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35
J. Biol. Today's World. 2016 Feb; 5 (2): 35-39
J o u r n a l o f B i o l o g y a n d T o d a y ' s W o r l d
ISSN 2322-3308
J o u r n a l h o m e p a g e : h t t p : / / j o u r n a l s . l e x i s p u b l i s h e r . c o m / j b t w
Received: 20 January 2016 Accepted: 27 February 2016
Short. C
doi:10.15412/J.JBTW.01050202
Interaction of embryonic chicken lung cell with
different strains of infectious laryngotracheitis virus
infections
Mohammad Bagher Ghadiri
1
, Shahla Shahsavandi
1*
, Gholam Ali Moradli
2
, Zahra Jamshidi-Navroud
1
1
Razi Vaccine & Serum Research Institute, Karaj, Iran
2
Saveh Azad University, Dr Beheshti Blv., Saveh, Iran
*Correspondence should be addressed to Shahla Shahsavandi, Razi Vaccine & Serum Research Institute, Karaj, Iran; Tell: +982634570038;
Fax: +982634552194; Email: s.shahsavandi@rvsri.ac.ir.
ABSTRACT
The economic losses of infectious laryngotracheitis virus (ILTV) are prevented using attenuated live vaccines. Differential
diagnosis of ILTV strains is still a critical problem in controlling programs. In this study, the embryonated chicken liver cell
(ECL) serves as a host model to study virulence characteristics of ILTV strains. The permissivity of ECL cells to ILTV
infection was investigated by assessing susceptibility of the cells to vaccine strain and virulent strain infections, analyzing
the impact of viral infection on cell viability, and determining the host cellular factor X (FX) and cyclophilin A (CypA) at three
passages. To evaluate the lytic replication dynamics of ILTV in infected cells the collected suspension of last passage of
each strain was inoculated onto the dropped chorio-allantoic membrane of specific pathogen free eggs then checked for
observing characteristic lesions. The results indicated that ECL cells are highly susceptible to attenuated vaccine strain ILTV
infection. Upon infection, the strain showed faster replication kinetics in cell culture and marked cytopathic effects. Virulent
strain was able to enter ECL cells but no infectious virus was produced at 3rd passage. The establishment of latency state
was not confirmed by reactivation assay. In contrast to vaccine strain, cellular FX was also traced following virulent strain
infection. The difference expression pattern of FX in ILTV strains-infected cells is most closely with the presence of
cytopathic effects in culture. The embryonated chicken lung cell system may potentiate the relevant tool for differential
diagnosis of ILTV strains.
Key words: Infectious laryngotracheitis virus, embryonic chicken lung cell, cyclophilin A, factor X
Copyright © 2016 Mohammad Bagher Ghadiri et al. This is an open access paper distributed under the Creative Commons Attribution License.
1. INTRODUCTION
nfectious laryngotracheitis virus (ILTV) is an
economically significant avian pathogen prevented by
attenuated live vaccines. ILTV belongs to the genus
Iltovirus of Herpesviridae has a linear double stranded
DNA genome consist of a unique long region and a unique
short region which flanked by two inverted repeats (1).
Entry of the virus into respiratory cells and mucosa
depends upon host cell surface receptors and viral surface
glycoproteins. The viral nucleocapsid is released into the
cytoplasm and migrated into the nucleus following fusion
of the viral envelope to the cell membrane (2, 3). ILTV
genome replication and transcription seems to be similar to
other alpha-herpesvirus. Three classes of genes include
immediate early, early, and late genes are expressed during
transcription of ILTV DNA. The immediate-early gene,
infected cell polypeptide 4 (ICP4), has regulatory functions
and involve in the cascade transactivation of other genes (4,
5). After assembly of viral particles, the enveloped virions
are released by cell lysis. ILTV strains are propagated
efficiently in embryonated chicken egg and primary cell
culture derived from the embryo tissues (1, 6). The viruses
can further diagnose using serological test including
fluorescent antibody technique, indirect
immunofluorescence, serum neutralization, and agar gel
immunodiffusion which are less sensitive and laborious.
Molecular techniques are preferred for rapid for detection
and quantitation of ILTV DNA in clinical samples
especially when the virus established lifelong latency state
in trigeminal ganglia (7-10). Reversion of virulence after
reactivation of the latent virus and evident of possible
I