HEK-293 Cell Lines and Derived cell lines ( contains left portion of Human Adenovirus 5 with nts 14344 is integrated into chromosome 19 of the HEK Cells)₁
Overall Risk Group₂
2
Agent Information
Family Adenoviridae - Genus Mastadenovirus - Human adenovirus Group C Human adenovirus 5 (HadV-5). Non –enveloped double stranded DNA virus.₇
Adenovirus has been completely sequenced and it includes a total of 35,937
nts (15 pp2359) The 1-4344 nts of HadV-5 represents approximately 12% of
the full genome. This agent is unusually stable to chemical , physical and
adverse pH conditions.₃
Proteins transcribed from this left portion of the Adenovirus include proteins
that are involved in Oncogenic transformation and in positive regulation of
transcription of the early genes of host infection.₇ as well as cell cycle genes
(14). However Group C viruses are not known to be tumirogenic (15 pp.2379)
It is well established that E1A proteins have mitogenic activity (14). In
mammalian cells, the human adenovirus type 5 early region 1A (E1A)
oncoprotein functions as a thyroid hormone (TH) dependent activator of the
thyroid hormone receptor (TR)₈ Transcription of viral early genes leads to
synthesis of some 17 early proteins, many of which perturb host cell or host
physiology (14). The Adenovirus DNA genome provides it with the advantage
that its DNA can persist in host cells as either a circular extra chromosome
(plasmid) or by integration into the host DNA after the complete viral
replication has stopped. It has been recently noted that adenoviral DNA from
the E1A gene is correlated with COPD.(10)
The DNA segment which induces transformation is located between 1 and 6%
from the left end of the HadV-5 DNA molecule. (11)
A 2002 study indicates that HEK-293 cells exhibit a pattern of intermediate
filament expression similar to that seen in early differentiating neurons as
opposed to cells derived from typical kidney epithelial or mesenchymal cells.
This fact could have significan implications for experiments that use these
cells as kidney cell controls or as non-neuronal control cells (16).
Primary Hazards
Ingestion via the fecal oral route₃ ,₆
Droplet exposure of the mucous membranes₃
Inhalation (through respiratory droplets)₆
Contact/hand-to-eye transfer₆
Special Hazard Notes
Generalized infections can occur in immunocompromised individuals₆
Exposure to this agent can cause serious infection in the congenitally
immunocompromised, in patients undergoing immunosuppressive treatment
for organ and tissue transplants and for cancers, and in human
immunodeficiency virus-infected patients. Adenovirus infections in these
patients tend to become disseminated and severe. In all
immunocompromised patients, generalized illness involving the central
nervous system, respiratory system, hepatitis, and gastroenteritis usually
have a fulminate course and result in death. Treatments for adenovirus
infections are of little proven value (9).
People who smoke may be at higher risk from exposure to HadV-5 (10)
Using Adenoviral vectors in conjunction with these cell lines will result in
replication competent viruses as the portion of HadV-5 incorporated into
chromosome 19 contain both the E1 and E3 regions that are generally deleted
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Containment
Requirements₃
Required PPE
Spills outside BSC₃
Effective
Decontamination₃
Post Exposure
Medical Surveillance
WSU Oversight
in commonly used Adenoviral vectors.(12)
Lab workers exposed to HEK-Cells may present an in vivo recombination
opportunity between the 12% Adenovirus and latent adenoviruses in the
human lab worker (host). Alternatively poor work technique can cause
propagation of adenovirus in the HEK cell lines by introduction of adenovirus
from the lab worker.(13)
Biosafety level 2 practices and containment facilities for all activities involving
HEK-293 cell lines.
Procedures that may generate droplets (aerosols, splashes, sprays) with HEK293 cell lines and their derivative should be handled in a Class II BSC
with BSL-2 PPE – (lab coat and gloves).₃
When open work with HEK-293 cell lines is performed outside the BSC
recommend wearing a full face shield to protect mucous membranes from
potential exposure.
Allow aerosols to settle; wearing protective clothing gently cover the spill
with absorbent paper towel and apply 1% sodium hypochlorite starting at the
perimeter and working towards the center; allow sufficient contact time (30
min) before clean up.
Susceptible to 1% sodium hypochlorite, 2%glutaraldehyde, 0.25% sodium
dodecyl sulfate.
This cell line is a transformed cell line and as such is classified as a Genetically
Modified Organism (GMO). The Human Embryonic Kidney cell lines have been
transformed with Human Adenovirus 5. Exposure to these cell lines
constitutes a reportable event to OBA “Office of Biotechnological Activities”
part of NIH (National Institute of Health)₄. Report all occupational exposures
to the WSU Biosafety Manager. Fill out an incident report and submit it.₅
Incubation Period: 1-10 days₃
There is a live vaccine against adenovirus that is administered to the military
but is not available for general use in part due to the concern with the live
vaccine’s Oncogenic potential.₉
Varies in clinical manifestation and severity; symptoms include fever, rhinitis,
pharyngitis, tonsillitis, cough and conjunctivitis.₃ Infections in
immunocompetent individuals are generally localized.₆
1. Submit a BAF (Biosafety Approval Form) to the IBC (Institutional
Biosafety Committee). This form can be downloaded at the following
website: http://www.bio-safety.wsu.edu/forms.asp .
2. Obtain approval from the IBC before starting work.
3. Submit a current BSL-2 Biosafety Manual that includes an Exposure
Control Plan for Established Cell Lines to the WSU Biosafety Manager
by the submission deadline date. The BSL-2 manual template can be
downloaded from the following website: http://www.biosafety.wsu.edu/forms.asp .
4. You will be contacted by EH&S to schedule a facility review.
Note: The Biosafety Manual and the facility review need to be completed
prior to the IBC meeting or the BAF will likely be deferred until these reviews
have been satisfactorily completed.
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REFERENCES:
1. http://www.atcc.org (ATCC Number CRL-1573)
2. HEK-293 cell line and derivatives WSU RG Risk Assessment located on the WSU Biosafety
Website: http://www.bio-safety.wsu.edu/biosafety/ under Agent Risk Assessments.
3. http://www.phac-aspc.gc.ca/msds-ftss/msds3e-eng.php
4. http://www4.od.nih.gov/oba/rac/guidelines/guidelines.html
5. WSU Policy as agreed to by signing and submitting a BAF (Biosafety Approval Form). See
Biosafety Approval Form Principal Investigator’s Certification. Can be downloaded at the
following site: http://www.bio-safety.wsu.edu/biosafety/
6. http://www.ebi.ac.uk/2can/genomes/viruses/Human_adenovirus.html
7. http://www.curehunter.com/public/keywordSummaryC084301.do
8. http://mend.endojournals.org/cgi/content/abstract/me.2002-0294v1
9. http://virus.stanford.edu/adeno/adeno.html
10. “Latent Adenoviral Infection in the Pathogenesis of Emphysema”, James C. Hogg and Shizu
Hayashi, University of British Columbia Pulmonary Research Laboratory, St. Paul’s Hospital
Vancouver, British Columbia, Canada V6Z 1Y6.
11. http://www.nature.com/nature/journal/v251/n5477/abs/251687a0.html
12. http://www.vectorbiolabs.com/vbs/page.html?m=61
13. “Using Viral Vectors in Animal Research” presented by Dr. Bruce Crise and Dr. Steve Hughes on
October 14th, 2006 at the American Biosafety Association National Conference. Power point
copy available by request from WSU Biosafety Manager courtesy of Bruce Crise.
14. http://genomebiology.com/2007/8/4/R58
15. “Virology” 5th ed. Fields, Vol. two, Lippincott Williams & Wilkins, 2007.
16. http://www.fasebj.org/gfi/content/full/16/8/869
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