COMMENTARY: GENETICS OF IMMUNITY

The Genetics of Immunity

Brian P. Lazzaro*,1 and David S. Schneider†
*Department of Entomology, Cornell University, Ithaca, NY 14853, and †Department of Microbiology and Immunology, Stanford
University, Palo Alto, CA, 94305

In this commentary, Brian P. Lazzaro and David S. Schneider inflammation-like phenotypes of increased lamellocyte (im-
examine the topic of the Genetics of Immunity as explored in mune cell) proliferation, appearance of melanotic masses,
this month’s issues of GENETICS and G3: Genes|Genomes| and induction of antimicrobial peptides. They found that
Genetics. These inaugural articles are part of a joint Genetics Toll activity in Drosophila larvae is negatively regulated by
of Immunity collection (ongoing) in the GSA journals. sumoylation controlled by the b-arrestin gene Kurtz. Loss of
Kurtz or the SUMO protease Ulp1 results in ectopic immune
D EFENSE against infection is increasingly recognized
to have a complex determination, shaped by the con-
tributions of multiple genes and a multitude of environ-
activity and inappropriate inflammation-like responses. No-
tably, however, the distinct immune reactions vary in their
relative magnitudes in the mutants, indicating that sumoy-
mental factors. While we may conventionally imagine
lation probably interacts with other elements of the cellular
defense against infection to be determined primarily by
machinery to balance the multiple activities of the highly
the activity of the host immune system, recent studies have
pleiotropic Toll pathway. Additionally, because Kurtz and
established diverse biological mechanisms for regulating
Ulp1 mutants result in global disruption of SUMO activity,
defense. The genetics of immunity is being studied using
there is probably dysregulation of other pathways that con-
a wide variety of approaches and organisms from agricul-
tribute to control of inflammation and immunity.
turally relevant plants to genetic models such as Drosoph-
De Arras et al. (2014) employed a clever cross-species
ila to humans. In this spirit of greater discourse among
mutant screen to identify a regulator that controls splicing
researchers across disciplines, the journals of the Genetics
of messenger RNA (mRNA) encoding the Toll pathway
Society of America—GENETICS and G3: Genes|Genomes|
adapter MyD88, and hence immune activity. They took ad-
Genetics—invite submissions that address the broad reach
vantage of high-throughput RNA interference (RNAi) screen-
and complexity of the genetics of immunity. Several such ar-
ing in Caenorhabditis elegans to scan the entire genome for
ticles are presented in the June issues of both journals. The
genes whose inhibition blocks immune induction. They found
call for papers is ongoing, and future articles will be high-
32 well-supported candidates, 20 of which have clear orthologs
lighted in the Genetics of Immunity collection.
in the mouse. Disruption of 8 of these genes in mice also yields
clear immune deficiency, and one of them, Eftud2, gives the
Complex Regulation of Immunity reciprocal phenotype of enhanced IL-6 expression in response
Several of the highlighted articles report studies of immune to lipopolysaccharide when overexpressed. Further investiga-
regulation in novel contexts. Anjum et al. (2013) examined tion revealed that Eftud2 protein mediates the relative balance
modulation of the Toll pathway, best known in Drosophila between a long (activating) and a short (inhibiting) spliced
for its role in activating synthesis of antimicrobial peptides form of MyD88. Loss of Eftud2 function results in a proportion-
in response to bacterial infection and in vertebrates for in- ally much larger decrease in the short form relative to the long
ducing inflammatory responses to microbial and viral elici- form, thus blocking Toll-pathway activity and immune defense.
tors. Anjum et al. (2013) focused on Toll pathway activity in This article nicely illustrates the power of comparative genomics
larval Drosophila and conclude that misactivation results in and immunology to uncover conserved biological functions.
In another dissection of pathway regulation, Stronach
et al. (2014) tackled the role of mitogen-activated protein
Copyright © 2014 by the Genetics Society of America kinases (MAPKs) in developmental vs. immunological cel-
doi: 10.1534/genetics.114.165449
1
Corresponding author: Department of Entomology, Cornell University, Ithaca, NY
lular contexts. MAPKs activate the Jun Kinase (JNK) path-
14853. E-mail: bplazzaro@cornell.edu way in response to infection and stress and are themselves

Genetics, Vol. 197, 467–470 June 2014 467

regulated by upstream kinases (MAPKKs and MAPKKKs, or effect an inflammatory response. Lawless et al. (2014) hy-
MAP3Ks). Stronach et al. (2014) posited that MAP3Ks are pothesize that this switch is mediated by a suite of miRNAs,
broken into a functional domains—some that receive stim- with upregulated miRNAs tending to target metabolic tran-
ulus or determine subcellular localization, plus a distinct script RNAs, presumably driving the switch to glycolysis. In a
protein kinase domain. Under this hypothesis, it should be twist, however, downregulated miRNAs are highly enriched
possible to swap the kinase domains on MAP3Ks that phos- for targeting immune genes, indicating that these miRNAs
phorylate the same substrate and recover full function in the probably function as inhibitors of inflammation in the ab-
chimeric proteins. The authors tested this idea with the sence of infection.
MAP3K Slpr, which is required for developmental signaling, In another example of agricultural disease, Connell et al.
and Tak1, which contributes to immune activation via the (2013) conducted a case-control study to determine why
JNK and Imd pathways. Swapping the kinase domains be- some birds in chicken flocks are genetically resistant to Cam-
tween these two proteins results in partial rescue of the plyobacter jejuni infection. Genome-wide mapping reveals
respective mutant phenotypes, but in neither reciprocal di- significant associations between resistance and polymor-
rection does the chimeric protein fully compensate for loss phism in the T-cadherin and calmodulin genes. These
of the native protein. Thus, it seems clear that the kinase associations make sense, as previous work has revealed
domains are not simply phosphorylating targets but are also infectious interaction between C. jejuni and a related E-
potentially involved in interactions with other protein part- cadherin, and calmodulin is a well-known modulator of
ners and certainly contribute to the inherent specificity of cadherin function. The phenotypic distributions of the
the proteins. C. jejuni load in birds carrying either variant are striking,
Two articles in this issue of GENETICS address the role of with some chickens absolutely resistant to infection and
reactive oxygen species (ROS) as signaling and defense mol- others carrying loads of up to 1010 bacteria after inoculation.
ecules. Oxidative radicals are highly reactive, and their cy- The mapped variants appear to be regulatory, and impor-
totoxicity can be harnessed in antipathogen defense. Tiller tantly, the resistant phenotype is determined by the rela-
and Garsin (2014) identified a novel peroxidase, skpo1, that tive—not absolute—expression levels of T-cadherin and
controls ROS production in the C. elegans hypodermal epi- calmodulin, with susceptible chickens expressing an approx-
thelia and correspondingly determines defense against in- imate 25% increase in the ratio of T-cadherin to calmodulin.
fection by the bacterium Enterococcus faecalis. Small et al. With these resistance factors identified, the genetic markers
(2014) found an unexpected pleiotropy between ROS pro- can be employed for selective breeding of resistant flocks.
duction and immune cell differentiation mediated by Notch In contrast, agricultural systems can sometimes be
signaling in Drosophila. Notch signaling promotes the em- engineered for disease resistance, as Subbaiah et al.
bryonic differentiation of crystal cells, which are responsible (2013) have shown by establishing stably transformed silk-
for ROS production in larvae attacked by parasitoid wasps. worms that are resistant to Bombyx mori nucleopolyhedro-
Infection by parasitoid wasps also results in proliferation of virus (BmNPV). The transformed silkworms carry RNAi
lamellocytes in the larval lymph gland. Unexpectedly, Small constructs that target four essential viral genes, generating
et al. (2014) found that Notch signaling regulates this lamel- .75% survival of experimental viral infection relative to
locyte differentiation. It appears that Notch signaling acts in ,15% survival in the parental strain. Even more promising,
a non-cell-autonomous manner in the lymphatic organ to the few viral occlusion bodies derived from the transformed
hold lamelloctye precursor cells in quiescence, but inhibition B. mori are impaired in their ability to infect naive silk-
of Notch by RNAi or parasitoid infection allows lamellocyte worms, further limiting disease spread. As BmNPV can result
differentiation to proceed. This surprising pleiotropy estab- in loss of .50% of commercial silk cocoon yield, the trans-
lishes Notch as a key regulator of alternative immune cellu- genic lines have great potential economic impact.
lar lineages in Drosophila, with important consequence for Natural systems do not lend themselves to easy manip-
the effectiveness of defense against infection. ulation to prevent or limit disease establishment and
spread, and we often struggle to understand what deter-
mines relative susceptibility and resistance in natural
Agriculturally and Ecologically Relevant Systems
settings. Batrachochytrium dendrobatidis (Bd) is a fungal
Infection and immunity are critical in agriculture. For pathogen that is devastating amphibian populations globally,
example, bovine mastitis (infection of the udders) costs and we have a poor understanding of why some species are
the U. S. dairy industry .$1.7 billion per year (Jones and more susceptible than others. Ellison et al. (2014) examined
Baily 2009). In the June issue of G3, Lawless et al. (2014) gene expression profiles in the highly susceptible Panama
correlate the inflammatory response to mastitis caused by Golden Frog after exposure to attenuated fungus. They found
Streptococcus uberis with microRNAs (miRNAs) that are an- strong immune activation, but that does not translate into
ticipated to alter mRNA expression profiles. Monocytes re- effective vaccination or protection from secondary infection.
leased from bone marrow are recruited to the site of Instead, the data suggest that inflammation of the skin might
infection by chemokine-mediated attraction, where they be pathological in the presence of infection and that reducing
switch from oxidative phosphorylation to glycolysis and inflammation could be protective. The transcriptional data

468 B. P. Lazzaro and D. S. Schneider

also reveal upregulation of antibacterial immune defenses in Drosophila, but that the effect is dependent on an intact
frogs infected with Bd, possibly indicating secondary infec- female germline. In an article published in G3, Short and
tion facilitated by the primary fungal infection. Intriguingly, Lazzaro (2013) report a transcriptional analysis of the re-
Ellison et al. (2014) hypothesize that Bd may suppress im- sponse to infection in virgin and mated female Drosophila
mune reactions in susceptible frog species based on their melanogaster with and without germlines. Confirming their
observation that expression of genes diagnostic for B cells previous study, they find that virgin females induce immune
and T cells is reduced at late stages of infection with fully genes more strongly than reproductively active females do,
active Bd, suggesting that the fungus might kill the progen- although the difference in expression patterns between
itors of these cells or block their differentiation. This inter- females with and without germlines is surprisingly small.
pretation is bolstered by the observation of reduced spleen A novel but logical finding is that egg production genes
size in infected frogs. The work of Ellison et al. (2014) sug- show reduced expression after infection in virgins but not
gests that the Panama Golden Frog attempts to resist Bd in mated females, suggesting that females prioritize egg lay-
infection, but that the immune defense is compromised ing over immune defense when given the choice. The tran-
and perhaps itself pathological, resulting in reduced toler- scriptional data also reveal other interesting patterns that
ance of infection. The effective defense observed in some provide fodder for follow-up study, such as signatures of
other amphibian species may then be due to more managed egg maturation in response to mating and altered feeding
immune activation and avoidance of pathogen mechanisms behavior in response to mating and infection.
for immune suppression. Although there is a tendency to treat immune capability
as a static property of the host, immune competence is
a dynamic trait that changes over the host life span.
Complexity in Immunity
Intuitively, we might expect that immune capacity would
Perhaps the most interesting frontier in the genetics of decline with host age, a process known as immunosenes-
immunity arises in the interaction between immune activity cence, and indeed this is observed at advanced ages. Felix
and other physiological or developmental processes. Such et al. (2012), however, have shown that capacity to clear
interactions must shape the balance of traits in an organism, nonpathogenic Escherichia coli infection may increase or
determining overall health in the context of infection. Evolu- decrease between young and middle ages (1 and 4 weeks
tionary and functional studies in this domain are of particular post-eclosion adults), although the magnitude of variation
interest in the GENETICS/G3 emphasis on the genetics of im- among genetic lines increases at the older age. They also
munity, and a few such articles are highlighted below. evaluated genome-wide gene expression in a panel of in-
Johnston et al. (2014) applied transcriptional profiling bred D. melanogaster lines at 1 and 4 weeks of adult age.
in a time series after immune challenge of the mealworm Consistent with previously published articles, Felix et al.
beetle Tenebrio molitor. They found that different compo- (2012) noted a tendency for immune gene expression to
nents of the immune response have different timings of increase with age, even in the absence of infection, and
expression, which indicates complex and specific regula- again found repressed expression of genes involved in lipid
tory control. The authors emphasize that expression of metabolism after infection of flies of both ages. Unexpect-
genes encoding the Toll pathway, antimicrobial peptide edly, however, they found that the genetic architecture of
genes, and iron sequestration processes remain upregulated immune capacity (the correlation structure between clear-
a week after infection, although their experiments do not ance of infection and genome-wide transcription) differed
exclude the possibility that residual bacteria may continue to between the two ages, with a stronger relationship between
stimulate immune activity. Most notably, however, infectious energy metabolism and immune capacity in older flies.
challenge in Tenebrio results in repression of genes involved
in glucose metabolism and biosynthesis of lipids, triglycer- Continued Emphasis on the Genetics of Immunity
ides, and vitamins. These data parallel an early Drosophila
microarray study in which benign immune challenge was GENETICS and G3 continue to seek submissions of manu-
also found to suppress transcription of basal metabolic genes scripts on the genetics of immunity, and both journals will
(De Gregorio et al. 2001) and support the hypothesis that continue to highlight articles published in this area. Topics
immunological activity is energetically costly. At the same of particular interest for future publication include:
time, insects have been reported to reduce feeding in re- Elucidation of genetic mechanisms leading to tolerance
sponse to infection (e.g., Adamo et al. 2007; Ayres and of, or resistance to, infection
Schneider 2009), and an alternative, and not mutually ex-
Genetic or signaling networks that affect immune system
clusive, interpretation is that hosts alter metabolism to se-
function
quester nutrients as part of a nutrient restriction strategy to
manage pathogen growth. Genetic responses to environmental factors that modu-
Reciprocally, immunity may be constrained by competing late resistance and tolerance
demands on the host. Short et al. (2012) have shown that Genetic interactions among hosts, pathogens, and sym-
mating and reproduction limit immune competence in bionts that shape infection outcomes

Commentary 469
 Defense responses stimulated by host tissue damage Felix, T. M., K. A. Hughes, E. A. Stone, J. M. Drnevich, and J. Leips,
Neuroimmunology and behavioral immunity 2012 Age-specific variation in immune response in Drosophila
melanogaster has a genetic basis. Genetics 191: 989–1002.
Mechanisms for specific recognition or memory in Johnston, P. R., O. Makarova, and J. Rolff, 2014 Inducible de-
invertebrates fenses stay up late: temporal patterns of immune gene expres-
sion in Tenebrio molitor. G3 4: pii: g3.113.008516v1.
The genetics of immunity is an area of considerable
Jones, G. M., and T. L. Bailey, Jr., 2009 Understanding the basics
research opportunity, particularly where conventional im- of mastitis. Virginia Coop. Ext. Available at: http://pubs.ext.vt.
mune pathways intersect with other aspects of host physi- edu/404/404-233/404-233.html.
ology and function. We look forward to reading about the Lawless, N., T. A. Reinhardt, K. Bryan, M. Baker, B. Pesch et al.,
next set of advances in the field. 2014 MicroRNA regulation of bovine monocyte inflammatory
and metabolic networks in an in vivo infection model. G3 4: pii:
g3.113.009936v1.
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470 B. P. Lazzaro and D. S. Schneider