1

Insect Immune Priming: Ecology and

Experimental Evidence

3
4

Jorge Contreras-Garduo1,4*, Humberto Lanz-Mendoza2, Bernardo Franco1,

Adriana Nava3, Mario Pedraza-Reyes1 & Jorge Canales-Lazcano3

6
71 Departamento de Biologa, Divisin de Ciencias Naturales y Exactas, Universidad de
8Guanajuato, campus Guanajuato. Noria Alta s/n, Noria Alta, 36050. Guanajuato, Guanajuato.
9
102 Centro de Investigacin Sobre Enfermedades Infecciosas, Instituto Nacional de Salud Pblica,
11Av. Universidad 655, C. P. 62508. Cuernavaca, Morelos, Mxico.
12
133 Posgrado en Ciencias Biomdicas, Instituto de Ecologa, UNAM. Ciudad Universitaria, Mxico
14D.F.
15
164 Present address: ENES, UNAM, unidad Morelia. Antigua Carretera a Ptzcuaro No.8701. Col.
17Ex-Hacienda San Jos de la Huerta. C.P. 58190. Morelia, Michoacn.
18
19Correspondence: jcont@ecologia.unam.mx
20
21Key words: immune priming, immune specificity, immune response, evolutionary immunology,
22ecoimmunology, immune memory.

3
23Abstract
24
251.

Immune priming refers to improved protection of host after a second

26encounter

with the same parasite or pathogen. This phenomenon holds

27similarities

to adaptive immunity in vertebrates.

282.

Evidence suggests that this improved protection can be species/strain

29specific

and protects organisms a lifetime. These two elements along with a

30biphasic

immune response are essential characteristics of immune priming and

31is

the basis for the effectiveness of resistance to parasites and pathogens.

323.

Consideration is given to the effect of immune priming within and across

33generations,
34and
354.

the influence of heterologous challenge during immune priming

the importance of testing the immune response with natural pathogens.

Current analysis encompasses the multifaceted nature of the invertebrate

36immune

response. We thus discuss the scarce evidence suggesting that

37bacterial

microbiome plays a complementary role on immune priming outcome.

385.

Finally, another scantly explored issue is the cost of immune priming, which

39might
40

explain why evidence for immune priming is not always supported.

5
41Introduction
42
43Organisms
44problems.
45but
46it

of different lineages have analogous solutions to confront similar

For example, to move from one place to another, organisms may fly

the structures to fly are different in bats, birds and insects. In the same way

is possible that phenomenological analogous aspects of immune memory in

47vertebrates
48structures

and invertebrates could evolve but with different molecules and

(Kurtz & Armitage 2006) to reach an optimal immune response (the

49highest

benefits with the lower costs). Hence, immune memory should be

50favored

in long-lived animals (Boots et al., 2013; Garnier et al., 2014) as well as

51in

short lived species exposed continuously to the same parasites or pathogens

52(Liu,

2000; Little et al., 2005).

53

Historical evidence that suggests the possible existence of memory as

54
55part

of the innate immune response of invertebrates dates back to the 1930s. In

56that

decade, studies revealed that insects exhibit enhanced survival after

57infection
58or

if they were previously challenged with the same or a similar pathogen

parasite species (see Brey, 1998). This observation remained dormant until

59empirical
601986;

evidence obtained in the1980s and early 1990s (Cooper & Roch,

Hartmann & Karp, 1989; Faulhaber et al., 1992) proposed that analogous

61mechanisms
62vertebrates
63the

of immune memory should be studied in invertebrates and

(Karp, 1990; Cooper, 1992). However, it was until the first decade of

21st century that new evidence supported this hypothesis and now has

64become

a theory. In a seminal paper, immune priming was clearly

65demonstrated

in the Maxillopoda Macrocyclop salbidus against the Cestoda

7
66Schistocephalus
67experimental
68latter

groups, were one was challenged with S. solidus and two days

challenged with either a genetically similar parasite (full-sibs) or non-

69genetically
70success
71than

solidus (Kurtz & Franz, 2003). Research conducted used two

similar parasite (non full-sibs), resulting in reduced reinfection

of S. solidus and less host intensity of reinfection in the former group

the latter (Kurtz & Franz, 2003).

72

This phenomenon is termed immune priming to distinguish the innate

73

74memory
75(Little

from the mechanistically different adaptive immunity in vertebrates

& Kraaijeveld, 2004). Initially, the lack of physiological and molecular

76evidence
77&

Smith, 2007). However, it was suggested that innate memory should not be

78mediated
79at

to support the immune priming concept generated skepticism (Hauton

through the same mechanisms in different taxa but should be similar

a phenomenological and organism level, since in both invertebrates and

80vertebrates
81&

self and non-self discrimination has a strong impact on fitness (Little

Kraaijeveld, 2004; Little et al., 2008). Now, immune priming is studied in

82vertebrates
832005;

(Sun et al., 2009; Paust & Andrian, 2011), invertebrates (Kurtz,

Schmid-Hempel, 2011; Masri & Cremer, 2014) and plants (Spoel & Dong,

842012).
85
86

Classic studies of immune priming have recorded host immune

87response,

parasite clearance and/or host survival within or across generations

88(Table

1 and 2). Here particularly we explore: (a) immune priming components

89within

and across generations, (b) the contribution of bacterial microbiota to

90immune

priming effectiveness, (c) costs of immune priming, and (d) the

9
91proposed
92in

lack of evidence for immune priming. In Box 1 we define key aspects

immune priming. Only a summary is herein given of the reported

93observations
94reviews
95insects

regarding mechanisms of immune priming due to the excellent

and research papers on this topic (Box 2). This review is focused on
because they are the most represented group in studies related to

96immune

priming (Tables 1 and 2).

97
98Immune

priming and experimental evidence

99
100For

considering immune priming fully analogous to immune memory in

101vertebrates,
102(Kurtz,

2005), (2) long-lasting protection (Kurtz, 2005), and (3) a biphasic

103immune
104levels

three attributes must be fulfilled: (1) specific immune resistance

response, increasing after the first challenge and returning to basal

and increasing again to a greater degree after the second challenge

105(Kurtz,

2005; Brehlin & Roch, 2008; Schmid-Hempel, 2011).

106

Regarding the first attribute, immune priming can be highly specific: host

107
108may

distinguish between parasite siblings or non-genetically related (Kurtz &

109Franz,

2003) or between strains or species (Table 1; Table 2). These scenarios

110reveal

that immune priming can be specific if the host is confronted twice with

111homologous
112a

heterologous challenge between two homologous challenges. Under natural

113conditions
114and

species are confronted by different immune challenges with parasites

pathogens (Schmid-Hempel, 2011) and if heterologous challenges occur

115between

10

challenges. However, a topic that remains untested is the effect of

homologous challenges, the increased immune response due to the

11
116second
117after

homologous challenge could be due to a sustained immune response

the heterologous challenge rather than a specific immune response. In the

118simplest

form, immune priming (specificity, long lasting and biphasic immune

119response)
120two

should occur even when a heterologous challenge occurs between

homologous challenges. This challenging condition will reveal the degree of

121specificity, plasticity, and

122For

example, the exposure to one pathogen twice, let us suppose Bacillus

123thuringiensis,
124marcescens
125the

128be

(B. thuringiensis-S. marcescens-B. thuringiensis), because during

and the memory of the innate immune system should better respond

the parasite or pathogen from homologous challenge. Some caution should

taken because the outcome of these results should depend on the time

129elapsed
130it

should not be affected by an intermediate challenge with Serratia

immune memory process different recognition molecules should be

126activated
127to

the ability to generate priming against multiple stimuli.

between challenge and parasite dose (i.e. Wu et al., 2015). In addition,

is possible that an heterologous challenge could interfere with the optimal

131response
132immune

of an homologous challenge and this could be a limitation for the

priming outcome.

133
134

Immune priming occurs not only within a generation but also across

135generations.
136immune
137protect
138there

priming across generations (Table 2). For example, parents may

their offspring against a pathogen that the parent has encountered and

is good evidence this phenomenon could be also highly specific, for

139example

12

This phenomenon is called transgenerational immune priming or

parasite species-specific (Roth et al., 2010). However, there is no

13
140evidence

about how multiple or single heterologous challenge between

141homologous

challenges can affect immune priming across generations.

142

The second attribute, a long-lasting enhanced resistance, have revealed

143
144that

an improved immune response may persist for a life-time in animals

145previously
146(Table
147effect

exposed to a pathogen thus spanning different developmental stages

1). Regarding immune priming across generations and its protective

on offspring in terms of immune response, host survival, and/or parasite

148clearance

has been found in eggs, larvae and adults; such protection could be

149inherited

from mothers and/or fathers to the offspring (Table 2). Regarding

150maternal

effect as a form of immune investment, mollusk females transfer

151antimicrobial
1522015).

peptides, lectins, and/or lytic enzymes into eggs (Wang et al.,

However, less is known about the role of fathers. A recent report

153demonstrated
154could

contribute to the offspring protection (Eggert et al., 2014). However, a key

155unresolved
156the

that spermatozoids and substances transferred in seminal fluid

question on this topic of long-lasting protection is how and where

immune response memorizes the information about a previous challenge

157(see

Box 2 for potential mechanisms). On this regard, one possibility is an

158epigenetic

mechanism involved in immune priming within and across

159generations
160evidence

(Jokela, 2010; Eggert et al., 2014; Ottaviani, 2015). There is

that epigenetic changes favor parental effects (Bossdorf et al., 2008;

161Norouzitallabet
162elicit
163as

14

al., 2014) and general stressors and not only parasites may

immune priming (Eggert et al., 2015). However, to our knowledge there are

yet no reports, either for males or females showing their role based on

15
164epigenetic

mechanisms related to immune priming within and/or across

165generations.
166

Another unsolved question within immune priming across generations is

167

168regarding
169last.
170virus
171In

to how many generations the transgenerational immune priming may

Plodia interpunctella challenged against Plodia interpunctella granulosis

protection was observed in F2 but not F3 generation (Tidbury et al., 2011).

Tenebrio molitor, priming generated against lipopolysaccharides (LPS) of

172Escherichia
173be

coli persisted for two generations (Moret, 2009). Differences could

explained because in the former study live pathogens were used and it is

174possible
175may

that under this conditions parents experienced an elevated cost which

lead to invest in the present generation causing costs to the following ones

176(Tidbuty

et al., 2011). However, Artemia challenged against live Vibrio

177campbellii,
178offspring
179studies

immune priming phenomenon last for 3 generations even that

were nave to bacterial challenges (Norouzitallab et al., 2015). More

are needed to investigate why immune priming last for one or more

180generations
181related

and the underlying molecular mechanisms and ecological traits

with this phenomenon.

182
183

Finally, regarding the third trait to be established as a functional

184mechanism
185memory

and immune priming, it is necessary to demonstrate a biphasic

186response
187a

and formally consider it as an analogy between vertebrate immune

(Kurtz, 2005; Brehlin & Poch 2008; Schmid-Hempel, 2011). Although

biphasic immune response could occur with both homologous (specific) and

188heterologous

16

(non-specific) challenges, only in the first case immune response

17
189should

be stronger and/or faster in the second challenge (Schmid-Hempel,

1902011).

This question has been practically unexplored in spite of being

191suggested
192about

10 years ago (Kurtz, 2005). As far as we know, the only evidence

a biphasic immune response during immune priming was found in

193Anopheles

albimanus against Plasmodium berghei (Contreras-Garduo et al.,

1942015). Authors
195and

found that levels of antimicrobial peptides (attaccin, cecropin

gambicin) increased after first exposure to pathogen, decreased to basal

196levels

with pathogen clearance and increased again after a second challenge

197(Contreras-Garduo
198existence

et al., 2015). This biphasic response suggests the

of a memory mechanism associated to subsequent exposures to a

199given

pathogen (Brehlin & Roch, 2008; Kurtz, 2005; Schmid-Hempel, 2011).

200More

studies are needed to answer if the biphasic response is maintained

201across

generations and whether this phenomenon is widespread in other host-

202parasite

interactions.

203

If specificity, long-lasting protection and a biphasic response are

204

205confirmed

in an insect species, its immune priming could be considered fully

206analogous

to vertebrate immune memory. If not all scenarios coexist in a single

207species,

we should ask for example how biotic or abiotic factors determine the

208immune

priming outcome (see below). It is very important to take into account

209that
210et

maximum immune response is not necessarily an optimal response (Viney

al., 2005), and for some species depending on their ecological circumstances

211and

internal physiology, immune priming could show more costs than benefits,

212favoring
213of

18

other optimal responses (i.e. sustained response, tolerance or the use

non-immunological barriers) than immune priming. Hence, those organisms

19
214that

exhibit immune priming or other strategies as well as the selective forces

215behind

them, will be evidence to understand why the effectiveness of immune

216priming

is low or absent.

217
218Bacterial

microbiota and its importance for immune priming outcome

219
220For

many years, bacteria have been widely studied from the host-parasite

221relationship
222provide
223Weiss

point of view. Recent studies have revealed that they may also

benefits to their hosts (Brownlie & Johnson, 2009; Feldhaar, 2011;

& Aksoy, 2011; Thaiss, 2014). For example, it was assumed that insects

224humoral

and cellular immune responses were the only mechanisms against

225parasites

and pathogens but recent studies have found a key role of bacterial

226microbiome
227(Feldhaar

to boost immune response and participate in killing hosts enemies

2011; Weiss & Aksoy, 2011; Eleftherianos et al., 2013).

228
229

Regarding adaptive immunity in mammals, an association has been

230made

between poor mechanistic diversity for elimination of bacteria and

231reduction
232should

of B cells, T cells or secretory IgA during exposure, which in turn

be different in invertebrates (Kato et al., 2014). Also, it has been

233proposed

that bacterial microbiome may play a key role on immune priming

234within

and across generations (Rodrigues et al., 2010; Freitak et al., 2014).

235Within

generations, in primed Anopheles mosquitoes treated with antibiotics, an

236increase

in Plasmodium parasites and lower phagocytic activity was reported in

237contrast

with primed controls without antibiotic treatment (Rodrigues et al.,

2382010).

20

One proposed mechanism by Rodrigues et al. (2010) is that midgut

21
239bacteria

could boost immune response. Hence, authors suggest that midgut

240bacteria

could play a key role on the immune priming outcome. However, no

241support

of specificity, long lasting and biphasic response will reveal that bacteria

242themselves
243pathogens

and not the immune priming could be responsible of parasites or

elimination.

244
245

In Anopheles albimanus challenged against Plasmodium berghei did not

246reveal

a key role of bacteria for immune priming, as the immune priming

247outcome
248of

was not affected by the elimination of midgut bacterial load by the use

antibiotics (Contreras-Garduo et al., 2015). Hence, the role of bacteria

249demand

further research to know how, why and in which species or host-

250parasite

interactions bacterial microbiome could play a role on the immune

251priming
252(NGS)

outcome. Using novel strategies such as Next Generation Sequencing

could provide novel insights into the bacterial population inside

253invertebrates

and their possible role in immune priming (Morgan et al., 2014).

254

Two reports address transgenerational immune priming, and both

255

256suggest

a key role of bacterial microbiome on its outcome. Firstly, in S. exigua

257offspring,
258against

B. thuringiensis was provided by mothers that vertically transmitted

259bacteria
260larval

(Enterococcus) to their eggs, thus favoring protection that lasted until

stages (Hernndez-Martnez et al., 2010). In this same study, the

261treatment
262load,
263et

22

the main contribution to the enhanced immune response (lytic activity)

of females with antibiotics decreased their eggs and larvae bacterial

lytic activity and resistance against B. thuringiensis (Hernndez-Martnez

al., 2010). Secondly, glorverine and prophenoloxidase were induced in eggs

23
264laid

by G. mellonella females challenged with P. entomophila or S. entomophila

265but

were absent or reduced in laid eggs derived by control mothers (Freitak et

266al.,

2014). Interestingly, authors also discovered these bacteria were able to

267penetrate
268body

in pupae. Once females were adults, these bacteria were deposited in the

269chorion

or the egg yolk, thus allowing the eggs to increase their immune

270response
271load

the gut and enter larvae haemocoel and then were attached to the fat

(Freitak et al., 2014). This last study provides evidence that bacterial

acts as a co-adjuvant inside the invertebrate body and could boost the

272immune

response suggesting a little effect on the three key hallmarks of

273immune

priming.

274

Another important avenue for research could be the contribution of

275

276primary

and secondary endosymbionts for immune priming. As reported by

277Hernndez-Martnez
278for

and collaborators (2010) a role of primary endosymbionts

immune priming is found. Primary endosymbionts are those vertically

279transmitted,
280years

(Eleftherianos et al. 2013). Secondary endosymbionts are those

281transmitted

horizontally, vertically or via the environment that include

282commensal,
283hemocoel

285immune

evolved relationships that are more recent and can be found in the

(Eleftherianos et al. 2013). Some questions that arise from this

284observation

286than

obligate, and with which the insects have co-evolved for millions of

are: how do primary, secondary or both microbes contribute to

priming? Are primary microbes more important for immune priming

secondary? Which is the contribution of bacteria living in the haemocele or

287intestine

for immune priming and this effect vary accordingly to the way the

288parasite

or pathogen invade the host? For example, does gut microbiota only

24

25
289affect
290by

the priming response to orally-infecting parasites or can it also be affected

hemocoelically-introduced priming parasites (or vice versa)? Perhaps being

291more

precise about these questions will reveal some general patterns about the

292host-parasite
293composition
294example

of microbiota is important for immune priming but it varies for

according to diet and among populations, this could also have

295implications
296of

interaction and microbiome during immune priming. Finally, if the

for priming and explain why it varies. Perhaps a core microbial set

commensals are permanently present that maintains homeostasis and allows

297immune

priming.

298

The greater diversity of bacteria observed under natural rather than

299

300laboratory

conditions could affect immune response (Hurst & Hutchence, 2010).

301Moreover,

hosts are confronted with a variety of enemy species (Schmid-

302Hempel,

2011) creating a multifactorial interaction in the gut between

303microbiome
304been

and parasites/pathogens. These complex interactions have not yet

analyzed in relation to immune priming. Insights in this sense should

305certainly

contribute to our understanding of the cross-talk between microbiome

306(parasitism,
307whether

mutualisms and commensalism) and immune priming. Also,

parasites or pathogens avoid the outcome of immune priming by

308interfering

with its mechanisms is an open question (see Evidence against the

309existence

of immune priming).

310
311

Finally, the most challenging difficulty to study any microbiome is the

312capability
313or

26

to isolate and grow under laboratory conditions microorganisms inside

outside any living organism. However with novel bioinformatic and statistical

27
314tools

(Morgan & Huttenhower 2014; West et al., 2015), becomes easier

315identifying
316immune

dysbioses (changes in the normal microbiota content) between

primed insects with respect with control. Analyses that identify

317perturbations
318their

in molecules related with immune priming can be used to interpret

physiological role, interaction with truly pathogenic organisms or integrate

319sequence

data with whole-community relationships (i.e. Morgan & Huttenhower,

3202014).
321
322The

cost of immune priming

323
324Evolutionary
325organisms
326which

biologists have alerted that immune response could vary between

and species due to evolutionary costs (negative impacts on fitness),

may limit immune response performance (Rolff & Siva-Jothy, 2003;

327Schmid-Hempel,
328immune
329well

2003, 2005). For example, activation and development of

response (Rolff & Siva-Jothy, 2003; Schmid-Hempel, 2003, 2005) as

as better resistance across generations (Kraaijeveld & Godfray, 1997)

330could

be limited by costs and may also depend on costs generated by food

331resources

and parasites (Valtonen et al., 2010; McKean & Lazzaro, 2011).

332Regarding

adaptive immunity there are few studies documenting cost. For

333example,

it is known that two weeks of food restriction in mammalian

334Peromyscus
335for

immune memory (Martin II et al., 2008). In fish (Poecilia reticulata) acquired

336memory
3371998).

was traded-off with courtship behavior and female preference (Lpez,

In invertebrates there are few studies reporting the cost of immune

338priming

28

maniculatus resulted in the reduction of several factors important

(Little & Kraaijeveld, 2004; Tables 1 and 2) and there is no information

29
339indicating
340within
341role

whether the effectiveness (i.e. parasite killing) of immune priming

and across generations is influenced by the quantity/quality of food, the

of sexual selection (i.e. courtship) or parental care (i.e., if cryptic parental

342care

could vary between males and females: Jokela, 2010).

343

Mathematical models have suggested that immune priming within a

344

345generation
346but

is costly (Tate & Rudolf, 2012; Tidbury et al., 2012; Bets, et al., 2013)

to our knowledge there are only two studies that have supported this

347assumption

within generations because improved immune priming impaired

348development
349that

and reproduction (Table 1). However, one of such studies revealed

not all traits regarding reproduction were impaired (egg number was not

350affected

but egg hatching was affected) suggesting a hidden trade-off between

351immune

priming and egg production (Contreras-Garduo et al., 2014), and that

352different

components of life history and fitness should be taken into account to

353support

or discard how costly the immune priming is. Further research is

354required

to provide greater insight into how costs determine immune priming

355occurrence

and maintenance within a generation.

356

Immune priming across generations may incur into costs for

357

358development,
359Zanchi

et al., 2011) but the impact of such costs for the offspring sexual

360selection
361costs
362the
363a

30

body size and reproduction (Sadd & Schmid-Hempel, 2009b;

has not been tested or the contribution of mothers or fathers to such

has been rarely tested despite a differential effect of mothers or fathers on

cost paid by their offspring (Zanchi et al., 2011). Another study did not reveal

lower developmental time, but the opposite in offspring derived from priming

31
364versus
365the

control parents (Tate & Graham, 2015) and this effect was dependent on

offspring birth order after their parents were challenged under laboratory

366and

wild conditions (Tate & Graham, 2015). This study highlights the question of

367whether

offspring may confront differential costs (and benefits) depending of

368birth

order after inducing into their parents immune priming. Another important

369point

of this study is that authors found that if hosts confronted a different

370parasite

(gregarines) during their immune priming with B. thuringiensis their

371offspring

do not show enhanced resistance against the same bacteria

372compared

to offspring whose parents only received B. thuringiensis. This means

373that

gregarines inhibited the immune priming or that offspring paid their parents

374cost

for confronting multiple enemies during immune priming (Tate & Graham,

3752015).

These hypotheses grant further research.

376

Hence, evidence shows that immune priming across generations is

377

378costly, although
379Understanding
380pathogens
381species

this line of research is only beginning to show definite patterns.

both costs and benefits as well as virulence of parasites and

will give insights into the distinct effects of priming in different

and how organisms respond optimally against immune challenges. In

382the

case of species in which immune priming is absent, it may be found that it is

383too

costly or that hosts are not constantly affected by the same parasitic species

384and

according with the latter idea it would be likely to find immune priming in

385colonial
386is

species than in solitary and wide dispersed species, if immune priming

too costly, the optimal response could not involve immune priming, but a

387sustained
388

32

response or tolerance.

33
389Evidence

against the existence of immune priming

390
391Within

generations

392
393Several

recent studies support the idea of immune priming concept, some

394evidence
395et

is apparently contradictory for some but not all challenges (i.e. Pham

al., 2007; Roth et al., 2009), but also there are some research papers that

396claim

no support at all for immune priming (Table 1 and 2). Here we review the

397available

evidence of immune priming within generations (Table 1). For

398instance,

it has been suggested that immune priming does not occur in male

399damselflies
400insects

primed with heat-killed Micrococcus lysodeiktikus did not show better

401survival
402M.

(Hetaerina americana). In this study (Gonzlez-Tokman et al., 2010)

than nave animals or controls after all groups were challenged with live

lysodeiktikus. However, in this study it was also shown that wound repair and

403manipulation
404study

did not incorporate natural parasites (i.e., gregarines or mites, Corbet,

4051992),

and thus did not consider the hypothesis that immune priming could be

406expected
407This

is harmful, so immune priming cannot be discarded. Moreover, this

using natural enemies but not novel pathogens (Roth et al., 2009).

hypothesis has not been explicitly tested and papers within and across

408generation
409been

that has used both natural parasites and novel challenges have not

able to show clear response patterns (Tables 1 and 2). However, studies

410should

take into account this possibility if their results may not to support the

411immune

priming hypothesis and it is necessary to do more field-based immune

412priming

studies with natural parasites to understand the importance of immune

413priming

in nature (Tate & Graham, 2015).

34

35
414
415

There is a controversy about the role of bacteria on immune priming

416within

generations. For example, adult ants of Formica selysi confronted with

417Beauveria
418tested

bassiana, a natural enemy, did not show immune priming under the

conditions (Reber & Chapuisat, 2012). However, another study found

419support

of immune priming in larvae of Camponotus pennsylvanicus

420(Rosengaus
421immune
422studies

et al., 2013). This may suggest that for some insect species

priming occurs in larvae but not in adults. However, there are no

testing this in within single species. Finally, in G. mellonella priming

423larvae

with dead Photorhabdus luminescens or B. thuringiensis protected the

424larvae

against these same live bacteria in homologous or heterologous

425challenges

as well as against the nematodes Heterorhabditis bacteriophora or

426Steinemema
427was

carpocapsae (Wu et al., 2014). This suggests that immune priming

not specific and hence should not be considered as evidence of innate

428memory
429from

12 h to 7 days, but not for 14 days and was influenced by initial priming

430doses.
431using

This report is important because alert about the possibility that even

natural pathogens in the experimental designs, priming could be non-

432specific
433seen

but immune enhancement. This immune enhancement was effective

and that the time elapsed between a first and second challenge (as

in vertebrates) as well as doses at the first challenge may affect the

434immune

priming outcome (Wu et al., 2015).

435
436Across
437

36

generations

37
438Evidence
439found

against the existence of immune priming across generations has been

in recent articles (Table 2). In Anopheles coluzzii, primed mothers with P.

440falciparum
441(Vantaux
442immune
443primed

et al., 2014) and authors proposed that mothers that invested for

response had a cost shown by their daughters resistance or that

females produced descendants of lower quality than control females

444(Vantaux
445show

et al., 2014). In another study, mothers of Myzus persicae did not

immune priming against the parasitoid Diaeretiella rapae (Vorburger et al.,

4462008).

Mothers that survived the attack had fewer offspring than controls

447suggesting
448been

that the resistance was very costly and that females may not have

able to devote enough resources for enhanced offspring resistance.

449Although
450Via,

did not favor their daughters resistance against the same parasite

the immune response is not well understood in this taxon (Henter &

2005), it is possible that an evolutionary arms race is occurring between the

451parasitoid

and host, meaning that the successful parasites may avoid or

452suppress

host resistance. From an evolutionary point of view, it is assumed that

453virulence

factors could favor overriding host immune priming. Hence, the

454virulence

of parasites and pathogens as well as its implications for the costs of

455immune

priming should be taken into account (Schmid-Hempel, 2005a; Tate &

456Graham,
457and

2015). In this sense, it may be useful to control parasite species used

test different levels of virulence. In vertebrates theory predicts that

458increased
459in

invertebrates it has been suggested that virulence favors such acquisition

460(Best
461has
462

38

virulence impairs acquired memory (Boots & Bowers, 2004) whereas

et al., 2013). As far as we know, the role of parasite or pathogen virulence

yet to be tested providing another important area for further research.

39

The lack of evidence of immune priming could be due to a bias towards

463

464publishing

and probably much more evidence have been found but is not

465published

yet. We encourage the publication of negative results to have a better

466understanding

of boundaries and effectiveness of primed immunity.

467
468Concluding

remarks

469
470There

are still only few studies that have tested natural enemies or at least

471parasites
472of

affecting the same taxa and these have been carried out in a handful

species (Tables 1 and 2). The results suggest that there are analogous

473aspects
474which

of immune memory in vertebrates and immune priming in invertebrates,

is consistent with the widespread evidence that organisms of different

475lineages
476(Kurtz

have often developed analogous solutions to face similar problems

& Armitage 2006).

477
478

Three phenomenological components have been established for immune

479priming:

the specificity of enhanced resistance, a long-lasting protection, and

480biphasic

response. On this regard, a key question is whether invertebrates

481present
482the

a biphasic or sustained immune response, which in turn may depend on

particular species and a variety of other influences. For example, recent

483mathematical
484individuals)
485parasite

could be influenced by specific parasite-host relationship, multiple

interactions, season of the year, sex, developmental stage and

486virulence

of specific parasites or pathogens, the proportion of susceptible

487individuals

40

models indicate that immune priming for populations (versus

and the population age and size (Tate & Rudolf, 2012; Tidbury et al.,

41
4882012).

However, few studies have addressed these questions experimentally

489(Thomas
4902014;
491of

et al., 2010; Tate et al., 2012; Rosengaus et al., 2013; Garbutt et al.,

Tate & Graham, 2015). Additionally, there are scant reports on the costs

immune priming, which when taken into account may not only shed light on

492the
493and

variations found experimentally between species, but also on its population

epidemiological consequences (Tate & Graham, 2015). Finally, in

494laboratory
495strong
496affect

diet is provided ad libitum but in nature the scarce of resources is a

selective pressure that impose variability and it is possible that may

the immune priming outcome.

497

There are several factors found in nature that are often not adequately

498

499reproduced

experimentally and hence, studies in laboratory should be validated

500in

nature and this is not only the case of immune response in general (Lazzaro

501et

al., 2004) but also for immune priming (Tate & Graham, 2015). For example,

502in

nature it is presumably common for invertebrates to encounter a great

503quantity

and diversity of parasites and pathogens and they have a diverse

504microbiome,
505immune

which may favor a tripartite relationship between microbiome, host

priming, and pathogen virulence.

506
507

Another important factor in nature is the presence of natural parasites or

508pathogens,

which could be more likely to stimulate immune priming than novel

509challenges

(Roth et al., 2009), but this hypothesis has not been formally tested

510and

the present studies do not show clear evidence to answer this question

511because

natural parasites or pathogens and novel challenges seems to favor

512immune

priming. Additionally, in nature it is presumably common for

42

43
513heterologous
514only

challenges to occur between homologous challenges, and there is

limited information about multiple infections on immune priming outcome

515(Sadd

& Schmid-Hempel, 2009b; Tate & Graham, 2015). Future research

516employing
517natural

invertebrates and their natural pathogens under laboratory and

conditions, taking the aforementioned variables into account, should

518certainly

provide greater insights into the possible functional analogy between

519immune

priming in invertebrates and immune memory in vertebrates.

520

Finally, immune priming may lead to the use of new vaccines to reduce

521

522infection
5232014;
524e.g.,
525a

in economically important invertebrates used as food source (Lin et al.,

Valdez et al., 2014) and/or to test better strategies for biological control,

using more than one natural enemy per season and/or avoiding the use of

pathogen with a relatively low virulence that could increase resistance in

526target
527pest

species, because this phenomenon illustrates that for example insect

will develop resistance within and across generations.

528
529Acknowledgements
530
531To Dr. Klaus
532reviewers
533priming

kindly improved our manuscript, particularly the section of immune

components and the contribution of bacterial microbiota to immune

534priming.
535review.

To Evan Fairn whose helpful comments improved and early draft of this

JCG was financed by CONACyT (grant no. 19660) and the Apoyo

536Institucional
537

44

Reinhardt for his kind invitation and patience. Two anonymous

para la Investigacin (Universidad de Guanajuato 2014).

45
538References
539
540Armitage,
541immune

S. A., Peu, R., & Kurtz, J. (2015). Dscam and pancrustacean

memory: a review of the evidence. Developmental & Comparative

542Immunology,
543Barrangou,

R. & Marraffini, L. A. (2014). CRISPRCas systems:

544prokaryotes
545Best, A.,
546of

48, 315-323.

upgrade to adaptive immunity. Molecular Cell 54, 234-244.

Tidbury, H., White, A., & Boots, M. (2013). The evolutionary dynamics

within-generation immune priming in invertebrate hosts. Journal of the Royal

547Society, Interface,
548Bossdorf,
549Ecology
550Bolte,

6, 1742-5662.

O., Richards, C. L., & Pigliucci, M. (2008). Epigenetics for ecologists.

Letters, 11, 106-115.

S., Roth, O., Philipp, E. E., Saphrster, J., Rosenstiel, P., & Reusch, T. B.

551(2013).

Specific immune priming in the invasive ctenophore Mnemiopsis leidyi.

552Biology

Letters, 9, 20130864.

553Boots,

M. & Bowers, R. G. (2004). The evolution of resistance through costly

554acquired
555271,

715-723.

556Boots,
557of

immunity. Proceedings of the Royal Society B: Biological Sciences

M., Donnelly, R., & White, A. (2013). Optimal immune defence in the light

variation in lifespan. Parasite immunology, 35, 331-338.

558Brehlin,
559immune

M., & Roch, P. (2008). Specificity, learning and memory in the innate

response. Invertebrate Survival Journal, 5, 103-109.

560Brey, P.T. (1998).

561Molecular
562D.

46

The contributions of Pasteur school of insect immunology.

mechanisms of immune responses in insects. Brey, P.T. & Hultmark,

(eds.). Chapman & Hall.

47
563Brownlie,
564insect

J. C., & Johnson, K. N. (2009). Symbiont-mediated protection in

hosts. Trends in microbiology, 17, 348-354.

565Buchon,

N., Silverman, N., & Cherry, S. (2014). Immunity in Drosophila

566melanogaster
567Nature

Reviews Immunology, 14, 796-810.

568Cannistra,
569function

S. A., & Griffin, J. D. (1988). Regulation of the production and

of granulocytes and monocytes. Seminars in Hematology, 25, 173-88.

570Cirimotich,
571Mulenga,
572to

[mdash] from microbial recognition to whole-organism physiology.

C.M., Dong, Y., Clayton, A.M., Sandiford, S.L., Souza-Neto, J.A.,

M., & Dimopoulos, G. (2011). Natural microbe-mediated refractoriness

Plasmodium infection in Anopheles gambiae. Science, 332, 855-858.

573Cong,

M., Song L., Wang, L., Zhao, J., Qiu, L., Li, L., & Zhang, H. (2008). The

574enhanced

immune protection of Zhikong scallop Chlamys farreri on the

575secondary

encounter with Listonella anguillarum. Comparative Biochemistry

576and

Physiology B., 151, 191-196.

577Contreras-Garduo,
578Alvarado,
579Meza,

J., Rodrguez, M. C., Martnez, S. H., Barnetche, J. M.,

A., Izquierdo, J., Herrera-Ortiz, A., Moreno-Garca, M., Velazquez-

M.E., Valverde, V., Argotte-Ramos, R., Rodrguez, M.H., & Lanz-

580Mendoza,

H. (2015). Plasmodium berghei induced priming in Anopheles

581albimanus

independently of bacterial co-infection. Developmental &

582Comparative

Immunology. 52, 172-181.

583Contreras-Garduo
584&

Lanz- Mendoza H. (2014). Cost of immune priming within generations: trade-

585off

between infection and reproduction. Microbes and Infection, 16, 261-267.

586Cooper, E.L.
587119-128.

48

J., Rodrguez M.C., Rodrguez M.H., Alvarado-Delgado A.,

(1992). Overview of immunoevolution. Bolletino di Zoologia, 59,

49
588Cooper, E.L.,
589earthworm
590Corbet,

& Roch, P. (1986). Second-set allograft responses in the

Lumbriculus terrestris. Transplantation, 4, 514-520.

P.S. (1999). Dragonflies behavior and ecology of Odonata. Cornell

591University

Press.

592Christofi,

T., & Apidianakis, Y. (2013). Drosophila immune priming against

593Pseudomonas
594humoral
595Dong,

immunity. F1000Research, 2.

Y., Taylor, H. E., & Dimopoulos, G. (2006). AgDscam, a hypervariable

596immunoglobulin
597immune
598Dong,

aeruginosa is short-lasting and depends on cellular and

domain-containing receptor of the Anopheles gambiae innate

system. PLoS Biology. 4, e229.

Y., Manfredini, F., & Dimopoulos, G. (2009). Implication of the mosquito

599midgut

microbiota in the defense against malaria parasites. PLoS Pathogens, 5,

600e1000423.
601Du

Pasquier, L. (2005). Meeting the demand for innate and adaptive immunities

602during

evolution. Scandinavian Journal of Immunology, 62, 39-48.

603Dubovskiy, I.M.,
604Kryukov, V.Y.,
605(2013).
6068:

Grizanova, E.V., Mukherjee, K., Vilcinskas, A., & Glupov, V.V.

Can insects develop resistance to insect pathogenic fungi? PLoS ONE

e60248.

607Eggert,

H., Kurtz, J., & Diddens-de Buhr, M. F. (2014). Different effects of

608paternal
609and

trans-generational immune priming on survival and immunity in step

genetic offspring. Proceedings of the Royal Society B: Biological Sciences,

610281,

50

Whitten, M.M.A., Yaroslavtseva, O.N., Greig, C., Kryukov, V.Y.,

20142089.

51
611Eggert,
612lead

H., Diddens-de Buhr, M. F., & Kurtz, J. (2015). A temperature shock can

to transgenerational immune priming in the Red Flour Beetle, Tribolium

613castaneum.

Ecology and evolution, 5, 1318-1326.

614Eleftherianos,
615bacteria
6164,

I., Atri, J., Accetta, J., & Castillo, J. C. (2013). Endosymbiotic

in insects: guardians of the immune system?. Frontiers in physiology,

46.

617Elrod-Erickson,
618cellular

M., Mishra, S., Schneider, D. (2000). Interactions between the

and humoral immune responses in Drosophila. Current Biology, 10,

619781-784.
620Faulhaber, L.M.,
621bacteria

in the American cockroach. Immunology, 75, 378-381.

622Feldhaar,
623traits

& Karp, R. D. (1992). A diphasic immune response against

H. (2011). Bacterial symbionts as mediators of ecologically important

of insect hosts. Ecological Entomology, 36, 533-543.

624Fisher, J.

J., & Hajek, A. E. (2015). Maternal exposure of a beetle to pathogens

625protects

offspring against fungal disease. Plos One, 10, e0125197.

626Freitak,

D., Schmidtberg, H., Dickel, F., Lochnit, G., Vogel, H., & Vilcinskas, A.

627(2014).

The maternal transfer of bacteria can mediate trans-generational

628immune

priming in insects. Virulence, 5, 547-554.

629Garbutt,

J. S., O'Donoghue, A. J., McTaggart, S. J., Wilson, P. J., & Little, T. J.

630(2014).

The development of pathogen resistance in Daphnia magna:

631implications

for disease spread in age-structured populations. The Journal of

632experimental

biology, 217, 3929-3934.

633Garnier, R.,

Boulinier, T., & Gandon, S. (2013). Evolution of the temporal

634persistence

of immune protection. Biology letters, 9, 20130017.

52

53
635Gonzlez-Tokman,
636Aguilar, A.
637the

D.M., Gonzlez-Santoyo, I., Lanz-Mendoza, H. & Crdoba

(2010). Territorial damselflies do not show immunological priming in

wild. Physiological Entomology, 35, 364-372.

638Goren,

M., Yosef, I., Edgar, R., & Qimron, U. (2012). The bacterial CRISPR/Cas

639system

as analog of the mammalian adaptive immune system. RNA biology, 9,

640549-554.
641Hartmann,
642memory

R. S., & Karp, R. D. (1989). Short-term immunological

in the allograft response of the American cockroach,

643Periplaneta
644Hauton,
645house
646He,

C. & Smith. V. J. (2007). Adaptive immunity in invertebrates: a straw

without a mechanistic foundation. BioEssays, 29,1138-1146.

X., McLean, J. S., Edlund, A., Yooseph, S., Hall, A. P., Liu, S. Y., Dorrestein,

647P.C.,

Esquenazi, E., Hunter, R.C., Cheng, G., Nelsonc, K.E., Lux, R., & Shi, W.

648(2015).

Cultivation of a human-associated TM7 phylotype reveals a reduced

649genome
650of

americana. Transplantation, 43, 920-922.

and epibiotic parasitic lifestyle. Proceedings of the National Academy

Sciences, 112, 244-249.

651Henter, H.J.
652system.

& Via, S. (1995). The potential for coevolution in a host- parasitoid

I. Genetic variation within an aphid population in susceptibility to a

653parasitic

wasp. Evolution, 49, 427-438.

654HernndezLpez

J., Schuehly W., Crailsheim K., Riessberger-Gall U. (2014).

655Trans-generationalimmune
656Society

B: Biological Science, 281, 20140454.

657Hernndez-Martnez,
658&

54

priming in honeybees. Proceedings of the Royal

P., Naseri, B., Navarro-Cerrillo, G., Escriche, B., Ferr, J.

Herrero, S. (2010). Increase in midgut microbiota load induces an apparent

55
659immune

priming and increases tolerance to Bacillus thuringiensis.

660Environmental
661Huang,

Microbiology, 12, 2730-2737.

W. S., Duan, L. P., Huang, B., Zhou, L. H., Liang, Y., Tu, C. L., Zhang,

662F.F.,

Nie, P., & Wang, T. (2015). Identification of three IFN- inducible lysosomal

663thiol

reductase (GILT)-like genes in mud crab Scylla paramamosain with distinct

664gene

organisations and patterns of expression. Gene.

665Hurst,
666Help

G.D.D., & Hutchence, K.J. (2010) Host Defence: Getting By with a Little

from Our Friends. Current Biology, 20, R806-R808.

667Hutter, T.,
668gut

Gimbert, C., Bouchard, F., & Lapointe, F. J. (2015). Being human is a

feeling. Microbiome, 3, 9.

669Johnson,
670shrimp

K. N., van Hulten, M. C., & Barnes, A. C. (2008). Vaccination of

against viral pathogens: phenomenology and underlying mechanisms.

671Vaccine,

26, 4885-4892.

672Jokella,

J. (2010). Transgenerational immune priming as cryptic parental care.

673Journal

of Animal Ecology, 79, 305-307.

674Karp,

R. D. (1990). Cell-mediated immunity in invertebrates. Bioscience, 732-

675737.
676Kato,

L. M., Kawamoto, S., Maruya, M., & Fagarasan, S. (2014). The role of the

677adaptive
678260,

immune system in regulation of gut microbiota. Immunological reviews,

67-75.

679Klein,

J. (1989). Are invertebrates capable of anticipatory immune responses?

680Scandinavian

Journal of Immunology, 29,499-505.

681Kraaijeveld, A.
682resistance
683389,

56

& Godfray, H. C. J. (1997). Trade-off between parasitoid

and larval competitive ability in Drosophila melanogaster. Nature,

278-280.

57
684Kurtz,

J. & Armitage, S. A. (2006). Alternative adaptive immunity in

685invertebrates.
686Kurtz,

Trends in immunology, 27, 493-496.

J. & Franz, K. (2003). Innate defense: Evidence for memory in

687invertebrate
688Kurtz,

immunity. Nature, 425, 37-38.

J. (2005). Specific memory within innate immune systems. Trends in

689immunology, 26,

186-192.

690Lazzaro,

B. P., Sceurman, B. K. & Clark, A. G. (2004). Genetic basis of natural

691variation

in D. melanogaster antibacterial immunity. Science, 303, 1873-1876.

692Lin,

Y. C., Chen, J. C., Morni, W. Z. W., Putra, D. F., Huang, C. L., Li, C. C., &

693Hsieh,

J. F. (2013). Vaccination enhances early immune responses in white

694shrimp

Litopenaeus vannamei after secondary exposure to Vibrio alginolyticus.

695PloS
696Little,

one, 8, e69722.
T.J., O'Connor, B., Colegrave, N., Watt, K., & Read, A.F. (2003). Maternal

697transfer

of strain-specific immunity in an invertebrate. Current Biology, 13, 489-

698492.
699Little,
700of

T. J. & Kraaijeveld. A. R. (2004). Ecological and evolutionary implications

immunological priming in invertebrates. Trends in Ecology and Evolution, 19,

70158-60.
702Little,

T. J., Hultmark, D. & Read, A. F. (2005). Invertebrate immunity and the

703limits

of mechanistic immunology. Nature Immunology, 6, 651-654.

704Little,

T. J., Colegrave, N., Sadd, B. M. & Schmid-Hempel, P. (2008). Studying

705immunity
706Liu,

at the whole organism level. Bioessays, 30, 404-405.

K. J. (2000). Confidence intervals of the simple difference between the

707proportions
708infection.

58

of a primary infection and a secondary infection, given the primary

Biometrical Journal, 42, 59-69.

59
709Longdon,
710an

B., Cao, C., Martinez, J. & Jiggins, F. M. (2013). Previous exposure to

RNA virus does not protect against subsequent infection in Drosophila

711melanogaster.

PloS One, 8(9), e73833.

712http://doi.org/10.1371/journal.pone.0073833
713Lpez,

S. (1998). Acquired resistance affects male sexual display and female

714choice

in guppies. Proceedings of the Royal Society B: Biological Sciences,

715265,

717-723.

716Martin,

II L. B., Navara, K. J., Bailey, M. T., Hutch, C. R., Powell, N. D.,

717Sheridan,
718memory

J. F., & Nelson, R. J. (2008). Food restriction compromises immune

in deer mice (Peromyscus maniculatus) by reducing spleen-derived

719antibody-producing

B cell numbers. Physiological and Biochemcal Zoology, 81,

720366-372.
721Masri,

L. & Cremer, S. (2014). Individual and social immunisation in insects.

722Trends

in immunology, 35, 471-482.

723McNamara,
724and

K. B., Lieshout, E., & Simmons, L. W. (2014). The effect of maternal

paternal immune challenge on offspring immunity and reproduction in a

725cricket.

Journal of evolutionary biology, 27, 1020-1028.

726McTaggart,
727reduced

S. J. Wilson, P. J. & Little T. J. (2012). Daphnia magna shows

infection upon secondary exposure to a pathogen. Biology Letters, 8,

728972-975.
729Mikonranta,
730immunity:
731protects

60

L., Mappes, J., Kaukoniitty, M., & Freitak, D. (2014). Insect

oral exposure to a bacterial pathogen elicits free radical response and

from a recurring infection. Frontiers in zoology, 11, 23.

61
732Milutinovi,
733beetle

B., Fritzlar, S., & Kurtz, J. (2013). Increased survival in the red flour

after oral priming with bacteria-conditioned media. Journal of innate

734immunity,

6, 306-314.

735Miyashita

A., Kizaki H., Kawasaki K., Sekimizu K., & Kaito C. (2014). Primed

736immune
737in

responses to gram-negative peptidoglycans confer infection resistance

silkworms. The journal of biological chemistry, 289, 14412-14421

738Moreau,

J., Martinaud, G., Troussard, J.-P., Zanchi, C. & Moret, Y. (2012).

739Trans-generational
740response

in an insect. Oikos, 121, 1828-1832.

741Moreno-Garca,
7422014.

immune priming is constrained by the maternal immune

M., Recio-totor, B., Claudio-Piedras, F., & Lanz-Mendoza, H.

Injury and immune response: applyng the danger theory to mosquitoes.

743Frontiers

in Plant Science, 5, 451-451.

744Moreno-Garca,

M., Vargas, V., Ramrez-Bello, I., Hernndez-Martnez, G. &

745Lanz-Mendoza,

H. (2015). Bacterial Exposure at the Larval Stage Induced

746Sexual
747Plos

One, 10(7), e0133240. http://doi.org/10.1371/journal.pone.0133240.

748Moret,
749the

Immune Dimorphism and Priming in Adult Aedes aegypti Mosquitoes.

Y. (2009). Trans-generational immune priming: specific enhancement of

antimicrobial immune response in the mealworm beetle, Tenebrio molitor.

750Proceedings

of the Royal Society of London Series B, 273, 1399-1405.

751Morgan,

X. C., & Huttenhower, C. (2014). Meta'omic analytic techniques for

752studying

the intestinal microbiome. Gastroenterology, 146, 1437-1448.

753Ng,

T. H., Hung, H. Y., Chiang, Y. A., Lin, J. H., Chen, Y. N., Chuang, Y. C., &

754Wang,

H. C. (2014). WSSV-induced crayfish Dscam shows durable immune

755behavior.

62

Fish & shellfish immunology, 40, 78-90.

63
756Norouzitallab,
757Bussche,
758induces

P., Baruah, K., Vandegehuchte, M., Van Stappen, G., Catania, F.,

J. V., Sorgeloos, P., & Bossier, P. (2014). Environmental heat stress

epigenetic transgenerational inheritance of robustness in

759parthenogenetic
760Norouzitallab,
761immune
762Vibrio

Artemia model. The FASEB Journal, 28, 3552-63.

P., Biswas, P., Baruah, K., & Bossier, P. (2015). Multigenerational

priming in an invertebrate parthenogenetic Artemia to a pathogenic

campbellii. Fish & shellfish immunology, 42, 426-429.

763Ottaviani,
764changes

E. (2015). Invertebrate immunological memory: could the epigenetic

play the part of lymphocytes?. ISJ, 12, 1-4.

765Pancer, Z., Amemiya,

766M.D.

(2004). Somatic diversification of variable lymphocyte receptors in the

767agnathan
768Paust,

sea lamprey. Nature, 8,174-80.

S., & von Andrian, U. H. (2011). Natural killer cell memory. Nature

769immunology,
770Pham,

772PLoS

12, 500-508.

L. N., Dionne, M. S., Shirasu-Hiza, M. & Schneider, D. S. (2007). A

771specific

primed immune response in Drosophila is dependent on phagocytes.

Pathogens, 3, e26.

773Pham,
774the

L. N. & Schneider, D. S. (2008). Evidence for specificity and memory in

insect innate immune response. In Insect Immunology. Beckage N. E. (ed).

775Academic
776Pigeault,

Press.

R., Vzilier, J., Nicot, A., Gandon, S., & Rivero, A. (2015).

777Transgenerational
778Biology

64

C.T., Ehrhardt, G.R., Ceitlin, J., Gartland, G.L., & Cooper,

effect of infection in Plasmodium-infected mosquitoes.

letters, 11, 20141025.

65
779Pope,

E. C., Powell, A., Roberts, E. C., Shields, R. J., Wardle, R., & Rowley,

780A.F. (2011).

Enhanced cellular immunity in shrimp (Litopenaeus vannamei) after

781vaccination.

PLoS ONE, 6: e20960.

782Portela,
783G.,

J., Duval, D., Rognon, A., Galinier, R., Boissier, J., Coustau, C., Mitta,

Thron, A., & Gourbal, B. (2013). Evidence for specific genotype-dependet

784immune
785Innate

priming in the Lophotrochozoan Biomphalaria glabrata snail. Journal of

Immunity, 5, 261-276.

786Rantala,

M. J., Kortet, R., Kotiaho, J. S., Vainikka, A., & Suhonen, J. (2003).

787Condition

dependence of pheromones and immune function in the grain beetle

788Tenebrio

molitor. Functional Ecology, 17, 534-540.

789Ramirez,

J. L., de Almeida Oliveira, G., Calvo, E., Dalli, J., Colas, R. A., Serhan,

790C.

N., Ribeiro, J. M. & Barillas-Mury, C. (2015). A mosquito lipoxin/lipocalin

791complex

mediates innate immune priming in Anopheles gambiae. Nature

792Communications.
793Reber, A.,
794exposed

& Chapuisat, M. (2012). No evidence for immune priming in ants

to a fungal pathogen. PLoS ONE, 7, e35372.

795Rodrigues,
796Hemocyte
797gambiae
798Rolff,

J., Brayner, F. A., Alves, L. C., Dixit, R. & Barillas-Mury, C. (2010).

differentiation mediates innate immune memory in Anopheles

mosquitoes. Science, 329, 1353-1355.

J. & Siva-Jothy, M. T. (2003). Invertebrate ecological immunology.

799Science,

301, 472-475.

800Rosengaus,
801larvae:
8029,

66

6: 7403.

R. B., Malak, T. & MacKintosh, C. (2013). Immune-priming in ant

social immunity does not undermine individual immunity. Biology letters,

20130563.

67
803Roth,

O., Sadd, B. M., Schmid-Hempel, P., & Kurtz, J. (2009). Strain-specific

804immune
805the

priming in the red flour beetle, Tribolium castaneum. Proceedings of

Royal Society of London Series B, 276, 145-151.

806Roth,

O., Joop, G., Eggert, H., Hilbert, J., Daniel, J., Schmid-Hempel, P., &

807Kurtz,

J. (2010). Paternally derived immune priming for offspring in the red flour

808beetle,
809Sadd,

Tribolium castaneum. Journal of Animal Ecology, 79, 403-413.

B. M., & Schmid-Hempel, P. (2006). Insect immunity shows specificity in

810protection
811Sadd,

upon secondary pathogen exposure. Current Biology, 16, 1206-1210.

B. M., & Schmid-Hempel, P. (2009a). Ecological and Evolutionary

812implications

of specific immune responses. In insect infection and immunity.

813Evolution,

ecology and mechanism. Rolff, J. & Reynolds, S.E. (eds). Oxford

814University

Press.

815Sadd,

B. M., & P. Schmid-Hempel, P. (2009b). A distinct infection cost

816associated
817bees.

Biology Letters, 5, 798-801.

818Saeed,
819F.,

with trans-generational priming of antibacterial immunity in bumble-

S., Quintin, J., Kerstens, H. H., Rao, N. A., Aghajanirefah, A., Matarese,

Cheng, S. C., Ratter, J., Berentsen, K., van der Ent, M. A., Sharifi, N.,

820Janssen-Megens,
821Burden,

F., Downes, K., Frontini, M., Kumar, V., Giamarellos-Bourboulis, E. J.,

822Ouwehand,
823J.

W. H., van der Meer, J. W., Joosten, L. A., Wijmenga, C., Martens,

H., Xavier, R. J., Logie, C., Netea, M. G., & Stunnenberg, H. G. (2014).

824Epigenetic
825innate

68

E. M., Ter Huurne, M., Mandoli, A., Van Schaik, T., Ng, A.,

programming of monocyte-to-macrophage differentiation and trained

immunity. Science, 345, 1251086.

69
826Salmela,
827mother

H., Amdam, G. V. & Freitak D. (2015). Transfer of immunity from

to offspring is mediated via egg-yolk protein vitellogenin. PLOS

828Pathogens.

11:e1005015.

829Schmid-Hempel,
830evolutionary

ecology. Proceedings of the Royal Society B., 270, 357-366.

831Schmid-Hempel,
832Annual

P. (2003). Variation in immune defense as a question in

P. (2005). Evolutionary ecology of insect immune defenses.

Review of Entomology, 50, 529-551.

833Schmid-Hempel,
834priming

P. (2005a). Natural insect host-parasite systems show immune

and specificity - puzzles to be solved. BioEssays, 27, 1026-1034.

835Schmid-Hempel,

P. (2011). Evolutionary Parasitology. Oxford: Oxford University

836Press.
837Shi

Z.H., Lin Y.T., & Hou Y.M. (2014). Mother-derived trans-generational

838immune

priming in the red palm weevil, Rhynchophorus ferrugineus Olivier

839(Coleoptera,

Dryophthoridae). Bulletin of Entomological Research 104, 742-

840750.
841Schulenburg,
842generate
84344,

a highly specific innate immune response?. Molecular immunology,

3338-3344.

844Spoel,

S. H., & Dong, X. (2012). How do plants achieve immunity? Defence

845without
846Sun,

848Sun,

specialized immune cells. Nature Reviews Immunology, 12, 89-100.

J. C., Beilke, J. N., & Lanier, L. L. (2009). Adaptive immune features of

847natural

killer cells. Nature, 457, 557-561.

J., & Deng, W.M., (2007). Hindsight mediates the role of notch in

849suppressing
850431-442.

70

H., Boehnisch, C., & Michiels, N. K. (2007). How do invertebrates

hedgehog signaling and cell proliferation. Developmental Cell. 12,

71
851Tate,

A.T., & Rudolf V. H. W., (2012). Immune priming across life stages and

852generations:

Implications for infectious disease dynamics in insects. Oikos,

853121,1083-1092.
854Tate,
855wild
856is

A. T., & Graham, A. L. (2015). Transgenerational priming of resistance in

flour beetles reflects the primed phenotypes of laboratory populations and

inhibited by coinfection with a common parasite. Functional Ecology. 29,

85710591069.
858Thaiss,

C. A., Zeevi, D., Levy, M., Zilberman-Schapira, G., Suez, J., Tengeler, A.

859C., Abramson,
860Gilad,

S., Harmelin, A., Shapiro, H., Halpern, Z., Segal, E., & Elinav, E. (2014).

861Transkingdom
862homeostasis.
863Tidbury, H.
864of

L., Katz, M. N., Korem, T., Zmora, N., Kuperman, Y., Biton, I.,

control of microbiota diurnal oscillations promotes metabolic

Cell, 159, 514-529.

J., Best, A., & Boost, M. (2012). The epidemiological consequences

immune priming. Proceedings of the Royal Society of London Series B, 279,

8654505-4512.
866Tidbury, H.
867immune
868of

J., Pedersen, A.B., & Boots, M. (2011). Within and transgenerational

priming in an insect to a DNA virus. Proceedings of the Royal Society

London Series B, 278, 871-876.

869Thomas,

A. M., & Rudolf, V. H. W. (2010). Challenges of metamorphosis in

870invertebrate
871Ecological
872Thornhill,
873Harvard

Entomology, 35, 200-205.

R., & Alcock, J. (1983). The evolution of insect mating systems.

University Press.

874Valdez, A.,

Yepiz-Plascencia, G., Ricca, E., & Olmos, J. (2014). First

875Litopenaeus

72

hosts: Maintaining parasite resistance across life-history stages.

vannamei WSSV 100% oral vaccination protection using

73
876CotC::Vp26
877of

fusion protein displayed on Bacillus subtilis spores surface. Journal

Applied Microbiology, 117, 347-357.

878Valtonen

T. M., Kleino A., Rmet M., & Rantala M. J. (2010). Starvation reveals

879maintenance
880Vantaux, A.,
881offspring
882Malaria

cost of humoral immunity. Evolutionary Biology 37, 4957.

Dabir, K. R., Cohuet, A., & Lefvre, T. (2014). A heavy legacy:

of malaria-infected mosquitoes show reduced disease resistance.

journal, 13, 442.

883Viney, M.

E., Riley, E. M., & Buchanan, K. L. (2005). Optimal immune

884responses:

immunocompetence revisited. Trends in Ecology & Evolution, 20,

885665-669.
886Voordouw, M.
887stimulation
888aegypti.

of the melanization response in the yellow fever mosquito Aedes

Oikos, 117, 1269-1279.

889Vorburger, C.,
890scope

J., Lambrechts, L., & Koella, J. (2008). No maternal effects after

Gegenschatz, S. E., Ranieri, G., & Rodriguez, P. (2008). Limited

for maternal effects in aphid defence against parasitoids. Ecological

891Entomology,
892Wang,

33, 189-196.

L., Yue, F., Song, X., & Song, L. (2015). Maternal immune transfer in

893mollusc.

Developmental & Comparative Immunology, 48, 354-359.

894Watson,

F. L., Puttmann-Holgado, R., Thomas, F., Lamar, D. L., Hughes, M.,

895Kondo,

M., Rebel, V. I., & Schmucker, D. (2005). Extensive diversity of Ig-

896superfamily
897Weiss,

proteins in the immune system of insects. Science, 309, 1874-1878.

B., & Aksoy, S. (2011). Microbiome influences on insect host vector

898competence.
899West,
900P.,

74

Trends in parasitology, 27, 514-522.

C. E., Renz, H., Jenmalm, M. C., Kozyrskyj, A. L., Allen, K. J., Vuillermin,

& Prescott, S. L. (2015). The gut microbiota and inflammatory

75
901noncommunicable
902therapies.
903Wilson,

Journal of Allergy and Clinical Immunology, 135, 3-13.

K., & Graham, R. I. (2015). Transgenerational effects modulate density-

904dependent
905Biology

diseases: Associations and potentials for gut microbiota

prophylactic resistance to viral infection in a lepidopteran pest.

Letters 11: 20150012.

906Witteveldt

J., Cifuentes C.C., Vlak J. M., & van Hulten M. C. W. (2004).

907Protection

of Penaeus monodon against White Spot Syndrome Virus by Oral

908Vaccination.
909Wu,

Journal of Virology. 78: 2057-2061.

G., Yi, Y., Lv, Y., Li, M., Wang, J., & Qiu, L. (2015). The lipopolysaccharide

910(LPS)

of Photorhabdus luminescens TT01 can elicit dose-and time-dependent

911immune

priming in Galleria mellonella larvae. Journal of invertebrate pathology,

912127,

63-72.

913Wu,

G., Zhao, Z., Liu, C., & Qiu, L. (2014). Priming Galleria mellonella

914(Lepidoptera:
915Enhanced
916Role

Pyralidae) Larvae with Heat-Killed Bacterial Cells Induced an

Immune Protection Against Photorhabdus luminescens TT01 and the

of Innate Immunity in the Process. Journal of economic entomology, 107,

917559-569.
918Yue,

F., Zhou, Z., Wang, L., Ma, Z., Wang, J., Wang, M., Zhang, H., & Song, L.

919(2013).

Maternal transfer of immunity in scallop Chlamys farreri and its trans-

920generational

immune protection to offspring against bacterial challenge.

921Developmental
922Zanchi,

C., Troussard, J-P., Martinaud, M., Moreau, J., & Moret, Y. (2011).

923Differential
924immune
9251183.

76

& Comparative Immunology, 41, 569-577.

expression and costs between maternally and paternally derived

priming for offspring in an insect. Journal of Animal Ecology, 80, 1174-

77
926Zhang,

S. M., Adema, C. M., Kepler, T. B., and Loker, E. S. (2004).

927Diversification

of Ig superfamily genes in an invertebrate. Science, 305, 251

928254.
929Zhang

T., Qiu L., Sun Z., Wang L., Zhou Z., Liu R., Yue F., Sun R., & Song L.

930(2014).

The specifically enhanced cellular immune responses in Pacific oyster

931(Crassostrea

gigas) against secondary challenge with Vibrio splendus.

932Developmental
933Zhao,

and Comparative Immunology, 45,141-150

Z., Wu, G., Wang, J., Liu, C., & Qiu, L. (2013). Next-generation

934sequencing-based
935immune-primed

78

transcriptome analysis of Helicoverpa armigera larvae

with Photorhabdus luminescens TT01. PLOS ONE 8, e80146.

79
936Legends
937
938Table

1. Studies showing immune priming within generations. Studies shown

939exhibit

the following: (a) tested the importance of bacteria playing a role on

940immune

priming outcome (one out of 27); b) its costs (two out of 27); (c) do not

941support

the priming (one out of 27); (d) use of novel parasites or pathogens

942(three
943(21

out of 27); (e) shown strain-specific (four out of 27); or (f) species-specific

out of 27); or (g) that have tested the biphasic response (one out of 27).

944ROS

= Reactive Oxygen species, PO = Phenoloxidase, SOD = Superoxide

945dismutase,

RBs = Respiratory bursts, PepG= Peptidoglycans.

946
947Table

2. Studies that tested immune priming across generations. Table shows

948studies
949tested
950(two

that: a) tested the importance of bacteria for priming (two out of 22); b)

its costs (seven out of 22); c) showed the role of strain-specific response

out of 22); or d) species-specific (16 out of 22), e) use of unnatural

951parasites
952out

or pathogens (three out of 22) or (f) do not support the priming (three

of 22). No studies have tested the biphasic immune response. PO =

953Phenoloxidase.
954
955Box

1. Key definitions regarding the immune priming theory.

956
957Box

2. Mechanism of immune priming. These mechanisms include recognizing

958and

effector molecules.

80