Summary
The inactivation of foot-and-mouth disease virus by formaldehyde was studied under different conditions, both as free virus and (as in routine vaccine production) after adsorption of the virus to aluminium hydroxide gel (alhydrogel). In the latter case infectivity was monitored after elution of the virus from the gel by isopycnic ultracentrifugation of the virusalhydrogel mixture in CsCl. By this method good virus recoveries were obtained. Adsorption of the virus to alhydrogel (without formaldehyde) did not reduce infectivity significantly. Both adsorbed and non-absorbed virus lost infectivity at a rate of about one log10 per day (at pH 8.5, 25° C—no formaldehyde).
Kinetics of formaldehyde inactivation of adsorbed and non-adsorbed virus were also identical, with a fast reduction in the initial phase (in case of O1 and A10-virus approximately one log10/hour). After this initial phase inactivation became linear and rather slow (for O1 and A10-virus 0.2 log10/hour). No “tailing-off” was observed. Under standard conditions (0.04 per cent formaldehyde, pH 8.5, 25° C) CD-virus was inactivated approximately 1.5 times faster than O1 and A10-virus. At 4° C the inactivation of the three strains continued at about one log10/day.
Increased lactalbumin hydrolysate concentrations reduced the inactivation rate, especially at the formaldehyde concentration of 0.02 per cent, which was originally applied. Quaternary amines like Tris strongly inhibited formaldehyde activity. These findings might explain some data of others who observed “tailing off”.
Analysis of formaldehyde inactivated antigen by SDS-PAGE and electrofocusing showed that extensive cross-linking occurs especially of VP1, probably with other virus proteins but also with non-virus proteins from the medium. VP2 and VP3 are less affected. Cross-linking was enhanced when the virus had been adsorbed to alhydrogel during inactivation.
Progressive cross-linking was observed during storage of the vaccine at 4° C, which also indicated that inactivation continued at this temperature.
These data show that formaldehyde inactivated adsorbate vaccines can be safe.
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References
Bahnemann, H. G.: Binary ethyleneimine as an inactivant for foot-and-mouth disease virus and its application for vaccine production. Arch. Virol.47, 47–56 (1975).
Barteling, S. J.: The use of frozen cells in the agar-cell suspension titration technique. Arch. ges. Virusforsch.38, 271–273 (1972).
Barteling, S. J.: Innocuity testing of foot-and-mouth disease (FMD) vaccines. II. Aziridin inactivated antigen produced in baby hamster kidney (BHK) cells. J. Biol. Stand.11, 297–304 (1983)
Barteling, S. J., Meloen, R. H.: A simple method for the quantification of 140 S particles of foot-and-mouth disease virus (FMDV). Arch. ges. Virusforsch.45, 362–364 (1974).
Barteling, S. J., Meloen, R. H., Wagenaar, F., Gielkens, A. L. J.: Isolation and characterization of trypsin-resistant O1 variants of foot-and-mouth disease virus. J. gen. Virol.43, 383–393 (1979).
Barteling, S. J., Wagenaar, F., Gielkens, A. L. J.: The positively charged structural virus protein (VP1) of foot-and-mouth disease virus (type O1) contains a highly basic part which may be involved in early virus cell interaction. J. gen. Virol.62, 357–361 (1982).
Barteling, S. J., Woortmeyer, R., Visser, N.: Innocuity testing of FMDV vaccines: I. Formaldehyde inactivated aluminium hydroxide gel vaccines. J. Biol. Stand.11, 305–312 (1983).
Brown, F., Cartwright, B., Stewart, D. L.: The effect of various inactivating agents on the viral and ribonucleic acid infectivities of foot-and-mouth disease virus and on its attachment to susceptible cells. J. gen. Microbiol.31, 179–186 (1963a).
Brown, F., Hyslop, N. St. G., Crick, J., Morrow, A. W.: The use of acetylethyleneimine in the production of inactivated foot-and-mouth disease vaccines. J. Hyg. (Camb.)61, 337–344 (1963b).
Capstick, P. B., Garland, A. J. M., Chapman, W. G., Masters, R. C.: Factors affecting the production of foot-and-mouth disease virus in deep suspension cultures of BHK 21 clone 13 cells. J. Hyg. (Camb.)65, 273–280 (1967).
Fontaine, J., Favre, D., Roulet, C., Dupasquier, M.: Inactivation du virus aphteux. Influence des agents inactivants sur la conservation du virion. Bull. Off. Int. Epizoot.81, 1089–1113 (1974).
Frenkel, H. S.: Research on foot-and-mouth disease. III. The cultivation of the virus on a practical scale in explantations of bovine tongue epithelium. Am. J. Vet. Res.12, 187 (1951).
Girard, H., Mackowiak, C.: Le vaccin antiaphteux du type Schmidt-Vallée-Waldmann est un vaccin vivant inactivé. Bull. Acad. Vét.22, 285–290 (1949).
Graves, J. H.: Formaldehyde inactivation of foot-and-mouth disease virus as applied to vaccine preparation. Am. J. Vet. Res.24, 1131–1135 (1963).
Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophae T4. Nature, Lond.227, 680–685 (1970).
O'Farrell, P. Z., Goodman, H. M., O'Farrell, P. H.: High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell.12, 1133–1142 (1977).
Schmidt, S.: Die Bedeutung der Adsorption für die aktive Immunisierung gegen Viruskrankheiten. Arch. ges. Virusforsch.1, 215–246 (1939).
Vallée, H., Carré, H., Rinjard, P.: Sur l'immunisation antiaphteuse par le virus formolé. Revue Gen. Med. Vét.35, 129–139 (1926).
Visser, N., Woortmeyer, R.: Quantification, characterization and safety testing of foot-and-mouth disease virus antigens eluted from alhydrogel vaccines. Dev. Biol. Stand.50, 277–283 (1982).
Waldmann, D., Köbe, K., Pyl, G.: Die aktive Immunisierung des Rindes gegen Maul- und Klauenseuche. Zentbl. Bakt. I. Orig.138, 401–412 (1937).
Waldmann, O., Pyl, G., Hobohm, K. O., Möhlmann, H.: Die Entwicklung des Riemser Adsorbatimpfstoffes gegen Maul- und Klauenseuche und seine Herstellung. Zentbl. Bakt. I. Orig.148, 1–15 (1941).
Wesslén, T., Dinter, Z.: The inactivation of foot-and mouth disease virus by formalin. Arch. ges. Virusforsch.7, 394–401 (1957).
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Barteling, S.J., Woortmeyer, R. Formaldehyde inactivation of foot-and-mouth disease virus. Conditions for the preparation of safe vaccine. Archives of Virology 80, 103–117 (1984). https://doi.org/10.1007/BF01310652
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DOI: https://doi.org/10.1007/BF01310652