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F L HORSFALL

Publications and source records attributed to F L HORSFALL.

At least 37 records · Page 2Linked to original sources
PubMed1954

On the reproduction of influenza virus; quantitative studies with procedures which enumerate infective and hemagglutinating virus particles.

Procedures which make possible the enumeration of both infective and hemagglutinating influenza A virus particles have been developed and used in a quantitative investigation on the reproduction of the agent. Infective particles were found to be highly unstable and their half-life was only 147 minutes in allantoic fluid at 35 degrees C. both in vitro and in vivo. The instability of infective particles provides an explanation for the rapid accumulation of non-infective particles which retained the hemagglutinating property. The number of non-infective (N) particles was determined from the difference between the number of hemagglutinating (H) particles and the number of infective (I) particles as indicated by the relation: [N] = [H]- [1]. When the half-life of infective particles was taken into account, both infective and hemagglutinating particles were found to disappear from the allantoic fluid; i.e., were adsorbed by the allantoic membrane, at the same logarithmic rate after inoculation. Inoculation of any number of particles up to 3 x 10(7) was followed by a constant and progressive decrease in the proportion of unadsorbed particles from 0 to 4 hours. Approximately 20 per cent of particles were unadsorbed at 2 hours and about 5 per cent at 4 hours. Inoculation of 3 x 10(8) or more particles led to a larger proportion of unadsorbed particles at 4 hours. The maximum number of particles adsorbed was computed to be about 1.6 x 10(9). The concentration of both infective and hemagglutinating particles increased rapidly in the allantoic fluid after 4 hours when any number of infective particles up to 3 x 10(7) was inoculated. With such inocula, the rate of increase during the logarithmic period was constant and the time to double the concentration of infective or hemagglutinating particles was 46 minutes. With larger inocula, i.e. 3 x 10(8) particles, the concentrations of infective and hemagglutinating particles did not increase until after 8 hours and the rate of increase was much slower. The time to double the concentration of either then became 92 minutes. The number of infective particles was approximately equal to the number of hemagglutinating particles during the logarithmic increase period when any number of infective particles up to 3 x 10(6) was inoculated and no more than 10(6) non-infective particles were included in the inoculum. This finding was taken to indicate that all or almost all particles produced and released under these conditions were infective. That such particles became inactivated rapidly and led to the accumulation of an increasing number of non-infective particles after the logarithmic period can be explained by the short half-life of infective particles. The number of infective particles was no larger than one-tenth the number of hemagglutinating particles during the logarithmic increase period after 3 x 10(7) or more infective particles had been inoculated or when smaller inocula were used which also contained 3 x 10(7) or more non-infective particles. Non-infective particles prepared in vitro at 35 degrees or 22 degrees C. were as effective as those which accumulated in vivo in diminishing the proportion of infective particles in the yield. The extent of the reduction in the proportion of infective particles was directly related to the number of non-infective particles included in the inoculum. The yield of hemagglutinating particles was diminished when the inoculum contained 3 x 10(7) or more non-infective particles. The rate of increase was reduced so that the time to double the concentration became 92 minutes when the inoculum contained 3 x 10(8) non-infective particles. It appears from these findings that the single condition which will lead to the emergence of non-infective particles during the logarithmic period is a high initial particle-cell ratio. Because non-infective particles are equally as effective as infective particles in producing this result, it seems probable that the appearance of non-infective but hemagglutinating particles is not a necessary accompaniment of the reproductive process.

Germ Cells↗
PubMed1954

Disruption of influenza virus; properties of degradation products of the virus particle.

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 degrees 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 degrees 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.

Animals↗
PubMed1954

Inhibition of influenza virus multiplication by N-glycosides of benzimidazoles-N.

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-beta-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 B(12), 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.

Animals↗
PubMed1953

Neutralization of viruses by homologous immune serum. I. Quantitative studies on factors which affect the neutralization reaction with Newcastle disease, influenza A, and bacterial virus T3.

Neutralization experiments with Newcastle disease, influenza A, or bacterial virus, T(3), reveal, under all conditions studied, a linear relationship between the logarithm of the serum dilution end point and that of the quantity of virus used. With Newcastle disease and influenza A, the slope of the neutralization line varies markedly with the host-cell system used and in the chick embryo is strikingly affected by the route of inoculation. The other variables examined have no definite effect upon the slope. Reactivation of neutralized influenza virus is demonstrable in the chick embryo on dilution of the mixture. There appears to be an inverse relationship between the degree of susceptibility of a host to infection with influenza A virus and the neutralizing titer of a serum as measured in that host. With the T(3)-serum system, comparable results are obtained when the number of unneutralized virus particles chosen as the end point is varied widely.

Animals↗
PubMed1953

Inhibition of influenza virus multiplication by alkyl derivatives of benzimidazole. I. Kinetic aspects of inhibition by 2,5-dimethylbenzimidazole as measured by infectivity titrations.

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.

Benzimidazoles↗
PubMed1953

Inhibition of influenza virus multiplication by alkyl derivatives of benzimidazole. II. Measurement of inhibitory activity by hemagglutination titrations.

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 10(5.5) EID(50) 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).

Benzimidazoles↗
PubMed1953

Inhibition of influenza virus multiplication by alkyl derivatives of benzimidazole. III. Relationship between inhibitory activity and chemical structure.

The degree of inhibition of multiplication of influenza B virus, Lee strain, in membrane cultures in vitro appears to be directly related to the concentration of the inhibitory compounds used in this investigation. With each of the alkyl derivatives of benzimidazole, evidence for such a relationship was obtained in the range between 60 and 90 per cent inhibition of virus multiplication. Alteration of the structure of benzimidazole by substitution of alkyl radicals at various positions in either the benzene or the imidazole ring resulted in diverse differences in the capacity to inhibit influenza virus multiplication in vitro. Minor increases in inhibitory activity resulted when one to three methyl groups were introduced at certain positions in the molecule. Marked increases in inhibitory activity were achieved by more extensive substitution in either the benzene or the imidazole ring. The position and nature of substituent groups appeared to be of decisive importance. Among the more highly active compounds were 2,4,5,6,7-pentamethyl-benzimidazole, 5,6-diethylbenzimidazole, and 2-ethyl-5-methylbenzimidazole. Further extension of the alkyl chain at position 2 caused no significant change in the inhibitory activity of the derivative. The most active compounds studied caused 75 per cent inhibition of Lee virus multiplication in membrane cultures in vitro at concentrations of approximately 0.0002 M. Some of the implications of these findings are discussed.

Benzimidazoles↗
PubMed1952

A mucoprotein derived from human urine which reacts with influenza, mumps, and Newcastle disease viruses.

A mucoprotein, present in normal human urine, has been isolated and obtained in a state of a high degree of purity. A number of the biological, chemical, and physicochemical properties of the substance have been studied. From the results obtained in the present investigation and those reported in succeeding papers (34, 35) it appears that the mucoprotein has a high molecular weight, i.e., of the order of 7.0 x 10(6), consists of thread-like molecules which have axial ratios of approximately 100, and is specifically antigenic. This substance, which appears to be free of contaminating material, possesses in extraordinary degree the capacity to react with influenza, mumps, and Newcastle disease viruses. At equilibrium, with influenza virus, the minimal amount of the substance capable of giving a demonstrable reaction with one hemagglutinating unit of virus appears to be about 0.0003 microg. The mucoprotein is altered by preparations of influenza viruses and its capacity to react with these agents or others is lost. The kinetics of the inactivation process brought about by influenza viruses is in accord with those of well known enzyme-substrate systems. With the exception of the capacity to react with viruses, altered mucoprotein did not differ from the native substance relative to any of the properties examined in the present study. That certain physicochemical properties of the altered mucoprotein are different from those of the native substance is demonstrated in succeeding papers (34, 35).

Animals↗
PubMed1952

Quantitative aspects of the multiplication of influenza A virus in the mouse lung; relation between the degree of viral multiplication and the extent of pneumonia.

Influenza A virus, PR8 strain, increases in amount in the infected mouse lung at a relatively constant rate. When more than 25 M.S.50 doses of virus is inoculated, the rate of multiplication appears to be independent of the amount of virus introduced; has a value of 1,100-fold increase per day. The rate of increase in the pulmonary lesions induced by infection of the mouse lung with PR8 also appears to be relatively constant and independent of the amount of virus inoculated; has a value of 8.5-fold increase per day. The essential variables in the PR8-mouse lung system appear to be equated satisfactorily by functions which were derived previously (4) during a similar quantitative investigation on pneumonia virus of mice (PVM). Evidence in support of the hypothesis that the processes of multiplication of PR8 and PVM are different in the mouse lung is presented.

Animals↗
PubMed1952

Effect of sodium monofluoroacetate on the multiplication of influenza viruses, mumps virus and pneumonia virus of mice (PVM).

Sodium fluoroacetate, given after virus inoculation in doses of 3 to 4 mg. per kg. in mice or 2 mg. in chick embryos, caused only a slight delay in the multiplication of the PR8 strain of influenza A virus in the mouse lung and of PR8 or the Lee strain of influenza B virus in the allantoic sac. The quantities of the compound used were sufficient to cause approximately 10 to 20 per cent mortality in mice and 100 per cent in chick embryos. The use of small virus inocula did not markedly increase the effect of sodium fluoroacetate on the multiplication of PR8 or Lee in the chick embryo and maximal titers were obtained in all cases. In contrast to the findings in the chick embryo, sodium fluoroacetate caused a definite delay in the multiplication of Lee virus in the mouse lung but did not affect the final virus titer. Sodium fluoroacetate in like amounts caused only a minimal delay in the multiplication of pneumonia virus of mice (PVM) in the mouse lung or of mumps virus in the chick embryo. With both PVM and mumps virus, maximal titers were obtained almost simultaneously in fluoroacetate and control animals. When three daily injections of the compound were given to mice infected previously with PVM, a definite diminution in the virus titer was demonstrable. However, pretreatment with three daily injections of the compound caused no alteration in the capacity of mice to support the multiplication of PVM. From the results of these experiments, it appears that the cellular metabolic processes blocked by sodium fluoroacetate are not essential for the multiplication of influenza viruses, mumps virus, or pneumonia virus of mice (PVM).

Animals↗