses of the vaccine, CD4-positive cells fell in anti-HIV positive individuals by 22.4%. A similar fall in CD4-positive cells of an average 24.9% was noted in 17 matching, but nonvaccinated, anti-HIV-positive homosexuals. The study indicates that the efficacy of vaccination in anti-HIV-positive individuals is questionable. There is, however, no evidence that vaccination against hepatitis B might be harmful to anti-HIV-positive subjects.

The natural killer (NK) cell activity of mice in the peritoneal cavity is very low or undetectable and testing peritoneal NK cells is a useful model for studying the influence of activating substances upon local injection. Injection of tumor necrosis factor (TNF) at doses of 10–200 ng caused a marked activation of NK cell activity which was maximal after 24 h and declined rapidly on day 2. A similar effect was observed when interferons alpha and beta were injected, and there were additive results when interferon was injected together with TNF. The NK cell nature of the effector cells activated by TNF was substantiated by the finding that previous injection with anti-asialo GM 1 antibody prevented activation. Interferon could not be detected in the peritoneal wash fluid after injection of TNF suggesting interferon-independent activation. In further experiments after i.p. injection of TNF peritoneal exudate cells (PECs) only killed YAC-1 targets in a 4-h assay. There was no additional killing in an 18-h assay towards neither YAC-1 cells or P815 cells, suggesting that macrophages were not involved. Furthermore TNF was also active in vitro by activating NK cells in isolated human peripheral blood cells. However in the PECs stimulated in vitro no significant induction of cytotoxic capacities by TNF was measured. Our data suggest that the action of TNF is not restricted to the lysis of tumor cells but can also induce immunological properties in the host defense against virus infections and neoplasms.

Avarol is a cytostatic and anti-human immunodeficiency virus (HIV) agent. In this study, the avarol caused induction of gamma-interferon (IFN-gamma) in buffy coat cells (human peripheral blood lymphocytes) is demonstrated by immunological and molecular biological techniques. IFN-gamma production was detected after a 24-hr incubation period with avarol; maximal production was obtained after 5 days in the presence of the optimal avarol concentration of 0.75 microgram/ml. Blotting experiments using human IFN-gamma cDNA and beta-actin cDNA containing plasmids showed that in the absence of avarol no IFN-gamma transcripts were present in lymphocytes. Already after a 24-hr incubation with avarol, IFN-gamma gene induction was detected, and maximal induction was found after a 5-day incubation period. The enhanced IFN-gamma production seems to be caused by a change at the transcriptional and/or post-transcriptional level, but not during subsequent nucleocytoplasmic transport of mRNA. This molecular event is specific, at least in relation to the expression of the beta-actin gene. Our studies demonstrate that avarol displays, besides its potential anti-tumor and anti-HIV activity, a potential immunomodulating effect.