bluuebunny/crossref_metadata_2025 huggingface dataset:
From virus to vaccine: Recombinant mammalian cell lines as substrates for the production of herpes simplex virus vaccines
Authors: Lasky Laurence A.
Published: March 2007

The use of mammalian cells as substrates for the production of various vaccines is a time‐honored procedure. With the advent of recombinant DNA technology, various investigators soon realized that these techniques could be applied to mammalian cells to convert them to reliable, safe producers of a variety of vaccines. While none of these types of recombinant vaccines have, as yet, been widely utilized, many individuals believe that these technologies are the wave of the future in vaccine production. In this paper our work on the use of mammalian cells for the development of subunit vaccines for the prevention of infection by herpes simplex virus (HSV) types 1 and 2 is reviewed. A surface glycoprotein of these viruses, gD, was chosen as a likely candidate for a subunit vaccine based upon its primary sequence conservation between the two viral types as well as upon data suggesting that antibodies against this glycoprotein were highly efficient at neutralizing virus infectivity. Stable mammalian cell lines were constructed that expressed secreted forms of this protein, and the protein could be purified from cell‐conditioned supernatants to near‐homogeneity. Vaccination of mice with this glycoprotein was found to protect them form a lethal intraperitoneal infection by either the type 1 or type 2 forms of this virus. In a more realistic study, guinea pigs vaccinated with either the type 1 or type 2 forms of this virus. In a more realistic study, guinea pigs vaccinated with either the type 1 or type 2 form of gD were found to be protected form a genital form of HSV 2 infection. Finally, analysis of these guinea pigs revealed that the establishment of latency by HSV 2 was also inhibited by vaccination with gD. These preliminary animal studies strongly suggest that gD produced in a recombinant mammalian cell expression system might induce protective immunity in humans vaccinated with this glycoprotein.
Hepatitis D virus infection among intravenous drug abusers in Taiwan: Analysis of risk factors and liver function tests
Authors: Hsu Hsu‐Mei , Wang Yue‐Fen , Lo Shu‐Hui , Sun Hui‐Chih , Yip Kwok‐Kee , Chen Jui‐San , Chuang Cheng‐Hua , Chen Ding‐Shinn
Published: March 2007

To investigate the prevalence of hepatitis D virus (HDV) and hepatitis B virus (HBV) infection among intravenous drug abusers in Taiwan, a total of 761 male prisoners, including 680 intravenous drug abusers, were studied for serological markers of HBV and HDV. Questionnaires were distributed to evaluate the risk factors for HDV infection and also to estimate the strength of association among HDV infection and the risk factors.

HBV infection was common, and the positive rates of HBV markers between intravenous drug abusers and non‐drug abusers were not statistically different. However, the positive rate of the antibody to HDV was significantly higher among intravenous drug abusers than among non‐drug abusers (21.3% vs. 8.6%). Of 131 chronic HBV carriers with intravenous drug abuse, 119 (91%) were anti‐HD positive. Using multiple logistic regression models, we found that the most important risk factor for HDV infection was hepatitis B surface antigen (HBsAg) carriage, and intravenous drug addiction the next. A matched case‐control study also was conducted to compare liver function tests among both anti‐HD‐ and HBsAg‐positive group anti‐HD‐negative, and HBsAg‐positive group as well as those with neither positive. Statistically significant difference in liver function tests was not found.

It is concluded that the HBsAg carriers with intravenous drug abuse in Taiwan are commonly HDV infected with and that the infection does not seem to affect the liver as assessed by liver function tests.
Antiviral strategies in chronic hepatitis B virus infection: II. Inhibition of duck hepatitis B Virus in vitro using conventional antiviral agents and supercoiled‐DNA active compounds
Authors: Civitico Gilda , Wang Yanyan , Luscombe Carolyn , Bishop Naomi , Tachedjian Gilda , Gust Ian , Locarnini Stephen
Published: March 2007

Primary duck hepatocyte (PDH) cultures, congenitally infected with the duck hepatitis B virus (DHBV), were grown on feeder cell layers of irradiated human embryonic lung fibroblasts and then exposed to a number of compounds with recognized or potential antiviral activity. These compounds included conventional antiviral agents, reverse transcriptase inhibitors, compounds with activity to supercoiled‐DNA, and DNA‐binding agents. Twenty‐three compounds were evaluated, and 13 were found to inhibit significantly viral DNA replication. Seven of these compounds (ellipticine, amsacrine, coumermycin A1, Adriamycin, mitozantrone, chloroquine, and neocarzinostatin) acted at the level of viral SC DNA and significantly inhibited production of duck hepatitis B surface antigen (DHBsAg). Conventional agents that inhibited DHBV DNA replication included ganciclovir, acyclovir, bromovinyldeoxyuridine, ribavirin, phosphonoformate, and dideoxyadenosine. Except for dideoxyadenosine, these inhibitors of viral DNA synthesis did not significantly inhibit DHBsAg production. Two additional compounds, novobiocin and nalidixic acid, altered the pattern of viral DNA replication, especially the generation and processing of viral SC DNA, and also inhibited the production of DHBsAg. Several compounds acting at the level of viral SC DNA have now been identified and may offer potential for the management of chronic hepatitis B virus infection.
A rapid and simplified micromethod for subtyping varicella‐zoster virus
Authors: Gharabaghi Farhad , Aymard Michèle , Trotemann Pierre , Gerdil Catherine
Published: March 2007

A simplified and rapid micromethod based on restriction endonuclease analysis of radiolabelled varicella‐zoster virus (VZV) DNA is described and applied to strains for comparison. This procedure is cheaper and less time consuming than that requiring viral nucleocapsid purification (macromethod). The micromethod is suitable for routine DNA analysis of VZV isolates, allows differentiation of vaccine strain from wild strains, and provides evidence for variability of wild strains.
Further studies of 48‐1 antigen in serial liver biopsies from chimpanzees infected with hepatitis delta virus
Authors: Sugitani Masahiko , Shimizu Yohko K. , Hanawa Mie , Azumi Jun‐Ichi , Abe Kenji , Shikata Toshio
Published: March 2007

The 48‐1 antibody, initially reported to react specifically with non‐A, non‐B infected liver tissue, has been found to react also with liver specimens from chimpanzees infected with hepatitis delta virus (HDV). To clarify further the relation between HDV and appearance of the antigen reacting with the 48‐1 antibody (48‐1 Ag), immunoperoxidase studies were carried out on serial liver specimens from chimpanzees infected with HDV. Immunohistochemical and serological findings suggested that the appearance of 48‐1 Ag paralleled that of HDV. Double immunoperoxidase staining revealed HDAg in the nucleus and 48‐1 Ag in the cytoplasm of the same hepatocytes as well as in different hepatocytes separately. The course of appearance of microtubular aggregates paralleled that of 48‐1 Ag. The present results suggested that expression of 48‐1 Ag was related to infection with HDV, probably because expression of this antigen is induced from the host genome.
Inhibition of hepatitis A virus replication in vitro by antiviral compounds
Authors: Crance J. M. , Biziagos E. , Passagot J. , van Cuyck‐Gandré H. , Deloince R.
Published: March 2007

Forty antiviral compounds were screened for inhibitory effect on hepatitis A virus (HAV) antigen expression in the human hepatoma cell line PLC/PRF/5. Ribavirin, amantadine, glycyrrhizin, and pyrazofurin were selected in this screening test and were studied further. The selectivity indices of these four compounds, calculated as the ratio of 50% cytotoxic dose (determined by the trypan blue exclusion and by inhibition of [3H] leucine incorporation) to the 50% effective dose (determined by the viral antigen expression), were 4.6 and 3.0 with ribavirin, 5.3 and 5.9 with amantadine, 15.2 and 16.9 with glycyrrhizin, and 45.4 and 74.6 with pyrazofurin. All four compounds resulted in concentration‐dependent reductions of HAV antigen expression and HAV infectivity. Ribavirin, amantadine, pyrazofurin, and glycyrrhizin emerged, from the present study, as promising candidates for chemotherapy of acute hepatitis A.
Hepatitis B virus transcripts and surface antigen in human peripheral blood lymphocytes
Authors: Lobbiani A. , Lalatta F. , Lugo F. , Colucci G.
Published: March 2007

The presence of hepatitis B virus (HBV) genome, transcripts, and antigens (HBsAg, HBcAg, HBeAg) was examined in peripheral blood lymphocytes (PBL) from 12 patients with HBsAg‐positive (B) chronic active hepatitis (CAH) and 8 normal donors by Southern and Northern blot techniques and enzyme‐linked immunoassays (ELISA). HBV DNA was detected in 5 patients with B‐CAH as extrachromosomal, full‐length monomers of 3.2 kb. In 3 of these patients Northern blot analysis revealed the presence of the 3.6–3.8 kb RNA species, which were accompanied in one case by the HBsAg‐specific 2.4 kb transcript. An ELISA performed on cell lysate obtained from this patient showed low but detectable amounts of HBsAg as compared to control PBL incubated with up to 50 μg/ml of the viral antigen.

Serum HBV DNA was found in 3 patients with B‐CAH, whereas all individuals positive for cellular HBV DNA had circulating HBeAg. These data indicate that lymphocytes from some patients with hepatitis B can harbor a transcriptionally and translationally active HBV genome.
Simplified test for detecting the resistance of herpes simplex virus to acyclovir
Authors: Agut Henri , Aubin Jean‐Thierry , Ingrand Didier , Blanc Sylvie , Clayton Anne‐Louise , Chantler Shireen M. , Huraux Jean‐Marie
Published: March 2007

The detection of herpes simplex virus (HSV) antigen by means of an enzyme amplified ELISA was investigated for rapid screening of acyclovir (ACV) resistance. Vero cell monolayers were inoculated with various strains of HSV and incubated in the presence of different concentrations of ACV. When cytopathic effect was present, the culture supernatants were tested by ELISA. The absorbance values were found to correlate with the results of virus yield and plaque reduction assays. The comparison between absorbance values obtained in the presence of 10 μM ACV and in the absence of drug provided the basis for a simplified sensitivity test. The use of a single ACV concentration allowed discrimination between ACV‐resistant and ACV‐sensitive reference strains, the detection of ACV‐resistant virus mixed in the proportion of 10% with ACV‐sensitive virus, and a study of the emergence of an ACV‐resistant virus population in serial samples taken from experimental rabbit keratitis. The simplified susceptibility assay is a sensitive and convenient method for rapid screening of HSV resistance to ACV.
Detection of human immunodeficiency virus‐1 by polymerase chain reaction and virus cultivation
Authors: Sönnerborg Anders , Abens Janis , Johansson Bo , Strannegård Örjan
Published: March 2007

Peripheral blood of 57 patients with antibodies to human immunodeficiency virus 1 (HIV‐1) and of five HIV‐1 seronegative subjects at risk for HIV‐1 infection were analysed by polymerase chain reaction (PCR) and virus isolation. The virus was recovered from peripheral blood cells in 89% and from plasma in 75% of the HIV‐1 seropositive cases. In contrast, proviral HIV‐1 DNA was detected in all HIV‐1 seropositive patients by dot blot hybridization of the amplified fragments. The intensities of the dot blot reactions were less pronounced in asymptomatic HIV‐1 seropositive individuals than in patients with acquired immunodeficiency syndrome (AIDS) or AIDS‐related complex (ARC), suggesting an increase in proviral DNA with advancing disease. Three of five seronegative patients with signs or symptoms suggesting HIV‐1 infection, but none of the controls, were positive for HIV‐1 DNA by one or two primer pairs. These results show a high sensitivity of the PCR for detecting HIV‐1 DNA in patients of all stages of HIV‐1 infection. Proviral DNA can also be detected in some individuals without detectable antibodies to the virus. The virus load in peripheral blood, as determined by virus cultivation and PCR, seems to increase with progression of the infection.
Different forms of hepatitis B virus DNA and expression of HBV antigens in peripheral blood mononuclear cells in chronic hepatitis B
Authors: Bouffard P. , Lamelin J.‐P. , Zoulim F. , Pichoud C. , Trepo C.
Published: March 2007

The presence of both hepatitis B virus (HBV) DNA and HBV antigens (HBsAg, HBeAg) was assessed in peripheral blood mononuclear cells (PBMC) from 32 patients chronically infected with HBV. Three different molecular forms of HBV DNA were observed: free monomers (5), high‐molecular‐weight free concatemers (11), and integrated HBV DNA (9). The HBV DNA patterns in the PBMC were different from those found in liver and did not correlate with any specific profile of serum HBV markers. When the same PBMC were assayed for HBsAg, 22 of the 25 HBV DNA positive samples, but only three of the seven HBV DNA negative samples, were positive. By contrast, none of the PBMC samples from five healthy HBV vaccine recipients gave any positive signal in the HBV DNA or HBsAg assays. In some patients, T and B cells, monocytes, and polymorphonuclear (PMN) cells were assayed separately, showing that the DNA pattern was similar for these different leucocytes subsets and ruling out the possibility that these patterns might reflect PMN cell contamination. Thus, in chronic HBV infection, 87.5% (28/32) of patients were found to contain at least one HBV marker in their PBMC, and a strong correlation was found between the presence of HBV DNA and viral antigens, suggesting a specific expression of HBV encoded proteins.
Virus excretion and mutation by infants following primary vaccination with live oral poliovaccine from two sources
Authors: Dunn Glynis , Begg Norman T. , Cammack Nicholas , Minor Philip D.
Published: March 2007

The excretion of virus by 30 children was followed over a period of 28 days after their first vaccination with live oral poliovaccine. The type 1 and type 2 strains were isolated at similar high frequencies throughout this period, while the type 3 strain was not excreted by most children after day 2 post vaccination. Mutations in the 5′ non‐coding region associated with the attenuated phenotype reverted most rapidly for type 3 and least rapidly for type 1. The data are consistent with different degrees of selection against the attenuating mutations in the three serotypes in the gut, but imply that reversion is required for prolonged excretion. The findings also provide a possible explanation for the reported distribution of the rarely occurring vaccine associated cases between recipients and contacts for the three serotypes. A statistically significant difference was not observed between vaccine from the two main suppliers to the UK market.
Antibody responses of adults, adolescents, and children to a plasma‐derived hepatitis B vaccine in a rural African setting
Authors: Tabor Edward , Gerety Robert J. , Cairns James , Bayley Anne C.
Published: March 2007

A field trial of a plasma‐derived hepatitis B vaccine in five rural villages in Zambia was analyzed to determine if adults in a rural African setting respond to this vaccine as well as adults in Western countries and to determine the immunogenicity of fewer than the recommended three doses; 255 residents, including 171 who were susceptible to hepatitis B, were vaccinated. Among those who received three vaccine doses, protective levels of antibody to hepatitis B surface antigen (anti‐HBs) developed in 67% of adults (ages 21 to 70 years), 87% of adolescents (ages 12 to 19 years), and 100% of children (ages 0 to 11 years). The 67% of vaccinated adults who developed anti‐HBs at the protective level was lower than the 96% reported among adults receiving the same vaccine at the same dose and dosage schedule in studies in Western countries. No difference was seen in the response of those receiving two doses compared with those receiving three doses among adults and adolescents, suggesting that a two‐dose regimen may be acceptable in these age groups in developing countries to reduce costs and improve compliance. Use of hepatitis B vaccine in a region where prevaccination hepatitis B serologic screening was not available did not appear to increase the number or severity of adverse reactions.
Immunogenicity of hepatitis B vaccine simultaneously administered with the expanded programme on immunisation (EPI)
Authors: Tsega E. , Tafesse B. , Nordenfelt E. , Wolde‐Hawariat G. , Hansson B‐G. , Lindberg J.
Published: March 2007

To assess the immunogenicity of hepatitis B vaccine simultaneously administered with the Expanded Programme on Immunisation (EPI) vaccines, a total of 300 consecutive infants, ranging in age between 6 weeks and 6 months, were alternatively assigned to the revised EPI and to EPI plus plasma‐derived hepatitis B vaccine (Hevac B Pasteur). Four dosages (2 mcg or 0.5 ml each) of hepatitis B vaccine were given intramuscularly. HBsAg, anti‐HBc, and anti‐HBs were determined at day 0 and at day 210 in both groups by radioimmunoassay methods. The anti‐HBs titre levels and geometric mean titres (GMTs) were determined at day 180 and day 210. There were three new hepatitis B virus infections in the control group and none in the study group by day 210. The seroconversion rates in the study group were 66% a day 180 and 84% at day 210, the respective GMTs being 116 and 940. Although the duration of observation is short (7 months) and the difference in the rate of infection between the two groups is not statistically significant (P>0.05), the immunogenicity of the vaccine appears to be good. The simultaneous administration of hepatitis B vaccine and the EPI vaccines is logistically ideal. However, we suggest a further study using a larger dose of the Hevac B Pasteur vaccine to find out if a higher seroconversion rate than the 84% observed in this study could be achieved.
Prevalence of hepatitis C in South Africa: Detection of anti‐HCV in recent and stored serum
Authors: Ellis L. A. , Brown D. , Conradie J. D. , Paterson A. , Sher R. , Millo J. , Theodossiadou E. , Dusheiko G. M.
Published: March 2007

The prevalence of anti‐HCV was studied in a cohort of 2,072 South Africans. The results were compared in selected recently collected sera and in stored sera. The serum ALT and anti‐HBc were also studied as surrogate markers in this population. The following groups were tested: (a) 948 urban, black blood donors (b) 500 white blood donors (c) 500 Asian blood donors (d) 216 rural hospitalized patients (e) 358 rural mineworkers. Sera found positive by the original ELISA were retested, and reproducibly positive tests in rural black men (group d) were confirmed both by recombinant immunoblot assay and by a second ELISA. An anti‐HCV prevalence of 1.2%, 0.8%, and 0.6% in urban blacks, Asians, and whites was found. Antibodies to hepatitis B core antigen were found in 42.9%, 3.4%, and 1.2% of black, Asian, and white donors, respectively; 76% of donors positive for anti‐HCV were anti‐HBc negative. In rural African men, 17% of stored serum samples and 9.2% of recently collected serum samples were positive for anti‐HCV. In this cohort 3.84% were positive by all three assays. These results suggest that the prevalence of anti‐HCV in low and high‐risk South African urban blood donors is comparable to high and low prevalence areas in Europe, the United States, and Japan, but indicates a relatively high degree of exposure to hepatitis C in rural African men. The reactivity of stored, frozen sera in this population requires further investigation. In South African urban blood donors, surrogate marker testing will not expedite HCV screening.
Diagnosis of herpes encephalitis via Southern blotting of cerebrospinal fluid DNA amplified by polymerase chain reaction
Authors: Klapper Paul E. , Cleator Graham M. , Dennett Carol , Lewis Alfor G.
Published: March 2007

Herpes simplex virus thymidine kinase gene specific polymerase chain reaction (PCR) amplification of DNA extracted from lumber cerebrospinal fluid (CSF) and Southern blotting (SB) were evaluated as a method for the diagnosis of herpes simplex encephalitis (HSE). Positive PCR‐SB results were obtained with CSF samples from 9 of 10 patients (11 of 12 CSF specimens) with proven herpes encephalitis as early as 2 days after onset of neurological illness. Our data support the suggestion that PCR techniques may provide a clinically relevant “non‐invasive” method for the diagnosis of HSE.
Detection of specific IgA antibodies against BK virus by ELISA
Authors: Nilsen Ingard , Flægstad Trond , Traavik Terje
Published: March 2007

The development and evaluation of an ELISA for analysis of anti‐BKV specific IgA antibodies in human sera are described. All children with cancer with a primary BKV infection developed specific IgA antibodies, without any specific symptoms during the infection. Specific IgA was found in 61% of sera from healthy persons containing BKV IgG antibodies, using the chosen cut‐off value, and BKV IgM in 4%. These results indicate that IgA production is more persistent than IgM. The high frequency of specific IgA antibodies could either be explained by frequent reactivations or long‐lasting persistence of anti‐bodies.
Hepatitis B virus infection in Cameroon: A seroepidemiological survey in city school children
Authors: Chiaramonte M. , Stroffolini T. , Ngatchu T. , Rapicetta M. , Lantum D. , Kaptue L. , Chionne P. , Conti S. , Sarrecchia B. , Naccarato R.
Published: March 2007

A pilot survey conducted in Cameroon comprising 702 children of primary school age revealed that 58.5% of the children were positive for at least one HBV marker. An overall HBsAg prevalence of 19.9% was observed. Both any HBV marker and HBsAg prevalences increased with age (P < 0.01). Of the HBsAg‐positive subjects, 28.4% were positive for HBV‐DNA; none was positive for anti‐HDV. There was no significant difference among sexes for either any HBV marker or for HBsAg prevalence. However, a significant difference by sex was found in the percentage ratio HBsAg+/and any marker + (41.0% in males against 28.2% in females; P < 0.05). No significant difference with respect to the father's occupation and family size was observed. When the zone of location of school was taken into consideration it was observed that suburban school children had a higher prevalence of any HBV marker as compared to central town school children (73.9% vs. 56.6%; P < 0.001). These results place Cameroon among countries of high endemicity for HBV. The results also indicate an early acquisition of infection, a high infectivity, and probably continuous infection even in the primary school ages.
Incidence of hepatitis B viraemia, detected using the polymerase chain reaction, after successful therapy of hepatitis B virus carriers with interferon–α
Authors: Carman William F. , Dourakis Spyros , Karayiannis Peter , Crossey Mary , Drobner Rita , Thomas Howard C.
Published: March 2007

Thirty‐nine patients (62 sera) who, after interferon‐α therapy for chronic hepatitis B virus (HBV) infection, were seronegative for HBeAg and HBV‐DNA by dot blot hybridisation, were tested using the polymerase chain reaction (PCR) for residual viraemia. Overall, 59% of the HBsAg‐positive sera and 43% of the HBsAg‐negative sera were positive by PCR. All except one of the HBsAg‐negative patients had seroconverted to anti‐HBs. Between 13 and 18 months after therapy, 33% of the HBsAg‐positive and 20% of the HBsAg‐negative patients remained viraemic. Eighteen months after the end of treatment, no patient tested was positive. Twenty‐three patients were tested sequentially over periods from 1 to 43 months: Thirteen lost HBV‐DNA by PCR, three remained positive, five remained negative, and two patients relapsed. The merits and disadvantages of PCR for assessing interferon treatment of HBV carriers are discussed.
Natural seroconversion from hepatitis be antigen to antibody among hepatitis B virus carriers in Okinawa islands
Authors: Sakugawa Hiroshi , Ohwan Tomohisa , Yamashiro Akihiro , Oyakawa Tominori , Kadena Keizo , Kinjo Fukunori , Saito Atsushi
Published: March 2007

In the Okinawa Islands, the great majority of hepatitis B surface antigen (HBsAg) carriers have already acquired antibody to hepatitis Be antigen (anti‐HBe) by the age of 30 years (preliminary cross‐sectional data). To elucidate natural seroconversion from hepatitis Be antigen (HBeAg) to anti‐HBe among HBsAg carriers found in the islands of Okinawa Prefecture, 34 HBeAg‐positive HBsAg carriers were followed for 1–6 years with serial measurements of aminotransferase levels, HBeAg, and anti‐HBe. The 34 subjects included 24 patients with chronic hepatitis (group 1) and ten asymptomatic HBsAg carriers (group 2). During the follow‐up period, HBeAg disappeared from 14 subjects in group 1 with the cumulative clearance rate of HBeAg of 56.3% within the first 2 years and with 10 of the 14 subsequently developing anti‐HBe. Moreover, the aminotransferases in 12 of the 14 spontaneously seroconverted fell into the normal range. The annual clearance rates of HBeAg among group 1 and group 2 were 25.6% and 9.3%, respectively. The tendency for early disappearance of HBeAg during a carrier's life time or in the course of chronic hepatitis may lead to the low death rate from hepatocellular carcinoma (HCC) particularly HCC associated with hepatitis B virus infection in this area.
Preparation of monoclonal antibodies to JC virus and their use in the diagnosis of progressive multifocal leucoeneephalopathy
Authors: Knowles W. A. , Sharp I. R. , Efstratiou L. , Hand J. F. , Gardner S. D.
Published: March 2007

Seven monoclonal antibodies (MAbs) to polyomavirus JC were produced, and one was selected for use in immunofluorescence (IF) tests on brain material from patients with suspected progressive multifocal leucoencephalopathy (PML). The selected MAb (5. 12.2) reacted by IF with JC‐infected primary human foetal glial (PHFG) cell cultures (titre 1/200,000) but not with BK‐infected human embryo lung (HEL) fibroblasts (titre <1/20). Its haemagglutination‐inhibition (HI) titre was high (>1/5 × 106) against JC virus but low (<1/5)against BK virus. A diagnosis of PML was confirmed on brain biopsy or at postmortem in four patients, two of whom were also infected with human immunodeficiency virus (HIV). In one of the patients, specific detection of JC virus antigen had not been possible using our routine high titred JC and BK human sera due to interference by JC antibody present in the cerebrospinal fluid (CSF). Viral antigen was, however, detected with the MAb 5. 12.2.
Evaluation of a chimpanzee colony for antibodies to hepatitis C virus
Authors: Lanford Robert E. , Notvall Lena , Barbosa Luiz H. , Eichberg Jorg W.
Published: March 2007

The chimpanzee is the only species other than man that is generally susceptible to infection by hepatitis C virus (HCV). Aspects of future studies on vaccines and therapeutics for HCV may continue to depend on the chimpanzee. In an attempt to determine the HCV status of the animals in a chimpanzee colony, the recently developed enzyme immunoassay (EIA) for antibodies to HCV was used. The results of the assay indicated that only 31.3% of the animals that had previously been inoculated with a non‐A, non‐B hepatitis agent were currently positive in the assay. A retrospective analysis suggested that an additional 20% of the chimpanzees had been positive at some time following infection. Seroconversion to an anti‐HCV antibody response using this assay did not appear to correlate with the severity of the initial disease or the development of chronic liver damage. Examination of the EIA‐positive samples using the second generation recombinant immunoblot assay (RIBA) containing four HCV antigens suggested that chimpanzees responded differentially to these antigens. Examination of serum samples from 139 uninoculated animals by EIA revealed seven positive samples and ten samples with borderline values. The nature of the reactivities in most of the positive samples could not be resolved, but analysis by RIBA indicated that at least one animal in the breeder colony had been exposed to HCV. Due to the low seroconversion rate and the uncertainties surrounding many of the positive reactions, this assay cannot be used to determine the initial source or extent of spread of HCV infections in the uninoculated animals.
Genital human papilloma virus infection in oslo studied by dot blot DNA hybridization and the polymerase chain reaction
Authors: Gjøen Kirsti , Siebke Jens C. , Flikke Magne , Háger Renate , Ertzeid Gudvor , Halsos Arne , Ekgren Josef , Norling Berit , Grinde Bjørn , Ørstavik Ivar
Published: March 2007

Samples from patients with genital condyloma acuminata or with cervical condylomas and/or dysplasia and from women without cytological/ clinical evidence of cervical affection were examined by dot blot DNA hybridization or the polymerase chain reaction (PCR). The PCR was much more sensitive than dot blot, more than doubling the human papilloma virus (HPV) findings. HPV DNA, mainly HPV 6/11, was detected in 18 of 19 biopsies of condyloma acuminata, whereas HPV 16 was most frequently detected in the 21 cervices (76%) with condyloma and/or dysplasia. HPV 16 was detected in eight of 103 cervical smears with no signs of infection. The prevalence of HPV 16 in cervical samples was somewhat higher than expected. This suggests that, in Oslo, HPV 16 is a common HPV type in women with cytologically normal cervices. HPV 18 was relatively rare and was detected only in combination with other HPVs.
Enzymatic amplification and sequence analysis of precore/core DNA in HBsAg‐positive patients
Authors: Ljunggren Karin , Kidd Alistair H.
Published: March 2007

Serum or plasma from 69 HBsAg‐positive patients was tested for the presence of precore/core gene specific DNA by the polymerase chain reaction (PCR). In both healthy individuals (n = 26) and chronic carriers (n = 25), there was a strong correlation between presence of circulating anti‐HBe and the absence of detectable HBV genome in serum. In 18 serum samples where HBsAg was the only detectable marker, i.e., anti‐HBc‐negative specimens, HBV DNA could be detected in three samples. HBV strains from 21 of the 24 PCR‐positive samples were sequenced over the precore/core junction. A stop codon at the end of the precore region, described by other workers, was found in strains from two blood donors, one of whom had detectable HBeAg in serum. Conversely, HBV strains from the three anti‐HBc‐negative patients where DNA of the HBV precore region could be amplified and who had no detectable serum HBeAg or anti‐HBe did not have this stop codon. The study indicates that further investigations are required before lack of HBeAg can be correlated with evidence of mutations in the precore region.
Current seroepidemiology of hepatitis A in Hong Kong
Authors: Chin Kenneth P. , Lok Anna S. F. , Wong Loretta S. K. , Lai Ching‐Lung , Wu Pui‐Chee
Published: March 2007

The current seroepidemiology of hepatitis A in Hong Kong was examined by testing stored sera from 702 healthy subjects, collected between 1987–1989, for antibody to hepatitis A virus (anti‐HAV). The overall prevalence of anti‐HAV antibody was 45.6%. There were significant increases in prevalence of anti‐HAV antibody with every 10‐year increase in age up to age 40. The prevalence of anti‐HAV antibody was 24% for subjects below age 30 and 89.2% for those above age 30 (P ≤ 0.0001). Socioeconomic factors did not appear to have any influence on the prevalence of anti‐HAV antibody. In comparison with another study conducted in Hong Kong 10 years ago, the prevalence of anti‐HAV antibody in the current study was significantly lower in every age group from 0 to 30 years.

In summary, it was shown that HAV infection is no longer highly endemic in Hong Kong. In view of the changing epidemiology, postexposure prophylaxis will be necessary for young adults and children, and hepatitis A vaccine may be indicated for high risk groups when it is generally available.
Hepatitis C virus: Buoyant density if the factor VIII‐derived isolate in sucrose
Authors: Bradley D. , McCaustland K. , Krawczynski K. , Spelbring J. , Humphey C. , Cook E. H.
Published: March 2007

Physicochemical and molecular characterization studies of hepatitis C virus (HCV), the major causative agent of parenterally transmitted non‐A, non‐B hepatitis (PT‐NANBH), strongly suggest that it is a pesti‐/flavivirus‐like virus. Additional studies show that the buoyant density of plasmaderived HCV in sucrose is significantly lower than that of most tissue culture‐derived flaviviruses (1.20g/cm3). Our finding suggests, but does not prove, that at least one physicochemical property of HCV is more similar to that of the pestiviruses, bovine viral diarrhea virus (BVDV) and hog cholera virus (HogCV), than that of the flaviviruses.
Prevalence and significance of antibodies to hepatitis c virus among Saudi haemodialysis patients
Authors: Ayoola E. A. , Huraib S. , Arif M. , Al‐Faleh F. Z. , Al‐Rashed R. , Ramia S. , Al‐Mofleh I. A. , Abu‐Aisha H.
Published: March 2007

Seventy‐four patients who were maintained on chronic haemodialysis in King Khalid University Hospital, Riyadh, Saudi Arabia were tested using the recently available ELISA to determine the prevalence of antibodies to hepatitis C virus (anti‐HCV) in a haemodialysis unit.

The prevalence rate of anti‐HCV antibodies of 41.9% in the haemodialysis patients was significantly higher than the rate of 3.9% detected in 488 asymptomatic blood donors who were similarly tested. In the haemodialysis patients, anti‐ HCV positivity was related to previous blood transfusion ( >5 units of blood) and to the duration of haemodialysis (> 4 years); but was unrelated to sex, age, positive HBV markers or to past or current elevation of serum ALT. The results indicate a relatively higher prevalence of anti‐ HCVantibodies in our patients compared to rates of 1‐20% reported from Europe and the U.S.A. An effective control strategy for HCV infection in this high risk group is urgently indicated.
Similar subclass antibody responses after intranasal immunization with UV‐inactivated RSV mixed with cholera toxin or live RSV
Authors: Reuman Peter D. , Keely Scott P. , Schiff Gilbert M. , Gamble James N.
Published: March 2007

To determine the effect of cholera toxin as a mucosal adjuvant on the class and subclass antibody response to RSV, mice were immunized intranasally with different doses of live RSV or V‐inactivated RSV mixed with cholera toxin. A single lo6 pfu dose of live RSV and a single 50 ú dose of UV‐inactivated RSV mixed with cholera toxin produced comparable serum IgG and respiratory secretion IgG and IgA antibody titers. Subclass antibody titers to whole RSV were also comparable between these two immunizing regimens. A predominance of IgG2a subclass to whole RSV was found for both reg‐ imens. The quantitiy of serum total IgG antibody to glycoprotein F or glycoprotein G did not differ between these regimens. The serum IgG subclass antibody response to both glycoprotein F and G was also not significantly different between regimens. Cholera toxin as a mucosal adjuvant can stimulate class and subclass antibody responses to UV‐inactivated RSV that are similar in quantity and distribution to those after live RSV infection.
Inhibition of Lymphocyte Proliferation Induced In Vitro by Microbial Antigens in HIV‐Infected Subjects
Authors: Maria Vasco A. J. , Pinto Lígia A. , Victorino Rui M. M.
Published: March 2007

In the course of a study of lymphocyte responses to microbial antigens in HIV‐infected patients, we detected a previously unrecognized phenomenon of inhibition of lymphocyte baseline proliferation, induced by the presence of tetanus toxoid and Escherichia coli in the cultures. The effects of tetanus toxoid and Escherichia coli on lymphocyte proliferation in vitro were assessed by comparing the 3H‐thymidine uptake by lymphocytes cultured without stimulant with the uptake of lymphocytes cultured in the presence of the antigens. Twenty‐six patients with HIV infection (20 asymptomatic/persistent generalized lymphadenopathy, 2 AIDS‐related complex, 4 AIDS) were investigated and the controls were 33 healthy individuals without evidence of HIV infection. Eight out of 22 asymptomatic/PGL and ARC patients progressed to full‐blown AIDS in the mean follow‐up of 26 months. The inhibition of proliferation was considered to be significant when the uptake of 3H‐thymidine was reduced by 20% in the presence of the antigens. Using these criteria, 50% of the patients studied with tetanus toxoid and 36% of those studied with E. coli had evidence of the inhibitory phenomenon. Seven of the eight patients who developed AIDS during the observation period had the inhibitory phenomenon. In the group of patients without the inhibitory signs only one evolved to AIDS during the follow‐up. The possibility of this phenomenon being related to an induction of suppressor cell activity by the antigens is discussed.
Antibodies to hepatitis C virus in uremic patients on continuous ambulatory peritoneal dialysis
Authors: Ng Yee‐Yung , Lee Shou‐Dong , Wu Shiao‐Chi , Yang Wu‐Chang , Chiang Shoou‐Shan , Huang Tung‐Po
Published: March 2007

The prevalence of antibody to hepatitis C virus (anti‐HCV) among 101 uremic patients receiving continuous ambulatory peritoneal dialysis (CAPD) was evaluated using a synthetic peptidebased HCV antibodies enzyme immunoassay. Thirty (29.7%) were found anti‐HCV positive. This is significantly higher than 500 unselected paid blood donors (4.2%, P < 0.0001). Among CAPD patients, anti‐HCV positivity was found more frequently in patients who had received frequent and longer duration of hemodialysis previously (40.4% vs. 20.4%, P < 0.05). These findings suggest that hemodialysis patients have a higher risk of HCV infection. At present, CAPD may be a suitable way to reduce the incidence of HCV infection in uremic patients.
Humoral immunity to respiratory syncytial virus infection in the elderly
Authors: Falsey Ann R. , Walsh Edward E.
Published: March 2007

The relationship between serum immunoglobulins and the severity and risk of Respiratory Syncytial Virus (RSV) infection in the institutionalized elderly was prospectively assessed during the winter of 1989‐1990 at a 591 bed nursing home. Forty RSV infections were identified out of 149 respiratory illnesses by isolation of the virus or by a ≥fold rise in RSV‐specific IgG by EIA. Acute serum RSV IgG levels were similar in those with RSV infection and those with non‐RSV illness. Additionally, among the RSV‐in‐fected elderly there was no correlation between severity of clinical symptoms and level of acute IgG titers by EIA or virus neutralization. The results of this study suggest that humoral immunity does not play a major role in reducing the risk of infection nor modulating the clinical severity of illness in elderly persons with RSV infections.
Hepatitis C virus in the etiology of chronic hepatitis and liver cirrhosis: Possibility of mixed viral infections due to parenteral transmission
Authors: Favorov M. O. , Fields H. A. , Yashina T. L. , Goldberg E. Z. , Yeramishantsev A. K. , Rakchimova H. K. , Burkov A. N. , Margolis H. S. , Lvov D. K.
Published: March 2007

Sera obtained from 381 patients with chronic liver disease from four cities within the USSR were studied for HBV, HDV, and HCV markers of infection. Anti‐HCV activity was detected in 41.2% of non‐A, non‐B cases. The etiological distribution of chronic hepatitis in Moscow and Dushanbe was similar with an approximate 20% prevalence for HBV, HDV, and HCV infections, whereas in Yakatsk 40% of cases were caused by HDV infections. The etiology of disease remained unrecognized in approximately 40% of patients with chronic liver disease in Moscow and Dushanbe and in 15% in Yakutsk. Anti‐HCV activity was detected in 18.8% of patients with chronic HBV infections and in 8.3% of patients with chronic HDV infections. Anti‐HCV activity was detected in 41% of patients without markers of HBV or HDV infections. The reasons for the observed differences in HCV prevalence among patients chronically infected with HDV are discussed.
Persistent rubella‐specific IgM reactivity in the absence of recent primary rubella and rubella reinfection
Authors: Thomas H. I. J. , Morgan‐Capner P. , Roberts A. , Hesketh L.
Published: March 2007

Between two and seven sera from cases of persistent detection of rubella‐specific IgM for periods in excess of 2.5 months, but in the absence of recent primary rubella or rubella reinfection, were examined for rheumatoid factor, heterophile antibody, and IgM reactivity against toxo‐plasma and a number of viruses. The relative avidity of the rubella‐specific lgG, has been assessed in all the sera by two methods.

None of the sera contained rheumatoid factor or heterophile antibody, nor did any contain detectable concentrations of IgM specific for any of the panel of antigens apart from five sera which contained low concentrations of IgM specific for some coxsackieviruses B. No sera were positive for low avidity specific IgG, although three did give equivocal results with one avidity test and one gave equivocal results with the second avidity test.
Virus safety of human immunoglobulins: Efficient inactivation of hepatitis C and other human pathogenic viruses by the manufacturing procedure
Authors: Nowak Thomas , Gregersen Jens‐Peter , Klockmann Ulrich , Cummins Larry Bill , Hilfenhaus Joachim
Published: March 2007

Human immunoglobulins are plasma derivatives with a low risk of transmitting viral infections. To the present, no proven case of human immunoglobulins transmitting human immunodeficiency viruses has been reported. However, there have been a few reports on the transmission of hepatitis C virus by these plasma proteins. To improve further the safety of both 5s iv human immunoglobulins and 7s im immunoglobulins, we introduced a 10‐hour heat treatment of the aqueous solutions a t 60°C (i.e., pasteurization) into the manufacturing procedure. This treatment was not added to the manufacturing procedure of 7s iv immunoglobulin that already contained the S‐sulfonation as a virus inactivating method. We now report on experimental data that show that the whole manufacturing procedures of the above immunoglobulins inactivate efficiently hepatitis C virus and that the specific virus inactivation methods alone, namely, pasteurization or S‐sulfonation, also inactivate completely viruses of the flavivirus family, to which the hepatitis C virus belongs. The inactivation of the Flaviviridae bovine viral diarrhea virus, tick‐borne encephalitis virus, and yellow fever virus by pasteurization or S‐sulfonation was at least 105. The clearance of HCV achieved by the entire manufacturing process of each of these immunoglobulins was also at least 105. The experiments therefore show that pasteurization or S‐sulfonation provides a high margin of safety to human immunoglobulins regarding the transmission of hepatitis C virus.
The specificity of antibodies induced by infection with rhinovirus type 2
Authors: Carey B. S. , Barclay W. S. , Russell S. M. , Tyrrell D. A. J.
Published: March 2007

Mouse monoclonal antibodies (MAbs) against three distinct antigenic sites on rhinovirus type 2 have been obtained and the sites identified. We describe how these MAbs were used in a blocking test to detect antibodies in human sera directed against the same three defined sites. Sera from twelve volunteers were studied. All had been exposed to rhinovirus type 2 by intranasal inoculation, four had been uninfected, eight were infected of whom four developed a cold while four did not. Blocking antibodies were high and did not increase in the resistant volunteers, and were lower and increased in the infected volunteers. The antibodies were almost as sensitive as other antibody assays for detecting infection. The responses to all three sites were similar. Correlations between the results of all tests were calculated and the results are summarised.

Tests were also devised to measure the lg subclass of antibodies against the whole virus particle. The A1, G1, and G4 classes showed most frequent rises in response to infection. Correlations between these results and other antibody assays were found and are presented.
Prevalence of four blood‐borne viruses (HBV, HC V, HTLV‐I, HIV‐1) among haemodialysis patients in Japan
Authors: Tamura Ikuo , Koda Tetsuzo , Kobayashi Yasunori , Ichimura Hiroshi , Kurimura Osamu , Kurimura Takashi
Published: March 2007

To investigate the prevalence of four blood‐borne viruses among a cohort of haemodialysis (HD) patients in Japan, hepatitis B surface antigen (HBsAg), antibody to hepatitis C virus (anti‐HCV), antibody to human T‐cell lymphotropic virus type‐I (anti‐HTLV‐I), and antibody to human immunodeficiency virus type‐1 (anti‐HIV‐1) were studied in the sera from 393 consecutive HD patients and in the sera from 786 age‐ and sex‐matched healthy individuals from the general population (controls). The prevalence of anti‐HCV and anti‐HTLV‐I was significantly higher in HD patients than in the controls (17.8% vs. 1.1% and 3.8% vs. 0.5%), but the prevalence of HbsAg showed no significant difference. No patients or controls were positive for anti‐HIV‐1. In HD patients with no history of blood transfusion, anti‐HCV was detected in only one (2.1%) of 48 patients undergoing HD treatment for less than 3 years, and there was no significant difference between the prevalence of anti‐HCV in these patients and in the controls. In HD patients who had received blood transfusion, anti‐HTLV‐I was detected in only one (1.0%) of 103 patients undergoing HD treatment for less than 3 years, and there was no significant difference between the prevalence of anti‐HTLV‐I in these patients and in the controls. These findings suggest that in recent years, the risk of HCV transmission by routes other than blood transfusion in HD patients is low, and that of HTLV‐I transmission by transfusion is very low or non‐existent.
Comparison of cervical cytology and the polymerase chain reaction for HPV 16 to identify women with cervical disease in a general practice population
Authors: Bavin P. J. , Giles J. A. , Hudson E. , Williams D. , Crow J. , Griffiths P. D. , Emery V. C. , Walker P. G.
Published: March 2007

A comparison of the ability of cervical cytology and the polymerase chain reaction (PCR) for human papilloma virus type 16 (HPV 16) to identify women with cervical disease has been performed in a general practice population of 249 women, none of whom were believed to have current cervical disease prior to examination. Within this population, 29 women were found by colposcopy and subsequent histopathology to have evidence of cervical disease [5 with cervical intraepithelial neoplasia (CIN) 3; 8 with CIN 2; and 16 with CIN 1]. The prevalence of HPV 16 in this population was 18.9% (CIN 3, 80%; CIN 2, 50%, CIN 1, 12.5%, normal, 16.8%). Women with severe disease (CIN 2 and CIN 3) had a significantly higher incidence of HPV 16 DNA than those with mild cervical disease (CIN 1) or no cervical abnormality (P = 0.001). There was no significant difference in the ability of either PCR for HPV 16 or cytology to identify women with cervical disease. The combination of screening by cytology and the presence of HPV 16 DNA resulted in the identification of a higher proportion of the women with disease, but this observation did not reach statistical significance. Although the failure to detect disease by the two screening methods was similar, HPV 16 DNA positivity was associated with a higher false‐positive rate for disease detection than cytology (P < 0.03). The PCR assay for detecting HPV 16 in this investigation was shown to have a false‐positive rate of 2.4% and a false‐negative rate of 10.4%. The prospect of screening women for cervical disease using PCR for HPV 16 is discussed. © 1992 Wiley‐Liss, Inc.