Virus B3 Research Articles

Infections with coxsackie B viruses in the cameroons

1. 1) The isolation of a strain of Coxsackie B4 from a case of illness is described, in which it was probably the aetiological agent. 2. 2) Although circulating in the community, it was not responsible for a group of other cases of illness associated with a rash and enlarged glands. The agent responsible for these was not identified; it was probably not an arthropod-borne virus, and may have been another enterovirus. 3. 3) A serological survey of the native population showed that infection with the Coxsackie viruses B1, B2, B3, B4, and B5, was very common. 4. 4) There was no evidence of serological overlap between these viruses.

A study on dissociation of Coxsackie B4 virus-antibody complex

Dissociation of neutralizing antibody from Coxsackie B4 virus was achieved by acid treatment; pH 2.5 was found to be optimal for the reaction. Reactivated virus could be reneutralized by the dissociated antibody, but at a slower rate than the initial neutralization. Simple dilution was an effective method for recovering infectious virus from virus-antibody mixtures only if these mixtures were first adjusted to an acid pH. In the presence of excess antibody, recovery of infectious virus increased five- to sevenfold if the acidified virus-serum mixture was diluted in hypertonic rather than isotonic saline.

L-CYSTINE REQUIREMENT FOR PRODUCTION OF COXSACKIE B3 VIRUS IN CULTURED MONKEY HEART CELLS.

Tyndall, R. L. (The Pennsylvania State University, University Park) and E. H. Ludwig. l-Cystine requirement for production of Coxsackie B3 virus in cultured monkey heart cells. J. Bacteriol. 85:1339-1345. 1963.-The requirement for l-cystine in both the production of Coxsackie B3 virus in cultured monkey heart cells and in prolonging the survival of the cultured monkey heart cells is demonstrated. d-Cystine, dl-homocystine, dl-allo-cystathionine, dl-homocysteine thiolactone, 2-mercaptoethlyamine, allylglycine, 3,3'-dithiopropionic acid, and dl-ethionine could not replace l-cystine in either supporting Coxsackie B3 virus production or prolonging monkey heart cell culture survival time. By their ineffectiveness, these compounds suggest the specificity of the l-cystine requirement. Allylglycine, 3,3'-dithiopropionic acid, and dl-ethionine were likewise incapable of inhibiting the l-cystine effect in supporting both virus production and cell survival. Prolonged starvation of cell cultures prior to virus inoculation failed to reveal any additional marked nutritional requirements, but rather tended to accentuate the l-cystine requirement for virus production. Increased cell starvation did, however, lead to the establishment of a latent Coxsackie B3 virus infection of cultured monkey heart cells, the activation of which is l-cystine-dependent.

Über die Virusempfindlichkeit von Schilddrüsen-Zellkulturen

Infected tissue culture fluid of prototype strains of various viruses was passed five times in human thyroid tissue cultures. Comparative titrations were carried out with both original culture fluid and virus from the fifth cell passage in thyroid cells as well as in HeLa or monkey kidney cells respectively. By this means thyroid cells were found to exhibit an equal (or in some instances higher) susceptibility as the culture of origin to vaccinia virus, herpes simplex virus, parainfluenza viruses types 1, 2 and 3, adenoviruses types 3, 6, 9 and 12, two prototype strains of Coxsackie B3 virus, Coxsackie A9 virus, ECHO 9 virus [prototype Hill and an epidemic strain (Russ.)], an attenuated and a virulent strain of each of the three poliovirus types. The virus yield in thyroid and the original cell system was found to be in the same range. ECHO 16 virus, two prototype strains of Coxsackie B4 virus and B.V.A. 96 strain of Coxsackie B2 virus produced no cytopathic effect in thyroid cell cultures and no inapparent multiplication was demonstrable. The Ohio 1 strain of Coxsackie B2 virus had a somewhat lower infectivity for thyroid than for monkey kidney cells. Reovirus type 1 showed minimal cytopathic change and only a slight multiplication in thyroid cells. From the ECHO virus group, the prototype strains of the types 1, 2, 3, 5, 6, 7, 8, 11, 12, 13, 19, 20, 22 and 24 could be passed readily with CPE in thyroid cells, whereas ECHO types 18 and 23 were erratic and type 17 produced no CPE. The prototype strains of adenovirus types 1 to 28 as well as 24 other strains of various serotypes were cytopathic for thyroid cells without exception. Our experience with the culture and maintenance of human thyroid cells as obtained in the course of this study has been reported.

Experimental Coxsackie group B virus infections.

The ability of Coxsackie B3 virus to produce experimental infection in the mouse prenatally and post-natally has been examined. The embryos become infected only after intrauterine inoculation. The examined tissues show a negligible damage or none at all.

Heterogeneity of Coxsackie B4 virus: Two kinds of particles which differ in antibody sensitivity, growth rate, and plaque size

Two kinds of virus particles which are antigenically similar but which differ in several biological properties have been isolated from a strain of Coxsackie B4 virus. These virus particles have been designated a s and a i. The a s particles are sensitive to specific antibody and to an inhibitor present in agar, and form small plaques; the a i particles are relatively insensitive to antibody, insensitive to the agar inhibitor, and form large plaques. The addition of DEAE dextran to agar abolishes the inhibition of a s particles by the agar inhibitor and results in the formation of large plaques by these particles. The a s virus adsorbs, multiplies, is released more rapidly, and causes more complete cell destruction, than the a i virus. The properties which distinguish the two kinds of virus were stable on passage. Early passages of Coxsackie B4 strains consisted predominantly of virus particles which formed plaques resembling those formed by a i virus; whereas the Powers strain, after many passages, consisted predominantly of a s virus, presumably because of the rapid growth rate of a s particles. The presence of two kinds of particles in varying proportions in Coxsackie B4 virus strains provides an explanation for some of the problems encountered in previous immunological studies with these strains.

Interaction of enteroviruses with receptors from erythrocytes and host cells

Receptors for ECHO viruses 7, 11, 19, and Coxsackie B3 were found only in the ghosts of human red cells, not in ghosts from other species. The receptor was destroyed by proteolytic enzymes, a fact suggesting that it may be a protein. The attachment of virus to receptor occurred in two stages: the first was a non-temperature-dependent attachment which was spontaneously reversible at 37° for ECHO 11 and ECHO 19 and reversible after different treatments for ECHO 7 and Coxsackie B3 virus. In some cases this was followed by a second stage in which the complex became increasingly resistant to attack by chymotrypsin. Intact monkey kidney and HeLa cells showed a similar two-step interaction between virus and cells. With inhibitory material from disintegrated cells and subcellular fractions, only the first attachment step could be demonstrated.

Gel Double Diffusion Studies with Group B and Group A, Type 9 Coxsackie Viruses

Summary The demonstration by the gel double diffusion technique of “specific” precipitating antibodies for the Coxsackie viruses suggests that the method offers considerable promise for use in the laboratory diagnosis of infections due to virus types A9, B1, B3, B4, B5 and possibly B6. Patients with Coxsackie B2 virus infections, however, could not be shown to develop “specific” precipitating antibody. “Specific” precipitating antibodies for the infecting virus type were found in the sera of 97% of patients from whom a Coxsackie A9, B1, B3, B4, B5 or B6 virus was isolated and in 40 to 65% of these patients “specific” precipitating antibody developed over the course of the illness. However, since “specific” precipitating antibody disappears relatively rapidly during convalescence, its presence in any serum specimen may be evidence of a current or very recent infection. Heterotypic “specific” precipitating antibody was absent in 78% of the patients who yielded a Coxsackie virus, and when present would appear to be due to recent infection with the heterotypic virus, or to anamnestic response.

Effect of Westphal lipid A on viral activities in mice and hamsters.

SummaryWestphal's lipid A assayed in mice and hamsters enhanced non-specific host resistance against the activities of viruses such as was demonstrated earlier for bacterial endotoxins and for other materials of microbial origin. Lipid A, given intracerebrally into mice, suppressed neurotoxicity of concentrated influenza A virus and reduced the activities of neurotropic influenza (NWS) and herpes simplex viruses as measured by survival and by survival time. Given subcutaneously, it was active against influenza A virus and, following intraperitoneal injection, was active against encephalomyocarditis virus but showed slight if any activity against measles in hamsters or neurotropic (NWS) influenza virus in mice. It was inactive against Lansing type II poliovirus and Coxsackie B3 virus when given intracerebrally and intraperitoneally, respectively. Greatest activity was in the suppression of PVM virus, even when administered up to 72 hours following viral infection. The lipid A did not appear to suppress pro...

Virus-Tissue Union and the Pathogenesis of Enterovirus Infections

Summary A heat labile, sedimentable substance capable of inactivating Coxsackie B1 and B3 viruses has been demonstrated in noncultured mouse brain and in certain other murine and primate tissues. The tissue and age distribution of this factor appears, in general, to be correlated with the in vivo sites of multiplication of these viruses and with their virulence for newborn and adult mice. It has been postulated that this material in brain and other tissues is specific virus receptor-site and that the relative abundance of this substance in cells of various tissues is an important determinant of age and tissue specific susceptibility to these agents. Sequential loss, with age, of adsorptive capacity of mouse brain for Coxsackie B1 virus was demonstrated in quantitative studies. Amount of tissue employed was rate limiting, but inoculum size was not critical. Cortisone treatment did not increase adsorptive activity of adult mouse brain for this virus. Adsorption of Coxsackie B1 and poliovirus by brain homogenates was temperature dependent, being more marked in the case of the former. Adsorption of poliovirus by noncultured, extraneural rhesus monkey tissues was confirmed; adsorption of Coxsackie B1 virus was limited to rhesus monkey cerebral cortex, cerebellum and liver. Brain weight-virus adsorption curves demonstrated about equal activity of rhesus monkey brain against both virulent and avirulent polioviruses but marked differences were observed in similar studies utilizing newborn mouse brain against virulent and avirulent strains of Coxsackie B3 virus. These studies are presented as evidence that age specific susceptibility and mouse virulence of certain Coxsackie viruses are related to the relative abundance of receptor-like substance in host tissues.

Influence of Potassium on Coxsackie B2 Virus Growth in Tissue Cultures

SummaryPotassium ion augments the amount of infectious Coxsackie B2 virus produced from MKC and MTC. Production of infectious Coxsackie B2 virus was reduced 500 to 2,000 times in MKC and MTC by depletion of potassium from their medium. Similar results were not obtained with Coxsackie B3 virus. Potassium was not required for virus attachment, but did stimulate glycolysis of tissue culture cells. Potassium could be replaced in a deficient medium after 12 hours of infection and still obtain maximal yields of infectious virus in MTC.

Experimental infections with Coxsackie viruses

Nine cynomolgus monkeys were inoculated intravascularly with Group B, type 4 (Edwards strain) Coxsackie virus. Viremia developed in 6 monkeys by the 2nd day, but was not detected thereafter. B4 virus was recovered in CSF obtained from 2 monkeys. The specific antibody responses were equivocal for 8 monkeys; all showed a slight rise in antibody levels; only one monkey surviving 28 days had an unequivocal antibody response.

Finding of virus particles in cultivated monkey kidney cells infected with coxsackie virus B5

Finding of virus particles in cultivated monkey kidney cells infected with coxsackie virus B5

The mammalian cell-virus relationship. VI. Sustained infection of HeLa cells by Coxsackie B3 virus and effect on superinfection.

Sustained infection of HeLa cells by Coxsackie B3 virus, dependent on presence of viral inhibitor in culture medium, was achieved. Persistent treatment of carrier cultures with anti-Coxsackie B3 hyperimmune monkey serum eventually eliminated virus from carrier cultures indicating that a lysogenic virus-cell relationship was not operative. Free virus was produced continuously by carrier cultures despite washing and neutralization with antiserum to eliminate free virus temporarily. In carrier cultures, about 1.5 to 1.9 plaque-forming units of virus per cell were cell-associated; approximately 6 per cent of this cell-associated virus was not neutralizable by antiserum. In growth medium containing anti-B3 antibody, cells from carrier cultures formed colonies as efficiently as cells from B3-cured cultures. Assays of carrier cultures for infectious centers indicated that less than 1 per cent of cells produced free infectious virus. The Coxsackie B3 virus-carrier state appeared to represent surface residence of B3 virus on the majority of carrier cells with restriction of productive infection to a small proportion of the population. Coxsackie B3 carrier HeLa cultures, unlike control cultures, were not destroyed by challenge with Coxsackie B1, B3, or B5 viruses. The B3 carrier state did not interfere with superinfection by herpes, vaccinia, and types 1 to 3 polioviruses. In contrast to parental or B3-cured lines, B3-carrier HeLa cultures superinfected with Coxsackie B1 virus produced no significant virus, and cultures superinfected with B5 viruses produced new virus to a limited extent only. Specific interference with Coxsackie virus superinfection by the B3-carrier state of HeLa cells was shown to be attributable to failure of attachment in the instance of Coxsackie B1 virus, and failure of penetration and/or eclipse in the instance of B5 virus. The interfering effect was circumvented successfully by superinfection of carrier cells with ribonucleic acid extracted from Coxsackie B1 and B5 viruses.

Enteroviral ribonucleic acid. II. Biological, physical, and chemical studies.

Chromatographic behavior of whole type 1 poliovirus and phenol-extracted viral RNA on diethylaminoethyl cellulose columns, as revealed by assay of plaque-forming capacity, indicated that infectious RNA had surface properties markedly different from those of the intact virus. Infectious RNA of type 1 poliovirus and Coxsackie B1 virus was relatively resistant to heat inactivation as compared to intact virus. Kinetics of inactivation at elevated temperatures were multi-hit in character. The structure of infectious enterovirus RNA was investigated by treatment with chemical inactivating agents. Urea and guanidine as hydrogen bond-disrupting agents, and mercaptoethanol and thioglycolate as disulfide bond-disrupting agents, and combinations of these did not destroy RNA infectivity whereas hydrogen bond-disrupting treatment inactivated intact virus rapidly. RNA infectivity was not reduced by chloroform extraction alone, or by octanol extraction alone, but was reduced by chloroform-octanol extraction which failed to depolymerize RNA to an extent detectable by ultracentrifugal analysis. Infectivity of type 1 poliovirus and Coxsackie B1 virus RNA was destroyed in accordance with first order kinetics by very dilute solutions of pancreatic ribonuclease, and by purified snake venom phosphodiesterase, but not at all by bacterial alkaline phosphatase. Inactivation by venom diesterase was not accelerated by prior treatment of RNA with bacterial alkaline phosphatase. These results indicated that infectivity of enteroviral RNA resided in a single stranded structure, that a single break of a phosphodiester bond anywhere along the structure was sufficient to destroy infectivity, and that infectivity did not require a terminal phosphate group. Hydroxylamine, but not other carbonyl reagents, rapidly destroyed infectivity of intact type 1 poliovirus viral RNA without depolymerization of RNA-detectable by behavior in the analytical ultracentrifuge. With S(35)-methionine-labeled poliovirus a very small fraction of radioactivity remained in RNA preparations following phenol extraction. No evidence could be obtained to indicate that infectious enteroviral RNA was composed of subunits. RNA extracted with phenol during the course of infection of HeLa cells with type 1 poliovirus resembled RNA obtained from purified whole virus with respect to heat inactivation, hydroxylamine inactivation, chromatographic separation, susceptibility to protein denaturing agents, and ability to infect productively both naturally susceptible HeLa cells and naturally insusceptible L strain mouse cells. Intracellular production of infectious RNA paralleled intracellular maturation of whole virus and preceded it by a very short interval.

ON THE ROLE OF VIRUS SULFHYDRYL GROUPS IN THE ATTACHMENT OF ENTEROVIRUSES TO ERYTHROCYTES

Many animal viruses possess the ability to agglutinate erythrocytes. In most but not all cases hemagglutination is due to the virus particle itself, and appears to result from the mechanical bridging of two or more erythrocytes by virus particles which attach to receptor sites on each erythrocyte (1, 2). Thus, attachment of virus particles to erythrocytes is a prerequisite for hemagglutination, and prevention of absorption of virus prevents hemagglutination. Among the enteroviruses, many ECHO viruses and some strains of Coxsackie B3 virus agglutinate human erythrocytes (3-7), and the evidence indicates that hemagglutination is due to the virus particle itself (3, 5, 6). The precise mechanism of attachment of enteroviruses to cells is unknown. Chymotrypsin treatment of erythrocytes prevents the absorption of some ECHO viruses (8). This suggests that the receptor sites on the erythrocyte may be at least in part protein in nature. The present communication is concerned with the mechanism of attachment of enteroviruses to cells. It is shown that sulfhydryl reagents block the hemagglutinating activity of enteroviruses. Treated virus fails to absorb to erythrocytes. Thiol compounds restore the hemagglutinating activity of enteroviruses treated with mercaptide-forming sulfhydryl reagents. The effect of sulfhydryl reagents on the infectivity of some enteroviruses is also described.

CLINICAL AND EPIDEMIOLOGIC OBSERVATIONS ON ENTEROVIRUS INFECTION IN A CIRCUMSCRIBED COMMUNITY DURING AN EPIDEMIC OF ECHO 9 INFECTION

A clinical, epidemiologic and laboratory study was undertaken in a housing development during an epidemic of illness due to ECHO virus, type 9. The clinical manifestations varied with age. The younger children exhibited an exanthem while older persons frequently had evidence of irritation of the central nervous system. Multiple cases occurred within families, with the incidence of illness highest in the younger age groups. No serious consequences of the illness were seen. Echo 9 virus was recovered with greater frequency from the intestinal contents of young children with illness than from ill adults, although serologic evidence of recent infection in these adults could be demonstrated. The rate of inapparent infection with ECHO 9 virus was low. Virus rarely persisted in the feces beyond 14 days after onset of illness. Group B Coxsackie viruses, type 4 and type 5, were found also to be the cause of many infections within the same population at the same time. Illnesses due to these viruses were similar to those due to ECHO 9 virus, except that no eruption was observed. The rate of inapparent infection was higher than that observed with ECHO 9 virus. Simultaneous infection with ECHO 9 virus and Coxsackie B4 or B5 virus was rare.

Contribution to the study of diseases in Italy caused by the Coxsackie B group of viruses. II. Epidemiological, clinical and virological data obtained in the course of a summer outbreak caused by Coxsackie B 4 virus.

The clinical, epidemiological and virological data obtained from the study of an outbreak of summer grippe mixed with pleurodynia, which spread throughout a whole town in Northern Italy, are given. Several strains of a Coxsackie type B4 virus were isolated.

Epidemiological studies of aseptic meningitis caused by Coxsackie virus B5.

Epidemiological Studies of Aseptic Meningitis Caused by Coxsackie Virus B5 Tom D. Y. Chin, Patrick H. Lehan, Harry Rubin, Irene L. Doto, Ralph H. Heeren, Herbert A. Wenner, and Michael L. Furcolow CopyRight https://doi.org/10.2105/AJPH.48.9.1193 Published Online: August 29, 2011

Application of the “most probable number” method for estimating concentrations of animal viruses by the tissue culture technique

Justification for use of an MPN (most probable number) method for estimating concentration of Coxsackie B1 virus in monkey kidney tissue culture tubes was demonstrated by comparison of dosage-response data with a theoretical MPN curve based on one-particle infection and by variance analysis of 24 replicate MPN determinations. This method yields units comparable to plaque counts or plaque-forming units and affords unbiased estimates even when data do not bracket the 50% response dilution. It provides a means of testing for conformity to the one-particle infection theory and gives usable results from one dilution if both positive and negative tubes are present. For these reasons it is preferred to conventional LD 50 methods for virus quantitation where the dilution response data are known to follow the one-particle infection curve. A table of 5-tube, 4-fold dilution series and an approximation equation are presented for conversion of data to “most probable number of cytopathogenic units” (MPNCU).