I need to inject 5X10^7pfu vaccinia virus into one mouse. So I have to produce more than 10^9 pfu vaccinia virus. Now I can only produce 10^8 pfu (1ml) virus from 10X150mm tissue plates using Hu-TK143B cell line. I don't want to HeLa cells suspension method because I don't have related specific incubator. So is there any good protocol to produce more vaccinia virus using regular incubator, whichever cell line? And as a kind cell-associated virus, do I need to harvest supernatant when I harvest vaccinia virus infected cells? Thanks
Modified Vaccinia Virus Ankara (MVA) as Production Platform for Vaccines against Influenza and Other Viral Respiratory Diseases
Arwen F. Altenburg,1 Joost H. C. M. Kreijtz,1 Rory D. de Vries,1 Fei Song,2 Robert Fux,2 Guus F. Rimmelzwaan,1,* Gerd Sutter,2,* and Asisa Volz2
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Abstract
Respiratory viruses infections caused by influenza viruses, human parainfluenza virus (hPIV), respiratory syncytial virus (RSV) and coronaviruses are an eminent threat for public health. Currently, there are no licensed vaccines available for hPIV, RSV and coronaviruses, and the available seasonal influenza vaccines have considerable limitations. With regard to pandemic preparedness, it is important that procedures are in place to respond rapidly and produce tailor made vaccines against these respiratory viruses on short notice. Moreover, especially for influenza there is great need for the development of a universal vaccine that induces broad protective immunity against influenza viruses of various subtypes. Modified Vaccinia Virus Ankara (MVA) is a replication-deficient viral vector that holds great promise as a vaccine platform. MVA can encode one or more foreign antigens and thus functions as a multivalent vaccine. The vector can be used at biosafety level 1, has intrinsic adjuvant capacities and induces humoral and cellular immune responses. However, there are some practical and regulatory issues that need to be addressed in order to develop MVA-based vaccines on short notice at the verge of a pandemic. In this review, we discuss promising novel influenza virus vaccine targets and the use of MVA for vaccine development against various respiratory viruses.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4113791/
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