scholarly journals STUDIES ON HOST-VIRUS INTERACTIONS IN THE CHICK EMBRYO-INFLUENZA VIRUS SYSTEM

1951 ◽  
Vol 94 (4) ◽  
pp. 305-322 ◽  
Author(s):  
Werner Henle ◽  
Oscar C. Liu
Keyword(s):  
Chick Embryo ◽  
Virus Particle ◽  
Prolonged Exposure ◽  
Influenza Viruses ◽  
Infective Virus ◽  
Virus Particles ◽  
E Coli ◽  
Partial Inactivation ◽  
Host Virus Interactions ◽  
Virus Interactions

Evidence has been presented that influenza viruses both of type A and B partially inactivated by ultraviolet irradiation may regain their capacity to propagate in the allantoic membrane of the chick embryo. In using such irradiated preparations as inocula for growth curve experiments it could be shown that the development of hemagglutinins as well as of infectivity preceded at rates resembling those noted with more than 10 times the amount of infective virus actually found in the irradiated seed. Partial inactivation of the inocula by heating to 56°C. gave similar results. The phenomenon was observed only with seed irradiated for short periods of time so that the virus particles sustained only few hits of radiation. On prolonged exposure resulting in numerous hits per virus particle the capacity of reactivation was lost. Likewise, an irradiated preparation capable of reactivation in the allantoic membrane, could not be diluted more than about 30-fold and still clearly produce this phenomenon. This indicated that reactivation is obtained only when one host cell adsorbs more than one non-infective virus particle but not upon adsorption of a single particle. These data are in striking agreement with the phenomenon of "multiplicity reactivation" observed in the bacteriophage-E. coli system by Luria and Dulbecco.

scholarly journals STUDIES ON HOST-VIRUS INTERACTIONS IN THE CHICK EMBRYO-INFLUENZA VIRUS SYSTEM

1955 ◽  
Vol 101 (5) ◽  
pp. 461-478 ◽  
Author(s):  
Norman B. Finter ◽  
Oscar C. Liu ◽  
Werner Henle
Keyword(s):  
Influenza Virus ◽  
Chick Embryo ◽  
Host Cell ◽  
Infectious Virus ◽  
Chick Embryos ◽  
In Ovo ◽  
Virus Particles ◽  
Host Virus Interactions ◽  
Virus Interactions ◽  
Virus System

An analysis has been made of factors contributing to the von Magnus phenomenon; i.e., the emergence of increasing quantities of non-infectious hemagglutinins (NIHA) in successive passages in the allantois of chick embryos of undiluted allantoic fluids infected with influenza virus. Using the PR8 strain, the von Magnus phenomenon was pronounced when the serial seeds were obtained under conditions which permitted extensive inactivation of infectious virus during individual passages. Correspondingly, it was reduced but not abolished when precautions were taken to avoid accumulation of inactivated virus in the inocula. Thus, inactivated virus may be taken as a contributing factor. Preparations of infectious virus obtained under conditions largely excluding the presence of inactivated virus were capable of yielding some NIHA on passage as long as sufficient amounts were injected to permit each host cell to adsorb several infectious virus particles. However, the fact remains that more NIHA was found in the harvests when the inocula contained a large proportion of non-infectious virus material. Following injection of various types of seeds NIHA appeared in the allantoic fluids as soon as liberation of virus became detectable. This time relationship and the rates of release of non-infectious virus components seemed to exclude that the NIHA obtained consisted entirely of infectious virus which had been inactivated during incubation in ovo. It was apparent rather that NIHA other than that due to heat-inactivated virus was released. Correlations between the infectivities and hemagglutinating capacities of over 50 standard and undiluted passage seeds and the compositions of the harvests derived therefrom on passage without dilution indicated that the corresponding activities in the yields did not depend entirely upon the relative concentrations of infectious virus and non-infectious hemagglutinins in the inocula but that apparently different forms of NIHA were obtained in successive undiluted passages.

scholarly journals STUDIES ON HOST-VIRUS INTERACTIONS IN THE CHICK EMBRYO-INFLUENZA VIRUS SYSTEM

1955 ◽  
Vol 101 (5) ◽  
pp. 493-506 ◽  
Author(s):  
Kurt Paucker ◽  
Werner Henle
Keyword(s):  
Influenza Virus ◽  
Infectious Virus ◽  
Allantoic Fluid ◽  
Progressive Reduction ◽  
Virus Particles ◽  
Host Virus Interactions ◽  
Virus Interactions ◽  
Available Information ◽  
Virus System

An experimental analysis is here presented of the conditions that lead to the appearance of non-infectious hemagglutinins (NIHA) in the allantoic fluid of chick embryos injected with standard influenza virus (PR8 strain) which had been exposed to 37°C. in vitro for various periods of time. On progressive reduction of the infectivity of the undiluted inocula from about 109 to 103 ID50 (103.2 HA units) the yields of infectious virus in 24 hours decreased in straight correspondence 1 millionfold, but those of hemagglutinins only by a factor of 10. Thus the proportions of NIHA in the yields increased sharply but the total quantity obtained decreased gradually. The quantities of infectious virus produced per ID50 injected were the same throughout this range; i.e., between 50 and 100 ID50, regardless of increasing proportions of heat-inactivated virus in the seeds. This value agrees with previous estimates of yields under other conditions. Thus, initiation and completion of first cycles by the infectious virus remaining in the inocula were not, or at most, slightly inhibited. The inactivated virus, therefore, failed to establish immediate interference. It was capable, however, of holding the infectious process to one cycle. Upon 10-fold dilution of the seeds essentially similar results were obtained except that a slight loss in interfering activity could now be detected with an increase in exposure to 37°C. With further dilutions little or no interference was noted. The capacity to yield NIHA decreased slowly during exposure of the seeds to 37°C. over a period of 5 days, thereafter more rapidly. It could not be restored by addition of infectious virus. Furthermore, since NIHA was obtained when the seeds contained as little as 102 or 103 ID50, it is unlikely that it was derived from those cells which had adsorbed both infectious and inactivated seed virus. It is suggestive that multiple adsorption of inactivated virus particles per se will yield NIHA. The available information, as discussed, favors the view that the NIHA does not represent seed virus in some form but is newly produced.

scholarly journals STUDIES ON HOST-VIRUS INTERACTIONS IN THE CHICK EMBRYO-INFLUENZA VIRUS SYSTEM

1956 ◽  
Vol 103 (6) ◽  
pp. 799-822 ◽  
Author(s):  
Werner Henle ◽  
Oscar C. Liu ◽  
Kurt Paucker ◽  
Florence S. Lief
Keyword(s):  
Chick Embryo ◽  
Infectious Virus ◽  
Potassium Cyanide ◽  
Cold Room ◽  
The Status ◽  
The Media ◽  
Host Virus Interactions ◽  
Virus Interactions ◽  
Virus System ◽  
Made In

Studies have been reported concerning the relationships between virus materials found in the allantoic membranes and media of eggs deembryonated after injection of Standard (ST), heat-inactivated (37°C.) standard (ΔST), and undiluted passage (UP) seeds. It was found that the membranes always contained relatively more non-infectious hemagglutinins (NIHA) than the media and, correspondingly, the ratios between infectious virus and hemagglutinin units (ID50/HA) in the tissues were up to 1.5 log10 units lower than in the liberated progeny. These differences were seen not only following inoculation of undiluted ST, ΔST, and UP seeds, the progenies of which always contain considerable proportions of NIHA, but also when dilute ST inocula were employed which lead to the liberation of only infectious virus. Essentially similar differences in the ID50/HA ratios were observed also in the allantoic membranes and fluids obtained from growth curve experiments in the intact chick embryo employing the various types of seeds. In correlating the liberated virus materials in the media of deembryonated eggs to those in the membranes it was noted that in any given 2 hour interval during the phase of nearly constant production and release up to 10 times the quantity of infectious virus was shed as was present in the tissues at the onset of that period. In contrast, only about ¼ of the hemagglutinins were released during the same time. The viral (V) and soluble (S) complement-fixing antigens were found in the tissues but no detectable quantities were released during any 2 hour interval. The NIHA in the membranes apparently is located within the cells since it could not be released by the action of RDE. Intracellular inhibitors of hemagglutination were readily inactivated following inoculation of undiluted ST, ΔST, or UP seeds but not when ultraviolet-inactivated virus was used. The inhibitor activity decreased in proportion to the hemagglutinins produced. Transfer of infected deembryonated eggs to the cold room after production and liberation of progeny were well under way immediately halted further release but in the tissues the status quo was maintained and release was resumed on return to the 37°C. incubator. The addition of potassium cyanide to the medium of deembryonated eggs at 37°C. during the period of nearly constant production and release of virus material reduced immediately and to comparable extents the ID50 and HA titers in the tissues and liberation decreased in proportion. On removal of the cyanide 2 hours later, both titers in the tissues gradually returned to those of the untreated control eggs with a corresponding increase in liberation. The ID50/HA ratios were not affected by these manipulations. It is concluded that the NIHA in the membranes forms part of a dynamic process. An attempt has been made in the discussion to integrate the present results with previous observations concerning the formation of incomplete forms of virus and their nature and role in the infectious process.

scholarly journals DISRUPTION OF INFLUENZA VIRUS

1954 ◽  
Vol 99 (4) ◽  
pp. 321-342 ◽  
Author(s):  
David A. J. Tyrrell ◽  
Frank L. Horsfall
Keyword(s):  
Influenza Virus ◽  
Influenza A ◽  
Influenza Viruses ◽  
Influenza B ◽  
Infective Virus ◽  
Freezing And Thawing ◽  
Virus Particles ◽  
Homologous Virus ◽  
Repeated Freezing ◽  
Antigen Activity

1. The hemagglutinating capacity, enzymic activity, and infectivity of several influenza viruses were destroyed by repeated freezing and thawing of dialyzed allantoic fluids containing them. 2. Influenza virus degraded by freezing and thawing, by treatment with 5 M urea, or by heating at 65°C. still combined with homologous antibody and was demonstrable by blocking of the hemagglutination-inhibition and virus neutralization reactions. 3. After 50 cycles of freezing and thawing, much of the blocking antigen activity was not sedimented by centrifugation at 120,000 g for 2 hours, and electron microscopy showed complete disruption of the virus particles. So called soluble blocking antigen was obtained from four strains of influenza A, the Lee strain of influenza B, mumps, and Newcastle disease viruses. 4. Soluble blocking antigens from influenza A viruses were highly strain-specific; gave little or no reaction in complement-fixation tests; stimulated but little antibody production in rabbits and did not induce immunity in mice; caused reactivation of infective virus in neutral mixtures of homologous virus and immune serum. 5. Repeatedly frozen and thawed influenza virus preparations did not interfere with the propagation of infective virus in the allantoic sac. The blocking antigen activity they contained was precipitated by half saturated ammonium sulfate, destroyed by trypsin, chymotrypsin, or heating at 56°C. for 30 minutes, but was unaffected by desoxyribonuclease or ribonuclease. 6. These findings are in accord with the view that soluble blocking antigen obtained from influenza virus particles on disruption by repeated freezing and thawing is protein in nature and represents the essential antigenic material of the intact virus.

scholarly journals STUDIES ON HOST-VIRUS INTERACTIONS IN THE CHICK EMBRYO-INFLUENZA VIRUS SYSTEM

1949 ◽  
Vol 90 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Werner Henle
Keyword(s):  
Chick Embryo ◽  
Influenza A ◽  
Allantoic Fluid ◽  
Active Virus ◽  
B Virus ◽  
The Difference ◽  
Host Virus Interactions ◽  
Virus Interactions ◽  
Allantoic Cavity ◽  
Partial Success

Upon injection of active influenza A or B virus into the allantoic cavity of the developing chick embryo, an average of only 70 per cent of the agent was adsorbed onto the tissue, as measured by the difference between the quantity of virus injected and that found free in the allantoic fluid of the injected eggs during the constant period. The degree of adsorption was similar, regardless of whether 109 or 102 ID50 of active virus was injected. Attempts to demonstrate the adsorbed virus in suspensions of the infected tissue met with partial success only in that not more than 1 to 5 per cent of the amount calculated to be adsorbed was actually found. All efforts to increase the yield of virus have failed. These results led to the suggestion that the seed virus, which participates in the propagation, becomes altered in such a way that it no longer may be demonstrated by infectivity titrations, whereas the active virus found represents superficially adsorbed virus, which does not multiply.

scholarly journals CHEMICAL STUDIES ON HOST-VIRUS INTERACTIONS

1946 ◽  
Vol 84 (5) ◽  
pp. 525-533 ◽  
Author(s):  
Seymour S. Cohen ◽  
Thomas F. Anderson
Keyword(s):  
Synthetic Medium ◽  
Interference Effects ◽  
Oxygen Utilization ◽  
E Coli ◽  
Chemical Studies ◽  
Bacterial Viruses ◽  
Host Virus Interactions ◽  
Virus Interactions

5-methyl tryptophane inhibited the multiplication of E. coli B without apparently affecting the rate of its oxygen utilization or R. Q. in a synthetic medium. E. coli B, under conditions of inhibition in the presence of this compound, was infected with the bacterial viruses T2 or T4. Infected organisms, in the presence of this compound, were unable to reproduce virus, assayable by the plaque method. Indeed, the number of infectious centers disappeared at a logarithmic rate in the presence of 5-methyl tryptophane, although the compound did not reduce the titers of B, T2, or T4, when the bacteria or viruses were exposed separately to the agent. In contrast to the irreversibility of the interference effects induced by viruses, the effects induced by short exposures to 5MT appear to be reversible on removal of the compound.

scholarly journals STUDIES ON HOST-VIRUS INTERACTIONS IN THE CHICK EMBRYO-INFLUENZA VIRUS SYSTEM

1949 ◽  
Vol 90 (1) ◽  
pp. 13-22 ◽  
Author(s):  
Werner Henle
Keyword(s):  
Influenza Virus ◽  
Chick Embryo ◽  
Active Agent ◽  
Allantoic Fluid ◽  
Inhibitory Effect ◽  
Influenza Viruses ◽  
Host Cells ◽  
Active Virus ◽  
Homologous Virus ◽  
Host Virus Interactions

Experiments have been reported on the propagation of influenza viruses in the allantoic membrane of the developing chick embryo during the first infectious cycle. After adsorption of the seed virus onto the host cells, only a small percentage of it remains demonstrable by infectivity titrations. This amount remains constant for 4 hours in the case of infection with PR8 virus, and for 6 hours in that of infection with Lee virus. Thereafter, a sharp rise in infectivity occurs 2 to 3 hours before liberation of the new generations of active virus into the allantoic fluid can be detected. Injection of homologous virus, inactivated by ultraviolet irradiation, following infection prevents or delays the production of virus in the tissues, depending to some extent upon the number of ID50 of active virus used as inoculum. The smaller the dose, the more pronounced the inhibitory effect. With increasing delay in the injection of the inhibitor, progressively more virus is produced and liberated 6 and 9 hours after infection with PR8 and Lee virus, respectively. Thus, production of virus may be interrupted by the homologous inhibitor when given up to 3 hours after infection with PR8, and up to4½ hours after infection with Lee virus. Since no increase in infectivity can bedetected during these 3 and 4½ hour periods in the tissues, it is suggested that influenza virus propagates in at least two major stages: first, non-infectious, immature virus material is produced which, subsequently, is converted into the fully active agent. Presumably the first step can be interrupted by the homologous inhibitor, while the second cannot. Heterologous irradiated virus, injected after infection of the tissue, exerts only a slight inhibitory effect on the production of virus.

scholarly journals CHEMICAL STUDIES ON HOST-VIRUS INTERACTIONS

1946 ◽  
Vol 84 (5) ◽  
pp. 511-523 ◽  
Author(s):  
Seymour S. Cohen ◽  
Thomas F. Anderson
Keyword(s):  
Respiratory Rate ◽  
Cytoplasmic Granules ◽  
E Coli ◽  
T4 Bacteriophage ◽  
Chemical Studies ◽  
Host Virus Interactions ◽  
Virus Interactions ◽  
Relationship Of ◽  
The Relationship ◽  
Similar Substance

The addition of active or irradiated T2 bacteriophage and T4 bacteriophage to E. coli B stops bacterial multiplication. The respiratory rate and respiratory quotient of the inhibited bacteria remained at the values observed just before infection. A respiratory rate decrease which occasionally appears can be roughly correlated with change of turbidity of the suspension. An intracellular inhibitor of multiplication appears to be liberated into lysates. A similar substance has been separated from normal E. coli B after sonic disintegration. These bacteriostatic preparations contain cytoplasmic granules with lactic acid dehydrogenase activity. The relationship of these phenomena to the interference effect in this system and others has been considered.

scholarly journals The effects of UK 2054 on the multiplication of influenza viruses

1970 ◽  
Vol 68 (1) ◽  
pp. 151-158 ◽  
Author(s):  
R. D. Barry ◽  
Patricia Davies
Keyword(s):  
Influenza Virus ◽  
Chick Embryo ◽  
Inhibitory Effect ◽  
Virus Multiplication ◽  
Influenza Viruses ◽  
Host Cells ◽  
Virus Infectivity ◽  
Virus Growth ◽  
Virus Particles ◽  
Embryo Fibroblasts

SummaryThe isoquinoline compound UK 2054 prevents the uptake of influenza virus by susceptible cells. Pre-incubation of virus particles with 500μg./ml. UK 2054 at 37°C. for 2 hr. does not reduce virus infectivity. Host cells vary in their responsiveness to the inhibitory effect of UK 2054; virus multiplication is inhibited in chick allantoic cells by lower concentrations than those required to inhibit virus growth in chick embryo fibroblasts. The effectiveness of UK 2054 is reduced by the presence of serum.It is concluded that inhibition of influenza virus multiplication by UK2054 might result from interaction of the inhibitor with both virus and cells. Any direct combination between inhibitor and virus is completely reversible.

scholarly journals STUDIES ON HOST-VIRUS INTERACTIONS IN THE CHICK EMBRYO-INFLUENZA VIRUS SYSTEM

1956 ◽  
Vol 103 (6) ◽  
pp. 777-797 ◽  
Author(s):  
Oscar C. Liu ◽  
Kurt Paucker ◽  
Werner Henle
Keyword(s):  
Influenza Virus ◽  
Chick Embryo ◽  
Virus Production ◽  
Multiple Infection ◽  
In Ovo ◽  
Cell Receptors ◽  
The Media ◽  
Host Virus Interactions ◽  
Virus Interactions

Certain aspects of the formation of non-infectious hemagglutinins (NIHA) in the chick embryo infected with influenza virus have been analyzed. It was shown by the use of combined in ovo-deembryonation technics that little or no NIHA is released following infection with small doses of standard virus during the most active and constant growth periods of the virus extending to about the 36th hour of incubation in spite of the fact that multiple infection of cells must have taken place in the latter half of that period. A slight decrease in the ID50/HA ratios of the yields obtained after the 36th hour, coinciding with the falling off of virus production and release may possibly be explained in terms of inactivation of completed virus or leakage of as yet incompleted virus from damaged cells. Exposure of the entodermal cells of the allantois of eggs deembryonated shortiy after injection of saturation or near saturation inocula of standard seed to large quantities of infectious virus added to the media at various times after infection and not extending over more than 2 hours resulted in a decrease of the ID50/HA ratios of the progenies only during the first 2 or possibly 4 hours after the primary inoculation. Later addition did not influence the yields. As discussed, such sudden and heavy exposures of cells are not expected to occur during the infectious process induced by small inocula of standard seed. The possible role of destruction of cell receptors in NIHA production has been analyzed in several ways. The addition of receptor-destroying enzyme (RDE) to the media of deembryonated eggs after near saturation inocula of standard seeds, if anything, increased the ID50/HA ratios of the progenies, and that only when added during the first few hours following infection, presumably by reducing the changes for high multiplicity of infection of cells. In contrast, ultraviolet-inactivated virus, which retains its enzymatic activity, lowered, if anything, the ID50/HA ratios of the progenies, when present in the media of deembryonated eggs from the 2nd to 4th or possibly 6th hour after infection. Excessive amounts of irradiated virus may still cause some degree of interference under these conditions. Later addition of irradiated viruses were without effect with respect to NIHA production or interference. In attempts to alter the cell receptors prior to infection by potassium periodate (KIO4), it was noted that the addition of glycerol led to the appearance and partial retention for at least 24 hours of substances in the allantoic fluids which were capable of inactivating considerable proportions of standard virus. These data indicate that destruction of external cell receptors plays little if any role in NIHA production. The implications of these findings are discussed.

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