title: The expression of essential components for human influenza virus internalisation in Vero and MDCK cells.
abstract: MDCK and Vero cell lines have been used as substrates for influenza virus replication. However, Vero cells produced lower influenza virus titer yield compared to MDCK. Influenza virus needs molecules for internalisation of the virus into the host cell, such as influenza virus receptor and clathrin. Human influenza receptor is usually a membrane protein containing Sia(α2,6) Gal, which is added into the protein in the golgi apparatus by α2,6 sialyltransferase (SIAT1). Light clathrin A (LCA), light clathrin B (LCB) and heavy clathrin (HC) are the main components needed for virus endocytosis. Therefore, it is necessary to compare the expression of SIAT1 and clathrin in Vero and MDCK cells. This study is reporting the expression of SIAT1 and clathrin observed in both cells with respect to the levels of (1) RNA by using RT-PCR, (2) protein by using dot blot analysis and confocal microscope. The results showed that Vero and MDCK cells expressed both SIAT1 and clathrin proteins, and the expression of SIAT1 in MDCK was higher compared to Vero cells. On the other hand, the expressions of LCA, LCB and HC protein in MDCK cells were not significantly different to Vero cells. This result showed that the inability of Vero cells to internalize H1N1 influenza virus was possibly due to the lack of transmembrane protein receptor which contained Sia(α2,6) Gal.
authors.affiliation: Research and Development Division, Bio Farma, Jl. Pasteur No.28, Bandung, 40161, Indonesia, m.rani@biofarma.co.id.
authors.forename: Maharani
authors.initials: M
authors.lastname: Ugiyadi
authors.forename: Marselina I
authors.initials: MI
authors.lastname: Tan
authors.forename: Ernawati A
authors.initials: EA
authors.lastname: Giri-Rachman
authors.forename: Fawzi R
authors.initials: FR
authors.lastname: Zuhairi
authors.forename: Sony H
authors.initials: SH
authors.lastname: Sumarsono
references.citation: Microbes Infect. 2004 Aug;6(10):929-36
references.pmid: 15310470
references.citation: Nat Rev Mol Cell Biol. 2011 Jul 22;12(8):517-33
references.pmid: 21779028
references.citation: PLoS One. 2009 Oct 05;4(10):e7317
references.pmid: 19802388
references.citation: Glycobiology. 2004 Jan;14(1):39-49
references.pmid: 14514712
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 26;105(8):2800-5
references.pmid: 18287068
references.citation: Nat Struct Mol Biol. 2004 Jun;11(6):567-73
references.pmid: 15122347
references.citation: Glycobiology. 2009 Sep;19(9):936-49
references.pmid: 19494347
references.citation: Annu Rev Cell Dev Biol. 1996;12:575-625
references.pmid: 8970738
references.citation: J Biol Chem. 1989 Oct 25;264(30):17615-8
references.pmid: 2681181
references.citation: Proc Natl Acad Sci U S A. 2004 Dec 28;101(52):18153-8
references.pmid: 15601777
references.citation: Annu Rev Biochem. 2000;69:699-727
references.pmid: 10966473
references.citation: Nature. 2003 Mar 6;422(6927):37-44
references.pmid: 12621426
references.citation: J Virol. 1996 Aug;70(8):5519-24
references.pmid: 8764064
references.citation: Arch Virol. 2002 Jun;147(6):1197-208
references.pmid: 12111429