bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies by immune electron microscopy of hepatitis B surface antigen in PLC/PRF/5 Cells
Authors: Shibayama Takao , Watanabe Toshiaki , Kojima Hideo , Yoshikawa Akira , Watanabe Shozo , Kamimura Tomoteru , Suzuki Shiro , Ichida Fumihiro
Published: March 2007

Electron microscopic studies of the morphology of hepatitis B surface antigen (HBsAg) produced by PLC/PRF/5 cells in vitro were carried out. Aggregates of 20‐nm spherical particles in 3‐day culture supernatants were observed by immune electron microscopy (IEM). Aggregates of tubular structures were found with IEM in the extracts of the cells. Tubular structures 18 to 22 nm in diameter were seen by electron microscopy (EM) in the cisternae of the endoplasmic reticulum in 2–3% of the cells. The tubular structures in the cytoplasm and extracts of PLC/PRF/5 cells resembled those observed in the hepatocytes of human carriers of hepatitis B virus (HBV). Intracellular localization of HBsAg in PLC/PRF/5 cells by direct peroxidase‐conjugated antibody staining was observed on the tubular structures and the cisternal wall, which contained these structures. Rotation technique analysis indicated that the tubular structures were composed of 11 or 12 subunits.
Polyalbumin receptors on hepatitis B virus and on 22 nm hepatitis B surface antigen (H BsAg)2 particles
Authors: Pontisso P. , Falcieri E. , Schiavon E. , Alberti A. , Realdi G.
Published: March 2007

Receptors for polymerized human serum albumin (pHSA) were studied by solidphase radioimmunoassay on different hepatitis B surface antigen (HBsAg) particles subpopulations prepared both from hepatitis B e antigen (HBeAg) and from anti‐HBe‐positive sera. HBsAg particles in HBeAg‐positive serum showed higher expression of the receptor compared with HBsAg particles from anti‐HBe‐positive serum. Analysis of different morphological forms of virus particles was performed after separation by density‐gradient ultracentrifugation. Maximum receptor expression was detected in HBV particles containing fractions while the 22‐nm HBsAg particles had significantly lower receptor activity. These observations support the hypothesis of a pathogenetic role of the pHSA receptor in mediating virus access to hepatocytes. Indeed, the higher pHSA binding activity on HBV particles could allow selective attachment of the infectious virion to liver cells that bear similar albumin receptors on their surface.
Antibody assays for varicella‐zoster virus: Comparison of competitive enzyme‐linked immunosorbent assay (ELISA), Competitive radioimmunoassay (RIA), Complement fixation, and indirect immunofluorescence assays
Authors: Wreghitt T. G. , Tedder R. S. , Nagington J. , Ferns R. Bridget
Published: March 2007

The competitive ELISA test was compared with competitivc RIA, complement fixation, and indirect fluorescent antibody test for the detection of antibody to varicella‐zoster virus (VZV). ELISA and RIA were the most sensitive tests, being tenfold as sensitive as indirect immunofluorescence and 20 times as sensitive as complement fixation, which was surprisingly inadequate for detecting anti‐VZV in sera from regular haemodialysis patients. The ELISA test was found to be the most suitable assay, employing a routine complement fixation test (CFT) grade viral antigen and not requiring radiolabelling and counting facilities, which are unavailable in most virology laboratories.
Diagnostic value of anti‐HBc IgM in high hbv prevalence areas
Authors: Papaevangelou George , Roumeliotou‐Karayannis Anastasia , Tassopoulos Nicolaos , Stathopoulou Paraskevi
Published: March 2007

The diagnostic value of an anti‐μ‐capture immunoassay for the detection of IgM antibody against hepatitis B core antigen (anti‐HBc) was evaluated. Strongly positive results were obtained from the acute phase sera of the 25 acute hepatitis B patients who were hepatitis B surface antigen (HBsAg) positive and of the 18 confirmed acute hepatitis B patients who had already cleared HBsAg when symptoms developed. Negative results were obtained in 5 hepatitis A patients, 20 non‐A, non‐B acute hepatitis patients serologically susceptible to HBV, 22 patients with chronic hepatitis B liver disease, 15 asymptomatic HBsAg carriers, and 10 healthy patients immune from past HBV infection. Fourteen of the acute hepatitis patients remained HBsAg positive for a follow‐up period of at least 6 months, and 12 of these were found consistently anti‐HBc IgM negative. These were considered as chronic HBsAg carriers with a superimposed form of acute liver injury.

These data show that this assay can differentiate between acute from chronic (HBsAg positive) and recent from old (HBsAg negative) hepatitis B virus infection. Thus, it should be very useful in the complex diagnostic situations encountered commonly in areas with high prevalence of HBV infections.
Monoclonal antibodies to hepatitis A virus
Authors: Dawson George J. , Decker Richard H. , Norton Debra K. , Bryce Warren H. , Whittington Richard O. , Tribby Use I. , Mushahwar Isa K.
Published: March 2007

This paper describes the development of monoclonal antibodies generated against hepatitis A virus (HAV). Monoclonal antibodies (MCABs) from two murine hybridoma cell lines were found to bind to an epitope recognized in the sera of patients recovering from infection with HAV. Ascitcs fluids containing MCABs from one hybridoma (Hl C19) inhibited a maximum of 70% of the 125I‐labeled polyclonal human anti‐HAV from binding to HAV‐antigen in a competitive radioimmunoassay, indicating that the MCAB recognizes a major epitope of HAV. Monoclonal anti‐HAV that was coated onto polystyrene beads was as effective as polyclonal antibodies in capturing HAV antigen from extracts of human feces, marmoset liver, and cultured cells. Radiolabeled MCAB was used to screen sera for anti‐HAV. A collection of 117 sera was tested for total anti‐HAV by competitive radioimmunoassays utilizing either 125I‐labeled human polyclonal or mouse monoclonal antibody. Thirty‐four specimens were similarly reactive by both systems, while the remainder were negative. Likewise, 28 specimens were similarly positive for IgM anti‐HAV, and 12 specimens were negative using each of the two labeled antibodies. The data show that anti‐HAV induced during disease is directed against a common major epitope of HAV and that MC anti‐HAV can be used effectively as a reagent to detect these antibodies.
Evaluation of a reduced dose of hepatitis B vaccine administered intradermally
Authors: Zoulek Gert , Lorbeer Bernd , Jilg Wolfgang , Deinhardt Friedrich
Published: March 2007

Intradermal inoculation of hepatitis B vaccine (HBsAg subtype adw) caused no side effects, but the vaccine was less immunogenic than following intramuscular administration. Intradermal inoculation does not, therefore, offer a major advantage to the generally used intramuscular immunization. A single multisite intradermal administration of a reduced dose of vaccine did not result in a more rapid seroconversion compared to intramuscular inoculation. Although the antibody levels were similar after two intradermal or intramuscular injections given 1 month apart, the booster (third injection) at 6 months resulted in anti‐HBs levels that were about 10 times higher following intramuscular inoculation as compared to intradermal.

All persons immunized developed anti‐HBs. The levels of anti‐HBs (a and w) were about 30–40% of the total anti‐HBs, and the proportion did not change significantly during the course of immunization. Cross‐protection against all HBV strains is thus also assured after intradermal administration of vaccine containing only one HBsAg subtype (adw).

A skin reaction was elicited only in a small proportion of anti‐HBs‐positive individuals, and the reaction correlated roughly with the immune response.
Prevalence of delta agent among Chinese in Taiwan and Los Angeles
Authors: Govindarajan Sugantha , Lee Shou‐Dong , Tong Myron J. , Tsai Yang‐Te , Lo Kwang‐Juei
Published: March 2007

The prevalence of delta agent among 120 Chinese patients with HBsAg‐positive chronic active and hepatocellular carcinoma from Taiwan and Los Angeles were studied by serological assay for antibody to delta antigen and immunoperoxidase staining for delta antigen in the liver biopsies. Of 120 patients, only four with chronic active hepatitis and one with hepatocellular carcinoma from Taiwan had antibody to delta antigen in their sera, while none was detected in the patients from Los Angeles. Indirect immunoperoxidase staining for delta antigen in the liver tissue from three of the patients with circulating anti‐delta was positive. Thus, the overall prevalence of delta infection was 5.8% among the 86 patients from Taiwan, and 0% in the 34 Chinese patients from Los Angeles.
Immunoglobulin class‐specific antibody response in respiratory syncytial virus infection measured by enzyme immunoassay
Authors: Meurman Olli , Ruuskanen Olli , Sarkkinen Hannu , Hänninen Pentti , Halonen Pekka
Published: March 2007

Immunoglobulin class‐specific enzyme immunoassay (EIA) was used for determination of antibody responses in sera collected from 26 children with acute primary respiratory syncytial virus (RSV) infections. All 26 patients had IgG antibody responses with a significant titer increase in 24 (92%); an IgM antibody response was detected in 19 of the 26 (73%). From patients aged 6 months or less only 5 of 8 produced detectable IgM antibodies, whereas all patients aged 1–2 years did so. IgM antibodies appeared within 1 week after onset of illness and persisted from 20 days to 2–3 months. An IgA antibody response was observed in 20 of 26 (77%) patients with a significant titer increase detected in 17 of 26 (65%) patients. In some patients the persistence of IgA antibodies followed that of the IgM antibodies, but in others the IgA antibody titers remained stable up to the end of the follow‐up. The most sensitive assay system for serological diagnosis of acute RSV infection in children was the determination of titer increases by IgG antibody.
Detection of IgM antibodies against cytomegalovirus: Comparison of two radioimmunoassays, enzyme‐linked immunosorbent assay and immunofluorescent antibody test
Authors: Kangro Hillar O. , Booth Jim C. , Bakir Talal M. F. , Tryhorn Yvonne , Sutherland Sheena
Published: March 2007

The sensitivity and specificity of direct antibody radioimmunoassay (RIA), M‐antibody capture RIA (MACRIA), enzyme‐linked immunosorbent assay (ELISA), and the immunofluorescent antibody (IFA) test for the detection of CMV‐specific IgM was compared using 40 sera selected from different groups of patients.

RIA, MACRIA, and ELISA gave concordant results with thirty‐two sera but discordant results with eight sera, of which three were cord sera from congenitally infected babies, three were from immunocompromised patients with recurrent CMV infections, and two were from patients with lymphadenopathy and Paul‐Bunnell‐positive mononucleosis, respectively.

RIA, MACRIA, and ELISA were of similar sensitivity with sera from adult patients, but ELISA was apparently less sensitive than RIA and MACRIA for the detection of CMV IgM in cord serum. By comparison IFA was significantly less sensitive than the other three tests.

Rheumatoid factor is reactive in RIA, ELISA, and IFA but can efficiently be removed by absorption with latex‐IgG beads or cross‐linked human IgG.
Immunogenic effect of hepatitis B vaccine in children: Comparison of two‐and three‐dose protocols
Authors: Yvonnet Bernard , Coursaget Pierre , Petat Eric , Tortey Eric , Diouf Cheikh , Barin Francis , Denis Francois , Diop‐Mar Ibrahima , Chiron Jean‐Paul
Published: March 2007

The immunogenic effect of hepatitis B vaccine was evaluated in 183 seronegative infants from Senegal. Seventy‐two seronegative infants received two 5‐μg doses of vaccine at a two‐month interval and 111 seronegative infants received three 5‐μg doses at one—month intervals. All the children had a booster dose one year after the first injection of vaccine. No difference between the two groups was observed in the seroconversion rate (93.1% and 94.6%, respectively); in the proportion of high anti—HBs titer; or in the anti—HBs geometric mean titer (82 and 92 mIU/ml, respectively). These results demonstrate that two doses of 5μg of hepatitis B vaccine are sufficient in infants to obtain a high immunogenic effect.
Acute type A hepatitis during chronic hepatitis B virus infection: Association of depressed hepatitis B virus replication with appearance of endogenous alpha interferon
Authors: Davis Gary L. , Hoofnagle Jay H. , Waggoner Jeanne G.
Published: March 2007

Two patients with chronic type B hepatitis and intercurrent episodes of acute type A hepatitis are presented. Serological markers of hepatitis B virus replication decreased or became undetectable in both patients during the acute illness, while interferon activity was transiently detected in serum. The presence of serum leukocyte (alpha) interferon was confirmed by neutralization with specific antisera and tests of pH2 stability. These observations suggest a role for natural leukocyte (alpha) interferon in the modulation and control of hepatitis B virus infection and provide further evidence to support trials of exogenous leukocyte (alpha) interferon in the chronic infection.
Reverse radioimmunoassays of IgM and IgG antibodies to coxsackie B viruses in patients with acute myopericarditis
Authors: Frisk Gun , Torfason Einar G. , Diderholm Hans
Published: March 2007

A reverse radioimmunoassay (RIA) of antibodies to enteroviruses, previously developed for the detection of IgM antibodies to Coxsackie B1 (CB1) and B3 (CB3) and to Echo 11 (Ell) and 30 (E30) viruses, was extended in the present study for the detection of IgM antibodies to Coxsackie B2 (CB2), B4 (CB4), and B5 (CB5) viruses and of IgG antibodies to CB1‐CB5, Ell, and E30 viruses. After standardisation of the assays and application to a collection of serum specimens from patients with proven enterovirus infections, specimens from patients with diagnosed or suspected acute myo‐ and/or pericarditis (myopericarditis group), and control specimens from patients with nonenterovirus infections, were studied, as well as from apparently healthy subjects.

Of the patients with enterovirus infections, 29 of 30 (97%) were positive in the IgM RIA and 19 of 25 (76%) in the IgG RIA. In the myopericarditis group, 18 of 37 (49%) patients showed Coxsackie B (CB) virus‐specific IgM titres and 9 of 37 (24%) CB virus‐specific IgG titres. In the control specimens very few positive responses were detected. The RIAs appeared to be type specific or at least predominantly type specific, provided that the amount of labeled virus was carefully standardised. The sensitivity of the RIAs seemed to be rather high for IgM but low for IgG. In the neutralisation (NT) test no significant rise or fall in titre against CB viruses was demonstrated in the myopericarditis group. It is concluded that the reverse IgM RIA may be valuable for studies of the role of CB viruses in acute myo‐ and/or pericarditis.
Specific IgG and IgA antibodies to herpes simplex virus (HSV)‐lnduced surface antigen in patients with HSV infections and in healthy adults
Authors: Hadar Tuvia , Sarov Israel
Published: March 2007

Herpes simplex virus (HSV)‐specific IgG and IgA antibody response in patients with HSV infection and in healthy adults was studied by the immunoperoxidase antibody‐membrane antigen (IPAMA) technique. In all HSV infections in which specific IgM antibodies were detected by enzyme‐linked immunosorbent assay (ELISA), a significant rise in the titer of HSV IgG and IgA antibodies was found. In contrast, in patients with recurrent herpes labialis in which no specific HSV IgM antibodies were detected by ELISA, HSV IgG and IgA antibodies were not found to fluctuate significantly during the course of infection. A higher geometric mean titer (GMT) for HSV IgG and for IgA antibodies was found in seropositive individuals with a previous history of recurrent HSV than in seropositive individuals without a previous history of recurrent HSV infection. Nineteen of 26 HSV IgG seropositive healthy medical students without a previous history of recurrent HSV infection had HSV IgA antibodies to membrane antigen. The significance of this finding in understanding the mechanism of latency in healthy seropositive individuals without previous history of HSV recurrent infections is discussed.
The removal of hepatitis B virus from factor VIII concentrates by fractionation on ethylene maleic anhydride polyelectrolyte
Authors: Galpin S. A. , Karayiannis P. , Middleton S. M. , Thomas H. C.
Published: March 2007

Chromatography with a solid‐phase polyelectrolyte based on ethylene maleic anhydride polyelectrolyte (EMA PE) has been shown to remove a large proportion of hepatitis B surface antigen (HBsAg), including hepatitis B virus (HBV) particles, from contaminated cryoprecipitate. This process may reduce the risk of transmission of HBV infection when factor VIII concentrates are administered.
DNA of 21 clinical cytomegalovirus strains detected by hybridization to cloned DNA fragments of laboratory strain AD‐169
Authors: Chou Sunwen , Roark Linda , Merigan Thomas C.
Published: March 2007

Nine distinct DNA probes prepared from cloned DNA fragments of the cytomegalovirus (CMV) strain AD‐169, each representing a relatively small portion of the total genome, were able to detect all 21 epidemiologically distinct clinical isolates of CMV that had been passaged in tissue culture. All probes were of comparable sensitivity in the detection of CMV, and under the hybridization conditions used, no probe gave an unusually high background with uninfected host cells.
The heat sensitivity of hepatitis A virus determined by a simple tissue culture method
Authors: Parry J. V. , Mortimer P. P.
Published: March 2007

A method for estimating residual infectious hepatitis A virus (HAV) after heat treatment of suspensions of the virus was devised. It made use of a readily maintained cell line (FRhK‐4) in which the rate of release of HAV antigen into the tissue culture medium was directly proportional to the size of the inoculum. Loss of viral infectivity after heating could be estimated by inoculating heat‐treated HAV suspensions into cell monolayers and measuring antigen released from the cultures during 5 weeks of incubation.

An inoculum of 105 tissue culture infecting doses of HAV suspension in phosphate‐buffered saline retained 1.0% of its infectivity after treatment for 2 min, and 0.01 % after treatment for 4 min at 65°C. The virus was fully inactivated within 4 min at 70°C, 30 sec at 75°C, 5 sec at 80°C, and virtually instantaneously at 85°C. When HAV was suspended in milk and subjected to statutory pasteurisation conditions, 0.1% of its infectivity remained after 30 min at 62.8°C and 1.0% after 15 sec at 71.6°C.

The implications of these results for the heat treatment of food, water, and milk are discussed. It appears that recommended heat processes for milk and shellfish may not be adequate to inactivate HAV.
Immunogenicity of wild and attenuated varicella‐zoster virus strains in rhesus monkeys
Authors: Asano Yoshizo , Albrecht Paul , Behr Dorothy E. , Neff Beverly J. , Vickers James H. , Rastogi Suresh C.
Published: March 2007

Thirty susceptible rhesus monkeys were inoculated with cell‐free varicella‐zoster virus strain OKA or strain KMcC. Both wild and attenuated strains were used. No clinical signs characteristic of human varicella were seen in any of the animals. Virus was not isolated from throat swabs, blood, or cerebrospinal fluid. Antibodies were measured by an enhanced plaque neutralization test. The wild and attenuated OKA strains produced comparable levels of antibodies for 3 months after inoculation. Attenuated KMcC strains produced lower titers than the wild strain. On rechallenge 3 months after primary inoculation animals boostered with the attenuated OKA strain developed significantly higher antibody titers than animals receiving the wild strain. Animals primed and challenged with the attenuated KMcC strains showed significantly lower antibody titers than animals which received the wild strain. The results indicate that the immunogenicity of attenuated OKA and KMcC strains in rhesus monkeys parallels the experience obtained with these strains in humans.
The effects of human cytomegalovirus challenge in vitro on subpopulations of T cells from seronegative donors
Authors: Sing Garwin Kim , Garnett Helen M.
Published: March 2007

Purifed T cells from seronegative donors were challenged with human cytomegalovirus (CMV) and assayed for response to the mitogen phytohemagglutinin (PHA) administered at various times after virus challenge. Cells were hyporesponsive to PHA added up to 6 h postinfection, while cells challenged with ultraviolet (UV)‐inactivated CMV showed normal PHA responses. T‐cell populations showed a complete inversion of helper/suppressor ratios by 6 h of infection, as determined by indirect immunofluorescence using the monoclonal antibodies OKT4 and OKT8. This increased proportion of suppressor cells was functionally demonstrable in an autologous one‐way mixed lymphocyte reaction (AMLR).
How useful are serum and CSF interferon levels as a rapid diagnostic aid in virus infections?
Authors: Flowers Dianne , Scott Geoffrey M.
Published: March 2007

Specimens from a total of 162 patients were studied prospectively using a rapid nucleic acid synthesis inhibition assay and a plaque‐reduction assay to detect interferon levels in serum and cerebrospinal fluid. The specificity, consistency, and diagnostic applications of the rapid assay were evaluated by intensively investigating each patient for possible viral illness. Forty‐one percent of patients with proven acute virus infections (excluding those with acute hepatitis and infectious mononucleosis) had unequivocal detectable interferon in serum or cerebrospinal fluid, compared with only two patients with bacterial infections (7%) who had equivocal interferon in the serum. Retrospective analysis showed the test to have high specificity for virus infections, but in patients with clinical syndromes of uncertain aetiology the test was not a particularly sensitive indicator of the presence of subsequently detected virus. The presence of interferon was not related to the time during the illness at which the sample was taken, nor to the numbers of lymphocytes in the cerebrospinal fluid, nor to the patients' general symptoms.
Presence of antibodies to human lymphoma‐leukemia virus (HTLV‐I) in Germans with symptoms of the acquired immunodeficiency syndrome (AIDS)
Authors: Born M. , von der Helm K. , Wernicke D. , Deinhardt F.
Published: March 2007

Sera from German patients exhibiting symptoms compatible with the acquired immune deficiency syndrome (AIDS) or the lymphadenopathy syndrome (LAS) were assayed for antibodies against human T‐cell lymphoma/leukemia virus (HTLV‐I)‐related antigens by enzyme imniunoassay, indirect immunofluorescence, and radioimmunoprecipitation. Antibodies against HTLV were detected in 3 out of 31 sera.
Immunogenicity of a five‐microgram dose of hepatitis B vaccine
Authors: Papaevangelou George , Roumeliotou‐Karayannis Anastasia , Vissoulis Charalambos , Richardson Steven‐Clive , Krugman Saul
Published: March 2007

The immunogenicity of a 5‐μg dose of vaccine (H‐B‐Vax, MSD) was evaluated in 50 young adults (17–19 years). Results were compared to our previous studies using similarly prepared vaccines using 20 μg and 10 μg per dose with the same trial protocol in a comparable population. Seroconversion rates for the 5‐μg doses of vaccine were 80% after the first dose and 98% after the second dose. The remaining participants did not develop anti‐HBs in the course of the trial. These results are not significantly different from those observed in the 10‐μg and 20‐μg studies. The increase of anti‐HBs titers was slower for the 5‐μg group. High geometric mean titers were observed after booster vaccination, but lower for the 5μg (3,591 mIU/ml) than for 10 μg (9,277 mIU/ml) and 20 μg (12,975mIU/ml) doses. It is concluded that 5‐μg dose of the vaccine is effectively immunogenic for young adults.
Horizontal transmission of hepatitis B virus among children and adults in five rural villages in Zambia
Authors: Tabor E. , Bayley A. C. , Cairns J. , Pelleu L. , Gerety R. J.
Published: March 2007

To determine whether horizontal transmission of the hepatitis B virus contributes to the high prevalence of infection with this virus in an endemic region, residents of five villages in Zambia were tested for hepatitis B serologic markers. The prevalence of hepatitis B was determined by testing samples from 620 residents. By examining paired serum samples from 79 children and 80 adults, it was determined that new infections occurred during the five years of this study in at least 14 children (18%) (aged 4–17 years) and ten adults (12%) (aged 23–65 years). These 24 new infections were distributed among 20 households and were not associated with active HBV infections in the mother or, in most cases, other family members. Intervention to prevent hepatitis B in regions such as rural Zambia will require vaccination of susceptible children and adults as well as newborn infants.
Genetic heterogeneity of the human papovaviruses BK and JC
Authors: Beckmann Anna Marie , Shah Keerti V. , Mounts Phoebe
Published: March 2007

We have examined the structure and infectivity of BKV and JCV genomes from prototype strains after cell culture passage and of BKV genomes from primary isolates. Genomic structures were determined by restriction endonuclease analysis of molecularly cloned DNA. Infectivity was determined by transfection of the cloned genomes into urine‐derived epithelial cells and assaying for viral proteins and virus production. Prototype BKV DNA, which was cloned after 14 passages in three different cell lines, contained no alterations in restriction enzyme sites and was infectious. In contrast, prototype JCV acquired changes in the late region of the genome during passage in cell culture and the cloned DNA was not infectious. Urine‐derived cells were used to isolate virus from the urine of two renal transplant patients and one asymptomatic individual. The genome of the virus isolated from the normal individual was indistinguishable from prototype BKV except for a 60‐base pair deletion, which was localized between 0.62 and 0.72 map units. Two isolates from transplant patients differed from each other and from prototype BKV at a number of restriction enzyme cleavage sites located in the early region and were infectious. Genomes containing deletions from 100 to 600 base pairs were also cloned but were not infectious.
AN6520 Ag: An antigen purified from liver with non‐A, non‐B hepatitis
Authors: Tohmatsu Jun‐Ichi , Morimoto Tomiaki , Katsuhara Norimichi , Abe Kenji , Shikata Toshio
Published: March 2007

An extract prepared from the liver of a patient with chronic non‐A, non‐B (NANB) hepatitis was found to produce a precipitin line in immunodiffusion with a serum from a multiply transfused patient and those from patients convalescent from NANB hepatitis. The antigen was purified by gel filtration and density gradient centrifugation. The antigen had a buoyant density of 1.16–1.20 g/cm3 in cesium chloride, a sedimentation coefficient (S20, w) of 51.5s, and a molecular weight of larger than 1.5 × 106daltons. Electron microscopic examination revealed particles 29–34 nm in diameter (average 31.5 nm), which could be agglutinated by the specific antiserum. We developed a reverse passive hemagglutination (R‐PHA) and a passive hemagglutination (PHA) technique for detection of the new antigen and antibody, respectively, and applied these to human sera. Antibody to the antigen was detected in 19/28 (67.9%) convalescent sera of NANB hepatitis. This prevalence was significantly higher than those found in convalescent sera of type A hepatitis patients (2/17 = 11.8%), type B hepatitis patients (2/15 = 13.3%), and normal blood donors (9/129 = 7.0%) (p < 0.01); and the prevalence in hepatitis A and B patients did not differ significantly from that of normal donors. Furthermore, most (66.7%) of the cases of NANB hepatitis endemic in Shimizu City, Japan, showed clear seroconversion with respect to this antibody. These results suggest that the new antigen/antibody system is associated with NANB hepatitis.
Detection of HBV‐DNA by in situ hybridization using a biotin‐labeled probe
Authors: Negro Francesco , Berninger Mark , Chiaberge Elisabetta , Gugliotta Patrizia , Bussolati Giovanni , Actis Giovanni C. , Riuetto Mario , Bonino Ferruccio
Published: March 2007

A biotin‐labeled DNA probe specific for hepatitis B virus (HBV) nucleotide sequences was hybridized in situ to liver tissue of 20 patients; 16 were chronic carriers of hepatitis B surface antigen (HBsAg) and 4 had no markers of HBV infection. HBV‐DNA was also analyzed in the serum and the liver of these patients by spot and Southern blot hybridization, respectively.

Liver specimens from six carriers were positive for HBV‐DNA both by in situ and Southem blot hybridization; ten carriers were negative by in situ hybridiza‐ tion, and two of these were positive by Southern blot technique. The staining was granular, mainly cytoplasmic, limited to liver specimens containing replicative forms of HBV‐DNA, and associated with detection of HBcAg in hepatocytes by immunofluorescence. The sensitivity of this technique was not sufficient to detect few copies of integrated HBV‐DNA.

The hybridization procedure was specific, as results were constantly negative in liver specimens of patients without markers of HBV infection, and no reaction was observed using DNA probes lacking HBV‐DNA sequences.

Detection of HBV‐DNA by in situ hybridization, using a biotinylated probe, is a rapid, reproducible, and specific histochemical method. Currently available biotinylated probes are advantageous when absolute sensitivity is not the limiting factor, and they also facilitate studies of the cellular and subcellar distribution of HBV nucleic acids.
Polyalbumin receptors, hepatitis B surface antigen (HBsAg), and HBsAg/IgM complexes in HBsAg positive patients with and without delta superinfection
Authors: Craxì A. , Magrin S. , Greco J. , Vinci M. , Tinè F. , Raimondo G. , Longo G. , Pagliaro Luigi
Published: March 2007

Receptors for polymerized human albumin are found at high litres during high‐level hepatitis B virus (HBV) replication and in small amounts in chronic low‐level infection. Complexes between hepatitis B surface antigen (HBsAg) and IgM without specificity for HbsAg are expressed in a pattern similar to that of receptors. Anti‐albumin antibodies could be involved in their formation. Delta infection depresses the synthesis of gene products of HBV. To assess whether delta modifies the expression of receptors on HBsAg and the level of HBsAg/IgM complexes, and if anti‐albumin antibodies are actually part of the complex, we tested sera from 86 subjects with acute and chronic HBV infection. Our findings show that the amounts of circulating receptors and HBsAg/IgM are proportional to the concentration of HBsAg and thus probably to the degree of viral replication. We did not find any correlation between circulating anti‐albumin antibodies of the IgM class and HBsAg/IgM complexes. Delta infection depresses HBsAg synthesis and causes a related decrease in receptors and HBsAg/IgM titres if superimposed on the more active stage of HBV infection. When HBsAg, receptors, and HBsAg/IgM are at low levels, no further depression is caused by delta infection. HBsAg/IgM titre is not enhanced by presence of anti‐delta antibodies, thus excluding a role of the latter in forming these complexes.
A final report on safety and immunogenicity of a bivalent aqueous subunit HBV vaccine
Authors: Prince A. M. , Brotman B. , Purcell R. H. , Gerin J. L.
Published: March 2007

The bivalent form of an aqueous formalin‐inactivated hepatitis B vaccine was evaluated for safety and immunogenicity in chimpanzees. To evaluate safety five animals were inoculated intravenously with vaccine containing 500 μg HBsAg and two animals with 50 μg. None of these animals developed hepatitis or any serologic marker indicative of the presence of residual live vims in the vaccine.

Twenty‐four animals were used to evaluate immunogenicity and protective efficacy. Seven of these immunized animals produced weak or no anti‐HBs responses. Two doses of 50 μg HBsAg given subcutaneously 1 month apart protected each of four animals that were challenged with 103.5 CID50 HBV at 6 and 12 months after immunization and protected three of four animals challenged at 24 months against development of hepatitis or HBsAg. Three of 4 animals in each group immunized with two doses of 20, 10, or 5 μg HBsAg were similarly protected when challenged 6 months after immunization.

Thirteen of 20 immunized animals that did not develop HBsAg after challenge with HBV developed anamnestic anti‐HBs or anti‐HBc responses between 2 and 18 months after challenge, indicating minimal replication of challenge virus. The time of onset and frequency of occurrence of these delayed responses was related to the titer of anti‐HBs at the time of challenge.

False positive Ausab test results were observed in quarantined chimpanzees. These were neither preceded by appearance of HBsAg nor accompanied by development of anti‐HBc. In most cases these reactions were due to a reactant having a sedimentation coefficient and an electrophoretic mobility resembling that of IgM. This reactant generally did not appear to confer resistance to challenge with HBV.

The humoral immune response was characterized as being entirely of the IgM class 2 weeks after immunization and switched entirely into the IgG class by 10–12 weeks after vaccine administration. At the time of challenge all animals with antibody had anti‐HBs of subtype a.
Direct detection of the human papovavirus BK in urine of bone marrow transplant recipients: Comparison of DNA hybridization with ELISA
Authors: Arthur Ray R. , Beckmann Anna Marie , Li Chou Chi , Saral Rein , Shah Keerti V.
Published: March 2007

Urine specimens from bone marrow transplant (BMT) recipients and from controls were directly tested for BK virus (BKV) DNA sequences by dot hybridization and for BKV antigen by a double‐antibody indirect ELISA. A total of 158 specimens from 55 BMT patients (57 collected prior to or at the time of transplantation and 101 in the posttransplant period) and single urines from 125 control subjects were examined by both methods. A molecularly cloned, 32P‐labelled BKV probe was hybridized with urine sediments that were spotted directly on nitrocellulose filters and denatured in situ. BKV DNA sequences were detected in 1 (1.8%) pretransplant and 22 (21.8%) posttransplant urines of BMT patients, and in none of control urines. In ELISA of urine supernatants, BKV antigen was detected in 1 (1.8%) pretransplant and 21 (20.8%) posttransplant urines of BMT patients and in 1 (0.8%) of the control urines. The results of the two tests correlated as follows: 16 urines were positive and 253 urines negative by both methods; seven specimens were positive by DNA hybridization only and seven were positive by ELISA alone. Virus excretion in urine was demonstrated in 20 (36.4%) patients by DNA hybridization, in 19 (34.5%) patients by ELISA, in 15 (27.3%) patients by both methods, and in 24 (44%) patients by at least one of the two tests.
Retrovirus‐like particles in hepatocytes of patients with transfusion‐acquired Non‐A, Non‐B hepatitis
Authors: Iwarson Sten , Schaff Zsuzsa , Seto Belinda , Norkrans Gunnar , Gerety Robert J.
Published: March 2007

Retrovirus‐like particles 60–85 nm in diameter were observed in the cytoplasm of hepatocytes in liver biopsies obtained during the acute and chronic phases of non‐A, non‐B hepatitis (NANBH) in three patients with transfusion‐acquired disease. The particles appeared in dilated endoplasmic reticulum cisternae as well as in enlarged Golgi vesicles. No such particles were seen in hepatocytes in liver biopsies similarly obtained during the acute or chronic phases of NANBH from 11 additional patients with NANBH who did not acquire their disease following blood transfusion. Particle‐associated reverse transcriptase activity (peak activity at a density of 1.14 gm/ml) was present in the sera of all three “particle‐positive” patients and also in 42% of the “particle‐negative” patients. The retrovirus‐like particles described here were apparently unrelated to the previously described human T cell lymphocytotropic retroviruses (HTLV), since none of the 14 patients studied had antibodies in their serum directed against antigens of any of the three known HTLVs.
Human‐enteric‐coronaviruslike particles (CVLP) with different epidemiological characteristics
Authors: Sitbon Marc
Published: March 2007

One hundred fifty‐six diarrheic and 115 control stools collected throughout a year from nonhospitalized children were examined by electron microscopy in Haut‐Ogooué, Gabon; 65.2% of the controls and 38.5% of the diarrheics were found to contain coronaviruslike particles (CVLP). In both diarrheic and control groups the CVLP prevalences showed a seasonal variation whereas significant variation of prevalence with age was observed only in the controls. Thus, the CVLP in controls were significantly more abundant in children over 2 years old (76% vs 48%, P < .01) and more frequently observed during the months of rainy seasons (75% vs 54%, P < .02). On the other hand, the higher prevalence of CVLP in diarrheics over 2 years old was not significant (48% vs 36%, P = .20), whereas a significantly lower prevalence of CVLP during the months of rainy seasons was observed in this group (27% vs 50%, P < .01). Studies of the climatological factors in this equatorial climate showed a parallel cyclical variation of parameters representing rainfall, temperature, as well as relative humidity. We were not able to distinguish which of these factors was influencing more directly the prevalence of CVLP.
Inactivation of the hutchinson strain of non‐A, non‐B hepatitis virus by combined use of β‐propiolactone and ultraviolet irradiation
Authors: Prince Alfred M. , Stephan Wolfgang , Dichtelmüller Herbert , Brotman Betsy , Huima Tellervo
Published: March 2007

A b̃‐propiolactone/ultraviolet irradiation procedure (βPL/UV) has been evaluated for its ability to inactivate 30,000 chimpanzee infectious doses of the Hutchinson strain of non‐A, non‐B (NANB) virus. The chimpanzees were inoculated with plasma to which this dose of the titrated virus had been added prior to application of the βPL/UV process in accordance with a procedure used for licensed blood derivatives in Germany. Neither animal developed hepatitis. When subsequently challenged with the same contaminated plasma, which had not been sterilized, both animals promptly developed typical NANB hepatitis.

This study extends the high (∼107‐fold) process efficiency of the βPL/UV procedure previously reported for hepatitis B virus to a blood‐borne NANB virus.
The immune response to herpes simplex virus: Comparison of the specificity and relative titers of serum antibodies directed against viral polypeptides following primary herpes simplex virus type 1 infections
Authors: Eberle R. , Mou Siao‐Wen , Zaia John A.
Published: March 2007

Employing an immunoblotting technique, the polypeptide specificity and relative titers of anti‐HSV IgG reactive with denaturation‐resistant epitopes on HSV proteins were determined in patients experiencing primary HSV‐1 infections at various anatomical sites. Early sera from previously seronegative patients with primary HSV‐1 infections were found to have comparatively low levels of antibody directed against the major viral glycoprotein antigens (gB, gC, and gD) relative to titers present in sera of individuals with long‐standing, latent orofacial HSV‐1 infections. Patients with primary infections did however have high titers of antibody directed against a series of low molecular weight HSV polypeptide antigens. These antigens were found to be antigenically related to a structural component of virion nucleocapsids. At later times postinfection, titers of antibodies directed against other viral polypeptides including the major glycoproteins increased to levels more closely approximating those observed in latently infected individuals. These results indicate that the anti‐HSV IgG detected by immunoblot analysis which appears earliest following primary infection is not directed against the known major infected cell or virion glycoprotein surface antigens but rather against an internal capsid protein of HSV.
Relapsing viral hepatitis type A
Authors: Jacobson Ira M. , Nath Barbara J. , Dienstag Jules L.
Published: March 2007

Three patients with a relapsing course of serologically confirmed viral hepatitis type A are presented. In addition, one patient had aminotransferase elevations for over 6 mo. In all three, hepatitis ultimately resolved. A relapsing or protracted course does not appear to alter the benign prognosis of hepatitis A.
Horizontal transmission of hepatitis b virus infection in household contacts, pune, india
Authors: Dhorje Sanjay P. , Pavri Khorshed M. , Prasad Subba R. , Sehgal Ashok , Phule Dnyaneshwar M.
Published: March 2007

The study was undertaken to detect the risk of infection, if any, among 193 household contacts of 40 hospitalised hepatitis patients (group I) with hepatitis B surface antigen (HBsAg) in their blood. As a control group, 103 household contacts of 27 hospitalised hepatitis patients who were negative for HBsAg (group II) were investigated. The family contacts of the former group had a significantly higher prevalence of HBV infection than those of the latter group (P < .001). Significant differences were observed both in the prevalence of HBsAg (P < .05) and antibody to HBsAg (anti‐HBs) (P < .025) between the two groups. IgM antibody to hepatitis B core antigen (anti‐Hbc‐IgM) was detected in 32 out of the 39 (82%) sera tested from the patients of group I with HBsAg. A statistically significant difference (P < .005) of HBV prevalence was also found in the contacts of these 32 patients suffering from acute hepatitis B as compared to the contacts of the patients of group II.

Overall, the children of the first group showed a significantly higher prevalence of HBsAg as compared to the second group. All the children with HBsAg were positive for HBeAg also, but were negative for anti‐HBc‐IgM. Anti‐HBs was detected in a significantly larger number of adult females. Spouses were found to be affected more than other relatives. A significant difference (P < .025) was noted in the number of families having HBV markers in group I (80.0%) as compared to those in group II (48.1%). In group I, contacts of male patients had a significantly higher (P < .025) prevalence of HBV infection than those of the female patients (46.4% vs 29.3%).
Comparison of various serological methods and diagnostic kits for the detection of acute, recent, and previous rubella infection, vaccination, and congenital infections
Authors: Enders Gisela , Knotek Frank , Pacher Uta
Published: March 2007

The antibody development after natural rubella infection and rubella vaccination has been followed in 802 sera from 493 patients and 71 sera from 22 vaccinees. Also examined were 67 sera from 28 infants with rubella embryopathy and sera from 50 children with presumed prenatal infection. In addition, 777 sera from 641 patients tested for routine rubella diagnosis were studied. Anamnestic information was available from all these patients. These sera were assayed for IgM antibody detection by sucrose density gradient (SDG), the commercial ELISAs (Enzygnost IgG Rubazyme), and the noncommercial anti‐my‐hemadsorption immunosorbent technique (HIT). For the determination of IgG antibodies the hemagglutination inhibition test (HAI), the commercial ELISAs (Enzygnost IgG, Rubazyme), and a single radial hemolysis test (SRH) were used. The SDG and HIT were less sensitive for IgM antibody detection than the two ELISAs, particularly when IgM concentrations were low. In total 26.5% of the IgM results with the newer tests were discordant with SDG, but only 0.5–3% of these results were not explicable when the clinical data was considered. Problems were encountered with all IgM assay systems used. For the detection of rubella antibodies after acute infection and vaccination the ELISA Enzygnost IgG was as sensitive as the HAI whereas the ELISA Rubazyme and SRH detected antibodies with some delay. Corresponding results with all tests were found more than 25 days after acute infection and more than 50 days after vaccination. All methods can be used for detection of antibodies in infants with rubella embryopathy. The results of this study suggest that certain combinations of tests can be used for the reliable detection of rubella infection.
Inhibition of in vitro HBsAg production by amphotericin B and ketoconazole
Authors: Pottage John C. , Kessler Harold A.
Published: March 2007

The effects of amphotericin B, ketoconazole, and adenine arabinoside on production of hepatitis B surface antigen (HBsAg) particles by the human hepatoma cell line PLC/PRF/5 were examined. In addition, the effects of these drugs on cellular protein synthesis were determined. These drugs caused a dose‐dependent decrease in HBsAg production that was paralleled by a decrease in cellular protein synthesis. Ketoconazole was the most active of these drugs and the most specific, causing a 72% reduction in HBsAg production with only a 38% reduction in protein synthesis. These data suggest that further studies evaluating ketoconazole for the treatment of chronic hepatitis B virus infection in animals are warranted.
Detection by ELISA of IgG‐Subclass‐Specific Antibodies in Primary Respiratory Syncytial (RS) Virus Infections
Authors: Hornsleth Allan , Bech‐Thomsen Niels , Friis Birgitte
Published: March 2007

The IgG‐subclass specific antibody response was investigated in primary RS‐virus infections in infants and small children by using an ELISA with monoclonal antibodies against the four human IgG subclasses. When 78 serum samples obtained from 21 patients during the first 3–4 mo following the onset of illness were studied, only IgG‐1 and IgG‐3 antibodies could be detected. Both of these subclass specific antibodies appeared during the second week following the beginning of illness. Approximately 4 wk after the onset of illness, IgG‐1 antibodies reached maximum levels, which persisted during the following months. Approximately 3 wk after the onset of illness, IgG‐3 antibodies reached maximum levels, which decreased during the following months. Both IgG‐1 and IgG‐3 antibodies showed neutralizing capacity.
Detection of markers of hepatitis B virus infection in urine of chronic carriers
Authors: Di Bisceglie A. M. , Dusheiko G. M. , Kew M. C.
Published: March 2007

Urine from 19 chronic hepatitis B virus (HBV) carriers was tested for markers of HBV infection. Hepatitis B surface and e antigens were excreted in the urine of about one‐half of the patients, but not in any consistent pattern. HBV DNA, a much more sensitive indicator of infectivity, was found in only one specimen and then in very low concentration. We conclude that urine of chronic HBV carriers is not a major vehicle for the transmission of hepatitis B.
Quantitative and qualitative detection of cytomegalovirus‐specific antibodies using two types of enzyme‐linked immunosorbent assay
Authors: Kinane Kevin A. , Hillary Irene B.
Published: March 2007

An indirect ELISA and an inhibition ELISA were developed for the detection of cytomegalovirus (CMV)‐specific immunoglobulin G (IgG) and CMV‐specific total immunoglobulin, respectively. Both assays were more specific than the complement fixation (CF) test, and titres of positive sera were 660 times higher by IgG ELISA and 6 times higher by inhibition ELISA than titres by the CF test. Titres by IgG ELISA were reliably determined using the absorbance obtained at a single serum dilution of 1/1,000 in conjunction with a standard graph. Both ELISAs compared favourably with each other in sensitivity and specificity in determining CMV immune status. The inhibition ELISA, in particular, provides a simple and reliable method of screening sera, which requires no control antigen or predilution of sera. It should prove useful for large‐scale screening procedures, such as blood donor testing.