Harold S. Ginsberg

The reaction of influenza viruses with guinea pig polymorphonuclear leucocytes. II. The reduction of white blood cell glycolysis by influenza viruses and receptor-destroying enzyme (RDE).

Abstract Influenza viruses, PR8 and Lee, markedly inhibited anaerobic glycolysis when glucose served as substrate. Inhibition of CO2 evolution corresponded to a reduction in lactic acid produced. Reduction of glycolysis was accomplished by the viral particles and was proportional to the viral concentration employed. Whereas receptor-destroying enzyme of Vibrio cholera (RDE) also inhibited glycolysis of white blood cells, neither heat-inactivated RDE nor virus inhibited this metabolic process. Calcium was not essential for adsorption of virus to white blood cells, but this cation was required for inhibition of glycolysis. Inhibition of glycolysis by influenza virus was obtained when glucose or glucose-6-phosphate was employed as substrate, but not when fructose-6-phosphate or fructose-1, 6-diphosphate was utilized. Periodate also inhibited glycolysis of white blood cells but the metabolic pattern was not similar to that obtained with influenza viruses or RDE.

Tryptose phosphate broth as supplementary factor for maintenance of HeLa cell tissue cultures.

Summary A mixture of tryptose phosphate broth, 15 to 25%, maintenance solution (MS), 67.5 to 77.5%, and chicken serum, 7.5%, maintained HeLa cells in tissue culture for at least 10 days. During this period HeLa cells increased 3 to 5 fold in number. Furthermore, smaller quantities of ARD, AD and Type I poliomyelitis viruses could be detected and significantly more ARD virus could be propagated in HeLa cells maintained in this medium than in cells supported with maintenance solution and chicken serum alone.

Reactions of influenza viruses with guinea pig polymorphonuclear leucocytes. I. Virus-cell interactions.

Abstract Influenza A and B viruses, PR8 and Lee strains respectively, adsorbed to guinea pig polymorphonuclear leucocytes (WBC) at the same rates, and at room temperature (22° to 23°) little or no virus eluted spontaneously even when incubated as long as 24 hours. At 37° less virus was adsorbed by WBC, but elution of approximately 20% of the adsorbed virus occurred. The rate of viral adsorption was directly related to the number of WBC and concentration of virus employed. Reduction of electrolyte concentration of the reaction mixture decreased the rate of viral adsorption but did not totally prevent virus-WBC combination. The same number of viral particles saturated a WBC whether the cells were exposed to a high concentration of virus in a single reaction or the same quantity of virus was employed in four adsorptive reactions. The receptor-destroying enzyme of Vibrio cholera (RDE) altered WBC so that combination with influenza viruses could not be accomplished. RDE, however, initiated elution of no more than 10% of virus al ready adsorbed to WBC. Less than 1% of virus which did not elute from WBC could be detected by infectivity or hemagglutination titrations when the cells were completely disrupted. The loss of virus on or in WBC could not be explained by a soluble inhibitor of infectivity in cell homogenates. The WBC homogenates, however, did contain a Francis-type inhibitor of hemagglutination.

Accumulation of organic acids by HeLa cells infected with type 4 adenovirus.

Summary Fluids from cultures infected with type 4 adenovirus were markedly more acid than fluids from non-infected control cultures. The lowered pH of infected cultures resulted from an accumulation of lactic, pyruvic, acetic and alpha ketoglutaric acids. The acids were separated by chromatography on a celite column, and measured by colorimetric and acid-base titration technics. Confirmation of the identity of the organic acids was obtained by paper chromatography. Concomitant with the increased accumulation of acid in fluids of infected cultures, an increased utilization of glucose by cells of the virus-infected cultures occurred.

The reaction of influenza viruses with guinea pig polymorphonuclear leucocytes. III. Studies on the mechanism by which influenza viruses inhibit phagocytosis

Abstract The degree of phagocytosis of heat-killed yeast cells by guinea pig polymorphonuclear leucocytes was shown to depend on the presence in the reaction mixture of guinea pig serum and an added substrate such as glucose, glucose-6-phosphate, or fructose-6-phosphate. The concentration of carbohydrate was important in order to obtain maximum phagocytosis. Phagocytosis by polymorphonuclear leucocytes was reduced when the cells had reacted with partially purified Lee virus and were incubated in the presence of serum and glucose or glucose-6-phosphate. When fructose-6-phosphate served as the added substrate, there was no inhibition of phagocytosis. The observed inhibition of phagocytosis by leucocytes to which influenza virus had been adsorbed was postulated to be due to the block in the energy yielding mechanisms of virus-treated cells.

Characteristics of the New Respiratory Viruses (Adenoviruses∗) II. Stability to Temperature and pH Alterations.†

Summary Types 1-4 of the new respiratory viruses, termed adenoviruses, were relatively stable when exposed to temperatures ranging from 4°C to 36°C. Type 1-3 viruses were inactivated in 10 to 20 minutes at 50°C, but the infectivity titer of type 4 virus was reduced less than 2 log units (99%) in 20 minutes. The adenoviruses were also resistant to inactivation at pH 6.0 to 9.0. The patterns of inactivation of the 4 viruses by alterations of pH as well as incubation at 50°C were similar, although consistent and significant differences among the 4 types were measured. The excellent technical assistance of Miss Mary K. Dixon is gratefully acknowledged.

Biological and physical properties of the adenoviruses.

The isolation of new viruses not only initiates a “search for diseases” with which the recently recognized agents may be identified, but also stimulates the quest for answers to other important and formidable questions. These queries are related to the classification of the viruses, their epidemiological behavior, and the types of host-parasite relationships that these agents may establish. Information concerning the characteristics of the viruses may permit the development of hypotheses pertinent to the questions posed. This paper will summarize the results of investigations into properties that may bear upon the classification and the epidemiological and biological behavior of the new respiratory viruse~, l -~ in this report termed adenoviruses (previously called APC or RI viruses).

Newer Aspects of Adenovirus Infections

FOUR YEARS ago at a combined meeting of the Epidemiology and Laboratory Sections of the American Public Health Association, there was a discussion of recent data pertaining to a newly isolated group of viruses, alluded to then by a variety of names, but now called adenoviruses.1 The papers presented centered about means for classification of the newly discoverea viruses, the diseases which they probably cause, and their possible importance as etiologic agents of acute respiratory infections.2-4 The intervening years have allowed time for many investigators to gather data which bear upon the points discussed with uncertainty at that time and to permit a perspective to be gained on the role of the adenoviruses as agents of disease. This paper will attempt to review some of the pertinent information obtained in the past four years and to interpret these data relative to what we may term the natural history of adenovirus infections.

Hemagglutination with adenoviruses; nature of viral antigen.

Summary A rise in antibodies to types 1-4 of adenoviruses during illness can be detected in human serum by employing hemagglutination technics. Although the original method described does not measure an increase in antibodies directed against a single type of virus, type-specific antibodies can be titrated if serum is mixed with a pool of heterologous viruses before tanned rbc treated with homologous virus are added. This modification measured type-specific antibodies solely in 4 of the 8 sera tested. The antigen adsorbed to tannic acid-treated rbc is probably not the infectious viral particle and is separable from it by high speed centrifugation.

An antipyretic action of aureomycin; inhibition of febrile response to influenza virus in rabbits.

Summary Intraperitoneal or intravenous injection of aureomycin prevented the febrile reaction of rabbits to intravenous inoculation of influenza B virus, but administration of aureomycin by gavage did not consistently prevent fever. Although intraperitoneal injection of aureomycin reduced rectal temperature below normal, the data suggest that this hypothermic action of the drug was not responsible for the antipyretic effect observed. Aureomycin did not alter the development of tolerance to the pyrogen. Intraperitoneal aureomycin also modified the febrile response of rabbits to a bacterial pyrogen, typhoid bacilli.