چکیده :
The cold-adapted (ca) live attenuated influenza vaccine (LAIV) strains are manufactured in embryonated
hens’ eggs. Recently, a clonal isolate from Madin Darby Canine Kidney (MDCK) cells was derived and
characterized to assess its utility as a potential cell substrate for the manufacturing of LAIV [1]. Since
MDCK cells are a transformed continuous cell line [2], and low levels of residual cellular components
(DNA and protein) are found in the intermediates and final filled vaccine, we sought to characterize the
uptake and clearance of MDCK DNA from tissues in order to assess theoretical risks associated with
manufacturing LAIV in MDCK cell culture.
In order to address this concern, MDCK DNA uptake and clearance studies were performed in Sprague
Dawley rats. DNA extracted from MDCK Master Cell Bank (MCB) cells was administered via an intranasal
(IN) or intramuscular (IM) route. Tissue distribution and clearance of MDCK DNA were then examined in
fourteen selected tissue types at selected time points post-administration using a quantitative PCR assay
specific for canine (SINE) DNA.
Results from these studies demonstrate that the uptake and clearance of MDCK DNA from tissues vary
depending on the route of administration. When DNA was administered intranasally, as compared to
intramuscularly, detectable DNA levels were lower at all time points. Thus, the intranasal route of vaccine
administration appears to reduce potential risk associated with residual host cell DNA that may be
present in cell culture produced final vaccine products.
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