This article appeared in a journal published by Elsevier.

The attached
copy is furnished to the author for internal non-commercial research
and education use, including for instruction at the authors institution
and sharing with colleagues.
Other uses, including reproduction and distribution, or selling or
licensing copies, or posting to personal, institutional or third party
websites are prohibited.
In most cases authors are permitted to post their version of the
article (e.g. in Word or Tex form) to their personal website or
institutional repository. Authors requiring further information
regarding Elsevier’s archiving and manuscript policies are
encouraged to visit:
http://www.elsevier.com/copyright
 Author's personal copy
Available online at www.sciencedirect.com
Eat, kill or die: when amoeba meets bacteria
Pierre Cosson1 and Thierry Soldati2
The core function of the innate immune response,
phagocytosis, did not evolve first in metazoans but rather in
primitive unicellular eukaryotes. Thus, though amoebae
separated from the tree leading to metazoan shortly after the
divergence of plants, they share many specific functions with
mammalian phagocytic cells. Dictyostelium discoideum is by
far the most studied amoeba, and it is proving useful to analyze
phagocytosis and intracellular killing of bacteria. Since the
basic the basic mechanisms involved appear extremely
conserved, Dictyostelium provides novel insights into the
function of many new gene products. Bacterial pathogenicity
was certainly largely developed to resist predatory amoebae in
the environment, and this accounts for the fact that a large
number of bacterial virulence traits can be studied using
Dictyostelium as a host. This provides a particularly powerful
system to analyze the complex interactions between
pathogenic bacteria and host cells, where both the
Dictyostelium host and the bacteria can be manipulated
genetically with relative ease.
Addresses
1
University of Geneva, Faculty of Medicine, Centre Médical
Universitaire, Dpt de Physiologie Cellulaire et Métabolisme, 1 rue Michel
Servet, CH1211 Geneva 4, Switzerland
2
Department of Biochemistry, University of Geneva, 30 quai Ernest
Ansermet, Sciences II, CH1211 Genève 4, Switzerland
Corresponding author: Cosson, Pierre
(Pierre.Cosson@medecine.unige.ch) and Soldati, Thierry
(thierry.soldati@biochem.unige.ch)
Current Opinion in Microbiology 2008, 11:271–276
This review comes from a themed issue on
Techniques
Edited by Fred Ausubel and Bruno Lemaitre
Available online 10th June 2008
1369-5274/$ – see front matter
# 2008 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.mib.2008.05.005
For researchers studying immune functions, considering
Dictyostelium as a model system may seem inappropriate.
After all, this unicellular soil amoeba diverged from the
path leading to metazoans shortly after the plant–animal
split, and before fungi divergence. Yet, starting with the
seminal observations of Metschnikoff [1], it has been
evident that amoebae and human phagocytic cells share
several unique functions, such as the ability to crawl on a
substrate or to ingest microorganisms by phagocytosis.
This resemblance is also supported by the particularly
high degree of conservation between the Dictyostelium and
www.sciencedirect.com
human proteomes [2]. The basic tools and strategies of
phagocytic cells, invented in primitive protozoan, are still
used in both amoebae and mammalian phagocytic cells.
In strictly practical terms, Dictyostelium is a primitive
macrophage.
Besides the evolutionary interest of studying this long-
lost cousin, there are many experimental advantages in
working with Dictyostelium amoebae. They grow at room
temperature in a simple and cheap medium or in the
presence of bacteria, with no need for a CO2-enriched
atmosphere. They can be grown in a shaken suspension,
allowing large-scale cultures and biochemical analysis.
Their small, haploid, sequenced, and annotated genome
[2] makes them so easily amenable to genetic analysis that
functional analysis of any given gene product most often
includes characterization of the knockout mutant. Here,
we review two aspects of phagocyte biology for which the
study of Dictyostelium has proven particularly useful: pha-
gocytosis and related cellular processes, and the complex
relationships with pathogenic bacteria.
Born to kill: the professional phagocyte
It is classical in metazoan organisms to distinguish pro-
fessional from occasional, nonprofessional phagocytes,
but both pale in front of amoebae, which throughout
their life ingest, kill, and digest microorganisms at a rate
of at least one per minute. Initial studies of phagocytosis
focused on the dynamics of the actin cytoskeleton, a
crucial parameter during cell locomotion and phagocytic
engulfment. Acanthamoeaba and Dictyostelium played a
pioneering role in establishing the function of the actin
cytoskeleton in various cellular processes [3–5]. One
example is the discovery of the actin-binding protein
coronin [6]. Its role in phagocytosis was established
initially in Dictyostelium primarily on the basis of analysis
of knockout mutants [7], and later in mammalian cells [8].
More recently, other facets of phagocytosis have been
studied in Dictyostelium, in particular cellular adhesion
during phagocytosis, phagosome maturation, and intra-
cellular killing.
Phagocytosis and adhesion: integrins in
amoebae?
Adhesion to a particle is the first step of the phagocytic
process. At least three membrane proteins (Phg1, SadA,
and SibA) [9–11] are essential for cell adhesion and
phagocytosis, and SibA is probably the main adhesion
receptor [11]. Interestingly, SibA presents features of
integrin b chains, notably an extracellular VWA domain
and a conserved cytoplasmic domain. On the cytoplasmic
side, like integrins, SibA binds to a complex between
Current Opinion in Microbiology 2008, 11:271–276
 Author's personal copy
272 Techniques
talin and the FERM-containing myosin VII, both of
which are essential for adhesion and phagocytosis [12–
14]. The emerging picture is remarkably similar to integ-
rin-dependent phagocytosis in mammals (e.g. CR3-
mediated phagocytosis) and the parallel might extend
further, since Rap1 (Ras-related protein 1) also seems to
be involved in both cases [15]. Dictyostelium is thus
bringing a new perspective to our knowledge of integ-
rin-dependent adhesion, a phenomenon so far only stu-
died in metazoans. It is also offering a number of
promising leads, since several other genes essential for
adhesion have been identified in Dictyostelium (e.g. TM9
proteins [9]) that have not yet been studied in mamma-
lian systems.
A well-orchestrated maturation program
The endocytic and phagocytic pathways of Dictyostelium
exhibit marked similarities with mammalian cells [16],
and have been extensively studied [17,18]. A number of
studies have defined the exact composition of phago-
somes at each step. During the formation of phagosomes
some membrane proteins are selectively excluded [19],
while others are delivered in the first minutes following
phagosome closure [20]. This is then followed by suc-
cessive maturation steps involving sequential delivery
and retrieval of components to and from phagosomes.
Immunocytochemical profiling of signaling, cytoskeletal,
and trafficking proteins defined at least three distinct
maturation steps [21], while exhaustive proteomic
analysis established a dynamic record of phagosome
protein composition [22]. Today the phagosomal compo-
sition is established with better temporal resolution in
Dictyostelium than in mammals. The major focus of many
laboratories is now to link this extensive knowledge of
phagosome composition to its specific functions. From
that perspective, comparing phagosomal proteomes in
evolutionarily distant organisms might allow to dis-
tinguish the elements linked to its conserved functions
(e.g. bacterial killing) from those dedicated to more
organism-specific functions (e.g. antigen presentation in
mammals). The relative ease with which Dictyostelium
knockout mutants can be generated will certainly be
useful to test the putative role of specific gene products,
as was done for example to establish the role of trimeric G
proteins in phagocytosis [22].
Killing: Dictyostelium likes it raw
For amoebae as well as for mammalian phagocytes, pha-
gocytosis is a prelude to intracellular killing, a still largely
mysterious phenomenon. Lysosomal hydrolases are
delivered to the bacteria-containing phago-lysosomes
and were therefore initially believed to play a key role
in killing. Cathepsin G and elastase are involved in the
intracellular killing of S. aureus and C. albicans, respect-
ively [23], and secreted b-hexosaminidase has been
implicated in the killing of mycobacteria [24]. The study
of patients suffering from chronic granulomatous disease
Current Opinion in Microbiology 2008, 11:271–276
suggested an alternative killing mechanism: deficiencies
in the ability of NADPH-oxidase to produce reactive
oxygen species (ROS) lead to inefficient intracellular
bacterial killing. However, it has proven difficult to
distinguish experimentally between the direct bacteri-
cidal effects of ROS and their indirect effects through
changes in ionic composition of the phagosome lumen.
Despite rises and falls in the dominance of each theory,
the currently prevailing model is that lysosomal enzymes
are the key players, while NADPH-oxidase regulates the
ionic composition of the phagosomal milieu (see [25] for
an insightful review).
Dictyostelium avidly engulf and kill most bacteria
(Figure 1), offering a simple but genetically tractable
model to investigate intracellular killing. Remarkably,
Dictyostelium NADPH-oxidase knockout mutants are
unaffected in their efficiency of Klebsiella pneumoniae
killing [26], and they can grow on a wide variety of
bacterial strains, suggesting that killing is largely inde-
pendent of ROS production at least in the experimental
conditions used in these studies. What are then the
superefficient bactericidal strategies of Dictyostelium?
The characterization of mutants deficient in Klebsiella
killing identified only two genes so far: KIL1 and
PHG1 [26]. They encode a sulfotransferase and a TM9
(nine transmembrane domains) protein, respectively, but
their mode of action in Dictyostelium as well as in mammals
remains to be established. Interestingly, Phg1 and Kil1
are dispensable for the killing of a number of other
Figure 1
The various steps of phagocytosis are apparent in this electron
micrograph of a Dictyostelium cell in the process of internalizing
Klebsiella bacteria: adhesion of the amoeba to the bacteria, engulfment
of a bacteria in a phagosome, killing and digestion of internalized
bacteria. In Dictyostelium, undigested material is then exocytosed in the
extracellular medium. Imperfect fixation of the cytosol is common in
Dictyostelium and can be noticed in some regions of the cell.
www.sciencedirect.com
 Author's personal copy

Eat, kill or die: when amoeba meets bacteria Cosson and Soldati 273

bacteria, suggesting that different mechanisms are used

Downstream of Gram-peptidoglycans, NF-kappaB insect immune defence pathway

to kill various kinds of bacteria.

Kinase, mammalian MAPK pathway, positive regulator of NF-kappaB cascade

Secreted receptor for yeast polymeric beta-(1,3)-glucans, insect TLR pathway
Scavenger and pattern recognition receptor, mammalian co-TLR signaling
Today, in mammalian and in Dictyostelium cells, intra-
cellular killing is still a largely unexplored field. It is too

TNF receptor-associated factor 6, signaling downstream of TLR–NLR

Cytosolic co-chaperone, resistance (R) gene-mediated plant defence
early to know if Dictyostelium will help us understand how

Cytosolic chaperone, resistance (R) gene-mediated plant defence

mammalian phagocytes kill bacteria, or lead us to the
discovery of new mechanisms, unparalleled in mammals.

And then things turned ugly. . .

Imagine a planet populated with the ancestors of pre-
sent-day bacteria, fungi, and plants, all shyly hidden

Function

Transcription factor, plant immune response

within a rigid cell wall. Imagine amoebae irrupting in
this rather peaceful world, capable of engulfing and
killing virtually any other cell. What a feast it must have
been for the predators, and what a terrible fate for cells
ill-equipped to deal with such a situation. All microor-
ganisms must have been under strong selective pressure
to develop the traits needed to survive an encounter with
phagocytic cells. These include not only passive (resist-
ant capsule) or active (toxin secretion) defense mech-
anisms but also the ability to replicate directly within the
predator cell. This might account for one of the most
sinister paradoxes of microbiology, that many environ-
mental microorganisms, though they only exceptionally
infect mammals, have nevertheless apparently evolved
E-value

2E 19
3E 48

1E 17
2E 17

5E 04
sophisticated mechanisms to do so (for a more extensive

6E-63
0.044
review see [27]). One of the clearest examples is the
0

intracellular replication of Legionella. How can a bacter-

NCBI accession

ium that did not infect humans before the invention of

NP_001043620
NP_001063503
NP_001008162

air conditioning present such sophisticated strategies to

number

NP_178433

NP_000063
NP_573394
NP_523986
NP_976226

transform the harmful phagosomal environment into a

cozy replication niche? The answer is clearly related to
the observation that amoebae are the natural host of
Legionella. Indeed, the virulence traits of Legionella (and
D. melanogaster
D. melanogaster

of many other pathogens) were probably initially

Of species

X. tropicalis

selected to fight amoebae long before the appearance

H. sapiens

H. sapiens
A. thaliana
O. sativa
O. sativa

of metazoans. In many cases, amoebae represent the

Has two additional paralogues, DDB0191198 and DB0191189.

natural host of pathogens, and human infections are only

an accidental encounter.
Orthologous

A long list of pathogens

WRKY3

MEKK3
GNBP3
to

Not surprisingly, many pathogens have turned out to use

Hsp90
Traf-6
SGT1

CD36
Imd

similar virulence mechanisms to fight Dictyostelium and

Family greatly expanded in D. discoideum.

mammalian hosts (reviewed recently in [28,29]). These

include extracellular pathogens (e.g. Pseudomonas aerugi-
HATPase_c, HSP90

PB1, S_TKc, WD40

nosa), or bacteria capable of intracellular replication like

Conserved

TPR, p23, SGS

domains

Death-domain

Legionella. In several cases, Dictyostelium has been instru-

mental in analyzing bacterial virulence, allowing for
zf-TRAF
WRKY

example the discovery of the role of the type VI secretion

GH16
CD36

system of Vibrio cholera pathogenesis [30,31]. Many key

findings concerning Legionella pathogenicity have been
DDB0191500 b
DDB0252616 a

made in Dictyostelium (reviewed in [32,33]), particularly

Dictyostelium

DDB0220001
DDB0190154
DDB0191163

DDB0189971
DDB0206476
DDB0220118
dictyBase Id
discoideum

when traditional cell culture models had failed to deliver

Table 1

clear answers. For example, the use of Dictyostelium

knockout mutants demonstrated that autophagy is dis-
b
a

pensable for Legionella replication [34]. Dictyostelium has

www.sciencedirect.com Current Opinion in Microbiology 2008, 11:271–276

 Author's personal copy
274 Techniques
also been instrumental in revealing how Legionella manip-
ulates host cell phosphoinositide metabolism [35], and
facilitated the discovery of a nonlytic release mechanism
for spreading the infection [36,37]. The analysis of the
transcriptional response of Dictyostelium infected with
Legionella also provided new insights into the infectious
process [38].
Probably one of the most original approaches has been to
use amoebae as microbial hosts to identify new pathogens
[39], a strategy implicitly assuming that bacteria capable
of surviving within amoebae are also potential human
pathogens.
Surviving infection: resistance genes
Phagocytic cells are confronted frequently with bacteria,
and the result of this encounter largely determines the
pathogenic potential of any given bacteria. Little is
known today about cell-intrinsic resistance mechanisms:
only a few potential resistance genes are identified, and
their exact function often remains unclear. One of the
best studied examples is N-Ramp1, a metal transporter
present in the phagosomal membrane in mammalian
cells, whose loss leads to enhanced replication of some
ingested bacteria [40]. An N-Ramp1 knockout Dictyoste-
lium is also more prone to sustain intracellular bacterial
replication [41], providing an additional tool to study N-
Ramp1 function, and indicating that at least some resist-
ance mechanisms are conserved from amoebae to
humans.
This functional conservation suggests that resistance
genes could be identified in genetic screens in Dictyoste-
lium, then further characterized in mammalian cells. For
example, recent studies of Mycobacterium marinum, a close
cousin of M. tuberculosis, indicated that pathogenic myco-
bacteria use similar mechanisms to invade mammalian
macrophages and Dictyostelium [42,43]. Vacuolin (a Dic-
tyostelium flotillin-like protein) was shown to accumulate
at the membrane of the mycobacterial replication niche,
allowing direct observation of the vacuole growth and
rupture and of the release of bacteria into the cytosol [43].
The generality of these findings was confirmed in
infected human blood monocytes where selective
accumulation of flotillin-1 around M. marinum vacuole
followed by membrane rupture was also seen. Genetic
disruption of the vacuolin gene rendered Dictyostelium
more immune against infection, indicating that the pre-
sence of vacuolin is important for M. marinum to success-
fully arrest phagosome maturation. The same study also
revealed a host resistance factor [43], since the absence of
RacH rendered Dictyostelium more susceptible to M. mar-
inum proliferation. The RacH GTPase has been reported
to play a role in the regulation of the actin cytoskeleton, in
early phagosomal acidification and in the control of the
morphology and function of vacuolin-positive compart-
ments [44]. These results illustrate how Dictyostelium can
Current Opinion in Microbiology 2008, 11:271–276
Box 1 Is there a molecular machinery for microbe recognition in
Dictyostelium?
It is now well established that phagocytes of animal innate immune
systems can recognize commensal and pathogenic microbes with
which they interact. From an evolutionary perspective, it might seem
logical for protozoan to be also equipped with the molecular
equipment necessary to recognize microbe-associated molecular
patterns in order to distinguish food from pathogens. Surprisingly,
this has virtually not been studied so far in Dictyostelium, but a
number of likely orthologs and weaker homologs to proteins involved
in innate immunity can be recognized. Table 1 lists a few
Dictyostelium orthologs of genes that have been involved in
pathogen recognition in plants, insects, or vertebrates. In all these
instances, it remains to be seen if these gene products do participate
in pathogen recognition in Dictyostelium. In addition, many
Dictyostelium proteins exhibit domains conserved in proteins
involved in immune functions, such as TIR, LysM (involved in
peptidoglycan-binding, e.g. DDB0184330), Saposin B1 and B2
(involved in membrane lysis, e.g. DDB0169339), or C-type lectin
domains (involved in recognition of fungi and mycobacteria, e.g.
DDB0189466). Finally, proteins with leucine-rich repeats (LRR) such
as TLRs and NLRs form a large group of innate response genes in
various organisms. Among the 163 LRR-containing proteins in the
Dictyostelium genome, 45 have a transmembrane domain, but none
of them exhibits a cytoplasmic TIR domain such as found in TLRs.
Nevertheless, some have homologies to plant flagellin surface
receptors that bear homologies to mammalian TLR5 (DDB0204296)
or to other innate immune response receptors (DDB0232377).
be used for the study of both host defenses and pathogen
virulence strategies.
Since not all microorganisms are good to eat, Dictyostelium
would certainly benefit from the ability to recognize patho-
gens and to modulate its response accordingly, though this
is still largely a matter of speculation. At least one recent
study points to mechanisms allowing recognition of patho-
gens by amoebae [45]. In addition, the Dictyostelium gen-
ome exhibits a number of putative elements of pathogen
recognition pathways (see Box 1). The characterization of
mechanisms by which Dictyostelium could recognize diverse
microorganisms would certainly bring a new dimension to
the study of its encounter with pathogens.
Conclusion
How many ways are there for a phagocyte to kill bacteria?
How many ways for pathogenic bacteria to escape killing?
What determines phagocyte resistance to pathogens? In
Dictyostelium, the answer to each of these questions will be a
long list of gene products. As these lists start growing, we
will begin to discern how many different strategies phago-
cytes have evolved to survive in the great game of evolution.
Acknowledgements
We would like to thank members of our research groups for helpful
discussions, and Bianca Habermann (Bioinformatics, Max Planck Institute
of Molecular Cell Biology and Genetics, Dresden, Germany) for her help
with protein homology searches. This work was supported by grants from
the Fonds National Suisse pour la Recherche Scientifique (to PC and TS).
Both of our laboratories participate in the NEMO network, supported by
the 3R Foundation.
www.sciencedirect.com
 Author's personal copy
Eat, kill or die: when amoeba meets bacteria Cosson and Soldati 275
References and recommended reading
Papers of particular interest, published within the annual period of
review, have been highlighted as:
 of special interest
 of outstanding interest
1. Metschnikoff E: Sur la lutte des cellules de l’organisme contre
l’invasion des microbes. Annales de l’Institut Pasteur 1887,
1:321-336.
2. Eichinger L, Pachebat JA, Glockner G, Rajandream MA,
Sucgang R, Berriman M, Song J, Olsen R, Szafranski K, Xu Q et al.:
The genome of the social amoeba Dictyostelium discoideum.
Nature 2005, 435:43-57.
3. Noegel AA, Schleicher M: The actin cytoskeleton of
Dictyostelium: a story told by mutants. J Cell Sci 2000,
113(Pt 5):759-766.
4. Cvrckova F, Rivero F, Bavlnka B: Evolutionarily conserved
modules in actin nucleation: lessons from Dictyostelium
discoideum and plants. Review article. Protoplasma 2004,
224:15-31.
5. Soldati T: Unconventional myosins, actin dynamics and
endocytosis: a menage a trois? Traffic 2003, 4:358-366.
6. de Hostos EL, Bradtke B, Lottspeich F, Guggenheim R, Gerisch G:
Coronin, an actin binding protein of Dictyostelium discoideum
localized to cell surface projections, has sequence similarities
to G protein beta subunits. EMBO J 1991, 10:4097-4104.
7. Maniak M, Rauchenberger R, Albrecht R, Murphy J, Gerisch G:
Coronin involved in phagocytosis: dynamics of particle-
induced relocalization visualized by a green fluorescent
protein Tag. Cell 1995, 83:915-924.
8. Okumura M, Kung C, Wong S, Rodgers M, Thomas ML: Definition
of family of coronin-related proteins conserved between
humans and mice: close genetic linkage between
coronin-2 and CD45-associated protein. DNA Cell Biol 1998,
17:779-787.
9. Cornillon S, Pech E, Benghezal M, Ravanel K, Gaynor E,
Letourneur F, Bruckert F, Cosson P: Phg1p is a nine-
transmembrane protein superfamily member involved in
dictyostelium adhesion and phagocytosis. J Biol Chem 2000,
275:34287-34292.
10. Fey P, Stephens S, Titus MA, Chisholm RL: SadA, a novel
adhesion receptor in Dictyostelium. J Cell Biol 2002,
159:1109-1119.
11. Cornillon S, Gebbie L, Benghezal M, Nair P, Keller S, Wehrle-
Haller B, Charette SJ, Bruckert F, Letourneur F, Cosson P: An
adhesion molecule in free-living Dictyostelium amoebae with
integrin beta features. EMBO Rep 2006, 7:617-621.
12. Niewohner J, Weber I, Maniak M, Muller-Taubenberger A,
Gerisch G: Talin-null cells of Dictyostelium are strongly
defective in adhesion to particle and substrate surfaces
and slightly impaired in cytokinesis. J Cell Biol 1997,
138:349-361.
13. Tuxworth RI, Stephens S, Ryan ZC, Titus MA: Identification of a
myosin VII–talin complex. J Biol Chem 2005, 280:26557-26564.
14. Tuxworth RI, Weber I, Wessels D, Addicks GC, Soll DR, Gerisch G,
Titus MA: A role for myosin VII in dynamic cell adhesion. Curr
Biol 2001, 11:318-329.
15. Jeon TJ, Lee DJ, Merlot S, Weeks G, Firtel RA: Rap1 controls cell
adhesion and cell motility through the regulation of myosin II. J
Cell Biol 2007, 176:1021-1033.
16. Maniak M: Fusion and fission events in the endocytic pathway
of Dictyostelium. Traffic 2003, 4:1-5.
17. Lefkir Y, Malbouyres M, Gotthardt D, Ozinsky A, Cornillon S,
Bruckert F, Aderem AA, Soldati T, Cosson P, Letourneur F:
Involvement of the AP-1 adaptor complex in early steps of
phagocytosis and macropinocytosis. Mol Biol Cell 2004,
15:861-869.
www.sciencedirect.com
18. Neuhaus EM, Almers W, Soldati T: Morphology and dynamics of
the endocytic pathway in Dictyostelium discoideum. Mol Biol
Cell 2002, 13:1390-1407.
19. Mercanti V, Charette SJ, Bennett N, Ryckewaert JJ, Letourneur F,
Cosson P: Selective membrane exclusion in phagocytic
and macropinocytic cups. J Cell Sci 2006,
119:4079-4087.
20. Clarke M, Kohler J, Arana Q, Liu T, Heuser J, Gerisch G: Dynamics
 of the vacuolar H(+)-ATPase in the contractile vacuole
complex and the endosomal pathway of Dictyostelium cells. J
Cell Sci 2002, 115:2893-2905.
The best of in vivo imaging, and highly biologically relevant.
21. Gotthardt D, Warnatz HJ, Henschel O, Bruckert F, Schleicher M,
Soldati T: High-resolution dissection of phagosome
maturation reveals distinct membrane trafficking phases. Mol
Biol Cell 2002, 13:3508-3520.
22. Gotthardt D, Blancheteau V, Bosserhoff A, Ruppert T, Delorenzi M,
Soldati T: Proteomics fingerprinting of phagosome maturation
and evidence for the role of a Galpha during uptake. Mol Cell
Proteomics 2006, 5:2228-2243.
23. Reeves EP, Lu H, Jacobs HL, Messina CG, Bolsover S, Gabella G,
Potma EO, Warley A, Roes J, Segal AW: Killing activity of
neutrophils is mediated through activation of proteases by K+
flux. Nature 2002, 416:291-297.
24. Koo IC, Ohol YM, Wu P, Morisaki JH, Cox JS, Brown EJ: Role for
lysosomal enzyme beta-hexosaminidase in the control of
mycobacteria infection. Proc Natl Acad Sci U S A 2008,
105:710-715.
25. Segal AW: How neutrophils kill microbes. Annu Rev Immunol
 2005, 23:197-223.
An excellent review on intracellular killing mechanisms.
26. Benghezal M, Fauvarque MO, Tournebize R, Froquet R,
Marchetti A, Bergeret E, Lardy B, Klein G, Sansonetti P,
Charette SJ et al.: Specific host genes required for the killing of
Klebsiella bacteria by phagocytes. Cell Microbiol 2006,
8:139-148.
27. Casadevall A, Pirofski LA: Accidental virulence, cryptic
 pathogenesis, martians, lost hosts, and the pathogenicity of
environmental microbes. Eukaryot Cell 2007, 6:2169-2174.
A lot of fun to read, yet serious and thoughtful.
28. Kurz CL, Ewbank JJ: Infection in a dish: high-throughput
analyses of bacterial pathogenesis. Curr Opin Microbiol 2007,
10:10-16.
29. Steinert M, Heuner K: Dictyostelium as host model for
pathogenesis. Cell Microbiol 2005, 7:307-314.
30. Pukatzki S, Ma AT, Sturtevant D, Krastins B, Sarracino D,
Nelson WC, Heidelberg JF, Mekalanos JJ: Identification of a
conserved bacterial protein secretion system in Vibrio
cholerae using the Dictyostelium host model system. Proc Natl
Acad Sci U S A 2006, 103:1528-1533.
31. Pukatzki S, Ma AT, Revel AT, Sturtevant D, Mekalanos JJ: Type VI
secretion system translocates a phage tail spike-like protein
into target cells where it cross-links actin. Proc Natl Acad Sci U
S A 2007, 104:15508-15513.
32. Jules M, Buchrieser C: Legionella pneumophila adaptation
to intracellular life and the host response: clues from
genomics and transcriptomics. FEBS Lett 2007,
581:2829-2838.
33. Hilbi H, Weber SS, Ragaz C, Nyfeler Y, Urwyler S: Environmental
predators as models for bacterial pathogenesis. Environ
Microbiol 2007, 9:563-575.
34. Otto GP, Wu MY, Clarke M, Lu H, Anderson OR, Hilbi H,
 Shuman HA, Kessin RH: Macroautophagy is dispensable for
intracellular replication of Legionella pneumophila in
Dictyostelium discoideum. Mol Microbiol 2004, 51:63-72.
How Dictyostelium mutants can give the ‘final’ answer to a long debate.
35. Weber SS, Ragaz C, Reus K, Nyfeler Y, Hilbi H: Legionella
pneumophila exploits PI(4)P to anchor secreted effector
proteins to the replicative vacuole. PLoS Pathog 2006, 2:e46.
Current Opinion in Microbiology 2008, 11:271–276
 Author's personal copy
276 Techniques
36. Chen J, de Felipe KS, Clarke M, Lu H, Anderson OR, Segal G,
Shuman HA: Legionella effectors that promote nonlytic release
from protozoa. Science 2004, 303:1358-1361.
37. Chen J, Reyes M, Clarke M, Shuman HA: Host cell-dependent
secretion and translocation of the LepA and LepB effectors of
Legionella pneumophila. Cell Microbiol 2007, 9:1660-1671.
38. Farbrother P, Wagner C, Na J, Tunggal B, Morio T, Urushihara H,
Tanaka Y, Schleicher M, Steinert M, Eichinger L: Dictyostelium
transcriptional host cell response upon infection with
Legionella. Cell Microbiol 2006, 8:438-456.
39. Greub G, Raoult D: Microorganisms resistant to free-living
amoebae. Clin Microbiol Rev 2004, 17:413-433.
40. Nevo Y, Nelson N: The NRAMP family of metal-ion transporters.
Biochim Biophys Acta 2006, 1763:609-620.
41. Peracino B, Wagner C, Balest A, Balbo A, Pergolizzi B, Noegel AA,
 Steinert M, Bozzaro S: Function and mechanism of action of
Dictyostelium Nramp1 (Slc11a1) in bacterial infection. Traffic
2006, 7:22-38.
Current Opinion in Microbiology 2008, 11:271–276
View publication stats
This paper demonstrates elegantly that Nramp pumps iron out of the
phagosome.
42. Solomon JM, Leung GS, Isberg RR: Intracellular replication of
Mycobacterium marinum within Dictyostelium discoideum:
efficient replication in the absence of host coronin. Infect
Immun 2003, 71:3578-3586.
43. Hagedorn M, Soldati T: Flotillin and RacH modulate
the intracellular immunity of Dictyostelium to
Mycobacterium marinum infection. Cell Microbiol 2007,
9:2716-2733.
44. Somesh BP, Neffgen C, Iijima M, Devreotes P, Rivero F:
Dictyostelium RacH regulates endocytic vesicular trafficking
and is required for localization of vacuolin. Traffic 2006,
7:1194-1212.
45. Chen G, Zhuchenko O, Kuspa A: Immune-like phagocyte activity
 in the social amoeba. Science 2007, 317:678-681.
Who would have believed that Dictyostelium slugs had an immune
system?
www.sciencedirect.com