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Biomedical subjects

G Haukenes

Publications and source records attributed to G Haukenes.

At least 37 records · Page 2Linked to original sources

Radioimmunoprecipitation and immunoblot studies of antibodies to rubella virus in patients with chronic liver disease.

Patients with autoimmune chronic active hepatitis (AICAH) and some other chronic liver disorders often have very high titres of rubella HI antibodies. In the present study sera from 46 patients with chronic liver disease and controls were examined for rubella antibodies using radioimmunoprecipitation assay (RIPA) and Western blot. RIPA appeared to be more suitable than Western blot for the study of the individual antibody specificities provided that proteins (possibly actin) interfering with the resolution of the E2 glycoprotein band are identified. It was shown that patients with high rubella HI titres reacted strongly against the E1 glycoprotein and in general also against the core protein (C). Reactivity to the E2 glycoprotein was detected with all sera from patients with chronic liver disease but varied more in strength. Three patients with post-acute rubella showed very faint E2 reactivity, but strong E1 and C reactivities. Patients with primary biliary cirrhosis had normal HI titres and showed no increase in reactivity in RIPA. The present findings show that patients with chronic liver disease and high rubella HI antibody titres exhibit an enhanced specific antibody response to rubella virus structural proteins.

Animals↗

Viral antibodies in infectious mononucleosis.

Patients with Epstein-Barr virus (EBV) infectious mononucleosis (IM) usually develop heterophilic antibodies and some autoantibodies. Antibodies to rubella, measles, adeno-, entero-, herpes simplex, cytomegalo- and varicella-zoster viruses were titrated in sera from IM patients and matched healthy controls using the complement fixation test (CFT) and the haemagglutination inhibition test. Except for herpes simplex virus and cytomegalovirus, the IM sera had significantly higher arithmetical and geometrical mean antibody titres and showed in most cases higher antibody prevalences in the CFT. The titre rise was most pronounced for rubella and measles antibodies, between 2- and 3-fold. There were no cases of very high titres occasionally seen in IM. The IM sera had higher total IgG serum levels than the controls, 17.27 g/l and 11.8 g/l, respectively (P < 0.001). The present data show that in addition to previously reported high levels of some autoantibodies and of heterophilic antibodies, there is a more general increase in IgG antibodies to commonly occurring viruses. This increase is most likely due to the polyclonal activation of B-lymphocytes following the binding of EBV to the complement receptor CR2 (CD21). When due consideration is given to the possible occasional occurrence of a false positive rubella IgM test, the raised antibody-titres will most likely not interfere with routine diagnostics.

Antibodies, Viral↗

Subcellular distribution of human immunodeficiency virus type 1 Rev and colocalization of Rev with RNA splicing factors in a speckled pattern in the nucleoplasm.

The human immunodeficiency virus type 1 (HIV-1) Rev (regulator of virion protein expression) protein exemplifies a new type of posttranscriptional regulation. One main function of Rev is to increase the cytoplasmic expression of unspliced and incompletely spliced retroviral mRNAs from which viral structural proteins are made. In that way, Rev is essential in order to complete the retroviral life cycle. The biology of Rev in the host cell has remained elusive. In this study, a complex distribution of Rev in single cells was found. Rev was found in the cytoplasm, in a perinuclear zone, in the nucleoplasm, and in the nucleoli. In the nucleoplasm, Rev colocalized in a speckled pattern with host cell factors known to assemble on nascent transcripts. Those factors are involved in the processing of heterogeneous RNA to spliced mRNA in the nucleoplasm of all cells. The distribution of Rev was dependent only on Rev and host cell interactions, since neither the Rev target RNA nor other HIV proteins were expressed in the cells. Rev was found in the same subcellular compartments of cells treated for extended periods with cycloheximide, an inhibitor of protein synthesis. This finding implies that Rev shuttles continuously between cytoplasmic and nucleoplasmic compartments. The results suggest a potential role for Rev both in the RNA-splicing process and in the nucleocytoplasmic transport of Rev-dependent HIV mRNA.

Amino Acid Sequence↗

The human immunodeficiency virus type 1 Rev protein shuttles between the cytoplasm and nuclear compartments.

A retroviral regulatory protein, Rev (regulator of virion protein expression), is made in cells infected by human immunodeficiency virus (HIV). Rev is essential for the completion of the retroviral life cycle and interacts with the host cell at some posttranscriptional step in order to express the incompletely spliced HIV mRNAs from which HIV structural proteins are translated. Neither the host cell components nor the mechanisms responsible for this important regulation have been defined. We now report that Rev is a nucleocytoplasmic shuttle protein which is continuously transported between the cytoplasm, the nucleoli, and nucleoplasmic speckles enriched in RNA splicing and processing factors. The results show that Rev has the potential to interfere specifically with the splicing of the HIV pre-mRNA in the nucleoplasm and, next, guide such mRNAs to the cytoplasm for translation.

Amino Acid Sequence↗

Association of hepatitis B and human immunodeficiency virus infections in Tanzanian population groups.

A study was performed to determine whether there is a correlation between hepatitis B virus (HBV) and human immunodeficiency virus (HIV) infection in population groups in the Dar es Salaam area in Tanzania where HBV infection is endemic. A panel of 460 sera from army recruits, health personnel and pregnant women was tested. In the whole group seromarkers of HBV infection were found in 61.9% of 134 HIV positive subjects versus 51.5% of 326 seronegative subjects, a difference which was not statistically significant (p > 0.05). In the group of pregnant women, however, 66.7% of 120 HIV positive subjects had markers of HBV infection versus 49.0% of the 157 HIV seronegative subjects (p < 0.01). This shows that a considerable proportion of young females are also exposed to HBV at the time they acquire HIV infection.

Female↗

Detection of antibodies to HIV in homologous sets of plasma, urine and oral mucosal transudate samples using rapid assays in Tanzania.

The aim of this study was to evaluate the performance of commercially available anti-HIV assays when testing plasma, urine and oral mucosal transudate (OMT) samples for the presence of antibodies to HIV. Homologous sets of plasma, urine and oral mucosal transudate specimens were collected from 288 hospitalized patients in northern Tanzania and tested for antibodies to HIV using a routine enzyme immunoassay (Recombinant 3rd Generation EIA, Abbott) and two rapid assays (Testpack HIV-1/HIV-2; Abbott and SUDS HIV-1, Murex). Incubation times and/or sample volumes when testing OMT or urine were increased as compared to those recommended for plasma. The corresponding plasma specimens from all repeatedly reactive samples and samples presenting discordant results were subjected to confirmational testing by an HIV-1/2 Western blot. A total of 15.3% (44/288) of the plasma samples were anti-HIV-1 positive by Western blot. The sensitivity using plasma was 100% by all assays, 69.7-97.7% using urine, and 92.7-100% using oral transudate specimens. The sensitivity of both rapid assays was excellent and higher than the EIA when testing OMT. Specificities ranged from 98.8-100% for plasma, 99-100% for urine and were 100% by all assays using oral samples. The results obtained using oral mucosal transudate specimens and rapid assays were at least comparable to those obtained with plasma, while the use of urine specimens produced suboptimal sensitivities with two of the three assays. The testing of alternative body fluids for antibodies to HIV is yet another strategy that may be applicable, particularly in developing countries.

Journal Article↗

Diagnosis of infection with human T-lymphotropic virus type II (HTLV-II) in Norwegian HIV-infected individuals.

Sera from 298 HIV-infected individuals from Southern Norway were examined for antibodies against HTLV. 30 sera (10.1%) were HTLV-II positive and 1(0.3%) HTLV-I positive. 25 of the HTLV-II infected subjects were intravenous drug abusers (IVDAs), giving a prevalence of HTLV-II infection of 24.5% in this group. Examination of blood samples by polymerase chain reaction followed by restriction enzyme analysis or sequencing confirmed the serological diagnosis. To evaluate current screening and verification HTLV tests, 44 sera were examined using a gelatin particle agglutination test, 5 different enzyme-linked immunoassays (ELISA) and 4 Western blots (WB). While earlier ELISAs and WBs were inadequate, a recent ELISA and WB including recombinant envelope glycoproteins from both viruses permitted serological diagnosis and distinction between HTLV-I and HTLV-II. Thus, HTLV-II now spreads among IVDAs in a North-European country. Health authorities in other countries should estimate the magnitude of the problem to decide upon measures to avoid transmission through blood transfusion.

Journal Article↗

Evaluation of a rapid membrane enzyme immunoassay (Testpack HIV-1/HIV-2) at zonal, regional and district hospital laboratories in Tanzania.

The performance of a rapid and simple membrane enzyme immunoassay for antibodies to HIV-1 and HIV-2 (Testpack HIV-1/HIV-2) was evaluated by testing 1000 sera from the Kilimanjaro region of Tanzania. A sensitivity of 100% (118/118 positives) and specificity of 95.1% were obtained following the manufacturer's procedure. The specificity was significantly enhanced to 97.2% (P = 0.026) by modifying the Testpack procedure by including an extra was after serum adsorption to the unit membrane. The testing of a single specimen could be completed in 8 min and up to 10 individual tests could be run simultaneously. There was complete agreement in interpretation when the results were read independently by two trained technicians. A built-in control insured against incorrect procedures or inactive reagents. In a subsequent field trial including 450 sera, one strongly reactive sample failed to be detected at a participating field hospital for unknown reasons. The Testpack reagents proved stable for up to one year at room temperature (25-30 degrees C). The data indicate that Testpack is suitable for the detection of serum antibodies to HIV and is especially applicable in laboratories with limited facilities. When used to test African sera which are known to produce a high degree of false positivity, an extra wash of the membrane after serum adsorption is recommended.

Journal Article↗

Serum antibodies to viral pathogens and Toxoplasma gondii in HIV-infected individuals.

Sera from 38 HIV-infected individuals were examined longitudinally for antibodies to viruses that may increase morbidity in HIV infection, as well as commensal viruses and Toxoplasma gondii. HTLV infection was seen in Norway for the first time as four patients had antibodies to HTLV-II and one had antibodies to HTLV-I. Antibodies to hepatitis B virus (HBV) were found in 47.2%, while 21.6% of the patients had antibodies to hepatitis C virus (HCV). There was no evidence of acquisition of HBV or HVC during the mean observation period of 2 years. A titre increase in CMV antibody with time was observed for 7 out of 21 patients and a decrease for 2 patients. For Epstein-Barr virus, herpes simplex, varicella-zoster, rubella and measles viruses, human polyomavirus BK as well as for Toxoplasma gondii, antibody prevalences and titres were within the range seen in normal populations. Also, no longitudinal changes were observed in titres of these antibodies, indicating that humoral immunity remained intact during the study period. The high prevalences of HTLV-I/II, HBV and HCV antibodies in HIV-infected patients reflect common modes of virus transmission, and the fluctuations in CMV antibody titre are indicative of reactivations. Such coinfections may influence disease progression.

Adult↗

Prevalence of hepatitis B and C and HIV antibodies in children in a Romanian orphanage.

Two hundred and fifteen children in an orphanage in Romania were examined for serum markers of present or past hepatitis B and C virus and HIV infection. In total, 183 children (85.1%) had at least one marker (HBsAg, anti-HBs or anti-HBc) of hepatitis B virus infection. An HBsAg carrier state was diagnosed in 38 (20.8%) of the infected children. Among the carriers 24.3% were HBeAg carriers, 51.4% had anti-HBe and 24.3% had neither HBe antigen nor antibody. Nine children (4.2%) had antibodies to hepatitis C virus. All sera were negative in tests for HIV antibodies. False-positive reactions represented a considerable problem with these sera. Six percent of the sera gave false-positive reactions in indirect ELISA tests for hepatitis C and HIV. Sera giving false-positive reactions had rather high serum IgG levels. The results of this study indicate that these children have been heavily exposed to hepatitis B virus and to a certain degree to hepatitis C virus, while there were no cases of HIV infection in this orphanage.

Child↗

Rotavirus infection in Tanzania: a virological, epidemiological and clinical study among young children.

Rotavirus infection in the Dar es Salaam area of Tanzania was studied in 99 hospitalized children with acute diarrhoea and 99 hospitalized non-diarrhoea referents matched for sex and age. Of the diarrhoea cases 43.4% had rotavirus in the stools as opposed to 15.2% of the referents. The high carrier rate among the referents represents a serious risk of nosocomial transmission. More referents than cases had serum IgG antibodies to rotavirus, 52.5% and 35.4%, respectively (P < 0.02), while there was no correlation with serum IgM and IgA or faecal IgA antibodies. The latex agglutination test had a sensitivity comparable to that of electron microscopy (100%) and a specificity of 93.8%. The Slidex test appeared to be superior to the Rotalex test in that it gives very few false-positive reactions. The SDS-PAGE patterns of 11 RNA segments were compatible with the presence of group A strains with considerable heterogeneity among the strains. Symptoms and signs and some environmental data were recorded. None of them was clearly associated with rotavirus infection among the diarrhoea cases. It is concluded that rotavirus is a major cause of acute infectious diarrhoea in Tanzania.

Antibodies, Viral↗

The AIDS epidemic in Tanzania: rate of spread of HIV in blood donors and pregnant women in Dar es Salaam.

The prevalence of antibodies to HIV in population groups in Dar es Salaam, Tanzania has been monitored from late 1984 through 1991. In the pregnant women and blood donors the doubling time was about 2 years. If, however, the doubling time is calculated from reported cases of AIDS of the whole country, an average doubling time of 10 months is obtained. We point out factors that may bias calculations of the rate of spread of the virus and thus give rise to erroneous projections for the nearest years, especially the so-called transient effect. It is recommended that serosurveys of representative population groups are carried out at regular intervals.

Acquired Immunodeficiency Syndrome↗

Kinetics of the binding of immunoglobulins, antibodies and virus haemagglutination inhibitors to kaolin.

Kaolin is widely used in diagnostic virology, mainly to remove serum lipoproteins that may interfere with antibody assaying. The binding kinetics of antibody to kaolin at different pH values and with varying amounts of kaolin indicated a uniform and characteristic binding pattern for IgG with maximum adsorption at pH 5 and no adsorption at pH above 9. To avoid loss of IgG antibody adsorption with kaolin should therefore be performed at pH greater than or equal to 9. The amount adsorbed increased with the amount of kaolin used. The IgM pattern was less uniform with maximum adsorption of total IgM at about 7.0, the amount adsorbed being highly dependent on kaolin concentration. Serum lipoproteins were rapidly and strongly adsorbed independent of pH from 7 to 11 and independent of the lipoprotein content of the serum. The amount of kaolin used was, however, critical.

Adsorption↗

Heterogeneity in the reactivity of various groups of IgM antibodies with kaolin.

Kaolin strongly adsorbed rheumatoid factor (RF) and mono-nucleosis antibodies, while cold agglutinins and some antimicrobial IgM antibodies were poorly adsorbed. Maximum adsorption took place at a pH of about 7. The degree of adsorption also depended on the amount of kaolin. Rheumatoid factor could be eluted from kaolin at pH 11 after adsorption at pH 7. The reported heterogeneity with regard to reactivity with kaolin is suggested to be due to hydrophobic interactions.

Adsorption↗

Study of transcription in measles virus-infected Vero cells using cDNA probes prepared from poly(A)RNA from uninfected and infected cells.

From af primary plasmid cDNA library prepared from measles virus-infected Vero cell poly(A)RNA, 435 clones selected at random were used to examine the sensitivity and specificity of cDNA probes derived from total poly(A)RNA from uninfected and infected Vero cells. The correlation between the abundance level of a particular species in the cDNA probe and the hybridization signal strength generated by the corresponding cDNA clone on a filter was reliably determined only when at least three independently prepared filters were examined. Variation in the amount of target plasmid was the most important cause of spurious signals. Variation in cDNA insert length did not disturb the signal strength within certain limits. cDNA species with abundance levels down to 0.08-0.01% were able to produce a hybridization signal above background. Unspecific cross-hybridization was shown to define the sensitivity limit of mixed cDNA probes. Despite the many false signals present at different stages, cDNA probes provided valuable information: the cDNA probes were used to monitor relative RNA expression levels and to clone five different measles virus transcripts and 2 host cell transcripts more abundantly expressed in infected cells. The abundance levels of the measles virus nucleocapsid, phosphoprotein, matrix, fusion protein and haemagglutinin genes were 1.5%, 1.5%, 1%, 0.75% and 0.5%, respectively, of the total cDNA library.

Animals↗

Viral RNA species in cell lines persistently and lytically infected with measles virus.

Identical measles viral mRNA species were present in similar amounts in the persistently infected cell lines LU106, HEpPi and MaSSPE, and in lytically infected cells as determined from Northern blots. The attenuation of transcription with the gene order did not vary significantly between different infected systems. A previously described selective restriction of F protein production in Lu106 cells could not be explained by defective transcription of F mRNA. RNA synthesis also continued unimpeded at restrictive temperatures for the temperature-sensitive viruses in Lu106 and HEpPi cells. Northern blotting revealed a prominent band in HEpPi RNA and a weak band in Lu106 RNA with the characteristics of incomplete genomes. In all infected cells, previously unrecognized small RNA species, hybridizing with the F- and H-specific probes, were discovered.

Animals↗