bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of viral antigens in cerebrospinal fluid of patients with herpes simplex virus encephalitis
Authors: Bos Cees A. , Wilterdink Jan B. , Scheffer Albert J. , Olding‐Stenkvist Elisabeth
Published: March 2007

Thirty‐two cerebrospinal fluid (CSF) samples from eighteen patients with confirmed herpes simplex encephalitis (HSE) were assayed by an indirect enzyme‐linked immunosorbent assay (ELISA) for the presence of viral antigens. The results are expressed as an antigen ratio distinguishing between herpes simplex virus (HSV) antigens containing samples and negative samples. Judged by this criterion a positive result was obtained in 33 % of the patients.

Overall, 25% of the CSF samples from HSE patients were positive. In one out of 33 control patients with other neurological disorders a positive antigen ratio was found.

Two or more CSF samples were available from eleven patients. In six of these, the second or later samples showed a decreased antigen ratio when compared to the first CSF sample. An increase of the anti‐HSV antibody titer was seen in the CSF of five of these six patients. Five out of six patients with a decreasing antigen ratio had an unfavorable outcome of their encephalitis, while a favorable outcome was seen in four of the five patients with an increasing or steady antigen ratio. A decrease of the antigen ratio in the course of HSE can be explained by the presence of immune complexes in CSF and may indicate a poor prognosis.
Isolation of inoue‐melnick virus from human meningioma‐derived cell cultures and detection of antibody in patients with meningioma
Authors: Inoue Y. K. , Nishibe Y. , Takeuchi J. , Uchida Y. , Handa H. , Hollinshead A. C.
Published: March 2007

Inoue‐Melnick virus (IMV) was isolated from six of seven human meningioma‐derived cell cultures, while the virus was not isolated from six other brain tumor cell cultures. Sera of 145 consecutive neurosurgical inpatients were tested for IMV‐neutralizing antibody. Of 26 patients with meningioma, 22 were positive for IMV antibody (84.6%). Of the remaining 119 patients, 16 were positive.
Chronic, relapsing encephalomyelitis associated with experimental measles virus infection
Authors: Carrigan Donald R.
Published: March 2007

An experimental central nervous system (CNS) disease that bears some similarity to human multiple sclerosis and that is associated with infection by measles virus has been induced in hamsters. This disease has been described previously (Carrigan and Johnson: Proceedings of the National Academy of Sciences 77:4297‐4300, 1980). At that time, it was believed that the disease was restricted to the spinal cords of affected animals. The present work demonstrates that the disease can also affect the brain, describes in more complete detail the types of clinical disease that occur, illustrates the histopathological changes found in diseased CNS tissues, and documents more accurately the incidence of the disease in the virally infected animals. In summary, the disease, which has now been termed chronic, relapsing encephalomyelitis (CRE) is associated with neonatal CNS infection of hamsters with a particular strain of measles virus. CRE occurs in approximately 12% of animals that survive the acute viral infection, and of these affected animals about half develop detectable clinical signs of neurological disease. The balance of the animals have subclinical disease detectable only by histopathologic changes within the CNS. These lesions are composed of varying degrees of demyelination, necrosis, mononuclear cell inflammation, and gliosis.
Infection of a poikilothermic cell line (XL‐2) with eastern equine encephalitis and western equine encephalitis viruses
Authors: Morier L. , Cantelar N. , Soler M.
Published: March 2007

Eastern Equine Encephalitis (EEE) was in Cuba before the 1940s; the virus has been isolated from horses, birds, and rodents during epizootic as well as interepizootic periods. The only isolation of Western Equine Encephalitis (WEE) virus was from a sick pigeon found in the vicinity of Havana University. Both viruses can cause human disease; the isolation of WEE virus from the centre of an urban area emphasises the need for the prompt isolation and rapid identification of these agents.

The object of this work was to compare the sensitivity of a continuous cell line (XL‐2) from the toad, Xenopus laevis, with primary chick embryo cell cultures (CEC) routinely used for isolation as well as assay of these viruses. Both cell systems were infected with EEE virus isolated from horse brain and WEE virus isolated from a sick pigeon. A clear cytopathic effect (CPE) consisting of rounding and detachment of cells was observed in both cell cultures infected with the two viruses. By 18 hours post‐infection, there was partial destruction of the cell monolayer and by 24 hours the CPE was total. The infectious titre of EEE and WEE viruses in XL‐2 and CEC were similar. Both viruses produced small plaques (0.5‐1.0 mm diameter) in XL‐2 cells.

Studies on the sensitivity of the XL‐2 cells for direct isolation of the two viruses from field samples and for the detection of Cuban flaviviruses by the immunofluorescence test are in progress.
Use of monoclonal antibodies for the diagnosis of cytomegalovirus infection by the detection of early antigen fluorescent foci (DEAFF) in cell culture
Authors: Stirk P. R. , Griffiths P. D.
Published: March 2007

A pool of seven monoclonal antibodies, each reactive with cytomegalovirus (CMV) early antigens, was used in an indirect immunofluorescence method for the rapid detection of CMV‐infected fibroblasts following inoculation with clinical specimens. A total of 1,639 specimens were examined, and the results were compared with those of conventional isolation procedures. The detection of CMV by early antigen fluorescent foci (DEAFF) was found to be comparable, both in terms of specificity and sensitivity, to that of conventional cell culture. Its great advantage, however, is the rapidity with which results are achieved. Thus, results were available from the DEAFF test within 24 hours of receipt of the specimens as compared to a mean of 16 days for cell culture. This single rapid assay for the detection of CMV in clinical samples may be performed by any laboratory familiar with cell culture techniques and in our hands is the preferred diagnostic method for CMV.
Splenic replication of hepatitis B virus in the chimpanzee chronic carrier
Authors: Lieberman Harvey M. , Tung William W. , Shafritz David A.
Published: March 2007

Low levels of hepatitis B virus (HBV) replication and serum Dane particles have been commonly observed in chimpanzee chronic HBV carriers. To evaluate the possibility of extrahepatic sites of replication, DNA from various organs of a chimpanzee HBV carrier were evaluated by Southern blot analysis. With cloned, repurified HBV DNA of 3.2 kilobases (Kb) as a hybridization probe under stringent conditions, analysis of liver DNA revealed a diffuse hybridization pattern below 3.2 Kb and hll‐length double‐stranded HBV genomes at 3.2 and 1.8 Kb, the latter representing the supercoiled (CCC) form found in the nucleus. No HBV DNA was found in pancreas, muscle, renal, or adrenal gland. Analysis of splenic DNA revealed diffuse hybridization below 3.2 Kb within the cytoplasmic subcellular fraction, and full‐length HBV genomic forms in the nuclear fraction of splenic tissue. Use of (−) and (+) strand‐specific HBV DNA and RNA probes demonstrated asymmetric viral replication within the spleen cytoplasm as previously demonstrated in liver. Northern blot analysis of total RNA from chimpanzee spleen and liver revealed HBV RNA sequences in both of these tissues, suggesting active viral gene expression and/or replication in chimpanzee spleen as well as in liver. Elucidation of the splenic cell type supporting viral propagation may serve as a basis for development of a tissue culture system to study molecular events of HBV replication.
Age‐ and sex‐related study of hepatitis B virus chronic carrier state in infants from an endemic area (Senegal)
Authors: Coursaget P. , Yvonnet B. , Chotard J. , Vincelot P. , Sarr M. , Diouf C. , Chiron J. P. , Diop‐Mar I.
Published: March 2007

This report concerns hepatitis B virus (HBV) infections observed in 155 infants from Senegal. studied with a view to determining the factors involved in development of the chronic carrier state. A chronic carrier state was observed in 50.3% of the infants.

This study confirms that the risk of chronic carriage is linked to age. This risk declines very rapidly with age, falling from 82% in infants under 6 months old, to 15% in children between the ages of 2 and 3 years. Spontaneous elimination of hepatitis B surface antigen (HBsAg) is uncommon in HBsAg carriers during childhood.

The difference observed in chronic carriage between males and females is due to a difference in susceptibility of the two sexes to the development of the chronic carrier state: HBV infections (before 2 years of age) lead to a chronic carriage in 77% of males as against 50% of females.

These conclusions are important in view of the immunisation programs being carried out against hepatitis B virus in endemic areas. For a maximum efficacy, vaccination must be carried out at birth, or shortly afterwards.
Spontaneous and induced interferon production by peripheral blood leucocytes from control population and patients with herpes genitalis
Authors: Lannello D. , Taylor M. W. , Fife K. H. , Chen L.
Published: March 2007

The spontaneous and PHA induced levels of interferon were measured in peripheral blood leucocyte cultures of twenty‐eight individuals diagnosed as positive for herpes genitalis, and in a group of control subjects. As reported by Cunningham and Merigan [1983] for herpes labialis, leucocytes from individuals with herpes genitalis produced low levels of interferon spontaneously; however, similar results were found for individuals within the control population. No statistically significant difference could be found for PHA induced interferon levels, antigen induced interferon levels, or helper/suppressor cell ratios between the herpes genitalis population and control population. Our results indicate that interferon does not play a major role in the latency or recurrence of herpes genitalis.
Virus‐specific serum IgG, IgM, and IgA antibodies in cytomegalovirus mononucleosis patients as determined by immunoblotting technique
Authors: Porath Avi , Hanuka Negba , Keynan Arie , Sarov Israel
Published: March 2007

The immune response to individual human cytomegalovirus (CMV) structural polypeptides was studied in paired sera from 15 adult CMV mononucleosis (CMVMN) patients and healthy controls by immunoblotting technique (IB). IgM and IgG antibodies to at least 11 structural polypeptides with molecular weights of 28K, 49K. 55K, 57K. &‐7OK, 82K, 87K, IIOK, 150K. 205K, and 235K were detected in the patients sera in the serum sample obtained in the acute phase of the disease. IgA antibodies to polypeptides with molecular weights of 66‐70K. 82K. 11OK, and 150K were also detected in these sera. In healthy seropositive adults, IgG antibodies with the same molecular weight polypeptides, excluding the 205K and 235K polypeptides, were detected as in convalescent CMV‐MN patients. A prominent reactivity of IgM and IgA antibodies to the 66‐70K and ISOK polypeptides was noted in the acute sera from all the CMV‐MN patients examined, but not in a number of late convalescent sera. The potential implications of these findings in the development of specific serological tests are discussed.
Antibody to human immunodeficiency virus correlates with decreased T helper lymphocytes in asymptomatic individuals
Authors: Weiser Barbara , Burger Harold , Steimer Kathelyn , Lifson Jeffrey , Engleman Edgar , Grimson Roger , Robinson William S.
Published: March 2007

To examine the relationship between human immunodeficiency virus (HIV) seropositivity and T lymphocyte subsets in a clinically well population, we assayed HIV antibody and analyzed T lymphocyte subsets in 30 people at increased risk for acquired immunodeficiency syndrome (AIDS) who were clinically well. Seventy‐six percent of the HIV‐seropositive individuals had abnormally low numbers of T helper lymphocytes, and HIV seropositivity was strongly correlated with an abnormally low number of T helper cells (p<0.00002). Among these clinically well subjects at increased risk for AIDS, HIV‐sero‐positive individuals had a significant decrease in mean T helper lymphocytes and mean T helper: T suppressor ratios as compared to those who were seronegative (483 cells/mm3 vs 915 cells/mm3, p<0.002: and 0.80 vs 1.7. p<0.002. respectively). Because of the strong correlation of HIV seropositivity and abnormally low numbers of T helper lymphocytes in this asymptomatic population, these findings suggest that asymptomatic seropositive individuals should be followed closely for development of AIDS‐related disease and should be considered for future antiviral therapy when it becomes available.
Production and characterisation of a human monoclonal antibody to cytomegalovirus and its use in an early nuclear fluorescence assay
Authors: Sutherland S. , Crawford D. H. , Wilson S. A. , Morgan B. , Azim T. , Huehns E. R.
Published: March 2007

A human monoclonal antibody to cytomegalovirus (CMV) was produced by transforming peripheral blood mononuclear cells of a patient with recent CMV infection. It is directed against a late antigen located in the nucleus of CMV infected fibroblasts at 24‐72 hours post infection and immuneprecipitates 65K and 48K proteins from 35S‐labelled CMV infected cells. Results of its use in an early nuclear fluorescence assay for rapid diagnosis are presented.
Cloning and characterisation of a delta virus cDNA Sequence derived from a human source
Authors: Saldanha J. A. , Thomas H. C. , Monjardino J. P.
Published: March 2007

We report the extraction of delta virus RNA from the serum of a delta‐virus‐infected patient and the subsequent cloning and analysis of a 380‐nucleotide‐long cDNA (D380). The nucleotide sequence of D380 shows overall differences of approximately 20% when compared with previously published sequences and docs not include the viroid consensus sequence previously reported (Wang et al: Nature 323:508‐514, 1986). A potentially coding open reading frame extending over the whole length of the D.380 has been identified. Our results demonstrate the existence of genetic heterogeneity amongst different delta virus isolates.
Leucocyte hepatitis B virus DNA in acute and chronic hepatitis B virus infection
Authors: Davison F. , Alexander G. J. M. , Anastassakos Ch. , Fagan E. A. , Williams Roger
Published: March 2007

In the present study we have investigated 53 patients with a spectrum of acute and chronic hepatitis B virus (HBV) infection for the presence of leucocyte HBV‐DNA with the aid of molecular techniques. HBV‐DNA was detected in peripheral blood mononuclear cells of 31 of 45 (69%) of chronic HBsAg carriers and 2 of 8 (25%) patients with acute hepatitis B. Although HBV‐DNA was detected more frequently in leucocytes from those HBsAg carriers seropositive for HBeAg (79%), 50% of those with anti‐HBe in serum had leucocytes positive for HBV‐DNA independent of the presence of serum HBV‐DNA.

Examination of various leucocyte subpopulations showed the presence of HBV‐DNA in polymorphonuclear leucocytes as well as T‐ and non‐T‐enriched mononuclear cell fractions. The HBV‐DNA identified was predominantly 3.2‐kilobase (kb), while higher molecular weight sequences were rarely detected, and lower molecular weight sequences indicative of active viral replication were not observed.

These data indicate that although leucocytes do not actively support viral replication, they frequently harbour 3.2‐kb HBV‐DNA and may act as a reservoir for infection and, more importantly, since leucocytes contaminate several body secretions, may be involved in virus transmission.
Human cytomegalovirus (HCMV) enhances bovine papilloma virus (BPV) transformation in vitro
Authors: Goldstein Simoy C. , Byrne Janet C. , Rabson Alan S.
Published: March 2007

Infection of NIH 3T3 cells with a combination of HCMV and BPV resulted in more foci than infection with BPV alone. Foci were microscopically apparent at 4 days in the mixed infection and did not appear until 2 days later in the cultures infected with BPV alone. The enhancement was abolished by heat inactivation of the HCMV and also when the HCMV was replaced by a “mock inoculum.” Southern blot analysis of cellular DNA from transformed cells showed a similar amount of extrachromosomal BPV DNA in cells infected by BPV alone and in cells co‐infected with HCMV. No HCMV antigens could be found in these cells by immunofluorescence. The mechanisms of the enhancement are not known. Stimulation of host DNA synthesis by HCMV could possibly increase the transforming efficiency of BPV. Alternatively, the increase in BPV transforming efficiency could be due to a transient increase in BPV‐1 transcription by an HCMV transcriptional transactivation factor. Since both HCMV and human papillomaviruses are commonly found in the uterine cervix, HCMV may play a role in human cervical cancer by enhancing the carcinogenic potential of human papillomavirus.
Detection of human immunodeficiency virus (HIV) genome from the lymphocytes of homosexual men
Authors: Lee Yih Shyong , Baker Louis
Published: March 2007

Human immunodeficiency virus (HIV) is the primary etiological agent of acquired immunodeficiency syndrome (AIDS). This virus can be transmitted by blood or blood products. Homosexual males (HM) are regarded as one of the high‐risk groups for AIDS. Lymphocytes obtained from HM were used for cocultivation with fresh HIV antibody negative lymphocytes. The presence of virus particles was monitored by reverse transcriptase (RT) activity. Three out of eleven donors were found to have reverse transcriptase activity. All three donors were also found to be HIV antibody positive. HIV DNA from infected cells was detected by dot blot hybridization. The number of genome per cell is low. The correlation between RT activity and the cytolytic effect of HIV will be discussed.
Specific serum IgA in rotavirus gastroenteritis
Authors: Angeretti A. , Magi M. T. , Merlino C. , Ferrara B. , Ponzi A. Negro
Published: March 2007

Virus‐specific serum IgM, IgA, and IgG were detected by ELISA in sera obtained from children with rotavirus gastroenteritis and fractionated by gel filtration. Specific IgM and IgG could be easily demonstrated, whereas IgA were very low. Moreover, polymeric IgA (p‐IgA) were not present, whereas in the immune response against viruses causing systemic infections, they are synthetised in large amounts.
Circulating interferon in the guinea pig infected with the XJ, prototype Junín virus strain
Authors: Dejean C. B. , Oubiña J. R. , Carballal G. , Teyssié A. R.
Published: March 2007

The interferon (IFN) induction capacity of the XJ prototype strain of Junín virus (JV) was investigated in the guinea pig model. Circulating alpha IFN was detected in 50% of the animals from days 2 to 9 postinfection (pi) and in 100% at day 11 pi, when all animals were in the premortem stage.

Individual levels ranged from 20 to 1,280 guinea pig IFN units (GPIFNU)/ml. A correlation between XJ strain virulence and IFN titers was recorded. A possible role of IFN as a pathogenic factor in the outcome of the disease is discussed.
Differential effect of systemic acyclovir treatment of genital HSV‐2 infections on antibody responses to individual HSV‐2 proteins
Authors: Ashley R. , Mack K. , Critchlow C. , Shurtleff M. , Corey L.
Published: March 2007

Western blot and reflectance densitometry were used to evaluate the antibody response from patients treated with systemic acyclovir during their primary episodes of genital HSV‐2 infection. Of 39 patients studied, 10 received oral acyclovir, 10 received intravenous acyclovir, and 19 received placebo. Total antibody levels as well as levels of antibodies to individual HSV‐2 proteins (gB, gG, gC/gE, VP16, gD, and p45) were determined in convalescent phase sera. The median number of HSV‐2 proteins recognized and the total amount of HSV‐2 antibody were significantly lower in acyclovir than placebo treated patients (P ≤ 0.01). Levels of antibodies to individual proteins were also lower in sera from acyclovir versus placebo treated patients: gB (P = 0.013), gC/gE (P = 0.017), VP16 (P = 0.001), gD (P = 0.009), and p45 (P = 0.015). Antibody response to gG‐92 and to a newly described gG species, gG‐70, was not significantly different among treatment groups. A low number of proteins recognized by convalescent serum antibodies were associated with a higher number of lesions at first recurrence (P = 0.02) and with a longer duration of the first recurrent episode (P = 0.02). Low levels of total antibody were associated with shorter times to first recurrence (P = 0.05). Low levels of antibody to VP16 (P = 0.05) and gD (P = 0.01) were associated with longer duration of the first recurrence.
Assessment of HBV persistent infection in an adult population in Taiwan
Authors: Peng Hong‐Wen , Su Tsung‐Sheng , Han Shou‐Hwa , Ho Chi‐Kuan , Ho Chao‐Hung , Ching King‐Nien , Chiang Benjamin N.
Published: March 2007

In order to study the prevalence of hepatitis B virus (HBV) in the adult population of Taiwan, we screened for the presence of HBV DNA in 205 blood samples from adult (20–59‐year‐old) volunteers. According to the serological markers of HBV, samples were divided into three groups: group I (173 subjects) was negative for both HBsAg and HBeAg; group II (14 subjects) was positive for both HBsAg and HBeAg; and group III consisted of 18 subjects who were HBsAg‐positive but HBeAg‐negative. Plasma HBV DNA was not detected in group I, but it was found in 85.7% and 11.8% of samples in group II and group III, respectively. A free‐form HBV DNA was found in 14.3% of the leukocyte samples in group II. Furthermore, an integrated form of HBV DNA was detected in the leukocytes of two cases of group I who remained healthy based on clinical data. HBV DNA was also detected in the spermatozoa and liver cells of one of the cases.
HBeAg/Anti‐HBe seroconversion during and after protracted immunosuppressive treatment in type B chronic hepatitis
Authors: Tanno Hugo , Fay Oscar H. , Rojman José A. , Palazzi Jorge , Bessone Fernando
Published: March 2007

Seventy‐seven consecutive HBeAg‐positive chronic hepatitis patients were studied from 1971 to 1983 to establish the seroconversion rate in the e system. Patients with less than a year of follow‐up were not included in the study. Fifty‐six patients with chronic active hepatitis (CAH) received immunosuppressive treatment (corticosteroids combined with azathioprine). The remaining twenty‐one patients received no treatment, nine of them with chronic persistent hepatitis (CPH) and 12 with CAH. A retrospective study was performed with stored sera samples: HBeAg and anti‐HBe were determined by RIA, and results were correlated with alanine aminotransferase (ALAT) levels in the same samples. The linearized seroconversion rate from HBeAg to anti‐HBe was expressed as percent per patient‐year. It was 9.6% in CPH patients and 8.8% in CAH patients without treatment. In CAH patients under immunosuppressive drugs it was as low as 1.1% and increased to 28.7% when treatment was withdrawn. ALAT levels were significantly lower in total seroconverted patients when compared with nonseroconverted (NS) patients, but no difference was found between partial seroconverted (PS) and NS patients. The results suggest that although immunosuppressive drug withdrawal may enhance seroconversion rate in type B CAH, delayed seroconversion and reported side effects during treatment stand against protracted usage of these drugs.
Comparison of five elsa assays for IgG antibody against coxsackievirus B1
Authors: Torfason Einar G. , Galindo Rosalba , Keyserling Harry L.
Published: March 2007

Enterovirus type and group specificities of five different IgG ELISA methods were compared, using neutralization titration tests as an indicator of the presence or absence of antibodies to coxsackie B (CB) viruses.

One of the ELISA assays was a “standard” IgG assay, where the solid phase was coated directly with the purified virus, followed by incubations with human serum, biotinylated anti‐human‐IgG, streptavidin‐peroxidase, and the substrate/chromogen In a modified standard assay, blocking of common epitopes was attempted by incubating the CBI virus antigen on the solid phase with a rabbit antiserum to CB5 before the human serum was added. In another modification the serum dilution buffer contained heat‐denatured heterologous enteroviruses in an attempt to consume human antibodies reacting with common epitopes. In one assay the purified CBl virus was captured by purified horse anti‐CB1 IgG on the solid phase, before incubation with human serum. In the last of the five assays the serum specimen was incubated with CBl virus (in the liquid phase) before the virus or virus‐antibody complex was captured with purified horse anti‐CB1‐IgG.

Reactions against common antigens dominated in the first three assays. The antigen‐capture assay appeared to be at least predominantly type specific. Our data indicate that the liquid‐phase assay may be type specific, but more studies are needed. The method of virus purification was critical for the type specificity of the antigencapture and liquid‐phase assays.
Hepatitis B vaccine produced in yeast
Authors: Bitter Grant A. , Egan Kevin M. , Burnette W. Neal , Samal Babru , Fieschko John C. , Peterson Darrell L. , Downing Michael R. , Wypych Jette , Langley Keith E.
Published: March 2007

A gene encoding the 226 amino acid hepatitis B surface antigen (HBsAg), subtype adw, was cloned into a generalized vector for the expression of heterologous genes in Saccharomyces cerevisiae. The 5′ end of the genomic HBsAg gene was replaced with a chemically synthesized DNA segment that conserved the amino acid sequence of the protein but utilized DNA sequences that optimize translation initiation in yeast. High‐cell‐density fermentations of laboratory strains of Saccharomyces cerevisiae have been developed in which HBsAg production increases linearly with respect to cell mass. The HBsAg is present as a lipoprotein particle in cell lysates and has been purified to homogeneity. The evidence presented indicates that the HBsAg particles may be formed during lysis of the yeast cells. The purified HBsAg particles have a morphology similar to that of the 22 nm particles present in the serum of human chronic carriers of hepatitis B. The reactivity of the yeast‐derived HBsAg particles with a series of monoclonal antibodies is essentially identical to that of human plasma HBsAg. By this analysis, therefore, the structure of the HBsAg protein is similar in yeast and in human particles. The purified yeast HBsAg particles were formulated with alum adjuvant and subsequently were shown to confer immunity in chimpanzees to challenge with two heterologous serotypes (adr, ayw) of hepatitis B virus.
Symptomatic anti‐HBe positive chronic hepatitis B in taiwan with special reference to persistent HBV replication and HDV superinfection
Authors: Wu Jaw‐Ching , Lee Shou‐Dong , Tsay Shyh‐Haw , Tsai Yang‐Te , Chan Cho‐Yu , Huang Yi‐Shin , Lo Kwang‐Juei , Ting Ling‐Pai
Published: March 2007

Ninety‐four patients, who were admitted with symptoms of liver disease and found to be positive for hepatitis B surface antigen and antibody to hepatitis B e antigen (anti‐HBe), were examined for hepatitis B virus (HBV) DNA in serum and immunoglobulin antibody to hepatitis B core antigen and liver biopsies were stained for hepatic hepatitis B core antigen. Of 94 patients, 34 (36%) had evidence of HBV replication and 35 (37%) evidence of hepatitis D virus (HDV) superinfection. Most of the latter two groups of patients (> 70%) had evidence of chronic active hepatitis or active cirrhosis in their liver biopsies. The majority of these patients (> 80%) also had high levels of serum alanine aminotransferase (> 200 U/L) during the acute stage of their illness, and suffered from prolonged hepatic inflammation (> 1 year). Many of the patients (59%) also experienced frequent (1–6 episodes) relapsing exacerbations during a two‐year follow‐up period. Thus, persistent replication or reactivation of HBV and HDV superinfection were the two major causes of clinical exacerbations in anti‐HBe‐positive chronic HBV carriers in Taiwan, and also played an important role in the progression of their liver diseases and unfavorable outcomes.
Analysis of a major target of the human immune response to cytomegalovirus using monoclonal antibodies
Authors: Gold D. , Ashley R. , Abbo H. , Corey L.
Published: March 2007

Two murine monoclonal antibodies (C6D1 and D2B1) were found to react with a set of cytomegalovirus (CMV)‐infected cell polypeptides, which comprise a major target of the human immune response to CMV. C6D1 reacted with proteins of 50 kilodaltons (KD) and 40KD molecular weight; D2B1 reacted with these two proteins plus a third of 35KD. Western blot analysis demonstrated that these protein targets also react with serum antibody from patients with acute or latent CMV infection. Immunofluorescence staining of CMV‐infected diploid fibroblast cells by C6D1 and D2B1 showed that the protein targets are found in the nucleus throughout the course of viral infection. The proteins were shown to be late proteins dependent on viral DNA synthesis for their expression. Not all wild‐type CMV strains tested expressed proteins that react with C6D1 and D2B1. Using an immunofluorescence stain of diploid fibroblasts infected with CMV strains from infected patients, we found that 70 of 76 (92%) wild‐type strains reacted with C6D1 and 23 of 24 (96%) with D2B1. One strain was not reactive with either C6D1 or D2B1. Western blot analysis of 11 wild‐type strains revealed that two isolates either lack the C6D1 and D2B1 protein targets or have forms of these proteins that migrate at different molecular weights.
Spread of acute hemorrhagic conjunctivitis due to enterovirus‐70: Effect of air temperature and relative humidity on virus survival on fomites
Authors: Sattar Syed A. , Dimock Kenneth D. , Ansari Shamim A. , Springthorpe V. Susan
Published: March 2007

Enterovirus 70 (EV‐70) has caused at least two pandemics and several major epidemics of acute hemorrhagic conjunctivitis (AHC) in the past 18 years, with the largest number of cases occurring in the coastal areas of the tropics. The exact means of its spread are not known, but fomites and hands contaminated by them are the most likely vehicles. We, therefore, tested EV‐70 survival under different environmental conditions using stainless steel disks (1 cm diameter). Each disk received 10 μl of the virus in phosphate‐buffered saline (PBS). The disks were held at various temperatures with the relative humidity (RH) at the low (20 ± 5%), medium (50 ± 5%), high (80 ± 5%), or ultrahigh (95 ± 5%) level. The virus was eluted from the disks with tryptose phosphate broth and the eluate assayed in LLC‐MK2cells. Inactivation rates (Ki), expressed as hourly loss of virus plaque forming units (PFU) in log10, were then calculated. At 20°C, virus survival was proportional to the RH level, with the highest virus survival at the ultrahigh RH; at this level nearly 5% of the input virus was detectable even after 24 hr. Virus inactivation rates were only slightly higher at the ultrahigh RH when the temperature was raised to 33°C or 35°C. However, at 80% RH, increasing the temperature from 20°C to 33°C resulted in a dramatic rise in virus inactivation. The finding that high levels of RH prolong EV‐70 survival on fomites may help explain the epidemiology of AHC resulting from EV‐70.
Prognostic value of HBsAg/IgM complexes in hepatitis B patients: Nature of the proteins involved
Authors: Grangeot‐Keros Liliane , Pelletier Gilles , Briantais Marie‐José , Pillot Jacques
Published: March 2007

Hepatitis B surface antigen (HBsAg)/IgM complexes were measured using a solid phase radioimmunoassay in sera of patients with acute or chronic hepatitis B. These complexes were found in 6 out of 18 patients four weeks after the onset of the disease and only one of them developed chronic hepatitis. HBsAg/IgM complexes correlated with HBsAg, hepatitis B e antigen (HBeAg), and pre‐S2 concentration. The precipitation of HBsAg/IgM reactivity by polyethylene‐glycol (PEG) and the binding of this activity to the surface of certain uncoated enzyme linked immunosorbent assay (ELISA) plates indicates that HBsAg/IgM positivity may reflect the presence of circulating complexes in serum. HBsAg and pre‐S2 were found as components of the complexes but anti‐HBs, anti‐pre‐S2, and polymerized human serum albumin (pHSA) were not. An immune binding between HBsAg and IgM is still questionable. Whatever the nature of the HBsAg/IgM complexes, their detection does not seem to be an earlier indicator of prognosis than HBeAg and/or pre‐S2.
Enzyme‐Linked immunosorbent assay of core antigens for clinical diagnosis of influenza
Authors: Coonrod J. Donald , Karathanasis Paraskevi , Betts Robert F. , Donofrio James C.
Published: March 2007

A solid‐phase enzyme‐linked immunosorbent assay (ELISA) with monoclonal secondary antibodies was used to detect matrix protein and nucleoprotein of influenza A. The sensitivity of the ELISA for highly purified A/Brazil nucleoprotein and matrix protein was 0.05 and 1.0 ng, respectively. Nasal washes from 10 of 20 adult subjects with culture‐proven, naturally acquired infection caused by A/Brazil/11/78‐like influenza virus were positive in the test, and 2 of 13 subjects with rhinovirus infection were falsely positive. To determine if ELISA results could be improved, nasal washes were obtained from 21 adult volunteers who had been inoculated intranasally with wild‐type A/Korea/1/82 (five subjects) or A/Korea recombinants with matrix protein or RNA‐2 protein of A/Ann Arbor/6/60 (16 subjects), and the nasal washes were processed by a variety of methods. Prompt addition of sodium azide to the nasal washes to limit bacterial growth, avoidance of freezing, and the use of an antiproteolytic agent all failed to improve ELISA results noticeably. Under the best conditions, ELISA was positive in only 12 of the 21 experimentally infected subjects and in 1 of 15 uninfected controls. Positive ELISA results in experimentally infected subjects correlated significantly with the titer of live virus in the nasal washes (r = +0.506; P < 0.001). Detection of gradient‐purified whole influenza virus or isolated core antigen in ELISA was inhibited by prior incubation with nasal washes, and the inhibitory activity was only partly decreased by heat treatment of the secretions. At present, the use of ELISA for detection of influenza antigens in respiratory secretions is not sufficiently sensitive or specific for routine laboratory diagnosis of influenza.
Successful postexposure vaccination against hepatitis B in chimpanzees
Authors: Iwarson Sten , Wahl Martin , Ruttimann Eva , Snoy Philip , Seto Belinda , Gerety Robert J.
Published: March 2007

To study the effect of postexposure vaccination, four chimpanzees were vaccinated with hepatitis B (HB) vaccine 4, 8, 48, and 72 hr, respectively, after intravenous injection of an infectious hepatitis B virus (HBV) inoculum. The second and third vaccine inoculations were given 2 and 6 weeks later, i.e., at considerably shorter intervals than recommended either for ordinary prophylactic vaccination or for postexposure vaccination in combination with hepatitis B immune globulin (HBIG).

The chimpanzees were followed for 1 year. None showed HBs‐antigenemia, liver enzyme elevation (ALT), or histopathological alterations in liver biopsies. Late appearance of anti‐HBc was observed only in the serum of the animal whose series of vaccination started 72 hr after HBV inoculation. An unvaccinated control chimpanzee, which received the HBV inoculum only, developed clinical hepatitis B with ALT‐elevations and HBs‐antigenemia within 2 months of the experimental HBV inoculation.

These results indicate that postexposure vaccination against hepatitis B begun within 48 hr after HBV exposure, with short intervals between the vaccine injections, can protect against hepatitis B infection also when concomitant HBIG‐prophylaxis is not given.
Evaluation of an enzyme‐linked immunosorbent assay that measures rhinovirus‐specific antibodies in human sera and nasal secretions
Authors: Barclay Wendy S. , Callow Kathleen A. , Sergeant Marianne , Ai‐Nakib Widad
Published: March 2007

Rhinovirus‐specific antibodies have traditionally been detected by their ability to neutralise the homologous rhinovirus serotype in tissue culture. Recently, however, we have described an enzyme‐linked immunosorbent assay that detects rhinovirus‐specific antibodies in sera and nasal secretions [Barclay and Al‐Nakib, 1987]. Here we describe an evaluation of the ELISA in a study involving 71 adult volunteers inoculated intranasally with human rhinovirus type 2 (HRV‐2). Pre‐and post‐inoculation serum samples and pre‐inoculation nasal washings were tested for the presence of HRV‐2‐specific antibodies by ELISA. Such antibodies were associated with protection against infection when present locally in nasal secretions, but when also present in the serum they were associated with protection against both infection and the development of illness. The antibody concentrations showed strong correlation with each other and with that of antibodies detected by the neutralisation test. Following HRV‐2 infection, rises in HRV‐2‐specific IgA in sera detected by ELISA occurred more frequently than rises in neutralizing antibody. These results suggest that the ELISA is a sensitive and reliable indicator of recent infection, as well as a predictor of homologous immune status.
Accentuated antibody response to paramyxoviruses in individuals infected with human immunodeficiency virus
Authors: Kövamees Jan , Sheshberadaran Hooshmand , Chiodi Francesca , Schwartz Stefan , Örvell Claes , Fenyö Eva Maria , Norrby Erling
Published: March 2007

Sera from 31 human immunodeficiency virus (HIV)‐infected patients, representing different clinical stages of HIV infection, were assayed for antibodies against measles and mumps viruses by various serological tests and compared to 23 healthy controls. Sera from four patients (two primary, one asymptomatic, and one acquired immunodeficiency syndrome) exhibited a pronounced antibody response to measles as detected by haemagglutination inhibition and radioimmuno precipitation assay. The RIPA‐positive sera showed increased reactivity to all the viral components and in particular to the haemagglutinin (HA) protein of the virus (Fig. 1). Three of these positive patients also showed a similar response to mumps virus. One of the control sera also showed an increase in antibody titre in measles serological tests.

The measles antibodies were shown not to be anti‐HIV antibodies crossreacting with paramyxoviruses. The reactivity to haemagglutinin was still present when using nonglycosylated measles virus antigen grown in the presence of tunicamycin.

Whether the accentuated antibody response is due to polyclonal activation mediated by HIV or to reactivation of the viruses remains to be answered.
Purification and characterization of a strain of coxsackievirus b4 of human origin that induces diabetes in mice
Authors: Chatterjee Nando K. , Nejman Catherine , Gerling Ivan
Published: March 2007

A diabetogenic strain of coxsackievirus B4 of human origin has been purified to study its biochemistry and diabetogenicity. Tissue culture cells infected with the virus contain two distinct types of particles—virions and membrane‐bound virions (MBV). MBVs are lighter (ρ = 1.29) than virions (ρ = 1.34), and they contain relatively more protein than RNA. Virions contain four capsid proteins, VP1–4, of various molecular weights: VP1, 37,500; VP2, 36,000; VP3, 26,000; and VP4, 5,500. MBVs contain three of these proteins and several additional proteins of molecular weights 45,000 to > 92,500, possibly of host or viral origin. The RNA in each type of particle is a 35s molecule; T1 oligonucleotide fingerprint profiles suggest minor differences in the two RNAs. Hybridization experiments show a great deal of sequence homology between the RNA of the diabetogenic strain and the RNA of prototype CB4, which does not induce overt diabetes. MBVs are 10‐ 70 times less infective than virions, yet they are more pathogenic in mice and induce significantly higher glucose intolerance (hyperglycemia). The hyperglycemic response appears to be lower in mice infected with both types of particles than in mice infected with MBVs alone. Thus, the two subpopulations of virions present in the diabetogenic strain differ biochemically and in their ability to induce diabetes.
Recombinant DNA related to hepatitis B and human immunodeficiency viruses in mononuclear cells of patients with AIDS
Authors: Hirschman Shalom Z. , Zucker Marcia L.
Published: March 2007

Sixty‐eight of 73 patients with human immunodeficiency virus (HIV) infection were positive when tested for the presence of hepatitis B virus (HBV)‐related DNA sequences in peripheral blood mononuclear cells (PBMCs) by the dot blot method. Twenty‐two of the positive DNAs were examined by Southern hybridization and all exhibited a 3.2 kb extrachromosomal DNA fragment that hybridized to HBV DNA. This DNA was isolated from agarose gels and cloned into the EcoRI site of pBR322 DNA. The cloned DNA (pHBI) hybridized to both HBV DNA and HIV cDNA; HBV DNA did not hybridize to HIV cDNA under the same conditions. The results of restriction enzyme analyses indicated that pHBI contains: 1) a large deletion of HBV sequences spanning the 3′ end of the HBV surface antigen gene; 2) a small deletion near the 5′ end of the HBV core antigen gene; and 3) a region of homology to a one kb central section of the HIV pol gene. These data suggest that the 3.2 kb DNA found in the PBMCs is a natural recombinant between HBV and HIV DNAs raising the possibility not only that this DNA plays a role in the pathogenesis of AIDS but also that other viral recombinant DNAs may be pathogenic in human disease.
Analysis of virus and host factors in a study of A/Peking/2/79 (H3N2) cold‐adapted vaccine recombinant in which vaccine‐associated illness occurred in normal volunteers
Authors: Betts Robert F. , Douglas R. Gordon , Maassab Hunien F. , Deborde Dan C. , Clements Mary Lou , Murphy Brian R.
Published: March 2007

Live attenuated cold‐adapted influenza vaccine is undergoing evaluation in man. Several strains have proven to be safe, immunogenic, nontransmissible, and protective against experimental challenge. In this study of A/Peking/2/79(H3N2), with six internal genes from the cold‐adapted (Ca) parent A/Ann Arbor/6/60(H2N2), we encountered at the highest input multiplicity, 28% illness rate among individuals infected with vaccine. Reversion to wild type and excessive viral replication did not occur. Physical characteristics of the vaccine were similar to nonreactogenic vaccine A/Washington/897/80(H3N2). At ten‐ and 100‐fold lower input multiplicities, infection frequency was maintained, but reactions did not occur. We compared the observations in this study with those made in a similar study of A/Scotland/840/74(H3N2), a cold‐adapted vaccine with five genes from the Ca parent in which reactogenicity also was noted. The dose of vaccine virus in relation to tissue culture infectious doses required to infect 50% of susceptibles (HID50) was proportionally lower for both A/Peking/2/79(H3N2) and A/Scotland/80(H3N2). Hence, when the vaccine was undiluted the recipients were inoculated with more than 100 HID50. We concluded that the very high input could be avoided if vaccines were screened beginning at 1/1,000 of maximum titers. Ca vaccines must be safe before they undergo field trials.
Natural antibodies to HIV‐tat epitopes and expression of HIV‐1 genes in vivo
Authors: Krone Willy J. A. , Debouck Christine , Epstein Leon G. , Heutink Peter , Meloen Rob , Goudsmit Jaap
Published: March 2007

The tat regulatory protein of HIV‐1 was expressed as a fusion protein in E. coli and used as antigen to detect antibodies against HIV‐tat (anti‐tat) in the serum of HIV‐1 infected children and adults. HIV‐1‐infected children showed a higher frequency (55%) of anti‐tat than HIV‐1‐infected adults (36%). Anti‐tat were present in only 15% (3/20) of acutely infected individuals. Forty percent (10/25) of individuals with prolonged HIV‐1 infection but without antigen were anti‐tat positive. Only 13% (3/23) of HIV‐1‐antibody‐positive individuals with prolonged HIV‐1 antigenemia were anti‐tat positive and titers of anti‐tat antibodies declined with time. Pepscan analysis identified the amino terminus of HIV‐tat as the major antibody‐binding site. Antibodies to HIV‐tat occurred as a harbinger of HIV‐1 antigen expression and disappeared thereafter, possibly reflecting the transience of HIV‐tat expression. Because of the low antigenicity of HIV‐tat, antibodies to this regulatory protein are not a reliable marker for either early HIV‐1 infection or subsequent disease progression.
Protection against neonatal rotavirus infection by breast milk antibodies and trypsin inhibitors
Authors: Jayashree S. , Bhan M. K. , Kumar R. , Bhandari N. , Sazawal S.
Published: March 2007

The role of breast milk antirotavirus immunoglobulin A (IgA) and trypsin inhibitors in limiting the acquisition of rotavirus infection during the initial 5 days of life was evaluated among 42 exclusively breast‐fed hospital‐born infants, 22 of whom experienced rotavirus infection. The mean concentrations of antirotavirus IgA (ELISA Units) in the breast milk of mothers of the 22 rotavirus‐infected neonates was 130.4 ± 46.4; the corresponding value in 20 noninfected neonates was 384.3 ± 328.3 (P < 0.001). Similarly, the trypsin inhibitory capacity (μmols/mt/ml) of breast milk in the rotavirus‐infected group was significantly lower (0.109 ± 0.095) than that in the noninfected group (0.376 ± 0.191; P < 0.001). The trypsin inhibitory capacity of milk showed an inverse correlation with infant stool tryptic activity (P < 0.01).

Our results indicate that the acquisition of rotavirus infection during the early neonatal period depends on the concentrations of antirotavirus IgA and trypsin inhibitors in human milk and that protection is mediated by high levels of these antiviral factors.
Alteration of infection pattern of duck hepatitis B virus by immunomodulatory drugs
Authors: Fukuda Ryo , Okinaga Satoshi , Akagi Shuji , Hidaka Mitsunobu , Ono Naomi , Fukumoto Shiro , Shimada Yoshihiro
Published: March 2007

The relationship between host immune state and hepatic inflammation and infection pattern of the Duck hepatitis B virus (DHBV) was investigated by experimental transmission of DHBV into 98 Japanese 7‐day‐old ducklings that had been pretreated with immunoregulatory drugs including cyclophosphamide, OK 432, and a steroid hormone.

Immunosuppressive treatment with cyclophosphamide revealed an extension of the viremic period associated with an absence of inflammatory changes in the liver. Although immunostimulating treatment with OK 432 showed a remarkable accumulation of inflammatory cells in the liver, the viremic period was not shortened. Treatment with a steroid used as a immunosuppressant did not suppress the hepatitis; moreover, it increased viral DNA replication and extended the viremic period. This phenomenon of viral replication seemed to be caused by the direct effects of the steroid.

Alteration of DHBV infection by modifying the host immune state is quite similar to that of hepatitis B virus (HBV) in humans. In DHBV infection, the host immune state seemed to have a considerable role in determining the infection pattern and degree of hepatitis activity. DHBV may be a helpful model of HBV for studying host‐viral interaction and the immunological mechanism of viral hepatitis.