ISOLATION AND TRANSMISSION STUDIES WITH BOVINE PARAINFLUENZA-3 VIRUS.
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Circulating interferon was detectable in mouse serum within 1 hour after the intravenous injection of various types of virus and it reached maximum levels in about 4 hours. Rapidly produced interferon may play a role in the pathogenesis of viral infection and in viral interference.
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Cystic fibrosis is the most common lethal or semi-lethal genetic disease in Caucasians of Central European origin, among whom it is inherited as an autosomal recessive trait at a frequency approximately 10 times that expected from recurrent mutation alone. A decreased sialic acid content has been observed in cell surface glycoproteins on cystic fibrosis fibroblasts and in numerous soluble glycoprotein preparations from cystic fibrosis homozygotes. Sialic acid residues on cell surface glycoconjugates play an essential role in the binding and infectivity of myxoviruses and paramyxoviruses, including those causing pandemic influenza. It is suggested that increased resistance to these viruses conferred by similar but quantitatively smaller alterations in sialoglycoconjugate structure in cystic fibrosis heterozygotes may have provided a selective advantage to maintain the high frequency of the cystic fibrosis gene in Caucasian populations.
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A survey of cell culture systems for virus isolation is presented. Because there is not single cell culture system which will support the many viruses that may be encountered in clinical specimens, the characteristics and susceptibility of primary, diploid, and continuous cell cultures are reviewed. The primary cell culture system should be susceptible to infection with myxoviruses, paramyxoviruses, adenoviruses, and Coxsackie B viruses, since these are not consistently recognized in diploid cell cultures. Conversely, rhinoviruses, cytomegalovirus and varicella-zoster virus may be isolated only in diploid cell cultures. Upon consideration of expected viruses, and cell cultures available for isolation of these, one can choose a combination of a primary and a diploid cell system for virus isolation which will assure detection of most human virus groups known. The usefulness of continuous cell culture systems is generally limited to a very few specific viruses.
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Live Newcastle disease virus (NDV) was used to investigate the in vitro effects of a viral infection on phagocytosis, chemiluminescence generation, superoxide production, oxygen consumption, NADPH-oxidase activity, and intracellular killing of bacteria by Ficoll-Hypaque separated human neutrophils. Phagocytosis of oil red O particles by NDV-treated PMN was inhibited by 50%. Chemiluminescence by PMN was inhibited 79% after zymosan stimulation and 86% after tetradeconyl phorbol acetate stimulation. Superoxide generation was inhibited by 68%. Oxygen consumption was inhibited in the presence of NDV by 37% after stimulation with phorbol myristate acetate, while membrane-associated NADPH-enzyme activity was decreased by 19%. The percent of surviving intracellular S. aureus was significantly elevated in NDV-treated PMN after 60 and 120 min of incubation. Purified bacterial neuraminidase markedly suppressed chemiluminescence, while neuraminic acid blocked the effects of the virus. These observations suggest that infections with myxoviruses may suppress a number of vital neutrophil functions. It appears that the effects may be partly mediated by the interaction of viral neuraminidase with the external neutrophil membrane.
Surface antigens of HeLaHVJ cells, a cell line persistently infected with HVJ, were studied by fluorescent antibody staining. After absorption with concentrated HVJ virions and HeLa cells, anti-HeLaHVJ antiserum was able to demonstrate specific surface fluorescence on HeLaHVJ cells, while this serum no longer reacted with original HeLa cells nor with HVJ virions. During cytolytic infection of HeLa cells with HVJ, this specific surface antigen appeared at an early stage of infection prior to the appearance of newly synthesized HVJ viral antigens and moreover appeared in spite of the inhibition of viral protein synthesis. This antigen was detected neither on HeLa cells infected with other myxoviruses except HVJ nor on various other kinds of cells infected with HVJ. The specific surface antigen was still found on the HeLaHVJ cell surface after incubation at 38 degrees C for two days, while HVJ structural antigens on the cell surface no longer could be detected. Mild short-term treatment of HeLa cells with trypsin, neuraminidase from vibrio cholerae, phospholipase-C and hyaluronidase failed to expose specific antigen. The antigen was distinguishable from the Forssman and human blood type antigens. The mechanism of appearance of a new antigen on the surface of HeLaHVJ cells remains unclear.
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534 serum samples from 73 renal transplant recipients, 41 haemodialysis patients, and 99 blood and organ donors were examined serologically for antibodies against Cytomegalo, H. simplex (types 1 and 2), Varicella-zoster, Epstein-Barr, Adeno, Influenza, Parainfluenza, Respiratory syncytial, Measles, Picorna- and human Polyoma- Viruses. Serum specimens were stored in the lyophilized state until examined thus enabling a simultaneous testing of all samples belonging to one patient. All antigens, complement, and control antisera were prepared, lyophilized, and standardized in this laboratory. This has enabled the use of single batches of any preparation throughout the study. Serologic results with antigens of the Herpesvirus group (CMV, HSV and VZV) compared favourably with previous results showing that infections with these agents, especially with CMV, can frequently be encountered among transplant recipients. Our results have indicated a moderately increased incidence of infections with some Herpesviruses in haemodialysis patients as well. Infections with VZV, for instance, were as frequently demonstrated in these patients, as in transplant patients although the former received no immunosuppressive therapy. Serologic results with non-Herpesvirus antigens indicated an increased incidence of infections with Polyomavirus, Myxoviruses (Influenza, Parainfluenza and RS) and Picornaviruses among transplant recipients. The incidence of acute infections with RS virus among adults was unusually high and there is no evidence so far to indicate such a high frequency of RS infections in any other group of adults. We were unable to demonstrate acute infections with non-Herpesviruses among haemodialysis patients, even though most of the patients were followed over a period of more than 2 years. Virus isolation attempts were performed with samples of urines and biopsy or autopsy samples. 23 out of 28 cytopathic agents recovered from urines, throat-swabs and/or from organs of transplant recipients were identified as CMV. Two HSV type 1, 1 HSV type 2, and 2 Coxsackie B type 3 viruses were also isolated. No viruses were isolated from a series of 31 kidneys randomly selected among autopsy cases.
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