Quantitative Distribution of Phosphorus in Chorioallantoic Membrane as Affected by Infection with Influenza Virus.

1952 ◽  
Vol 79 (4) ◽  
pp. 566-568 ◽  
Author(s):  
Z. A. Cohn
Keyword(s):  
Influenza Virus ◽  
Chorioallantoic Membrane ◽  
Quantitative Distribution

scholarly journals SOME ENERGY RELATIONS IN A HOST-VIRUS SYSTEM

1953 ◽  
Vol 97 (3) ◽  
pp. 315-322 ◽  
Author(s):  
W. Wilbur Ackermann ◽  
R. Bernal Johnson ◽  
Keyword(s):  
Influenza Virus ◽  
Chorioallantoic Membrane ◽  
Host Tissue ◽  
Endogenous Respiration ◽  
Excellent Correlation ◽  
Viral Propagation ◽  
Twofold Increase ◽  
Energy Relations ◽  
Virus System ◽  
Stimulation Of

It was found that DNP (2,4-dinitrophenol) will inhibit completely the propagation of influenza virus in chorioallantoic membrane. This reagent did not permanently alter those metabolic processes required for the synthesis of virus and at the concentrations employed demonstrated no virucidal effects. In minced preparations of chorioallantoic membrane DNP was shown to have a pronounced stimulatory effect upon ATPase (adenosinetriphosphatase). When DNP was used with intact tissues, an excellent correlation was found between the inhibition of viral propagation and the stimulation of respiration and release of phosphate. Concentrations of DNP which permitted a twofold increase in the endogenous respiration of intact membranes allowed little or no viral synthesis. It is concluded that the energy required for viral synthesis derives from the oxidative phosphorylative activity of the host tissue.

scholarly journals INHIBITION OF INFLUENZA VIRUS MULTIPLICATION BY N-GLYCOSIDES OF BENZIMIDAZOLES

1954 ◽  
Vol 99 (3) ◽  
pp. 227-250 ◽  
Author(s):  
Igor Tamm ◽  
Karl Folkers ◽  
Clifford H. Shunk ◽  
Frank L. Horsfall
Keyword(s):  
Influenza Virus ◽  
Influenza A ◽  
Benzimidazole Derivative ◽  
Chorioallantoic Membrane ◽  
Inhibitory Effect ◽  
Virus Multiplication ◽  
Cellular Growth ◽  
Tissue Proliferation ◽  
Fresh Culture Medium

Chloro derivatives of benzimidazole were found to be 2 to 3 times more active than corresponding methyl derivatives in causing inhibition of Lee virus multiplication in chorioallantoic membrane cultures in vitro. The most active benzimidazole derivative thus far tested is 5,6-dichloro-1-ß-D-ribofuranosylbenzimidazole (DRB); it caused 75 per cent inhibition of Lee virus multiplication in membrane cultures at a concentration of 0.38 x 10–4 M. On the other hand, 5,6-dimethyl-1-alpha;-D-ribofuranosylbenzimidazole, the moiety present in vitamin B12, failed to inhibit Lee virus multiplication at a concentration of 35 x 10–4 M. Other N-glycosides of 5,6-dichlorobenzimidazole were considerably less active than DRB. In single cycle experiments, the degree of inhibition of Lee virus multiplication by DRB in membrane cultures was not dependent on the amount of virus in the inoculum. This compound did not inactivate the infectivity of extracellular Lee virus, had no effect on virus-erythrocyte interaction, did not interfere with the adsorption of the virus by the host tissue, nor affect the release of newly formed virus from the membrane. The inhibitory effect of DRB on Lee virus multiplication, in contrast to that of 2,5-dimethylbenzimidazole, persisted after transfer of infected membranes into fresh culture medium not containing the compound. Both DRB and the 2,5-dimethyl compound caused 99 per cent inhibition of Lee virus multiplication without affecting oxygen uptake of the membrane. Tissue proliferation of membrane pieces in roller tube culture was not significantly affected by DRB at inhibitory concentration, whereas at equivalent concentration the 2,5-dimethyl compound did restrict cellular growth. At higher concentrations, both compounds caused retardation of cell proliferation. This effect was reversible on removal of either compound from the medium. The multiplication of several strains of influenza A and B viruses, i.e. Lee, MB, PR8, and FM1, was inhibited to the same degree by each of the two compounds; DRB was 35 times more active than the 2,5-dimethyl compound relative to each of the strains. DRB caused inhibition of Lee virus multiplication in intact embryonated chicken eggs and in mice without causing significant signs of toxicity in either host. Some of the implications of these findings are discussed in relation to the mechanism of the inhibition of influenza virus multiplication.

scholarly journals GROWTH CHARACTERISTICS OF INFLUENZA VIRUS

1955 ◽  
Vol 102 (4) ◽  
pp. 393-402 ◽  
Author(s):  
W. Wilbur Ackermann ◽  
Hunein F. Maassab
Keyword(s):  
Influenza Virus ◽  
Latent Period ◽  
Growth Curve ◽  
Chorioallantoic Membrane ◽  
Growth Characteristics ◽  
Stages Of Development ◽  
Viral Inhibitor ◽  
Site Of Action ◽  
Two Stages

A further analysis of the growth curve obtained in vitro for influenza virus in chorioallantoic membrane has been made using the viral inhibitor p-fluorophenylalanine. It has been found that p-fluorophenylalanine is phase-specific, does not interfere with the initiation of infection but rather acts during the productive period of the infectious sequence. The site of action of this fluoroderivative has been described relative to the site of action of methoxinine. By the use of this inhibitor in combination with methoxinine, it has been possible to recognize two stages of development which were not discernible from the usual growth curve. In a multicellular culture these two reactions, A and B, occur for the most part simultaneously, during the latent and productive periods. Reaction A is inhibited by methoxinine, but not by fluorophenylalanine. It begins early in the latent period and can proceed independent of reaction B. Reaction B is inhibited by fluorophenylalanine, but not by methoxinine, and it cannot proceed unless reaction A is proceeding or has operated for some prior interval.

scholarly journals INHIBITION OF INFLUENZA VIRUS MULTIPLICATION BY ALKYL DERIVATIVES OF BENZIMIDAZOLE

1953 ◽  
Vol 98 (3) ◽  
pp. 219-227 ◽  
Author(s):  
Igor Tamm ◽  
Karl Folkers ◽  
Frank L. Horsfall
Keyword(s):  
Influenza Virus ◽  
Latent Period ◽  
Chorioallantoic Membrane ◽  
Virus Multiplication ◽  
Influenza B Virus ◽  
Influenza B ◽  
In Ovo ◽  
Alkyl Derivatives ◽  
Rate Of Increase

At a concentration of 0.0026 M, 2,5-dimethylbenzimidazole caused a number of alterations in the first cycle of multiplication of influenza B virus, Lee strain, in chorioallantoic membrane cultures in vitro. As determined by infectivity titrations in ovo on the membrane per se, the following alterations were observed: The duration of the latent period was increased by 80 per cent. The rate of increase in titer during the incremental period was somewhat decreased. The yield of virus was decreased by about 99 per cent. When the compound was added to membrane cultures at various periods before or after inoculation with the virus, the following findings were obtained: On addition before or along with the virus, the substance caused about 99 per cent inhibition of multiplication. When added during the first 2 hours after inoculation, the compound caused inhibition of a degree which was inversely proportional to the time of addition. When added 3 to 8 hours after inoculation, the substance caused about 80 per cent inhibition. When added after the end of the latent period, no definite inhibition was obtained in the first cycle of multiplication. These results are interpreted as indicating that 2,5-dimethylbenzimidazole acts by reducing the rate of biosynthetic mechanisms necessary for the reproduction of influenza virus particles.

scholarly journals INFLUENZA VIRUS MULTIPLICATION IN THE CHORIOALLANTOIC MEMBRANE IN VITRO: KINETIC ASPECTS OF INHIBITION BY 5, 6-DICHLORO-1-ß-D-RIBOFURANOSYLBENZIMIDAZOLE

1954 ◽  
Vol 100 (6) ◽  
pp. 541-562 ◽  
Author(s):  
Igor Tamm ◽  
David A. J. Tyrrell
Keyword(s):  
Influenza Virus ◽  
Chorioallantoic Membrane ◽  
Inhibitory Effect ◽  
Virus Multiplication ◽  
Benzimidazole Derivatives ◽  
Similar Degree ◽  
Virus Concentration ◽  
Virus Particles ◽  
Rate Of Increase

A procedure is described for kinetic studies on the multiplication of Lee virus in the chorioallantoic membrane in vitro employing the hemagglutination technique for measurement of virus concentration. A linear relationship was found between the logarithm of virus adsorbed and the amount of membrane used. Of the virus adsorbed less than 10 per cent could be recovered from the membrane. Of the recoverable virus 90 per cent was eliminated by specific immune serum. Lee virus was adsorbed by the allantoic and chorionic layers of the membrane to a similar degree. Multiplication occurred in both layers and to a similar extent. When 107.66 EID50 of Lee virus was inoculated per 2.9 cm.2 of chorioallantoic membrane, the ratio of infectivity to hemagglutination titer in the yield was low, although the rate of appearance of virus particles was not diminished despite the large inocula. Virus produced in membranes was liberated rapidly and continually into the medium. 5,6-Dichloro-1-ß-D-ribofuranosylbenzimidazole (DRB), 0.000055 M, prolonged the latent period by more than 100 per cent. The rate of increase during the period of rapid rise was similar in the presence or absence of DRB, but the yield was markedly reduced at the end of this period in the presence of DRB. The amount of the virus in the membranes continued to rise in the presence of DRB and eventually approached the maximal levels reached much earlier in the controls. Measurement of the amount of virus in the media indicated a greater degree of inhibition than did measurement in the membranes. Comparative studies with two benzimidazole derivatives on the dependence of the inhibitory effect on the time of addition of the compound showed that processes which could be inhibited by DRB were of shorter duration than those inhibited by 2,5-dimethylbenzimidazole (MB). With MB the relationship between the time of addition and the inhibitory effect was similar both for virus and for soluble complement-fixing antigen; with DRB the inhibitable processes were of shorter duration for the complement-fixing antigen than for virus particles. DRB was not only 35 times more active on a molar basis but also was more selective in its action than MB. DRB interfered with processes which preceded the emergence of either soluble complement-fixing antigen or virus particles. Some of the implications of these findings are discussed in relation to the mechanism of inhibition of influenza virus multiplication by benzimidazole derivatives.

scholarly journals The use of influenza virus labelled with radio-sulpher in studies of the early stages of the interaction of virus with the host cell

1957 ◽  
Vol 55 (2) ◽  
pp. 290-297 ◽  
Author(s):  
L. Hoyle ◽  
N. B. Finter
Keyword(s):  
Influenza Virus ◽  
Amino Acid ◽  
Envelope Protein ◽  
Medical Research Council ◽  
Chorioallantoic Membrane ◽  
Soluble Antigen ◽  
Virus Envelope ◽  
Virus Envelope Protein ◽  
Ether Treatment ◽  
Original Virus

1. Influenza virus can be labelled with 35S by cultivation in fertile eggs into which radioactive methionine has been introduced.2. When virus labelled with 35S is fractionated by ether treatment the nucleoprotein soluble antigen fraction contains 15 % of the 35S, the haemagglutinin also contains 15%, while the denatured envelope protein contains 70% of the 35S.3. When sulphur-labelled virus is introduced as a primary inoculum in fertile eggs and extracts of the chorioallantoic membrane are made 1–20 % of 35S is recovered as amino-acid, while the remainder is present as non-infective material of particle size similar to that of the original virus.4. It is suggested that on entry into the cell the virus nucleoprotein is hydrolysed with the release of amino-acid and free nucleic acid, while the virus envelope protein and haemagglutinin remain on the cell surface.The authors are indebted to the Medical Research Council for a grant in aid of expenses, and to Messrs. Eli Lilly and Co. for the generous provision of the Spinco centrifuge used in this work. The radioactive methionine was obtained from the Radiochemical centre at Amersham.

scholarly journals INHIBITION OF INFLUENZA VIRUS MULTIPLICATION BY ALKYL DERIVATIVES OF BENZIMIDAZOLE

1953 ◽  
Vol 98 (3) ◽  
pp. 229-243 ◽  
Author(s):  
Igor Tamm ◽  
Karl Folkers ◽  
Frank L. Horsfall
Keyword(s):  
Influenza Virus ◽  
Culture Medium ◽  
Chorioallantoic Membrane ◽  
Virus Multiplication ◽  
Influenza B Virus ◽  
Influenza B ◽  
Experimental Conditions ◽  
Virus Reproduction ◽  
B Virus

The activity of compounds which inhibit the multiplication of influenza virus can be measured in chorioallantoic membrane cultures in vitro by means of hemagglutination titrations on the medium. Studies on the reproducibility of virus reproduction in membrane cultures have revealed the major variables which affect the results and thus have led to the development of a precise technique. Under strictly controlled experimental conditions, the extent of reproduction of the virus in membrane cultures is predictable within narrow limits of variation. With 105.5 EID50 of influenza B virus, Lee strain, and 5.75 cm.2 of chorioallantoic membrane per ml., the ratio of infective virus particles to susceptible allantoic cells appears to be approximately 1:28. Under these conditions, the evidence indicates that two cycles of multiplication occur and nearly maximal hemagglutination titers are found with culture medium at 36 hours. The extent of the deviation in the absolute titer in different experiments was only 0.112 log unit. At a concentration of 0.0017 M, 2,5-dimethylbenzimidazole caused inhibition of the multiplication of influenza B virus, Lee strain, which persisted for at least 70 hours as measured by hemagglutination titrations on the culture medium. The degree of inhibition was closely comparable to that demonstrated by infectivity titrations on the membrane at the end of the first cycle of virus reproduction (1).

scholarly journals SYNERGISTIC ACTION OF HEMOPHILUS INFLUENZAE SUIS AND THE SWINE INFLUENZA VIRUS ON THE CHICK EMBRYO

1943 ◽  
Vol 77 (1) ◽  
pp. 7-20 ◽  
Author(s):  
Frederik B. Bang
Keyword(s):  
Influenza Virus ◽  
Chick Embryo ◽  
Synergistic Effect ◽  
Swine Influenza Virus ◽  
Chorioallantoic Membrane ◽  
Swine Influenza ◽  
Human Influenza ◽  
Human Influenza Virus ◽  
Hemophilus Influenzae ◽  
The Individual

The synergistic effect of Hemophilus influenzae suis and swine influenza virus in the pig can be reproduced by the inoculation of these agents on the chorioallantoic membrane of 9 to 10 day old chick embryos. Two strains of human influenza virus that were studied failed to substitute for the swine virus in the synergistic reaction. No loss of synergistic effect was noted when the swine influenza virus was put through 11 chick embryo passages. Recently isolated and old stock strains of Hemophilus were equally able to enhance the effect of the virus. Heat-killed cultures of H. influenzae suis can be substituted for the bacterial component of the reaction. Infection of the embryo with swine influenza virus predisposes to infection with H. influenzae suis. The combination of H. influenzae suis and swine influenza virus causes a selective destruction of the embryo lungs, not produced by the individual components. This pneumonia exhibits the essential features of the natural disease.

scholarly journals SYNERGISTIC ACTION OF HEMOPHILUS INFLUENZAE SUIS AND THE SWINE INFLUENZA VIRUS ON THE CHICK EMBRYO. II

1943 ◽  
Vol 78 (1) ◽  
pp. 9-16 ◽  
Author(s):  
Frederik B. Bang
Keyword(s):  
Influenza Virus ◽  
Swine Influenza Virus ◽  
Chorioallantoic Membrane ◽  
Swine Influenza ◽  
Allantoic Fluid ◽  
Blood Cultures ◽  
Synergistic Action ◽  
Spreading Factor ◽  
Spreading Effect ◽  
Hemophilus Influenzae

Blood cultures of embryos killed by the synergistic action of swine influenza virus and Hemophilus influenzae suis are consistently negative, and embryos infected with swine influenza virus may be killed both by filtered extracts of frozen and dried Hemophilus and by suspensions of heat-killed bacteria. The addition of Hemophilus to the chorioallantoic membrane of embryos infected with swine influenza virus causes the virus to spread from the membrane to the allantoic fluid and embryo. This spreading effect also obtains when a purified preparation of hyaluronidase is used instead of Hemophilus, but it is unaccompanied by a comparable increase in mortality. It is probable that the spread of the virus produced by the bacteria is only partly responsible for the development of the complex infection and that products of these organisms other than the spreading factor play a large part in the mortality of embryos receiving the combination of virus and bacterium.

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