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Title: Studies in influenza A virus induced apoptosis
Author: Pappworth, Isabel Yseult.
ISNI:       0000 0001 3467 1687
Awarding Body: University of Birmingham
Current Institution: University of Birmingham
Date of Award: 2006
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Influenza viruses induce apoptosis in tissue culture cells in vitro. In a study to examine the role of individual genes in apoptosis a twelve plasmid based reverse genetic system was used to produce a series of viruses, these being: rA/Victoria and rA/WSN, recombinant forms of the parent viruses A/Victoria/75 (H3N2) and A/WSN/33 (H1N1), rA/Vic/WSNNP and rA/Vic/WSNM, recombinant A/Victoria based viruses with the A/WSN nucleoprotein (NP) and matrix (M) gene respectively. These viruses were examined for their ability to induce apoptosis (determined morphologically), cell lysis (determined by lactate dehydrogenase release) and infection (determined by antibody labelling of nucleoprotein [NP]). Virus replication assays were also performed. The rA/WSN virus behaved differently to rA/Victoria and rA/Vic/WSNNP. A difference in the proportions of morphological apoptosis and cell lysis induced at 24 hours p.i. was observed. When these two measurements were taken over time it was seen that the rA/Victoria and rA/Vic/WSNNP infected cells progressed through apoptosis at a faster rate with more cells lysed at 24 hours post infection than cells infected with the rA/WSN virus. Other viruses that were thought to be reassortants of A/Victoria/75 and A/WSN/33 were found not to be, and were instead clone 7a (a reassortant of A/England/939/69 and A/Puerto Rico/8/34 parents) used in the laboratory. Clone 7a infected cells behaved similarly to rA/WSN infected cells in that progression through apoptosis was slower than for rANictoria and rANic/WSNNP infected cells. Clone 7a replicated to a higher titre and over a more prolonged period suggesting that the purpose of the delay may be to enhance replication. However, inhibition of apoptosis by the pan-caspase inhibitor (z-VAD-fmk) did not affect replication of clone 7a, rA/Victoria or rA/Vic/WSNNP.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available