bluuebunny/crossref_metadata_2025 huggingface dataset:
An enzyme‐linked immunosorbent assay for an antigen related to non‐a, non‐b hepatitis and its antibody: Partial characterization of the antigen and chimpanzee transmission
Authors: Duermeyer W. , Stute R. , Hellings J. A.
Published: March 2007

An enzyme‐linked immunosorbent assay (ELISA) was developed based on sera from patients convalescent from non‐A, non‐B hepatitis and haemophilia A patients who had been frequently treated with commercial blood products. Using this ELISA, an antigen was detected which appears to be related to non‐A, non‐B hepatitis. The antigen is provisionally designated as DS‐antigen (DS‐Ag). The serum of another patient with haemophilia A, which was strongly positive for the DS‐Ag, caused a typical case of non‐A, non‐B hepatitis in a chimpanzee. DS‐Ag could be detected in the serum of the chimpanzee during the acute phase of the infection.

The ELISA for DS‐Ag reacted with neither hepatitis A or B virus antigens, nor Epstein‐Barr virus or cytomegalovirus. The assay was provisionally evaluated using sera from different groups of patients. Out of 17 patients with posttransfusion hepatitis non‐A, non‐B, 11 were found positive in the ELISA for DS‐Ag (65%).

As expected, a relatively high prevalence of DS‐Ag (9%) was found in patients with haemophilia, who are regularly treated with blood‐clotting factor‐concentrates. Antibodies to DS‐Ag were found in 48% of these patients. The DS‐Ag was found in 8 of 1400 (0.6%) volunteer blood donors, and antibody to DS‐Ag in 3% of the sera. Remarkably, a high incidence (41%) of antibodies to DS‐Ag was found in prostitutes, suggesting that this antigen may be transmitted by a sexual route.

The DS‐Ag was pelleted by ultracentrifugation for four hours at 100,000g and was found to have a buoyant density of 1.32 g/cm3 in a CsCl gradient.
Naturally occurring antibodies to the human syncytial virus in west africa
Authors: Achong B. G. , Epstein M. A.
Published: March 2007

The seroepidemiology of the human syncytial virus has been investigated by means of an indirect immunofluorescence test on 113 sera from normal antenatal women in Nigeria and 233 sera from normal individuals of both sexes and all ages in the Ivory Coast. It was found that 3.5% of samples from Nigeria and 19.7% of those from the Ivory Coast were positive. The findings are discussed.
Comparative studies of the immunogenic activity of hepatitis B surface antigen (HBsAg) and hbsag polypeptides
Authors: Sanchez Yanuario , Ionescu‐Matiu Irina , Melnick Joseph L. , Dreesman Gordon R.
Published: March 2007

Three hepatitis B surface antigen (HBsAg) preparations were compared: Purified intact 22‐nm HBsAg particles; HBsAg‐derived, sodium dodecyl sulfate (SDS)‐denatured P25 + GP30 polypeptide pool; and nondenatured P25 + GP30 micelles. The micelles had the same polypeptide composition as the P25 + GP30 pool. The immunogenicity in mice of each preparation, administered either in saline suspension or adsorbed to aluminum gel, was compared. The SDS‐denatured polypeptides were less immunogenic than intact HBsAg particles, whereas the micelles were more immunogenic. High anti‐HBs titers were observed in mice immunized with micelle preparations in either saline suspension or adsorbed to aluminum gel for as long as 200 days after a booster inoculation, administered 26 days after the primary dose.
Hepatitis B infection in vietnamese families
Authors: Skinhøj P. , Kjersem M. , Aldershvile J. , Black F.
Published: March 2007

The occurrence of hepatitis B infection as measured by sensitive serological tests for HBsAg, HBeAg and hepatitis B‐specific antibodies was studied in Vietnamese refugee families. HBsAg was found in 10% of 301 children studied. Totally 74% of all HBsAg‐positive children had an HBsAg‐positive mother or father and an additional 7% had a positive sibling. The distribution of these cases indicated that the risk of HBs‐antigenemia was increased 11‐fold for those who had an HBsAg‐positive mother compared with all other children.

The presence of HBeAg in the HBsAg‐positive parents was associated with a threefold higher risk of HBs‐antigenemia in children as compared with the presence of anti‐HBe.

It was thus documented that HBV in this population is spread primarily by intrafamilial routes and prophylactic measures within risk families should largely be able to control the appearance of new HBsAg carriers.
Studies on human lacteal rotavirus antibodies by immune electron microscopy
Authors: Totterdell Barbara M. , Banatvala J. E. , Chrystie I. L.
Published: March 2007

In vitro studies carried out by immune electron microscopy (IEM) indicate that rotavirus aggregation detected in the stools of newborn breast‐fed infants with rotavirus infection is antibody‐induced. Aggregation of rotavirus particles occurred with the IgA‐containing fraction of expressed breast milk (EBM) obtained five days postpartum and with the IgA‐ and IgG‐containing fractions of a pool of EBMs containing samples collected 2–3 days postpartum. Bovine milk fractions also demonstrated this activity in the IgG‐ and IgA‐containing fractions. Studies on unfractionated EBMs from a mother who experienced a rotavirus infection during the 43rd week of lactation showed that following rotavirus infection all three major classes of rotavirus‐specific antibodies were present in breast milk, this being confirmed by enzyme immunoassay.
Non‐A, non‐b hepatitis in persistent carriers of hepatitis b virus
Authors: Tsiquaye K. N. , Tovey G. , Kessler H. , Hu Shanlian , Lu Xiao‐Zhen , Zuckerman A. J. , Craske J. , Portmann B. , Williams R.
Published: March 2007

There are reports in the literature that infection with hepatitis A virus in hepatitis B carriers can result in resolution of the carrier state. In an attempt to induce clearance of the carrier state of hepatitis B virus in two persistently infected chimpanzees, the chimpanzees were infused with documented non‐A, non‐B infectious material. Biochemical and histopathological evidence of hepatitis was accompanied by the unique abnormalities of endoplasmic reticulum associated with non‐A, non‐B hepatitis in the chimpanzees. Elevation of alanine aminotransferase was accompanied by fourfold reduction in one chimpanzee and sixfold reduction in the other in the plasma levels of HBV‐associated DNA polymerase activity and simultaneously by twofold reduction in the concentration of hepatitis B surface antigen in both chimpanzees. A mediator may account for these changes in markers of hepatitis B virus infection, and this mechanism may also explain the occurrence of spontaneous regression in some persistently infected carriers. The significance of transient red cell anaemia in non‐A, non‐B hepatitis, which was observed in one of the chimpanzees, is yet to be established.
Interference between non‐a, non‐b and hepatitis b virus infection in chimpanzees
Authors: Brotman Betsy , Prince Alfred M. , Huima Tellervo , Richardson Linda , Van Den Ende M. C. , Pfeifer U.
Published: March 2007

Pre‐existing chronic or concurrent acute non‐A, non‐B (NANB) virus infection has been found to interfere with hepatitis B virus (HBV) replication and to delay and moderate markedly the appearance of disease related to HBV infection in chimpanzees. The mechanism for this phenomenon remains unclear.

These findings are of practical significance for vaccine safety testing and evaluation of methods for virus inactivation in chimpanzees. The possible occurrence of dual NANB and HBV infection requires that prolonged follow‐up be carried out. Attempts to carry out inactivation studies on materials containing both types of virus may also provide misleading observations.
Immune responses in serum and respiratory secretions following vaccination with a live cold‐recombinant (cr35) and inactivated a/ussr/77 (h1n1) influenza virus vaccine
Authors: Zahradnik John M. , Kasel Julius A. , Martin R. Russell , Six Howard R. , Cate Thomas R.
Published: March 2007

One hundred adult volunteers were administered inactivated vaccine (20 m̈g/0.5 cc) intramuscularly (IM) or intranasally (IN), or 104.7 TCID50 of a live coldadapted vaccine (CR35) IN. Microneutralization (Nt) and radioimmunoprecipitation methods were employed to measure hemagglutinin antibody responses in sera, nasal washes, and in bronchopulmonary lavage fluids. In unprimed recipients, the relative frequency of serum antibody response and magnitude of rise was highest following the IM‐inactivated vaccine (100%) and lowest after IN‐live vaccine (29%). However, in individuals with pre‐existing antibody, the three vaccines given were comparably immunogenic. Occurrences of secretory IgA hemagglutinin antibody in nasal washings were more frequently associated with topical administration of live or inactivated vaccine, whereas, IgG hemagglutinin antibody responses occurred with equal frequency in nasal washings in all three vaccine groups. Analysis of the hemagglutinin antibody responses in the lower respiratory tract showed that the IN‐live vaccine favored the induction of secretory IgA hemagglutinin antibody and the IM‐inactivated vaccine stimulated a more frequent IgG hemagglutinin antibody response.
The use of monoclonal antibodies in (reverse) passive haemagglutination tests for herpes simplex virus antigens and antibodies
Authors: Cranage M. P. , Coombs R. R. A. , McLean C. S. , Buckmaster E. A. , Minson A. C. , Wildy P.
Published: March 2007

Three monoclonal antibodies against herpes simplex virus type 2 have been tested for their suitability as reagents in reverse passive haemagglutination. Two of these antibodies with specificity for virus glycoprotein D. when linked to red blood cells, were able to capture antigens without being agglutinated. but addition of immune serum subsequently led to agglutination. Haemagglutination using these monoclonal antibody‐linked, antigen‐captured red cells was readily applicable to testing human sera for antibodies to herpes simplex virus and the titres obtained correlated with those from virus plaque neutralisation tests. The procedure has been termed “Specific Antigen Capture Passive Haemagglutination.” A further monoclonal antibody with specificity for the major DNA‐binding protein of type 2 herpes virus‐infected cells (a nonstructural protein) showed conventional reverse passive haemagglutination when linked to red blood cells and was specific for type 2 herpes simplex virus.

The nature and potential uses of these simple reverse passive haemagglutination procedures using monoclonal antibody reagents are discussed.
Human venezuelan equine encephalitis virus infection and diabetes in zulia state, venezuela
Authors: Ryder Elena , Ryder Slavia
Published: March 2007

Venezuelan equine encephalitis (VEE) virus has been implicated as producing alterations in glucose metabolism in animals. We performed oral glucose tolerance tests and measured serum immunoreactive insulin responses in 13 patients who were infected by VEE virus during an epidemic in 1969, in Zulia State, Venezuela. No significant alterations in the glucose tolerance test were found. Sera of 86 diabetic outpatients and 98 control individuals with normal glycemia at a local hospital were tested for antibodies to VEE virus by hemagglutination inhibition. No statistically significant difference was found between the two groups; 10.4% of the diabetic patients had detectable antibodies against VEE virus, compared to 7.1% of controls. Seventy‐three percent of the diabetics with antibodies were individuals over 40 yr old, whose diabetes could be catalogued as insulin independent. The results of these studies indicate no relationship of VEE virus infection to subsequent diabetes.
Variation in frequency of natural reinfection with influenza A viruses
Authors: Frank Arthur L. , Taber Larry H.
Published: March 2007

To investigate the frequency of natural reinfection with influenza A viruses, 55 children followed from birth along with the 44 other children and 85 adults in their families were observed for varying periods between 1975 and 1981 in the Houston Family Study. Persons infected in 1976 or early 1977 with influenza A subtype H3N2 experienced a high rate of reinfection during the 1977–78 season (63% in one group of young children) and additional reinfections in 1980–81. Persons in whom H3N2 infection was first observed in 1977–78 had little or no reinfection in 1980–81; one reinfection was observed in 1980–81 among 19 young children at risk. Only one reinfection with influenza A subtype H1N1 was observed following introduction of the virus in early 1978 despite exposure in 1978–80 and a substantial outbreak in 1980–81. Statistically significant protection from reinfection in young children followed from birth was not noted for the 1977–78 H3N2 outbreak but was present in 1979–81 for both H3N2 and H1N1 viruses. Reasons for variations in reinfection suggested by this data include (1) socioeconomic and other factors influencing intensity of exposure, (2) age at primary infection, (3) varying degrees of antigenic difference between sequential variants of a major subtype, and (4) unrecognized immunogenicity or virulence differences between subtypes and variants.
Attenuated marmoset herpesvirus isolated from recombinants of virulent marmoset herpesvirus and hybrid plasmids
Authors: Kit Saul , Qavi Hamida , Dubbs Del Rose , Otsuka Haruki
Published: March 2007

Marmoset herpesvirus (MarHV) deletion mutants in the thymidine kinase (TK) gene were isolated after infection of OMK cells with TK+ MarHV DNA and the hybrid plasmid, pMAR401, which lacks a 2.6‐kb KpnI‐M fragment in the coding region of the MarHV TK gene. After plaque purification in TK− HeLa(BU25) cells, the DNA's from five recombinant araT‐resistant MarHV clones were analyzed with restriction nucleases to verify that the 2.6‐kb KpnI‐M fragment (and a 0.9‐kb Bg1II‐Q fragment) were deleted from the viral DNA's. Molecular hybridization experiments using 32P‐labeled pMAR4 probes and viral DNA fragments also showed that the recombinant viral DNA's lacked the KpnI‐M and Bg1II‐Q fragments, that BamHI‐I of parental virus was shortened, and that three new HindIII fragments replaced the parental virus HindIII‐G fragment. The recombinants did not induce TK activity in LM (TK−) cells. To study the relative virulence of the recombinants, 3‐week‐old Swiss mice were injected intracerebrally (Ic) or subcutaneously (Sc) in the sacro‐lumbar region with either parental or recombinant viruses. The LD50 for the parental virus was 3 p.f.u. (Ic) and 7,600 p.f.u. (Sc). The recombinant viruses were significantly less virulent than TK+ MarHV after Ic inoculation (LD50 of 62,000 and 32,000 p.f.u., respectively, for viruses 5D‐6B and 5D‐4B) and gave no fatalities after Sc inoculation. Mice surviving TK− MarHV infections were protected from challenge with TK+ parental MarHV. Recombinant TK− MarHV's may be useful as vectors for the expression of foreign genes in animal cells and as the starting material for the design of vaccines.
Detection of a human rotavirus strain different from types 1 and 2—A new subgroup? epidemiology of subgroups in a swedish and an ethiopian community
Authors: Tufvesson B.
Published: March 2007

A possible new subgroup of human rotavirus was found by crossed immunoelectrophoresis and enzyme‐linked immunosorbent assay. The epidemiology of the established subgroups 1 and 2 and this new variant was studied in two different communities. Of 398 rotavirus isolates from Malmö, Sweden, 26.8% were of type 1, 71.7% of type 2, and 1.5% of the new variant. Corresponding figures for 384 samples from Addis Ababa, Ethiopia, were 33.1% type 1 and 66.9% type 2. A total of 87% of the Swedish and 79% of the Ethiopian rotavirus‐positive samples could be classified. The yearly distribution of the subgroups was stable and similar in Sweden and Ethiopia. The new variant could only be found in one outbreak during 1979. Among children with sequential infections eight of ten primary infections were type 1, and no one shed the same type of rotavirus twice.
Inhibition by acyclovir of herpes simplex virus type 2 morphologically transformed cell growth in tissue culture and tumor‐bearing animals
Authors: Kucera Louis S. , Furman Phillip A. , Elion Gertrude B.
Published: March 2007

Rat embryo fibroblasts (REF) morphologically transformed by herpes simplex virus type 2 (HSV‐2) and tumor‐derived cells were tested for ability to grow in the presence of 9‐(2‐hydroxyethoxymethyl) guanine (acyclovir). Results indicated that the effective dose of acyclovir (ACV) required to inhibit HSV‐2‐transformed and tumor‐derived cell growth by 50% (ED50) compared to mock‐treated control cells averaged 15 to 75 μg/ml. In contrast, the ED50 of acyclovir was more than 250 μg/ml for nontransformed REF, retrovirus‐transformed rat cells, and human HEp‐2 cells. HSV‐2‐transformed and tumor‐derived cells after both low (less than 30) and high (greater than 30) serial passages expressed detectable levels of the virus‐coded thymidine kinase (TK) measured in cell extracts by serum neutralization assay. HSV‐2‐transformed or tumor‐derived cells converted two‐ to ten‐fold more acyclovir to phosphorylated forms than nontransformed REF cells. Preliminary data showed that the drug inhibited tumor development in newborn syngeneic rats inoculated with HSV‐2‐transformed cells. The inhibitory activity of acyclovir and presence of low levels of HSV‐2 TK activity appeared to correlate.
Disseminated neonatal echovirus 11 disease following antenatal maternal infection with a virus‐positive cervix and virus‐negative gastrointestinal tract
Authors: Reyes Milagros P. , Ostrea Enrique M. , Roskamp Jacqueline , Lerner A. Martin
Published: March 2007

An infant girl was born apparently well one week after her mother had had a mild illness with chills, fever, and diarrhea. On the third day of life, the infant became ill and died four days later with necrotizing hepatitis. On the same day, echovirus type 11 was recovered from the throat, rectum, and buffy coat of the infant and from the cervix of the mother. At this time, the mother had an IgM neutralizing antibody titer to echovirus type 11 of 1:128, but no IgG antibodies. The infant had no echovirus type 11 antibodies. The virus was also isolated from the baby's liver and adrenal at autopsy. These findings raise the possibility of enterovirus infection at delivery from a contaminated cervix.
Hepatitis A and hepatitis B virus infection in children and adolescents in North‐East Italy
Authors: Chiaramonte M. , Floreani A. , Silvan C. , Zampieri L. , Trivello R. , Renzulli G. , Moschen M. , Naccarato R.
Published: March 2007

The sera of 722 children and adolescents without overt liver disease were tested for hepatitis B surface antigen (HBsAg), antiHBs and anti‐hepatitis B core antiHBc; 658 of the sera were also tested for anti‐hepatitis A virus anti‐HAV. Except for the “passive” antibody peak observed in babies, the anti‐HAV age‐specific prevalence was negligible until the age of 3; it then increased, reaching 35% by the age of 15. Serological evidence of HBV was present in 16% of the subjects: this prevalence was almost constant at all ages. The HBsAg carrier rate was highest in children under 5 years of age (7.6%) and decreased with age. However, only one HBsAg carrier was under 1 year of age. Anti‐HBs age‐specific prevalence increased progressively from 2.7% to 11.4%. Anti‐HBc alone was present in 4.1% of the subjects. No significant sex differences were found in the prevalence of HBV serum markers or in the HBsAg carrier rate. Neither HAV nor HBV infection was significantly influenced by place of residence or socioeconomic status. It is concluded that in this area both HAV and HBV are endemic, but while HAV is mainly acquired at school, most of HBV infections occur within the household. The results suggest that not only perinatal transmission, but also intrafamilial horizontal infection, plays a role in HBV spread among infants.
Measles virus‐specific IgM antibodies in sera from patients with chronic active hepatitis
Authors: Christie K. E. , Haukenes G.
Published: March 2007

The occurrence of measles virus‐specific IgM antibodies in sera from patients with chronic active hepatitis not caused by Hepatitis B virus was examined by a specific enzyme‐linked immunosorbent assay (ELISA). Using whole serum, specific IgM antibodies were detected in 12 of 23 sera from patients with hepatitis B surface antigen (HBsAg)‐negative chronic active hepatitis. In nine of these sera the finding of specific IgM antibodies was verified by separation of IgM by density gradient centrifugation and examination of the fractions by ELISA.

Most of the sera from the patients with measles virus‐specific IgM antibodies had an elevated level of specific IgG antibodies compared to the level of IgG found in control sera.

The significance of these findings in view of a possible persistent measles virus antigen production in patients with chronic active hepatitis is discussed.
Mixed infection with human cytomegalovirus and human polyomavirus (BKV)
Authors: Goldstein Simoy C. , Tralka Tommie S. , Rabson Alan S.
Published: March 2007

Mixed infection, the infection of a single cell by two distinguishable viruses, has been demonstrated electron microscopically in human embryo lung (HEL) and human embryo kidney (HEK) cells infected with human cytomegalovirus (HCMV) and human polyomavirus (BKV). There was no evidence of enhancement of HCMV by BKV in either cell system; however, in HEL cells under certain conditions, HCMV enhanced the growth rate of BKV. Since both BKV and HCMV are often found in the urine of immunosuppressed patients, the enhancement of BKV by HCMV may be of clinical significance.
Detection of varicella‐zoster virus by dot‐blot hybridization using a molecularly cloned viral DNA probe
Authors: Seidlin Mindell , Takiff Howard E. , Smith Holly A. , Hay John , Straus Stephen E.
Published: March 2007

Varicella‐Zoster Virus (VZV) infection can be definitively diagnosed by isolation of virus in cell culture, a process that usually takes 7–14 days. In order to facilitate the more rapid detection of this virus, we developed a technique for hybridization of DNA from clinical specimens using an in vitro‐labeled mixture of cloned fragments of VZV DNA as a probe. The assay can be completed in 36–48 hr and can be successfully carried out in the range of 10 pg to 10 ng of viral DNA. In analyses of 38 specimens from patients with a clinical diagnosis of VZV infection, the results of viral isolation and this assay were highly concordant. The sensitivity of standard cell culture for detection of VZV was 58%, whereas the sensitivity of the assay was 76%, not significantly different (P = 0.14). The specificity of cell culture was 100%, whereas that of the assay was 94% (P = 0.49). The technique appears to be sensitive, specific, and useful for analyses of tissues and body fluids.
The behaviour of recent isolates of human respiratory coronavirus in vitro and in volunteers: Evidence of heterogeneity among 229E‐related strains
Authors: Reed Sylvia E.
Published: March 2007

Strains of human coronavirus (HCV) isolated between 1974 and 1976 have been studied in vitro and in volunteers. All strains caused colds in volunteers, and those cultivable in tissue culture (TC) produced significantly more coryza and less sore throat than strains growing only in organ culture (OC). The TC strains were serologically related to 229E, but these isolates produced colds with a frequency and severity that contrasted with the effects of 229E itself. Tests on volunteers' preinfection sera showed that the prevalence of antibody to 229E had increased during the period 1961–1979 and that during 1977–1979 only 11% of subjects had no neutralising antibody against 229E. Susceptibility to the 229E‐related isolates PR and TO was associated with low preinfection serum neutralising antibody against the homologous virus, and paired sera frequently showed fourfold or greater antibody rises, most commonly against the homologous strain. Volunteers infected with TO were immune when reinoculated with the same strain approximately 1 year later, but other similar volunteers were at least partly susceptible to infection with a heterologous 229E‐related virus after similar time intervals. Although the strains of HCV that were grown in tissue culture were all related to the prototype 229E, they appeared not to be identical with it, and this heterogeneity is probably a significant factor in the epidemiology of HCV infections.
Hemagglutination by human rotavirus strains
Authors: Kitaoka Setsuko , Suzuki Hiroshi , Numazaki Toshio , Sato Tetsuo , Konno Tasuke , Ebina Takusaburo , Ishida Nakao , Nakagomi Osamu , Nakagomi Toyoko
Published: March 2007

Human rotavirus isolates, KUN, MO, and Wa strains were found to agglutinate erythrocytes of the day‐old chicken and adult goose, optimally at pH 6.6. Only those fractions containing double‐shelled rotavirus particles isolated by isopycnic centrifugation in cesium chloride had hemagglutinating activity. Trypsin treatment decreased the hemagglutination titer of human rotavirus strains and conversely increased their infectivity. Hemagglutination inhibition tests with antisera against type‐specific antigens demonstrable by neutralization showed no type specificity.
Evaluation of cold‐adapted adn temperature‐sensitive mutants of parainfluenza virus type 3 in weanling hamsters
Authors: Crookshanks Frances K. , Belshe Robert B.
Published: March 2007

Cold‐adapted (ca) and temperature‐sensitive (ts) mutants of parainfluenza virus 3 were produced by serially passaging wild type (wt) parainfluenza virus 3 at 20°C 45 times. Previously, plaque‐purified viruses (clones) were selected from the wt parent and from cold passage levels 7, 12, 18, and 45 and characterized in vitro. In the present study, we evaluated at least one mutant from each cold passage level for attenuation in hamsters. Four of the five mutants tested were ts and ca (one each from cold passage levels 7, 12, 18, and 45) and one was ca but not ts (from cold passage 18). Groups of hamsters were inoculated intranasally with either parent wt or mutant virus. Four or five hamsters from each group were sacrificed prior to inoculation and daily on the first, second, third, fourth, and sixth or seventh day postinoculation and the amount of virus in nasal turbinates and lungs was quantitated. The quantity of virus recovered from the hamsters and the parainfluenza virus 3 antibody titers were inversely related to the cold passage level. Two of the mutants did not replicate in hamsters. Cold adaptation of parainfluenza virus 3 resulted in progressive attenuation of the virus in weanling hamsters. These highly attenuated mutants are suitable for evaluation in children as live virus vaccines.
Absence of IgG Fc receptors on varicella‐zoster virus‐infected cells
Authors: Ishak Ricardo , Andiman Warren A. , Tucker Grace
Published: March 2007

Herpesvirus‐infected cells usually express receptors for the Fc portion of immunoglobulin G. Varicella‐zoster virus has so far been the sole exception in the family. Both immunehemadsorption and immunofluorescence techniques failed to detect the expression of such receptors. This observation excludes the possibility of false‐positive results in serological tests for antibodies to this virus. It is possible that the function of these receptors early in infection is not needed in the subsequent reactivation(s) of the virus. No variation has been shown to occur with different isolates.
Altered hemolysis in single radial hemolysis from a single serum sample as an indicator of recent primary rubella virus infection
Authors: Hedman Klaus , Räihä Kirsti , Meurman Olli , Vaheri Antti
Published: March 2007

The quality of hemolysis in the single radial hemolysis (SRH) test was observed to be altered in a small proportion (5.7%) of Sera (N = 9628) studied for rubella antibodies. Three different types of altered hemolysis were identified. Two of these types, the “soft margin” (SM) or the “soft zone” (SZ), occurred singly or in combination in 97% of paired sera (N = 321) taken 3–30 days after primary rubella infection (diagnostic seroconversion). For comparison, diagnostic inercases of rubella antibodies (N = 77), including both primary and reinfections, contained these types of altered hemolysis in 80% of the cases. Of the remaining 20% of the samples (N = 15) rubella antibodies of IgM class were looked for in seven cases but not found. After primary rubella, SZ was always present in the first serum sample and disappeared rapidly within 20 days while SM persisted longer. The predictive value of the different types of hemolysis was estimated from 96 primary sera, which had altered hemolysis. In this material, SZ was followed by a diagnostic increase of antibodies in 87% of the cases; the prognostic value of the SM was significant but lower. These data show that the appearance of SM or SZ in a single serum sample is a useful marker of recent primary rubella. On the other hand, a normal hemolysis in SRH can be used to exclude recent primary rubella, but not reinfection, with a high degree of probability.
Immune responses to live attenuated and inactivated mumps virus vaccines in seronegative and seropositive young adult males
Authors: Ilonen J. , Salmi A. , Tuokko H. , Herva E. , Penttinen K.
Published: March 2007

Live attenuated mumps virus vaccine and Formalin‐inactivated mumps vaccine were compared for their ability to induce antibody responses and cell‐mediated immune responses as measured by a lymphocyte blast transformation test (LBT). The subjects studied were conscripts of the Finnish Defense Forces. In seronegative subjects antibody levels induced by attenuated virus were more variable than those induced by inactivated virus, including one failure with the live vaccine out of a total of nine subjects. IgM antibody class response was seen only in four of nine subjects vaccinated with live virus—in those subjects with the highest post vaccination antibody levels and strongest LBT responses. Increases of antibody levels in originally seropositive subjects were higher in subjects vaccinated with inactivated virus. Cell‐mediated immunity induced by both vaccines was relatively poor when measured by the LBT test. At a time of 6–8 months after vaccination, LBT responses in originally seronegative subjects were not observed.
Age‐specific prevalence and significance of hepatitis B e antigen and antibody in chronic hepatitis B virus infection in taiwan: A comparison among asymptomatic carriers, Chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma
Authors: Liaw Yun‐Fan , Chu Chia‐Ming , Lin Deng‐Yn , Sheen I‐Shyan , Yang Chaur‐Young , Huang Miau‐Ju
Published: March 2007

The age‐specific prevalence of hepatitis B e antigen (HBeAg) and its antibody (anti‐HBe) were studied by radioimmunoassay, and compared in a large series of patients with chronic hepatitis B virus (HBV) infection, including 268 asymptomatic carriers. 389 chronic hepatitis, 114 liver cirrhosis, and 278 hepatocellular carcinoma (HCC). The prevalence of HBeAg/anti‐HBe in asymptomatic carriers and patients with chronic hepatitis correlated closely with age as HBeAg prevalence decreased and anti‐HBe prevalence increased with increasing age (P < 0.0005), and is probably due to high infection rate at young age in Taiwan. The prevalence of HBeAg in patients with both cirrhosis and HCC are much significantly lower and had no correlation with age. Two peaks of age‐specific prevalence of HBeAg and anti‐HBe were observed in patients with HCC, implicating two patterns of HBV infection in these patients. The difference in the prevalence of HBeAg and anti‐HBe might indicate that asymptomatic carriers, chronic hepatitis, liver cirrhosis, and HCC are sequential sequelae of HBV infection.
Cytomegalovirus infection after bone marrow transplantation: Relation of pneumonia to postgrafting immunosuppressive treatment
Authors: Skinhøj P. , Andersen H. Kerzel , Møller J. , Jacobsen N.
Published: March 2007

Five fatal cases of CMV associated interstitial pneumonia occurred among 20 patients who had received allogenic bone marrow transplantation for acute leukemia or aplastic anaemia. This outcome was analysed in relation to prospectively obtained data on complement fixing (CF) and IgM anti‐CMV litres and excretion of CMV in urine or pharynx. Altogether 16 patients had primary or reactivated CMV infection. Five of these patients receiving corticosteroids at the time of infection, at a dose of more than 1 mg/kg/day, failed to mount a significant CF antibody response, excreted large amounts of virus, and died from pneumonia, while the remaining 11 infected patients obtained high CF antibody titres and showed no severe symptoms related to CMV infection. IgM antibodies occurred simultaneously with CF anlibodies in most cases of primary and reactivated infection. The results of the study suggest a protective role of the humoral immune response to CMV infection in BMT recipients, and passive immunisalion with CMV hyperimmune globulin should be attempted especially in patienls given additional immunosuppression with high dose corticosteroids.
Production of subgroup‐specific monoclonal antibodies against human rotaviruses and their application to an enzyme‐linked immunosorbent assay for subgroup determination
Authors: Taniguchi Koki , Urasawa Tomoko , Urasawa Shozo , Yasuhara Toshio
Published: March 2007

Nonneutralizing monoclonal antibodies were prepared against two strains, S2 and YO, of human rotaviruses isolated in cell culture. S2‐37 and YO‐5 antibodies had subgroup I and subgroup II specificities, respectively. The remaining antibodies (S2‐65, YO‐71, YO‐89, and YO‐156) reacted commonly with all the rotaviruses examined. All of the monoclonal antibodies agglutinated exclusively single‐shelled particles and immunoprecipitated 42,000‐dalton protein, a major component of inner capsid.

Using the three monoclonal antibodies (S2‐37, YO‐5, and YO‐156), an enzyme‐linked immunosorbent assay was developed for detecting and subgrouping human rotavirus isolates.
Isolation and identification of enteroviruses from faecal samples in a differentiated epithelial cell line (HRT‐18) derived from human rectal carcinoma
Authors: Patel J. R. , Daniel J. , Mathan M. , Mathan V. I.
Published: March 2007

A human rectal carcinoma‐derived differentiated epithelial cell line, HRT‐18, was inoculated with faecal samples in an attempt to grow coronavirus‐like particles (CVLP), which are widely prevalent in human stools in southern India. While CVLP did not grow in this cell line, a variety of enteroviruses were isolated from 48 of the 114 stool samples from healthy controls and patients with diarrhoea. The results suggest that human gastrointestinal tumour‐derived differentiated epithelial cells in continuous culture may be useful for the primary isolation of enteroviruses and merit further study.
Effect of environmental factors on aerosol‐induced lassa virus infection
Authors: Stephenson Edward H. , Larson Edgar W. , Dominik Joseph W.
Published: March 2007

Previous studies suggested that the most frequent means of transmission of Lassa virus was by either direct or indirect contact with infectious material. Aerosol stability and respiratory infectivity of the Josiah strain of Lassa virus were assessed to determine the effect of environmental factors on aerosol‐induced infection. The stability of the virus in aerosol, particularly at low relative humidity (30% RH), plus the ability of the virus to infect guinea pigs and monkeys via the respiratory route emphasize the potential for aerosol transmission of Lassa virus. Biological half‐lives at both 24 and 32°C ranged from 10.1 to 54.6 min, and were sufficient for aerosol dispersion of virus to considerable distances in natural situations. Infectivity of Lassa virus in small particle aerosol was demonstrated in outbred guinea pigs and cynomolgus monkeys using dynamic aerosol equipment. Monkeys exposed to inhaled doses ⪖465 PFU were infected and died. The median infectious dose (ID50) for guinea pigs was 15 PFU, yet a definitive median lethal aerosol dose (LD50) could not be established. Organ tropism of aerosol‐induced Lassa virus infections in outbred guinea pigs was similar to that previously reported for inbred guinea pigs infected by subcutaneous inoculation.
Murine forebrain anomalies induced by coxsackievirus B3 variants
Authors: Gauntt C. J. , Jones D. C. , Huntington H. W. , Arizpe H. M. , Gudvangen R. J. , Deshambo R. M.
Published: March 2007

Neonatal or 7‐day‐old mice inoculated intracranially with either of two temperature‐sensitive mutants (tsl, ts6) or the parent coxsackievirus B3 (CVB3) subsequently developed porencephaly or hydranencephaly. The forebrain anomaly induced depended upon age of the animal at inoculation and virus variant inoculated. Sections of brains from hydranencephalic mice revealed severe meningeal reactions, necrotizing encephalitis, and liquifactive necrosis in the cerebrum. No pathology was found in the pons, medulla, or cerebellum. Immunofluorescence studies with hyperimmune anti‐CVB3 antiserum showed a random distribution of virus‐infected cells in the cerebrum. Virus was recovered from several organs but little to no interferon and no anti‐CVB3 neutralizing antibody were present in brain tissues. Availability of cells for replication of virus at the time of inoculation and replicative properties of each virus likely contributed to the outcome. Thus, forebrain anomalies resembling those found in infants can be induced in a murine model by select variants of coxsackievirus B3.
Aetiology of acute sporadic hepatitis in adults in kenya
Authors: Greenfield C. , Karayiannis P. , Wankya B. M. , Shah M. V. , Tukei P. , Galpin S. , Jowett T. P. , Thomas H. C.
Published: March 2007

Markers for acute hepatitis A virus (HAV), hepatitis B virus (HBV), and hepatitis non‐A, non‐B (HNANB) infections were examined in the sera of 94 patients presenting with acute hepatitis in Kenya. Hepatitis B virus was responsible for 70% of cases, HNANB for 18%, and HAV for only 12%. The use of an IgM anti‐HBc assay increased the rate of diagnosis of acute HBV infection, thereby reducing the proportion of cases designated as NANB.
Hepatitis a virus: Growth characteristics of in vivo and in vitro propagated wild and attenuated virus strains
Authors: Bradley Daniel W. , Schable Charles A. , McCaustland Karen A. , Cook E. H. , Murphy Bert L. , Fields Howard A. , Ebert James W. , Wheeler Cosette , Maynard James E.
Published: March 2007

Serial passage of the MS‐1 strain hepatitis A virus (HAV) in marmosets was shown to increase the yield of virus and to shorten the incubation period from approximately 55 days in the first passage to 3–7 days in the ninth and higher passages. Intravenous inoculation of susceptible chimpanzees with MS‐1 HAV was found to result in a typical course of disease in two animals who had received eighth marmoset‐passage virus, including the occurrence of elevated ALT activity, presence of HAV antigen in liver and stool, and seroconversion to anti‐HAV. Two chimpanzees inoculated with 20th passage MS‐1 HAV (M001 liver homogenate) exhibited normal or nearly normal ALT activity and had no demonstrable or significant HAV in weekly liver biopsy specimens or in serial stool suspensions obtained during 64 days of observation. However, both animals seroconverted to anti‐HAV within 2 weeks after inoculation, as did the animals who had received eighth passage MS‐1 HAV. These findings suggest that subpassage of the MS‐1 strain of HAV in marmosets resulted in the generation of an attenuated virus strain that was still capable of inducing a vigorous antibody response in intravenously infected chimpanzees. Serial propagation of wild and attenuated strains of HAV (HAS‐15 and MS‐1/M001, respectively) in FRhK‐4 cells was associated with a significant decrease in the growth period for both viruses. Our studies have also shown that HAS‐15 HAV can be recovered in maximum yield in later passages as early as 2 to 3 days after inoculation.
Study of herpes simplex virus type 1 populations obtained from recurrences and primary infections
Authors: Mannini‐Palenzona Anna , Bartoletti Anna M. , Foa‐Tomasi Laura , Costanzo Franco , Borgatti Mariangela , Tognon Mauro , Cassai Enzo
Published: March 2007

The analysis of 23 clinical isolates of herpes simplex virus type 1 (HSV‐I) showed that 15 of 15 isolates that had undergone a few passages in tissue culture (fresh isolates) and two of eight isolates that had never been passaged (new isolates) were composed of a mixed population with respect to plaque morphology in Vero cells. Cloning and characterization of 10 large plaque viruses (L variants) and nine small plaque viruses (S variants), obtained from seven different isolates, showed the following. BamHI DNA restriction patterns of the L and the S variants from a single isolate differed only with respect to the electrophoretic mobility of the fragments that contain reiteration of specific sequences; they did not differ regarding the presence or the absence of restriction endonuclease cleavage sites. The L and the S variants differed with respect to the electrophoretic profiles of infected cell glycoproteins, thermosensitivity of growth and plaquing efficiency at 39°C, and, at least in the case of the two couples of variants that we tested, pathogenicity for the mouse. The hypothesis that the L variants might arise from the S variants during in vitro replication is discussed.
Hepatitis B virus markers in children with acute leukemia: The effect of chemotherapy
Authors: Locasciulli A. , Santamaria M. , Masera G. , Schiavon E. , Alberti A. , Realdi G.
Published: March 2007

Hepatitis B virus markers were tested in the serum of 49 children with acute leukemia on clinical presentation and during subsequent chemotherapy. Hepatitis B surface antigenemia was observed in only six patients (12%), none of whom progressed to chronic infection. Chemotherapy had a marked suppressive effect on the production of antibodies to hepatitis B virus antigens and overt infection occurred in two children after suppression of protective immunity. Evidence of liver damage was frequently observed and was largely independent of serologic data. These results indicate that active immunization with hepatitis B vaccine may not find a clear place in this clinical setting.
Detection of hepatitis B virus DNA in serum and relation with the IgM class anti‐HBc titers in hepatitis B virus infection
Authors: van Ditzhuijsen T. J. M. , Selten G. C. M. , van Loon A. M. , Wolters G. , Matthyssen L. , Yap S. H.
Published: March 2007

Sera from four groups of patients with different serologic markers of HBV infection were examined for HBV DNA using molecular hybridization technique and for IgM class anti‐HBc using an ELISA based on the antibody capture principle. Results of HBV DNA assay were generally in good agreement with the presence of HBeAg. However, HBV DNA was found in 13% of anti‐HBe+sera and in one patient with anti‐HBc as a sole marker.

IgM anti‐HBc was detected at high titers in acute hepatitis B patients and was also present during the “window‐period.” This marker was also found, though less frequently when other markers for HBV infectivity were absent, in chronic hepatitis B patients and healthy carriers. From these findings we conclude that the HBV DNA assay provides a reliable method of detecting the infectious agent, particularly in anti‐HBe+sera and sera with anti‐HBc as a sole marker.

The assay for IgM anti‐HBc is useful for establishing the diagnosis of recent infection in patients with anti‐HBc as a sole marker, and during acute hepatitis with very high aminotransferase values, a condition in which HBV DNA may be undetectable.
Pathogenesis of attenuated Junin virus in the guinea pig model
Authors: de Guerrero Lucía B. , Boxaca Martha C. , Malumbres Ester , Gómez María de las Mercedes
Published: March 2007

The purpose of this work was to elucidate the pathogenesis of attenuated Junin virus (JV) strains in the guinea pig model. Three groups of guinea pigs were infected by the IM route with 103 PFU of the XJC13 and XJO‐attenuated strains or with the XJ pathogenic strain of JV, respectively. Viremia was studied at 3, 5, 7, 9, 12, and 14 days postinfection (pi) (a) in serum samples of all animals and in washed cells from XJC13‐infected guinea pigs by conventional techniques and (b) in whole blood samples from XJC13 and XJO animals by coculture with Vero cells. Virus spread was studied at 14 days pi in brain, spleen, lymph nodes, and bone marrow by parallel suckling mouse inoculation or organ homogenates and coculture of cell suspensions with Vero cells. By coculture techniques of whole blood, an otherwise undetectable viremia was demonstrated for both attenuated strains throughout the observation period. In contrast, XT viremia was easily detected by direct techniques, as has already been shown. Attenuated virus was also shown to reach brain and bone marrow when coculture methods were employed. But titers were always markedly lower than those of the pathogenic strain. The sustained viremia demonstrated in guinea pigs infected with either attenuated strain explains the mode of viral dissemination and accounts for viral rescue and antigen detection from some organs.

These results suggest that attenuated strains do not differ greatly in their invasive capacity in guinea pigs, but later on viral replication is impaired. Therefore, these findings reveal potential risks and should be noted when developing human vaccines.
Adenoviruses in faeces of children with acute gastroenteritis in Rio De Janeiro, Brazil
Authors: Leite J. P. G. , Pereira H. G. , Azeredo R. S. , Schatzmayr H. G.
Published: March 2007

Faeces from 746 children less than 5 years old with acute gastroenteritis were screened for the presence of adenovirus particles or antigens by immunoelectron microscopy (IEM) and enzyme immunoassay (EIA). Thirty‐five samples were positive by both IEM and EIA, two only by IEM, and two only by EIA, giving a total of 39 (5.2%) samples with positive results. Of these, 25 could be propagated in HEp2 cells and were neutralized by one of the antisera to adenovirus types 1 to 18. The remaining 14 samples could be propagated only in the 293 permanent line of human cells transformed by adenovirus type 5 DNA [Graham et al, 1977] and were not neutralized by antisera to adenovirus types 1 to 31. An EIA carried out by the antibody‐capture technique, using antiserum specific for “enteric” adenoviruses [Johansson et al, 1979], gave positive results with all isolates that could be propagated only in 293 cells and with none of those capable of growing in HEp2 cells.
Identification and typing of herpes simplex virus types 1 and 2 by monoclonal antibodies, sensitivity to the drug (E)‐5‐(2‐bromovinyl)‐2′‐deoxyuridine, and restriction endonuclease analysis of viral DNA
Authors: Zimmerman Daniel H. , Mundon Francis K. , Croson Susan E. , Henchal Laraine S. , Docherty John J. , O'Neill Sean P.
Published: March 2007

With development of antiviral drugs, the need to identify a virus as to drug sensitivity becomes increasingly of importance. The compound (E)‐5‐(2‐bromovinyl)‐2′‐deoxyuridine (BVDU) has been shown to be much more inhibitory to the replication of herpes simplex virus type 1 (HSV‐1) and varicella‐zoster virus as opposed to herpes simplex virus type 2 (HSV‐2). We have typed over 170 isolates, using an immunofluorescent technique and sensitivity to the drug BVDU. These results were then compared to the typing of isolates by analysis of viral DNA after restriction endonuclease digestion (EcoRI). Without exception the results were in agreement between the monoclonal antibody results and sensitivity to the drug BVDU. Furthermore, the typing with monoclonal antibodies was also in excellent agreement with the DNA analysis. Only those isolates inhibited with BVDU showed DNA characteristics of HSV‐1 and reacted only with the S‐200 antibody. On the other hand, those isolates which reacted with the monoclonal antibody S‐141 were insensitive to BVDU, and again this was in agreement with the DNA analysis. These results could provide the basis for developing a diagnostic test using the two monoclonal antibodies to type either isolates or direct smears and to use the results as a basis for possible drug therapy.