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Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis

The rapid worldwide spread of the pandemic H1N1 2009 influenza A virus in the early 2009 has contributed to high morbidity and mortality especially in children, the elderly and immuno-compromised individuals. Current available influenza vaccines are effective in terms of providing immunity protectio...

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Main Author: Siaw, Stella Xiu Joan
Format: Thesis
Language:English
Published: 2014
Subjects:
H1N1 influenza
Lactococcus lactis
Hemagglutinin test
Online Access:http://psasir.upm.edu.my/id/eprint/70115/1/FBSB%202014%2039%20IR.pdf
http://psasir.upm.edu.my/id/eprint/70115/
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spelling my.upm.eprints.701152019-10-31T06:28:48Z http://psasir.upm.edu.my/id/eprint/70115/ Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis Siaw, Stella Xiu Joan The rapid worldwide spread of the pandemic H1N1 2009 influenza A virus in the early 2009 has contributed to high morbidity and mortality especially in children, the elderly and immuno-compromised individuals. Current available influenza vaccines are effective in terms of providing immunity protection but there are several limitations and disadvantages such as allergic reactions, biosafety issues and limited production capacity. This study uses Lactococcus lactis as a vehicle for cloning of the haemagglutinin (HA1) epitope of the pandemic H1N1 2009 influenza virus. The HA1 epitope was amplified and cloned into a L. lactis vector together with the usp45 signal peptide and nisP anchor protein (nisP-anc) in order to assemble a surface displayed HA1 recombinant L. lactis. The recombinant L. lactis were able to express HA1 epitope protein when induced with nisin and was further confirmed with Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The recombinant host cells were further examined by using whole cell ELISA, fluorescence microscopy and flow cytometry to detect the surface displayed HA1 epitope protein on their surfaces. Despite it could not be confirmed that HA1 epitope has been successfully surface displayed, the recombinant cells were fed to BALB/c mice host to observe for its capability to induce immunogenic response. From the analysis of antigen-specific serum antibody by sandwich ELISA, significant anti-HA1 sIgA antibody was produced in the mice fecal suspension of mice group NZ9000 (pNZ:HN) when compared to the negative control NZ9000 (pNZ8048) suggesting that high specific anti-HA1 sIgA antibody was produced in the mice rectum. This study was conducted to gain more insights of the potential of L. lactis as a vaccine vehicle and future optimization can be adopted to further enhance its capability in vaccine delivery. 2014-12 Thesis NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/70115/1/FBSB%202014%2039%20IR.pdf Siaw, Stella Xiu Joan (2014) Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis. Masters thesis, Universiti Putra Malaysia. H1N1 influenza Lactococcus lactis Hemagglutinin test
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
topic H1N1 influenza
Lactococcus lactis
Hemagglutinin test
spellingShingle H1N1 influenza
Lactococcus lactis
Hemagglutinin test
Siaw, Stella Xiu Joan
Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis
description The rapid worldwide spread of the pandemic H1N1 2009 influenza A virus in the early 2009 has contributed to high morbidity and mortality especially in children, the elderly and immuno-compromised individuals. Current available influenza vaccines are effective in terms of providing immunity protection but there are several limitations and disadvantages such as allergic reactions, biosafety issues and limited production capacity. This study uses Lactococcus lactis as a vehicle for cloning of the haemagglutinin (HA1) epitope of the pandemic H1N1 2009 influenza virus. The HA1 epitope was amplified and cloned into a L. lactis vector together with the usp45 signal peptide and nisP anchor protein (nisP-anc) in order to assemble a surface displayed HA1 recombinant L. lactis. The recombinant L. lactis were able to express HA1 epitope protein when induced with nisin and was further confirmed with Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. The recombinant host cells were further examined by using whole cell ELISA, fluorescence microscopy and flow cytometry to detect the surface displayed HA1 epitope protein on their surfaces. Despite it could not be confirmed that HA1 epitope has been successfully surface displayed, the recombinant cells were fed to BALB/c mice host to observe for its capability to induce immunogenic response. From the analysis of antigen-specific serum antibody by sandwich ELISA, significant anti-HA1 sIgA antibody was produced in the mice fecal suspension of mice group NZ9000 (pNZ:HN) when compared to the negative control NZ9000 (pNZ8048) suggesting that high specific anti-HA1 sIgA antibody was produced in the mice rectum. This study was conducted to gain more insights of the potential of L. lactis as a vaccine vehicle and future optimization can be adopted to further enhance its capability in vaccine delivery.
format Thesis
author Siaw, Stella Xiu Joan
author_facet Siaw, Stella Xiu Joan
author_sort Siaw, Stella Xiu Joan
title Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis
title_short Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis
title_full Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis
title_fullStr Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis
title_full_unstemmed Nisp as a potential surface anchor for pandemic H1N1 2009 Haemagglutinin (HA1) viral epitope in Lactococcus lactis
title_sort nisp as a potential surface anchor for pandemic h1n1 2009 haemagglutinin (ha1) viral epitope in lactococcus lactis
publishDate 2014
url http://psasir.upm.edu.my/id/eprint/70115/1/FBSB%202014%2039%20IR.pdf
http://psasir.upm.edu.my/id/eprint/70115/
_version_ 1651869116169977856
score 13.211869

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