Abstract
Fundamental experiments on biological aspects and physico-chemical properties of hemagglutinin of vaccinia virus were carried out. Inhibition of hemagglutination by specific antibody was also investigated, with special reference to its reliability as a diagnostic aid in small-pox case. The results obtained are summarized as follows.
A. Production of V. H. in fertile hen's egg
1) From 6 to 14 day egg was inoculated with seed virus through various routes and every organ was examined after 72 hours for presence of V. H. The most luxurious production of V. H. was observed in chorioallantoic membrane when seed virus was inoculated via air sac. The most adequate age of egg at the time of inoculation was found to be from 11 to 12 days. V. H. was located just around the inoculated area of membrane, whereas other part of the membrane than that although showing as much infectivity, contained little amount of hemagglutinin.
2) Tne chorioallantoic membrane of 12 day egg inoculated with seed virus via air sac showed a little rise of V. H. titer after 24 hours, reached to its maximal titer after 48 hours and then retained that titer for 72 hours.
3) It is necessary for V. H. production to use seed virus of higher virulence than minimum V. H. -producible.
B. Physico-chemical properties of V. H.
1) V. H. was partly destroyed rapidly at lower pH than 6.0 or higher than 9.0.
2) The optimal pH range for hemagglutination was 7.0 to 8.0.
3) Monoiodium acetic acid, sodium cyanide, sodium azide, cystein hydrochloride, sublimat, Merzonin (Takeda), * formaldehyde, benzaldehyde, hydrogen dioxide, potassium permanganate, phenol, urea, sodium sulphite, sodium thiosulphate or sodium sulphate hardly destroyed V. H. at the concentration of 5×10-3 to 5×10-5 M or 0.5 to 0.05%, even after treated at 37°C for 2 hours. Hydroxylamine was a little effective in lessening V. H. titter.
4) Methyl-bis-β-chlorethylamine (nitrogen mustard) destroyed V. H. almost completely at the concentration of 10-2 to 7.5×10-3 after treated at 37°C for 2 hours, whereas methylbis-β-chloretylamine-sodium-oxide did not destroy it in the same conditions.
5) Petroleum ether, ethylether, chloroform, benzene, ethyl acetate, toluene or butylalcohol showed no destructive function on V. H. at -6°C after 30 minutes.
6) V. H. emulsion was treated with petroleum ether, ethylether or chloroform and then left overnight at 2°C. Ethylether and chloroform were effective in V. H. destruction, and petroleum ether was less effective.
7) V. H. could be preserved with little change of its titer when kept in ice box after treated with monoiodium acetic acid at the concentration of 5×10-4 or heated at 50°C for 30 minutes.
C. Relation between V. H. and vaccinia virus particle
Vaccinia virus particle, thermostable and thermolable components of V. H. were separated from each other by means of either differential centrifugation or ultrafiltration. Thus, it was proved that their molecular sizes are different from each other.
