- Email: [email protected]
RESPIRATORY SYNCYTIAL VIRUS IN BRITAIN
466
Preliminary Communication RESPIRATORY SYNCYTIAL VIRUS IN BRITAIN IN 1956, a virus indistinguishable from chimpanzee coryza agent was isolated from human patients with bronchopneumonia and croup.1 Chanock and Finberg suggested that this group be renamed respiratory syncytial (RS) virus.2
prepared by Dekking against a virus (Boshardt) isolated by him in Holland in 1958.12 This virus was known to give rise to syncytia in tissue culture, to form cytoplasmic inclusions, and to be intermediate in its sensitivity to ether.13 serum,
DISCUSSION
A virus indistinguishable from the RS virus of Chanock has been isolated from two cases of bronchiolitis in infants. The American strain ofRS virus was not received
The xtiological role of Rs virus in lower-respiratory infections in infants and young children, and upperrespiratory infections in older children and adults, has been further elucidated by Chanock and his colleagues,— and by McClelland and her colleagues.78 The subject has recently been reviewed elsewhere.9 In the winter of 1961, an outbreak of bronchiolitis in infants occurred in Bristol, and we received throat swabs from several of the cases admitted to hospital. From two patients, Gwilliam aged 7 months and Nutt aged 6 months, viruses were isolated which were indistinguishable from RS virus. VIRUS
ISOLATION
Throat swabs were taken into 50% milk-saline, and primary isolation was effected in HeLa cells. In one case the specimen was frozen before being inoculated. The cytopathic picture consisted of large round cells and of syncytia, sometimes very large and striking. Numbers of small, round, refractile cells were also always present. The entire cell sheet was affected 4 or 5 days after the cytopathic effect first appeared.
Examination of stained preparations showed large numbers of eosinophilic cytoplasmic inclusions in affected cells (see figure). Intranuclear inclusions were originally described with RS virus1 but Chanock has since observed cytoplasmic inclusions." OTHER
PROPERTIES
Neither isolate produced haemadsorption or haamagglutination with human, guineapig, or chicken erythrocytes at 4°C, room temperature, or 37°C. The Gwilliam virus was intermediate in its sensitivity to ether. 20% ether for 20 hours at 4°C resulted in a 100-fold loss of infectivity compared with the control. SEROLOGICAL
STUDIES
of 20 sera, submitted to this laboratory for Wassermann reaction, fixed complement in the presence of tissue-culture fluid infected with Gwilliam virus. 10 of these sera were also tested against Nutt and against a strain of RS virus (Randall) isolated in Chicago by Beem et al.11 The three viruses showed an identical pattern of antibody titres. 17
out
A
paired
serum from an American patient of Dr. Hamre’s virus infection showed a rise in neutralising antibody titre to Gwilliam and Randall. Further, Gwilliam was neutralised to the same titre as Randall by a guineapig anti-
with
HeLa-cell tissue culture infected with Gwilliam virus showing intracytoplasmic inclusions. (Haematoxylin and eosin.)
an RS
1. Chanock, R. M., Roizman, B., Myers, R. Amer. J. Hyg. 1957, 66, 281. 2. Chanock, R. M., Finberg, L. ibid. p. 291. 3. Chanock, R. M., Kim, H. W., Vargosko, A. J., Deleva, A., Johnson, K. M., Gumming, C., Parrott, R. H. J. Amer. med. Ass. 1961, 176, 647. 4. Parrott, R. H., Vargosko, A. J., Kim, H. W., Cumming, C., Turner, H., Huebner, R. J., Chanock, R. M. ibid. p. 653. 5. Kravetz, H. M., Knight, V., Chanock, R. M., Morris, J. A., Johnson, K. M., Rifkind, D., Utz, J. P. ibid. p. 657. 6. Johnson, K. M., Chanock, R. M., Rifkind, D., Kravetz, H. M., Knight, V. ibid. p. 663. 7. McClelland, L., Hilleman, M. R., Hamparian, V. V., Ketler, A., Reilly, C. M., Cornfeld, D., Stokes, J. New Engl. J. Med. 1961, 264, 1169. 8. Reilly, C. M., Stokes, J., McClelland, L., Cornfeld, D., Hamparian, V. V., Ketler, A., Hilleman, M. R. ibid. p. 1176. 9. British Medical Journal, 1961, i, 1814. 10. Chanock, R. M. Unpublished. 11. Beem, M., Wright, F. H., Hamre, D., Egerer, R., Oehme, M. New Engl. J. Med. 1960, 263, 523.
in this laboratory until after most of the work reported in this paper was finished. The cytopathic effect in HeLa tissue cultures is quite striking. It is therefore not easy to understand why the virus has not previously been isolated in this country. In part, the explanation is that the virus is highly susceptible to freezing and thawing." Thus Chanock et al.3 point out that from October, 1959, After to February, 1960, they isolated only 4 strains. adopting the method of inoculating tissue cultures without first freezing specimens, they isolated 60 strains in the 5 months, March to July, 1960. The medium into which the throat swab is taken also seems to play a part.14 It may also be that strains of any particular cell line vary in their susceptibility to this virus. The finding that a high proportion of adult sera contain antibodies to RS virus suggests that infection with this virus may be as widespread in this country as it is in North America. We are grateful to Dr. D. A. J. Tyrrell for helpful suggestions and supplies of Boshardt antiserum, and to Dr. D. Taylor-Robinson for the Randall strain of RS virus and the paired sera from the case of RS infection.
D. B. PEACOCK Department of Bacteriology, University of Bristol
M.B.
Brist.
Lecturer in Bacteriology
SUZANNE K. R. CLARKE Public Health Laboratory, Bristol 12. 13. 14.
M.D. Brist. Senior Bacteriologist
F. Unpublished. Tyrrell, D. A. J. Personal communication. Hamparian, V. V., Ketler, A., Hilleman, M. R., Reilly, C. M., McClelland L., Cornfeld, D., Stokes, J. Proc. Soc. exp. Biol., N.Y. 1961, 106, 717.
Dekking,
Preliminary Communication RESPIRATORY SYNCYTIAL VIRUS IN BRITAIN IN 1956, a virus indistinguishable from chimpanzee coryza agent was isolated from human patients with bronchopneumonia and croup.1 Chanock and Finberg suggested that this group be renamed respiratory syncytial (RS) virus.2
prepared by Dekking against a virus (Boshardt) isolated by him in Holland in 1958.12 This virus was known to give rise to syncytia in tissue culture, to form cytoplasmic inclusions, and to be intermediate in its sensitivity to ether.13 serum,
DISCUSSION
A virus indistinguishable from the RS virus of Chanock has been isolated from two cases of bronchiolitis in infants. The American strain ofRS virus was not received
The xtiological role of Rs virus in lower-respiratory infections in infants and young children, and upperrespiratory infections in older children and adults, has been further elucidated by Chanock and his colleagues,— and by McClelland and her colleagues.78 The subject has recently been reviewed elsewhere.9 In the winter of 1961, an outbreak of bronchiolitis in infants occurred in Bristol, and we received throat swabs from several of the cases admitted to hospital. From two patients, Gwilliam aged 7 months and Nutt aged 6 months, viruses were isolated which were indistinguishable from RS virus. VIRUS
ISOLATION
Throat swabs were taken into 50% milk-saline, and primary isolation was effected in HeLa cells. In one case the specimen was frozen before being inoculated. The cytopathic picture consisted of large round cells and of syncytia, sometimes very large and striking. Numbers of small, round, refractile cells were also always present. The entire cell sheet was affected 4 or 5 days after the cytopathic effect first appeared.
Examination of stained preparations showed large numbers of eosinophilic cytoplasmic inclusions in affected cells (see figure). Intranuclear inclusions were originally described with RS virus1 but Chanock has since observed cytoplasmic inclusions." OTHER
PROPERTIES
Neither isolate produced haemadsorption or haamagglutination with human, guineapig, or chicken erythrocytes at 4°C, room temperature, or 37°C. The Gwilliam virus was intermediate in its sensitivity to ether. 20% ether for 20 hours at 4°C resulted in a 100-fold loss of infectivity compared with the control. SEROLOGICAL
STUDIES
of 20 sera, submitted to this laboratory for Wassermann reaction, fixed complement in the presence of tissue-culture fluid infected with Gwilliam virus. 10 of these sera were also tested against Nutt and against a strain of RS virus (Randall) isolated in Chicago by Beem et al.11 The three viruses showed an identical pattern of antibody titres. 17
out
A
paired
serum from an American patient of Dr. Hamre’s virus infection showed a rise in neutralising antibody titre to Gwilliam and Randall. Further, Gwilliam was neutralised to the same titre as Randall by a guineapig anti-
with
HeLa-cell tissue culture infected with Gwilliam virus showing intracytoplasmic inclusions. (Haematoxylin and eosin.)
an RS
1. Chanock, R. M., Roizman, B., Myers, R. Amer. J. Hyg. 1957, 66, 281. 2. Chanock, R. M., Finberg, L. ibid. p. 291. 3. Chanock, R. M., Kim, H. W., Vargosko, A. J., Deleva, A., Johnson, K. M., Gumming, C., Parrott, R. H. J. Amer. med. Ass. 1961, 176, 647. 4. Parrott, R. H., Vargosko, A. J., Kim, H. W., Cumming, C., Turner, H., Huebner, R. J., Chanock, R. M. ibid. p. 653. 5. Kravetz, H. M., Knight, V., Chanock, R. M., Morris, J. A., Johnson, K. M., Rifkind, D., Utz, J. P. ibid. p. 657. 6. Johnson, K. M., Chanock, R. M., Rifkind, D., Kravetz, H. M., Knight, V. ibid. p. 663. 7. McClelland, L., Hilleman, M. R., Hamparian, V. V., Ketler, A., Reilly, C. M., Cornfeld, D., Stokes, J. New Engl. J. Med. 1961, 264, 1169. 8. Reilly, C. M., Stokes, J., McClelland, L., Cornfeld, D., Hamparian, V. V., Ketler, A., Hilleman, M. R. ibid. p. 1176. 9. British Medical Journal, 1961, i, 1814. 10. Chanock, R. M. Unpublished. 11. Beem, M., Wright, F. H., Hamre, D., Egerer, R., Oehme, M. New Engl. J. Med. 1960, 263, 523.
in this laboratory until after most of the work reported in this paper was finished. The cytopathic effect in HeLa tissue cultures is quite striking. It is therefore not easy to understand why the virus has not previously been isolated in this country. In part, the explanation is that the virus is highly susceptible to freezing and thawing." Thus Chanock et al.3 point out that from October, 1959, After to February, 1960, they isolated only 4 strains. adopting the method of inoculating tissue cultures without first freezing specimens, they isolated 60 strains in the 5 months, March to July, 1960. The medium into which the throat swab is taken also seems to play a part.14 It may also be that strains of any particular cell line vary in their susceptibility to this virus. The finding that a high proportion of adult sera contain antibodies to RS virus suggests that infection with this virus may be as widespread in this country as it is in North America. We are grateful to Dr. D. A. J. Tyrrell for helpful suggestions and supplies of Boshardt antiserum, and to Dr. D. Taylor-Robinson for the Randall strain of RS virus and the paired sera from the case of RS infection.
D. B. PEACOCK Department of Bacteriology, University of Bristol
M.B.
Brist.
Lecturer in Bacteriology
SUZANNE K. R. CLARKE Public Health Laboratory, Bristol 12. 13. 14.
M.D. Brist. Senior Bacteriologist
F. Unpublished. Tyrrell, D. A. J. Personal communication. Hamparian, V. V., Ketler, A., Hilleman, M. R., Reilly, C. M., McClelland L., Cornfeld, D., Stokes, J. Proc. Soc. exp. Biol., N.Y. 1961, 106, 717.
Dekking,