Abstract
Examination of the virus-cell interactions is of both scientific and practical importance. Our study was aimed at comparative characterization of rabbit myxoma virus and Shope fibroma virus biological properties that manifested during the virus reproduction in RK-13/2-03 clonal continuous rabbit kidney cell culture. It was demonstrated that the viruses varied in infection development and cytopathic effect duration in RK-13/2-03 cell culture. Apparent lesions in cell monolayers infected by myxoma virus and fibroma virus at similar multiplicity of infection and cultivation temperature were observed on day 2 and day 3 of cultivation, respec- tively, as well as maximum cell lesions with evident degeneration were observed on day 3 and day 6 of cultivation, respectively. Myxoma virus was accumulated at titre of 6.25–6.50 lg TCID 50 /0,2 cm 3 , and Shope fibroma virusа was accumulated at titre of 5.50−5.75 lg TCID 50 /0.2 cm 3 . Shope fibroma virus demonstrated such infectivity during three passages and myxoma virus demonstrated such infectivity during twenty passages. Prepared cultures were identified as myxoma virus and Shope fibroma virus with molecular genetic analysis. Tests of the viruses for their antigenic relatedness showed that antibodies against myxoma virus were able to neutralize Shope fibroma virus also. NT titres of antibodies against both viruses were similar (1:8). RK-13/2-03 cell culture was found to be highly permissive to Shope fibroma virus that had been isolated from the diseased rabbit and not been an attenuated variant.
Highlights
Использование в качестве клеточного субстрата перевиваемых линий клеток (ПЛК) для накопления вирусного и антигенного сырья при разработке ветеринарных противовирусных вакцин с значительно повысило эффективность и стабильность биотехно логических процессов
Our study was aimed at comparative characterization of rabbit myxoma virus and Shope fibroma virus biological properties that manifested during the virus reproduction in RK-13/2-03 clonal continuous rabbit kidney cell culture
It was demonstrated that the viruses varied in infection development and cytopathic effect duration in RK-13/2-03 cell culture
Summary
В работе использовали: клон сублинии перевиваемых клеток почки кролика RK-13/2-03 Вирусы фибромы Шоупа и миксомы кроликов штамма В-82 культивировали в двухсуточной культуре клеток RK-13/2-03, выращенной в среде DМЕМ с 10% фетальной сыворотки крови КРС. После формирования через 2 сут конфлюэнтного монослоя клетки заражали 10-кратными разведениями (10-1−10-7) вирусов фибромы Шоупа и миксомы кролика и инкубировали при температуре (33,0 ± 0,5) °С в атмосфере с повышенным до 5% содержанием СО2 и 95%-й относительной влажности. Нормальную и специфическую к вирусу миксомы сыворотки крови кроликов раститровывали с двукратным шагом (1:2−1:64) в двух планшетах, затем в первый вносили 1000 доз вируса миксомы, во второй – 1000 доз вируса фибромы Шоупа. Смесь сыворотки и вируса выдерживали при температуре (33,0 ± 0,5) °С в атмосфере с повышенным до 5% содержанием СО2 и 95%-й относительной влажности, переносили ее в планшеты с культурой клеток и инкубировали при вышеуказанном режиме в течение 10 сут. Myxoma virus manifestations on day 2 (A), Shope fibroma virus manifestations on day 3 (B), myxoma virus-associated lesions on day 3 (C), fibroma virus-associated lesions on day 4 (D), fibroma virus-associated lesions on day 5 (E), intact RK-13/2-03 cell (F)
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