Virus B3 Research Articles

CSF Viral Antibodies

Serum and CSF from 48 patients with amyotrophic lateral sclerosis and six patients with late-onset postpoliomyelitis progressive muscular atrophy were investigated for the presence of antibody to poliovirus types 1, 2, and 3, coxsackie viruses B3 and B4, influenza A, measles, rubella, mumps, herpes simplex types 1 and 2, cytomegalovirus, varicella-zoster, and Toxoplasma gondii. These results were compared with those from 53 control patients with neuromuscular disease matched for age, sex, race, and poliovirus vaccine exposure. There was no difference either in distribution of serum or CSF antibody titers or the geometric-mean antibody titers. There was no evidence suggesting the presence of locally produced specific viral antibody within the CNS to any of the agents studied. In particular, there was no serological evidence to suggest an association between persistent infection with any poliovirus type and amyotrophic lateral sclerosis or late-onset postpoliomyelitis progressive muscular atrophy.

Studies on the frequency and associations of islet-cell antibodies in juvenile diabetes mellitus

Ninety-six juvenile onset type diabetics showed an increase in the frequency of HLA B8 and B15 and a decrease in frequency of HLA B7 antigens. Sixty-four maturity onset diabetics showed no disturbance in the frequency of these antigens. Fifty-four of the juvenile onset type diabetics, with an average duration of disease of 3.2 years were tested for the presence of islet cell antibodies (ICAs). Thirty-two percent were positive, the frequency decreasing from 70% in those patients tested within one year of diagnosis to 0.5% in those patients tested more than 5 years after diagnosis. No correlation was found between HLA type and the frequency of ICAs, but there was an increase in other autoantibodies in B15 positive patients. Preliminary absorption studies suggest a cross reactivity of ICAs and Coxsackie B4 virus.

Virus-induced Diabetes Mellitus: XI. Replication of Coxsackie B3 Virus in Human Pancreatic Beta Cell Cultures

The capacity of Coxsackie B3 virus to infect insulin-containing beta cells was studied in human pancreatic cell cultures. Antibody to Coxsackie B3 virus was labeled with fluorescein isothiocyanate, and antibody to insulin was labeled with rhodamine. By use of a double-label antibody technique, three populations of cells were identified: uninfected insulin-containing beta cells, which stained only with rhodamine-labeled anti-insulin antibody; Coxsackie-infected (noninsulin-containing) cells, which stained only with fluorescein-labeled anti-Coxsackie antibody; and Coxsackie-infected insulin-containing beta cells, which stained with both antibodies. Radioimmunoassay showed that intracellular immunoreactive insulin decreased rapidly beginning at 24 hours after infection, and the decrease in insulin roughly paralleled the increase in viral titer. It is concluded that, under in vitro conditions, human beta cells are susceptible to Coxsackie B3 virus.

Virus-induced diabetes mellitus. XI. Replication of coxsackie B3 virus in human pancreatic beta cell cultures

Virus-induced diabetes mellitus. XI. Replication of coxsackie B3 virus in human pancreatic beta cell cultures

Coxsackie B4 viral infection, anti-islet immunity and immunogenetics in insulin-dependent diabetes mellitus

The interrelationship between Coxsackie B4 (CB4) virus antibodies, islet-cell antibodies and HLA B8 was studied in 67 patients with juvenile-onset diabetes mellitus (JOD). In 41 JOD patients (61%) CB4 antibodies (greater than 1:4) were found. A high prevalence (69%) of significant CB4 antibody titers (greater than 1:64) was observed in patients with age at onset between 11 and 20 years compared with those with age at onset below or over that age-range (27% and 33%, respectively). Islet-cell antibodies (ICA) could be detected only in 11 JOD patients, which is probably due to the relatively long mean duration of disease. However, 8 of them presented with significant CB4 antibody production. Although no correlation between CB4 antibodies and the JOD-associated HLA-antigens (B8, Bw15, Cw3 and B7) could be detected in the entire group of JOD patients, a close association between ICA persistence and HLA B8 was found. Furthermore, in the ICA-positive, moderate or high CB4 antibody titers were prevalent. Thus, the present data point to a possible interrelationship between CB4 virus antibodies, persistent ICA production and HLA B8 in a minor proportion of JOD patients. However, it seems premature to assume direct implication of these factors in the pathogenesis of JOD.

A comparative electrophoretic examination of swine vesicular disease virus and Coxsackie B5 virus

Purified suspensions of Coxsackie B5 virus and swine vesicular disease virus (SVDV) were prepared by harvesting and purifying cell pack virus. Crossed immunoelectrophoresis was carried out with purified N and H antigen fractions (full and empty particles). Relative migration velocity (RMV) was calculated for the N antigen fraction of 3 SVD viruses (UKG72, HK71 and Italy 66) and 2 Coxsackie B5 viruses (Faulkner and 8068). The prototype strain of Coxsackie B5 virus (Faulkner) had a low RMV almost identical to that of the first isolated strain of SVDV, Italy 66. A recent isolate of Coxsackie B5 virus (8068, isolated in UK in 1973) had a relatively high RMV very close to that of the UKG72 strain of SVDV isolated in the UK in 1972. Also the Hong Kong strain of SVDV (HK71) had a high RMV value. These observations are considered in relation to the emergence of swine vesicular disease.

Effect of Immunosuppression on Dengue Virus Infection in Mice

Mean survival time following intracerebral inoculation of dengue virus was reduced and the titre of the virus in the brain of immunosuppressed mice was markedly increased. A single dose of cyclophosphamide given 24 h after dengue virus i.c. or i.p. substantially reduced the number of antibody forming cells in the spleen. Three doses of dengue virus, each followed by cyclophosphamide 24 h later, produced specific hyporesponsiveness to the dengue virus but not to a heterologous virus (Coxsackie B4), with a reduction in antibody forming cells in the spleen of such animals against dengue virus but not against Coxsackie B4 virus. Adoptive immunity by antiserum was abolished along with increased titres of the virus in the brain of immunosuppressed mice but the protection could be restored by a second dose of antiserum. Pre-treatment of mice with immune or normal spleen cells i.v. or reconstitution of immunosuppressed mice by such cells had no effect. Thus, humoral antibodies play a crucially important role in host defence mechanism in recovery of mice from primary dengue virus infection.

Molecular Aspects of the Antigenic Variation of Swine Vesicular Disease and Coxsackie B5 Viruses

SUMMARY The antigenic variation of swine vesicular disease virus (SVDV) and Coxsackie B5 virus (CB5) has been examined at the molecular level by analysing the protein and nucleic acid of the virus particles. The tryptic peptides of carboxymethylated 35S-methionine labelled virus particles were very similar, although some minor differences were apparent. Competition hybridization experiments confirmed that there is variation in the RNA sequence of antigenically distinct SVD viruses and some limited homology of these RNAs to the CB5 virus RNAs. Competition hybridization using mixtures of two RNAs as competitors showed that the sequences shared by the CB5 virus RNAs were largely the same as those shared by the CB5 virus RNAs and the SVDV RNAs. Similar experiments with the SVDV RNAs established that the homologous regions in these RNAs were also shared. Thermal denaturation curves of SVDV RNA-RNA hybrids generally supported the competition hybridization results but the hybrids between SVDV RNA and CB5 virus RNA were shown to be mismatched. Ribonuclease T1 oligonucleotide maps of the virus RNAs were also compared to obtain another measure of relatedness. A number of long oligonucleotides were shared by the SVDV RNAs but few of these were found in either of the CB5 RNAs. Further ways of investigating the antigenic variation of SVDV at the molecular level are discussed.

Pancreatitis and altered glucose tolerance in mice infected with coxsackie B4 virus.

CD1 mice infected with Coxsackie B4 virus showed an early polymorph infiltration of the pancreas, which later changed to a mononuclear exudate. A state of glucose tolerance developed concurrently and both events coincided with the inhibition of migration of spleen cells in the presence of the virus. The relevance of these findings to human juvenile-onset diabetes mellitus is discussed.

Coxsackie B2 virus infection and acute-onset diabetes in a child.

Coxsackie B2 virus infection and acute-onset diabetes in a child.

Enterovirus Infections in Acute Pancreatitis - a Possible Etiological Connection

All patients admitted with a preliminary diagnosis of acute pancreatitis during a one-year period were subjected to virus investigations. 142 patients were included in the study and of these 91 were found to have acute pancreatitis. Evidence of enteroviral infection was found in 18 of the 91 patients (19.8 per cent). From 17 of the 18 patients enterovirus was isolated from feces or urine, and in one a significant titer rise (CF-test) against Coxsackie virus B5 was found. A significant titer rise of antibodies against the virus type isolated could be demonstrated in 7 of the 17 patients. Thus in 8 cases there was obvious evidence of an acute enteroviral infection during the episode of acute pancreatitis. The etiological agents in these cases were Coxsackie viruses B2, B3, and B5, and ECHO-viruses 6, 11, 22, and 30.

Viral coronary arteritis and myocardial infarction

Two young male patients are described in whom a viral type of upper respiratory tract infection was followed by myocardial infarction. Based upon previously reported experimental studies in mice and monkeys in which Coxsackie B4 virus produced extensive coronary arterial and capillary injury, and upon the clinical data in these two patients, it is suggested that the myocardial infarcts in our two patients were the result of viral coronary arteritis.

A follow-up study of 15 cases of neonatal meningoencephalitis due to Coxsackie virus B5.

Summary o.1.Thirteen of 15 children who developed meningoencephalitis due to Coxsackie virus group B5 in the neonatal period had physical development, scores for intelligence and visual perception at the age of 6 years similar to those of 15 control children matched for sex, birth weight, gestational age, and socioeconomic status. 2.Two of three infants who were noted to be irritable or twitching in association with the meningoencephalitis in the neonatal period developed spasticity and their intelligence was below the mean for the group. 3.The three infants who also had myocarditis had no sequelae at the age of 6 years. Summary o.1.Thirteen of 15 children who developed meningoencephalitis due to Coxsackie virus group B5 in the neonatal period had physical development, scores for intelligence and visual perception at the age of 6 years similar to those of 15 control children matched for sex, birth weight, gestational age, and socioeconomic status. 2.Two of three infants who were noted to be irritable or twitching in association with the meningoencephalitis in the neonatal period developed spasticity and their intelligence was below the mean for the group. 3.The three infants who also had myocarditis had no sequelae at the age of 6 years.

Virus infections in infant mice causing persistent impairment of turnover of brain catecholamines

Newborn mice were inoculated with attenuated Coxsackie type B4 virus. Three-to-4-day old mice were infected with yellow fever virus vaccine. A number of mice survived the acute infections. Some of these demonstrated residual neurological symptoms, some showed recovery from symptoms while others survived the infection without revealing symptoms of disease. Determinations of dopamine, noradrenaline, 5-hydroxytryptamine, homovanillic acid and 5-hydroxyindoleacetic acid in the inoculated brains indicated an impaired turnover of neurotransmitters. Subnormal concentrations of catecholamines and homovanillic acid were encountered in the acutely-infected mice as well as among the survivors. Failure to synthesize catecholamines was observed not only in mice demonstrating symptoms of disease or in animals which recovered from their infection but also among a proportion of the mice which never demonstrated neurological symptoms. In contrast, 6-week-old Swiss albino mice infected with West Nile virus showed no effect on the turnover of brain monoamines either in acutely infected mice or in animals which survived the acute infection. Herpes simplex virus infection of 3-week-old mice induced during the acute infection an increased release of neurotransmitters. When these mice were “cured” of the infection by increasing the environmental temperature the elevated turnover of monoamine metabolism was normalized. Two months later there were no differences in concentrations of catecholamines or homovanillic acid between infected animals or uninfected controls. Thus, persistent impairment of brain monoamine metabolism was induced in mice infected when very young. The possible importance of the observations, in particular the findings of an impaired turnover after subclinical infection, is discussed.

Coxsackie B4 virus crystal formation in mouse pancreas.

For the first time, Coxsackie B4 virus crystals are demonstrated in tissues of animals inoculated with the virus. Until this report, Coxsackie B4 virus had never been found to form crystalline aggregates in tissues of any kind. The viral crystals which were located in the pancreas of newborn mice were associated with ultrastructural damage to the pancreas not observed in control animals, thereby indicating that viruses directly invade and damage these tissues. Thus, this virus must be considered among etiologic factors in pancreatic disease.

LACK OF CAUSAL ASSOCIATION BETWEEN COXSACKIE B4 VIRUS INFECTION AND DIABETES

An epidemic of Coxsackie B4 virus infection in an isolated group of islands in the Bering Sea in 1967 provided an opportunity to test the suggestion that infection with this virus might be associated with an increased incidence of diabetes. In 1973 islanders were tested by glucose-tolerance tests and their two-hour plasma glucose levels were analysed in the light of serological evidence of CB4 infection five years earlier. There was no evidence of any increased prevalence of diabetes in those who had been infected in 1967.

Coxsackievirus infection in mice

White mice of the NYA: NYLAR strain were infected with coxsackie B1 or B4 virus and studied for persistence of virus, antibody conversion, histologic changes, and glucose tolerance during periods up to 13 months. Pancreatitis was observed during the acute phase of infection. Suggestions of subclinical diabetes were found during the first six months after virus inoculation. A normal glucose tolerance at 10 to 13 months suggestes that the functional lesion is temporary.

Coxsackievirus Infection in Mice

White mice of the Nya: NYLAR strain were infected with coxsackie B1 or B4 virus and studied for persistence of virus, antibody conversion, histologic changes, and glucose tolerance during periods up to 13 months. Pancreatitis was observed during the acute phase of infection. Suggestions of subclinical diabetes were found during the first six months after virus inoculation. A normal glucose tolerance at 10 to 13 months suggests that the functional lesion is temporary.

Spherical Aggregates of Coxsackie B 4 Virus Particles in Mouse Pancreas

A high titer culture of Coxsackie B4 virus was used to induce pancreatitis in newborn mice. In animals sacrificed 1 or 2 days after intraperitoneal inoculation, we observed cytonecrosis consistent with picornaviral infection as well as necrosis indicative of pancreatitis. In addition, we observed aggregates of particles which seem to be Coxsackie B4 virus particles, some arranged in the typical picornaviral crystalloid lattice formation and others arranged into spherical masses approximately 102 nm in diameter. Depending upon the depth and orientation of section through the spherical aggregates, the particles were arranged into two patterns which were readily distinguishable. When the plane of section was through the center of the sphere, 10 particles circularly arranged around a dense particle core were observed. When the sphere was cut tangentially, the particles were arranged in a zig-zag pattern so that there were two concentric layers of at least 6 particles per layer, with no central core. Both crystalloid and spherical aggregates were observed free within acinocyte cytoplasm, and within autophagic vacuoles, cytosegresomes, and fine granular bodies of acinocytes, and within phagocytic vacuoles of macrophages. We conclude that the spherical aggregates represent a distinct crystalloid form of Coxsackie B4 virus during its replicative cycle, which may eventually develop into the more typical picornaviral crystalloid lattice configuration and that the spherical aggregates are located in foci of viral synthesis. Marked pathogenicity of Coxsackie B4 virus in the newborn mouse pancreas should be considered a factor in the observations noted.

野外豚における豚水胞病ウイルスに対する低力価抗体の解析

Swine vesicular disease (SVD) caused by enterovirus broke out for the first time in Japan in November, 1973. It occurred originally in a region producing breeding swine in Kanagawa Prefecture and prevailed up to the next month. Then an antibody survey was conducted to clarify the degree of infiltration of this virus among swine. Incidentally, antibody against SVD virus was detected in it from the sera of some swine which were considered to be free from the outbreak of SVD. These sera included those collected before the present outbreak and those harvested from swine produced and raised on farms in a region geographically apart from the site of the outbreak.SVD virus is antigenically very well identical with Coxsackie virus B5 (CB-5), which is human enterovirus. Serum samples were collected from swine in the field and examined for the presence of antibody against CB-5 virus. Then studies were made on the relationship between this antibody and that against SVD virus with the following results.1. Serum samples were collected from swine all over Japan at three points of time; that is, prior to the outbreak of SVD and 1 and 5 months after the termination of this outbreak. Serum positive for SVD antibody was detected at every point of time. The coefficient of correlation between the two types of antibody, however, was considerably low, or 0.34, 0.31, and 0.34 before and 1 and 5 months after the outbreak, respectively.2. When serum samples were collected from swine apparently infected on farms involved in the outbreak of SVD and examined for the two types of antibody, there was a very high correlation (r=0.74-0.85) between the antibody titers of these two types.3. The coefficient of correlation was low, or 0.44, in serum samples collected from swine manifesting only indistinct signs of infection and from those which had withstood the infection without showing any symptom on farms involved in the outbreak of SVD. It was also low, or 0.53, in serum samples (antibody titer, <44.0) harvested from swine which seemed to be in the ascending stage of antibody.From these results, it was suggested that there might be an inapparent infection of swine in Japan with such type of virus as mentioned above which would not cause the production of vesicles, but that of low-titer antibody. As such virus, CB-5 virus, which was antigenically identical with SVD virus, or any other unknown virus was presumed to exist.