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Journal of Clinical Microbiology, October 2000, p. 3729-3734, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular and Antigenic Characterization of a Highly Evolved Derivative of the Type 2 Oral Poliovaccine Strain Isolated from Sewage in Israel

Lester M. Shulman,1,* Yosef Manor,1 Rachel Handsher,1 Francis Delpeyroux,2 Michael J. McDonough,3 Tova Halmut,1 Ilana Silberstein,1 Jacklyn Alfandari,1 Jacqueline Quay,3 Tamar Fisher,1 Jana Robinov,1 Olen M. Kew,3 Radu Crainic,2 and Ella Mendelson1

Central Virology Laboratory, Public Health Laboratories, Chaim Sheba Medical Center, Tel-Hashomer 52621, Israel1; Molecular Epidemiology of Enteroviruses, Institut Pasteur, 75724 Paris, France2; and Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 303333

Received 9 November 1999/Returned for modification 18 February 2000/Accepted 26 June 2000

An unusual, highly diverged derivative of the Sabin type 2 oral poliovaccine (OPV) strain was recovered from environmental samples during routine screening for wild polioviruses. Virus was cultivated in L20B cells and then passaged on BGM cells at 40°C (RCT [reproductive capacity at supraoptimal temperature]-positive marker) to select against most OPV strains. All but 1 of 25 RCT-positive OPV-derived environmental isolates were antigenically and genetically (>99.5% VP1 sequence match) similar to the respective Sabin strains. However, isolate PV2/4568-1/ISR98 (referred to below as 4568-1) escaped neutralization with Sabin 2-specific monoclonal antibodies and cross-adsorbed sera, and had multiple nucleotide substitutions (220 of 2,646; 8.3%) in the P1 capsid region. Fourteen of the 44 associated amino acid substitutions in the capsid mapped to neutralizing antigenic sites. Neutralizing titers in the sera of 50 Israeli children 15 years old were significantly lower to 4568-1 (geometric mean titer [GMT], 47) than to Sabin 2 (GMT, 162) or to the prototype wild strain, PV2/MEF-1/EGY42 (GMT, 108). Two key attenuating sites had also reverted in 4568-1 (A481 to G in the 5' untranslated region and the VP1 amino acid I143 to T), and the isolate was highly neurovirulent for transgenic mice expressing the poliovirus receptor (PVR-Tg21 mice). The extensive genetic divergence of 4568-1 from the parental Sabin 2 strain suggested that the virus had replicated in one or more people for ~6 years. The presence in the environment of a highly evolved, neurovirulent OPV-derived poliovirus in the absence of polio cases has important implications for strategies for the cessation of immunization with OPV following global polio eradication.

* Corresponding author. Mailing address: Central Virology Laboratory, Chaim Sheba Medical Center, Tel-Hashomer 52621, Israel. Phone: 972-3-530-2341. Fax: 972-3-530-2457. E-mail: cvlsheba{at}netvision.net.il.

Journal of Clinical Microbiology, October 2000, p. 3729-3734, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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