A variety of markers to differentiate the P phase virus from the Q phase virus among the population of A2 viruses have been proposed from this laboratory. They are as follows: 1) P virus is more avid to immune sera than Q virus, particularly in hemagglutination inhibition test using chicken red cells. 2) P virus is more sensitive to α′-inhibitors detectable in human plasma or in normal guinea pig serum. 3) Heat stability of hemagglutinins is higher with Q virus than P virus. 4) Bovine red cell agglutinins are detectable only with Q virus but not with P virus. In this article, the fact that the activity of receptor destroying enzyme is higher with Q virus than P virus was added as the fifth marker. In the second place, higher avidity of P virus than Q virus to immune sera was studied further in critical manner. The result revea ed the fact that in hemagglutination inhibition test with human convalescent sera, when human O red cells were used instead of chicken red cells, hemagglutinin inhibitory antibody was detectable even with Q virus as an antigen. Secondly, in the complement fixation test withboth viral antigens, there was no difference between P and Q in the avidity to fix the antibody contained in human convalescent sera. Thus the idea to assume P virus as in a indicator state of Q virus was proposed. In fact, KIO4 treatment of various Q viruses gave rise to the virus which was succeptible to human antibody and various inhibitors in hemagglutination inhibition test. Treatment of Q virus with formalin or urea did not change the characteristics of Q virus as KIO4. Trypsin treatment of both P and Q virus revealed the fact that they are both fairly resistant to this enzymatic action. Bovine red cell agglutinin found with Q virus was assumed to be the marker associated to the virus particle, as a whole. Because it was easily lost with ether treatment and was not detectable with the small hemagglutinins derived from Q virus.
Both human and chicken sera were fractionated by means of zone electrophoresis, in order to separate α-inhibitor from antibody against influenza virus. First proposal of this work was made in order to separate α-inhibitor active against heated Lee virus from α′-inhibitor, specifically active against Asian P virus. However, this trial was unsuccessful so far examined in this experiment. Second purpose of this experiment was to see the effect of serum treatments whether with KIO4 or filtrate of V. comma, on the antibody titer. As a result, treatment by M/50 KIO4 was shown to destroy some fraction of the human antibody against influenza virus. On the other hand, treatment with the filtrate of V. comma did not impair human antibody titer as far as tested against influenza virus. Both treatments made under our standard condition destroyed α and α′ inhibitors. Thus the conclusicn entitled here, once obtained in the field trial was secured by separating human serum into various fractions, by means of zone electrophoresis.
Comparative analysis of antigens of vaccinia, ectromelia, fowl pox, pigeon pox and canary pox viruses were made by various precedures of complement fixation test with the following results: (1) The mammalian pox viruses, vaccinia and ectromelia, demonstrated so strong cross reaction that the differentiation of these viruses was difficult by this reaction. (2) The viruses of fowl pox, pigeon pox and canary pox were found to be closely related in the antigenic make-up. The extent of cross reaction indicated that fowl pox and canary pox viruses were relatively different and the pigeon pox virus was between them. (3) No antigenic relationship was revealed between the group of mammalian viruses and that of avian viruses. Based on these findings, the classification of these two groups of viruses was discussed.
Temperate phages P-22 and S-12 grown on S. typhimurzum strain LT-2 were found to produce plaques on S. enteritidis strain No. 11 with e. o. p. of about 10-4. These phages were further found to transduce prototrophy into auxotrophic mutants of LT-2 and No. 11 with about equal frequencies. The phage particles which lysed No. 11 cells were found to be host-range mutants rather than host-controlled variants, because the growth of the “No. 11-adapted” phage particles on LT-2 cells for one cycle did not convert the progeny phage particles to the original type with regard to their e. o. p. on LT-2 and No, 11. The wild type phage particles were found to induce the temporary synthesis of O (1) antigen in the No. 11 cells, wherease neither lysis nor lysogenization took place. The temporary synthesis of O (1) antigen can be taken as the evidence for the injection of phage DNA into No. 11 cells, because phage conversion is known to be brought about by phage genome.
Bacteriophage T2 can produce progeny phage in cells of E. coli which had been inactivated by ultraviolet irradiation. In this case, however, the latent period increases and the burst size decreases as compared to the infection in normal cells. The degrees of elongation in latent period and decrease in burst size depend on dose of ultraviolet irradiation. This phenomenon was examined with various doses of UV using both Iris-glucose and peptone-glucose media. When irradiation was performed at 30cm from the source of ultraviolet light (15w), the increase in latent period was about 20 minutes in tris-glucose medium and about 30 minutes in peptoneglucose medium. The burst size decreased by one third. When bacterial cells grown in tris-glucose medium were used, a stage, which might be called an incomplete infection, was observed during infection period. This phenomenon appears to enhance the increase of latent period with cells grown in this medium over that with cells grown in peptoneglucose medium. The precise reason for this is as yet unknown.
The synthesis of protein and nucleic acids in cells of ultraviolet irradiated Escherichia coli were studied with or without infection of bacteriophage T2r+ using 35S or 32P as tracer. The synthetic capacity of host cells was lost at considerably lower doses of irradiation than the synthetic capacity for bacteriophage particles. The synthesis of RNA by infected cells appeared to be little inhibited by irradiation of relatively high dose. The experiments were performed in which bacteriophage T2 was infected to either unirradiated or heavily irradiated (15w, 30cm, 5 minutes) bacteria and incorporation of isotopes was studied with the following results; (1) The RNA which is synthesized in irradiated bacteria-phage system is quite similar to that produced by normal infection in its quantity, turnover, and nucleotide composition. (2) The rate of synthesis of protein and DNA by irradiated coliphage system is reduced to about one half to one third of normal infection. (3) The amount of non-phage protein which is synthesized by irradiated cell-phage system is apparently less than that by normal infection, at least during the first 15 minutes' incubation. Later, however, the former exceeds the latter. (4) Nearly the same amounts of phage antigenic protein and of DNA are accumulated at the time of the appearance of intracellular bacteriophage (13 minutes by unirradiated cell-phage system and 35 minutes by irradiated cells-phage system). The following may be concluded from these results. (i) Phage maturation occurs after the accumulation of certain amounts of phage precursors in host cells. The pool size of phage precursors in bath systems are the same at that time. (ii) Since the sysnthesis of precursors does not occur in excess, inactive phage particle does not appear to be formed. (iii) Inhibition at the step of maturation cannot be expected by irradiated coli-phage system. (iv) The observed increase in latent period and decrease in burst sized by irradiation of host cells may result from the reduction in the rate of synthesis of phage constitutents. The reduction in the rate of synthesis of phage constituents has not been fully explained until now. This may be, however, attributed to either the reduction in the amount of nonphage protein synthesized in the early period or to the reduction in biological activity of this protein. This must be clarified by future studies.