See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/46580685

Bacterial vectors and delivery systems in cancer therapy

Article in IDrugs: the investigational drugs journal · October 2010

Source: PubMed

CITATIONS READS
56 1,961

2 authors, including:

Roman Gardlík
Comenius University Bratislava
76 PUBLICATIONS 1,504 CITATIONS

SEE PROFILE

All content following this page was uploaded by Roman Gardlík on 03 April 2014.

The user has requested enhancement of the downloaded file.

 IDrugs 2010 13(10):701-706
© Thomson Reuters (Scientific) Ltd ISSN 2040-3410

FEATURE REVIEW

Bacterial vectors and delivery systems in cancer therapy

Roman Gardlik 1,2 & Johannes H Fruehauf 2,3
Addresses
1
Comenius University, Institute of Molecular Biomedicine, Faculty of Medicine,
Sasinkova 4, 811 08, Bratislava, Slovakia
Email: [email protected]
2
Harvard Medical School, Skip Ackerman Center for Molecular Therapeutics, GI Cancer Laboratory,
Beth Israel Deaconess Medical Center, 330 Brookline Avenue, Boston, MA 02215, USA
3
ViThera Laboratories, 1 Kendall Square,
Suite 6101, Cambridge, MA 02139, USA
Email: [email protected]

*Correspondence may be addressed to either author

Live bacterial vectors may be useful tools for the development of novel cancer therapies that can be added to the repertoire of existing drugs.
Several bacterial strains effectively colonize solid tumors and act as antitumor therapeutics. The naturally occurring tumor-colonizing
characteristics of bacterial species such as Salmonella sp, Clostridium sp and Escherichia coli can be further modified by genetic
manipulations, making these bacterial systems excellent vehicles for the production and targeted delivery of therapeutic molecules into
cancer cells. This feature review summarizes recent research on cancer therapy using genetically modified bacteria. Different
approaches – bactofection, DNA vaccination, and bacterially mediated protein and RNAi delivery – in which modified bacteria are
used as anticancer therapeutics, are discussed.

Keywords Bacteria, bacterially mediated protein, bacterially mediated RNAi, bactofection, cancer, DNA vaccination, gene therapy,
transkingdom RNAi, vector delivery

Introduction One of the main reasons for the lack of efficacy of

radiotherapy and chemotherapy in many solid tumors
The first observations that bacterial infection may
is the presence of hypoxic (ie, poorly vascularized) areas
slow the growth of tumors in humans date from the
that are resistant to these interventions. However, this
19th century. The first attempts at using this
apparent limitation can be an advantage for alternative
phenomenon for therapeutic purposes were made more
approaches, such as with the use of obligate or
than 40 years ago, following the discovery that bacteria
facultative anaerobic bacteria. Several strains of
could replicate predominantly in solid tumors. These
Clostridia, Bifidobacteria and Salmonella sp selectively
findings remained generally unexplored until the end of
colonize the hypoxic areas of tumors and destroy tumor
the 20th century, when oncolytic bacteria capable of lysing
cells, thereby providing a more specific tumor-targeted
host cells began to be studied by various research groups.
therapy. Despite the selective colonization of hypoxic
areas in tumors, several studies indicate that these
Most research on gene therapy for cancer has focused on
hypoxia-specific strains are able to destroy various tumor
the use of viral vectors that exhibit high gene transfer cells, including non-hypoxic cells. Therefore, the off-target
efficiency. However, certain disadvantages of viral tissue toxicity observed for systemic cancer therapies
transduction methods have been noted, particularly and relapse as a result of residual tumor cell growth,
safety issues, as well as high cost, short bioactivity, size both of which represent two crucial deficiencies of
limitations for the DNA payload, and difficulties associated conventional cancer treatments, can potentially be
with immunogenicity and cytotoxicity. As a result, overcome by employing tumor-targeting bacteria.
alternative delivery vectors are being examined. A
framework for the use of bacterial carriers as vectors for The concept of bacterial oncolytic therapy has been
the delivery of eukaryotic expression plasmids was verified in various experimental studies, as well as
introduced in the mid-1990s, following research on a a limited number of clinical trials. More recently, a
variety of bacteria such as Shigella sp, Salmonella enterica tumor-targeting auxotrophic strain of Salmonella
Typhimurium, S enterica Typhi, Listeria monocytogenes Typhimurium was demonstrated to eradicate primary
and Escherichia coli. tumors, as well as cancer metastases, almost completely
702 IDrugs 2010 Vol 13 No 10

in various mouse models (J Cell Biochem (2009) However, the use of attenuated strains has not been
106(6):992-998; Cell Cycle (2009) 8(6):870-875). The demonstrated to be effective in clinical trials of such
use of bacterial systems for therapeutic purposes can be cancer therapy. Salmonella strains are the most widely
enhanced further by genetic modifications, providing a used systems in experiments to develop bactofection-
promising tool for the targeted delivery of genes and their mediated cancer therapy. For example, in one study, a
products. Advantages for the use of bacteria in anticancer Salmonella choleraesuis strain was engineered to carry
gene therapy include the natural oncolytic potential of a eukaryotic expression plasmid encoding the endostatin
some strains and species, the direct targeting of tumor gene. The expression of endostatin was limited exclusively
tissue, and the ease of positive regulation and eradication to tumor tissues colonized by bacteria, and significant
of the bacteria. Moreover, the naturally occurring inhibition of tumor growth (ie, 40 to 70%) with decreased
anticancer effect of tumor-targeting bacteria can also intratumoral microvessel density was observed (J Gene Med
be achieved after oral administration, circumventing the (2004) 6(12):1382-1393). Bactofection using auxotrophic
need for an intravenous route of delivery. Despite notable Salmonella as a vector to express plasmids carrying
successes obtained in studies using oncolytic bacteria for genes encoding various cytokines, such as IL-12,
the treatment of cancer, bacteriolytic therapy alone has GM-CSF, IL-4 and IL-18, as well as other molecules,
often been insufficient to eradicate tumors completely in such as Flt3 ligand, has also been employed effectively
experimental models. Therefore, the use of bacteria as in experimental tumor models. In addition, the dual
carriers of therapeutic molecules has been suggested in tumoricidal and antiangiogenic effect of S choleraesuis
order to augment the anticancer efficacy of the treatment. carrying an expression plasmid containing the
thrombospondin-1 gene under the control of a eukaryotic
This feature review summarizes recent findings on cancer promoter was observed in a murine model of malignant
therapy using genetically modified bacteria as vectors and melanoma (Cancer Gene Ther (2005) 12(2):175-184). The
delivery systems. Different approaches – bactofection, possibility of a treatment for non-solid tumors has also
DNA vaccination, and bacterially mediated protein and been described using a Salmonella Typhimurium-based
RNAi delivery – in which bacterial systems are used as oral delivery system encoding the CD40 ligand to target
anticancer therapeutic vehicles (Figure 1), are discussed. B-cell lymphoma. Despite the positive results obtained,
only a limited number of studies have been conducted
Bactofection using bactofection for the treatment of cancer. Results
Bactofection is a method in which bacteria are used to of other studies indicate that this approach may also be
transfer genes directly into the target organism, organ or suitable for targeting cells or tissues involved in other
tissue; this basic principle was first described 30 years ago. indications, such as the colonic mucosa for colitis and the
In the case of cancer therapy, bacteria deliver plasmids lungs for acute and chronic lung diseases (eg, acute
encoding the therapeutic (eg, anticancer) gene, under the respiratory distress syndrome and cystic fibrosis). For
control of a eukaryotic promoter, into tumor cells. After cancer therapy, a potential limitation of bactofection is
entering the target cell, the plasmid is released into the that the effector molecule will be expressed exclusively
cytoplasm and is subsequently transferred into the cell in cells infected by bacteria, leaving a potentially large
nucleus, where the therapeutic gene is expressed by the population of tumor cells untreated. However, if the
host cell's transcription and translation systems. The product of the transgene were secreted from the target
bacterially mediated transfer of plasmid DNA into cell, then the effector molecule might still have a good
mammalian cells thus represents a potent approach for therapeutic effect on non-infected tumor cells. However,
expressing plasmid-encoded heterologous proteins in a this proposed mechanism of action has not yet been
large set of different cell types. confirmed experimentally.

In cancer therapy, the therapeutic protein usually acts DNA vaccination

as a cytotoxic molecule toward the target cell in which The bactofection of plasmids encoding tumor-expressed
it is expressed, as well as toward surrounding cells, by antigens can lead to the induction of humoral and cellular
triggering suicide pathways. The bacteria used in such immune responses in the host, thereby providing a
therapy are engineered to undergo lysis after entry into protective defense against tumors. This approach is
the target cell and to release the plasmid into the termed DNA vaccination. Most experimental DNA
cytoplasm of the cell. This lytic mechanism prevents vaccination studies for anticancer therapy use attenuated-
the undesired long-term survival of the bacteria in the strain Salmonella Typhimurium, as the suitability of this
host organism. In addition, the products of bacterial lysis vector has been demonstrated in various preclinical
may act as potential foreign antigens and adjuvants, investigations. Oral Salmonella-based DNA vaccines
inducing cellular immune responses in the host organism against VEGFR-2 have exhibited efficacy in suppressing
and potentially adding to the tumor-ablating effects. tumor growth in animal models of malignant melanoma,
However, severe immune reactions (ie, cytokine storms) colorectal carcinoma, glioblastoma and lung cancer.
can cause unwanted toxicities and need to be avoided. Similar antitumor effects have been observed with oral
Therefore, most bacterial strains used as vectors have been bacterial vaccines against the cytokine IL-18, the apoptosis
genetically attenuated to avoid serious adverse effects, inhibitor survivin, the tumor endothelial marker 8 and
such as immune system activation and severe infections. the TGFβ1 coreceptor endoglin. Angiogenesis-related
 Bacterial vectors and delivery systems in cancer therapy Gardlik & Fruehauf�����
703

Figure 1. Antitumor effects of tumor tissue-colonizing bacteria.

A B Bactofection
Necrotic and
hypoxic area

Nucleus
Tumor

Tumor cell

C DNA vaccination
Tumor cell

APC

D Bacterially mediated protein delivery

Tumor cell

E Bacterially mediated RNAi delivery

Bacterium Therapeutic protein

Plasmid vector expressed by the bacteria
Therapeutic protein expressed RNA-induced silencing
by the cancer cell complex
Tumor antigen shRNA
Tumor cell

(A) Auxotrophic bacteria specifically colonize tumors with necrotic and hypoxic areas. The anticancer effect of bacteria can be exerted by
different strategies: bactofection, DNA vaccination, and bacterially mediated protein and RNAi delivery. (B) Bactofection: after escaping the
blood vessel and entering the target cell, bacteria disrupt and release a plasmid vector encoding the therapeutic gene. The plasmid is
transferred into the cell nucleus, and the therapeutic protein is expressed by the host cell's expression system. (C) DNA vaccination: bacteria
deliver therapeutic plasmids into the host cell (eg, APC) in a similar manner to bactofection. The plasmid encodes a tumor cell-expressed
antigen to help prime a T-cell response against the tumor antigen that is present on the surface of tumor cells, leading to the induction of
humoral and cellular immune responses against the tumor. (D) Bacterially mediated protein delivery: bacteria, either within the
extracellular environment or within tumor cells, express the therapeutic gene directly and serve as protein delivery vehicles. (E) Bacterially
mediated RNAi delivery: bacteria deliver plasmid-encoding shRNAs or express the shRNAs to induce RNAi against an oncogene or a
tumor-expressed factor. The RNA-induced silencing complex recognizes and cleaves the target mRNA according to homologous shRNAs
delivered by the bacteria.
(Adapted with permission from Roman Gardlik. © 2010 Roman Gardlik)
704 IDrugs 2010 Vol 13 No 10

factors are among the most commonly used target into its active compound. Thus, tumor-colonizing bacteria
antigens for bacterial anticancer DNA vaccination studies, express the enzyme (eg, cytosine deaminase) specifically
underlining the relevance of inhibiting tumor blood vessel in tumor tissue and, subsequently, the systemically applied
growth in cancer therapy. In addition to Salmonella, other inactive prodrug (eg, 5-fluorocytosine) is converted
bacterial species have also been used for DNA vaccination into the active cytotoxic substance through the activity
against cancer, including Pseudomonas aeruginosa. of the enzyme. This approach limits the toxicity of the
active compound to the targeted areas of tumor tissue.
Most therapeutic cancer vaccines that have been Attenuated Salmonella Typhimurium, Clostridia and
developed induce a protective CD8+ T-cell-mediated Bifidobacteria have all been used successfully for
immune response that likely reflects a breakdown of enzyme-prodrug therapy in preclinical models, and
peripheral immunological tolerance to self-antigens. several therapies have advanced into clinical trials. For
Therefore, selecting mainly T-cell epitopes from tumor example, attenuated Salmonella Typhimurium has been
antigens as targets of vaccination appears to be adequate used for the delivery of cytosine deaminase in several
for cancer therapy. However, the precise molecular phase I trials. These trials demonstrated that engineered
mechanism that regulates this phenomenon remains Salmonella Typhimurium strains could be administered
obscure. The need for further research and a better safely up to a certain dose; however, the delivered
understanding of these observations is highlighted by protein appeared to lack efficacy in the clinical setting.
the fact that similar vector strains are currently used for In preclinical models, the natural oncolytic activity of
bactofection-based approaches and for DNA vaccination. Salmonella Typhimurium has been further enhanced by
In bactofection and bacterial gene therapy approaches, genetic modification, leading to the production of various
immune activation may be an undesired side effect, antitumor molecules, such as IL-2, IL-18, CCL21 and
whereas in DNA vaccination, the immunogenic properties proapoptotic Fas ligand. In addition, Clostridia appear
of the carrier bacteria are a pivotal component of the to be particularly suited to bacterial protein delivery
therapeutic effect. into tumors because of their obligate anaerobic nature,
capacity to form spores and ease of transformation.
A main advantage of bacterial DNA vaccination as a Furthermore, the ability of Clostridia to colonize tumors
cancer treatment strategy is the potential for therapeutic in vivo has been enhanced by the deletion of genes
bacterial strains to be administered orally, as demonstrated associated with basal oxygen tolerance, such as the
in animal studies. Current research in the field is focused gene encoding superoxide dismutase. Conversely, the
on the development of clinically relevant oral bacterial main advantage of Bifidobacteria is a non-pathogenic
vaccines. However, significant knowledge gaps remain, nature. Various Bifidobacterium strains have been used in
and the mechanism of action for oral DNA vaccines has not preclinical studies as vectors for the expression of
been fully elucidated. Prior to proceeding to clinical trials, endostatin and TRAIL.
it is important to have a comprehensive understanding
of the underlying processes of bacterially mediated DNA A key aspect of the studies investigating in vivo
vaccination. bacterially mediated protein delivery is the ability of
this system to regulate the bacterial expression of the
Bacterially mediated protein delivery therapeutic protein specifically. Precise temporal and
Another approach for the use of bacteria in innovative spatial control is important to optimize the therapeutic
cancer therapies is the bacterially mediated delivery effect of the protein, and to minimize any potential
of proteins. This strategy is based on the transfer of adverse effects. One of the key tools in this area is
bacterially expressed therapeutic proteins to the host the use of inducible promoters that are activated only
organism using genetically modified (ie, transformed) in the presence of specific exogenously administered
bacteria. Unlike bactofection- and DNA vaccination-based compounds (eg, isopropyl thiogalactoside or arabinose).
methods, the persistence of bacteria in the target tissue A system based on an arabinose-inducible bacterial
may be a desired property for protein delivery. Persisting promoter has been used successfully for the specific
bacteria produce the therapeutic polypeptide in situ, thus expression of a therapeutic gene in tumor tissue using
providing the ideal situation of a drug that is distributed the bacterial strains Salmonella Typhimurium and E coli
locally, but without increasing systemic levels. Compared Nissle 1917. These strains, if administered intravenously,
with bacterially mediated gene delivery (bactofection), can selectively colonize tumors in vivo, but only
the bacterially mediated delivery of protein provides express the transgene upon the concurrent delivery of
the important possibility of regulating gene expression l-arabinose. Moreover, the exogenous regulation of
using low-molecular-weight inducers, depending on the therapeutic gene expression can be improved further
expression system used. Moreover, the effect of bacterial by the use of bacteria carrying inducible suicide genes.
vehicles can possibly be eliminated by using antibiotics.
Another approach to improve regulated gene expression,
A widely researched strategy that uses bacterially mediated as well as the targeting of bacteria to the hypoxic regions
protein delivery as an anticancer therapy is the enzyme- of tumors, is the use of hypoxia-responsive promoters in
prodrug approach, in which systemically administered tumor-colonizing bacterial strains. A subset of promoters
bacteria deliver an enzyme that converts the prodrug that are activated preferentially inside tumors has also
 Bacterial vectors and delivery systems in cancer therapy Gardlik & Fruehauf�����
705

been identified, suggesting that there are additional human and veterinary applications ranging from the
functional regulatory mechanisms that could be exploited treatment of colon cancer to gastrointestinal inflammatory
for targeted expression, in addition to those related to conditions.
hypoxia. In combination, recent data provide evidence
that intravenously administered, non-virulent genetically Unlike the tkRNAi delivery system, carrier bacteria in
modified bacteria may be efficiently exploited as vehicles the bmRNAi system do not produce shRNA, but instead
for the local generation of anticancer agents in tumors. transfer an shRNA expression plasmid to the host cell.
Additional research is required to address safety issues The transcriptional machinery of the host cell is then
and to enhance the efficacy of such bacteria without used to produce shRNA. Thus, bmRNAi acts analogously
inducing life-threatening systemic immune responses. to bactofection. Attenuated Salmonella Typhimurium has
been used for the in vivo delivery of eukaryotic
Listeria sp are being actively developed as therapeutic expression plasmids encoding shRNA against STAT3,
agents using the anticancer vaccination strategy. In this MDR1 and the anti-apoptotic protein Bcl-2 to various
approach, the bacteria are engineered to produce an mouse models of cancer, such as mouse prostate
antigen that will elicit an antitumor response by the carcinoma, human tongue squamous cell carcinoma and
immune system. This strategy has been demonstrated murine melanoma, respectively. While the bacterially
to be successful for the treatment of experimental mediated delivery of RNAi clearly provides a new area
mesothelioma and various other cancers in vivo, and is of cancer therapeutic research, additional studies are
being evaluated in clinical trials by several biotechnology required to uncover its potential applications.
companies, including Aduro BioTech Inc (formerly Anza
Therapeutics Inc) and Advaxis Inc. ADXS11-001 (Advaxis), Conclusion
a live attenuated L monocytogenes vector that secretes The application of new approaches using bacteria for
the Listeria protein listeriolysin O (LLO) fused to the the transfer of therapeutic genes or for the production of
HPV16 E7 antigen, was demonstrated to be safe in a therapeutic proteins or nucleic acids has the potential
phase I trial in patients with cervical cancer (Vaccine to advance cancer gene therapy significantly. Recent
(2009) 27(30):3975-3983). Phase II trials of the agent in studies indicate that treatments targeting a single
patients with cervical cancer and cervical intraepithelial molecule or pathway, even if such molecules or pathways
neoplasia are ongoing. have pleiotropic effects, are unlikely to eradicate tumors
completely; however, if the naturally occurring antitumor
Bacterially mediated delivery of RNAi activity inherent to some anaerobic bacteria strains
A promising new approach for bacterially mediated can be combined successfully with their ability to deliver
anticancer therapy is the combination of two distinct agents targeting tumor angiogenesis, apoptosis or
methodologies: bacteriotherapy and RNAi. As noted, the immune system, then this approach represents a
bacteria have been employed as a versatile gene delivery significant step toward achieving tumor eradication.
vector, and have been demonstrated to be an effective, Future directions that may progress the field include
safe and inexpensive strategy for delivering RNAi to improvements in the spatial and temporal regulation of
mammalian cells. Two systems that have been used to bacterial therapy, improved specificity (ie, tumor and gene
exploit bacteria for the delivery of RNAi are transkingdom targeting), and the inclusion of other novel therapeutic
RNAi (tkRNAi) and bacterially mediated RNAi (bmRNAi). payloads (eg, microRNA agonists or antagonists). All of
tkRNAi, developed at the Beth Israel Deaconess Medical these improvements could be combined with bacterial
Center, uses genetically modified bacteria to produce and delivery systems to achieve synergistic anticancer effects
deliver shRNA against oncogenes or tumor-expressed for combination therapy. Nevertheless, safety issues
factors into tumor cells. Thus, the invasive bacteria enter and public attitude will have a significant impact on the
the target tumor cell, release shRNA into the cytoplasm adoption of bacterially mediated anticancer therapy in the
and activate the RNAi pathway to induce gene silencing. clinical setting. Therefore, these considerations will need
In terms of the production of therapeutic molecules by to be addressed to progress bacterial-based therapies
bacterial carriers, tkRNAi resembles bacterially mediated from the bench to bedside. The careful selection of
protein delivery, but may have the advantage of not therapeutic indications for bacterial therapies, combined
requiring host/target cell-mediated transcription of a with the ability to minimize toxicities and side effects,
transgene. The delivery of shRNA into the cytoplasm will be crucial to the success of this approach. The use of
of the host cell is sufficient to induce RNAi. The invasive bacterial therapeutics that are specific for organs in
E coli has been used successfully as a vector for the which a local bacterial flora is present, and for which
delivery of shRNA against the human colon cancer strains derived from this flora may be used to construct
oncogene catenin β-1 and the multidrug-resistance the therapeutic agent, appears to be the optimal
gene MDR1. Such experiments have demonstrated the approach. For example, such therapeutic bacteria could
efficacy of orally administered or systemically injected be used in the gastrointestinal tract and the skin, as well
tkRNAi bacteria in mouse models of human cancer. as in the genitourinary tract. Various opportunities exist
tkRNAi is being developed as a novel method for with bacterial agents, including the prevention of colon
the delivery of RNAi-based therapeutics by Cequent cancer, the treatment of inflammatory conditions of
Pharmaceuticals and ViThera Laboratories, with various the gastrointestinal tract, luminal enzyme replacement
706 IDrugs 2010 Vol 13 No 10

treatment, and the prevention of sexually transmitted 5. Nemunaitis J, Cunningham C, Senzer N, Kuhn J, Cramm J,
Litz C, Cavagnolo R, Cahill A, Clairmont C, Sznol M: Pilot trial of
diseases, mucosal cancers and dysplasia caused by genetically modified attenuated Salmonella expressing the
viral infections. Focusing the development of bacterial E coli cytosine deaminase gene in refractory cancer
patients. Cancer Gene Ther (2003) 10(10):737-744.
therapeutics to these areas is expected to facilitate
clinical success and to prevent challenges caused by the 6. Niethammer AG, Xiang R, Becker JC, Wodrich H, Pertl U,
occurrence of major adverse events. Thus, the strategic Karsten G, Eliceiri BP, Reisfeld RA: A DNA vaccine against
VEGF receptor 2 prevents effective angiogenesis and inhibits
development of bacterial therapeutics should enable tumor growth. Nat Med (2002) 8(12):1369-1375.
bacteria to become more acceptable both as carriers of
7. Pálffy R, Gardlík R, Hodosy J, Behuliak M, Resko P, Radvánský J,
therapeutic molecules and as therapeutic 'factories' for Celec P: Bacteria in gene therapy: Bactofection versus
use in innovative therapies for cancer and other alternative gene therapy. Gene Ther (2006) 13(2):101-105.
debilitating diseases. 8. Stritzker J, Weibel S, Hill PJ, Oelschlaeger TA, Goebel W,
Szalay AA: Tumor-specific colonization, tissue distribution,

Further reading and gene induction by probiotic Escherichia coli Nissle 1917
in live mice. Int J Med Microbiol (2007) 297(3):151-162.
1. Agorio C, Schreiber F, Sheppard M, Mastroeni P, Fernandez M,
Martinez MA, Chabalgoity JA: Live attenuated Salmonella as a 9. Xiang R, Luo Y, Niethammer AG, Reisfeld RA: Oral DNA vaccines
vector for oral cytokine gene therapy in melanoma. J Gene Med target the tumor vasculature and microenvironment and
(2007) 9(5):416-423. suppress tumor growth and metastasis. Immunol Rev (2008)
222(1):117-128.
2. Buttaro C, Fruehauf JH: Engineered E coli as vehicles for
targeted therapeutics. Curr Gene Ther (2010) 10(1):27-33. 10. Xiang S, Fruehauf J, Li CJ: Short hairpin RNA-expressing
bacteria elicit RNA interference in mammals. Nat Biotechnol
3. Hoffman RM: Tumor-targeting amino acid auxotrophic (2006) 24(6):697-702.
Salmonella Typhimurium. Amino Acids (2009) 37(3):509-521.

4. Loeffler M, Le'Negrate G, Krajewska M, Reed JC: Inhibition of

tumor growth using Salmonella expressing Fas ligand.
J Natl Cancer Inst (2008) 100(15):1113-1116.

View publication stats