May 2016 | Volume 7 | A icle 1961
Mini Re iew
published: 19 May 2016
doi: 10.3389/ immu.2016.00196
F on ie s in Immunology | www. on ie sin.o g
Edi ed by:
Jason Paul Gigley,
Uni e si y o Wyoming, USA
Re iewed by:
Ma c S. Dionne,
Impe ial College London, UK
Philip Elks,
Uni e si y o She ield, UK
*Co espondence:
Henna Myllymäki
[email p o ec ed]
Special y sec ion:
This a icle was submi ed
o Mic obial Immunology,
a sec ion o he jou nal
F on ie s in Immunology
Recei ed: 29Feb ua y2016
Accep ed: 06May2016
Published: 19May2016
Ci a ion:
MyllymäkiH, Bäue leinCA and
Räme M (2016) The Zeb a ish
B ea hes New Li e in o he
S udy o Tube culosis.
F on . Immunol. 7:196.
doi: 10.3389/ immu.2016.00196
The Zeb a ish B ea hes new Li e
in o he S udy o Tube culosis
Henna Myllymäki1*, Ca ina A. Bäue lein1 and Mika Räme 1,2,3,4
1BioMediTech, Uni e si y o Tampe e, Tampe e, Finland, 2Depa men o Pedia ics, Tampe e Uni e si y Hospi al, Tampe e,
Finland, 3Depa men o Child en and Adolescen s, Oulu Uni e si y Hospi al, Oulu, Finland, 4PEDEGO Resea ch Uni , Medical
Resea ch Cen e Oulu, Uni e si y o Oulu, Oulu, Finland
Tube culosis (TB) is a global heal h eme gency. Up o one- hi d o he wo ld’s popula ion
is in ec ed wi h Mycobac e ium ube culosis, and he pa hogen con inues o kill 1.5 mil-
lion people annually. Cu en ly, he means o p e en ing, diagnosing, and ea ing TB a e
unsa is ac o y. One o he main easons o he poo p og ess in TB esea ch has been
a lack o good animal models o s udy he la ency, do mancy, and eac i a ion o he
disease. Al hough sophis ica ed in i o and insilico me hods sui able o TB esea ch
a e cons an ly being de eloped, hey canno ep oduce he comple e e eb a e immune
sys em and i s in e play wi h pa hogens and accines. Howe e , he zeb a ish has
ecen ly eme ged as a use ul al e na i e o mo e adi ional models, such as mice, ab-
bi s, guinea pigs, and non-human p ima es, o s udying he complex pa hophysiology o
a mycobac e ial in ec ion. The model is based on he simila i y be ween Mycobac e ium
ma inum–a na u al ish pa hogen–and M. ube culosis. In bo h zeb a ish la ae and
adul ish, an in ec ion wi h M. ma inum leads o he o ma ion o mac ophage agg e-
ga es and g anulomas, which esemble he M. ube culosis in ec ions in humans. In his
e iew, we will summa ize he cu en s a us o he zeb a ish model in TB esea ch and
highligh he ad an ages o using zeb a ish o dissec mycobac e ial i ulence s a egies
as well as he hos immune esponses elici ed agains hem. In addi ion, we will discuss
he possibili ies o using he adul zeb a ish model o s udying la ency, do mancy, and
eac i a ion in a mycobac e ial in ec ion.
Keywo ds: ube culosis, zeb a ish model sys em, accina ion, Mycobac e ium ma inum, Mycobac e ium
ube culosis, Mycobac e ium in ec ions, g anuloma, la ency
inTRODUCTiOn
Tube culosis (TB) is s ill he wo ld’s second deadlies in ec ious disease killing 1.5 million people and
wi h an es ima ed 9.6 million new cases epo ed o he WHO in 2015 (1). An es ima ed one- hi d o
he wo ld’s popula ion has been exposed o TB. 5–10% o hese la en ca ie s will e en ually de elop
he ac i e disease (1).
The causa i e agen o TB, Mycobac e ium ube culosis, sp eads h ough he ai (Figu e1A).
Al eola mac ophages phagocy ose he inhaled mycobac e ia and anspo hem in o he lung
issues (2). A cascade o p o- and an i-in lamma o y signaling leads o he ec ui men and accu-
mula ion o addi ional mac ophages and o he leukocy es in he pulmona y issues. E en ually,
he o ma ion o g anulomas, he hallma k o pa hological TB, is ini ia ed. The g anuloma is
a he e ogeneous, bu well-o ganized, and dynamic accumula ion o immune cells, including
FiGURe 1 | G anuloma o ma ion in human Mycobac e ium ube culosis in ec ion and in zeb a ish M. ma inum in ec ion.
(Con inued)
2
Myllymäki e al.
Zeb a ish as a Tube culosis Model
F on ie s in Immunology | www. on ie sin.o g May 2016 | Volume 7 | A icle 196
TABLe 1 | Cha ac e is ics o a mycobac e ial in ec ion in humans, adul zeb a ish, and zeb a ish emb yos.
Human Adul zeb a ish (>3mon hs) Zeb a ish emb yo (<6days) Re e ence
immune sys em Inna e and adap i e Inna e and adap i e Inna e only (6–8)
Mycobac e ial pa hogen Mycobac e ium ube culosis and a ypical
mycobac e ia including Mycobac e ium ma inum
Mycobac e ium ma inum Mycobac e ium ma inum (9–11)
Na u al suscep ibili y Yes Yes Yes (9, 10, 12)
In ec ion ou e and
in ec ious dose
na u al in ec ion
Ai ways ID50 <10 bacilli Diges i e ac ? (9, 10, 12)
expe imen al in ec ion
N/A Mul iple injec ion echniques,
in ape i oneal injec ion is he mos
commonly used; <30–10,000c u
Mul iple injec ion echniques,
caudal injec ion is he mos
commonly used; <10–>300c u
(8, 13–15)
In ec ion phases Acu e Acu e P og essi e? (10, 13–17)
La en La en
Reac i a ion Reac i a ion
G anuloma ypes Ea ly Ea ly P imi i e? (4, 5,
13–18)Fib ous Fib ous
Nec o ic Nec o ic
Cell ypes in ol ed in
g anuloma o ma ion
Mac ophages Mac ophages Mac ophages (4, 5,
13–18)Neu ophils Neu ophils Neu ophils
Dend i ic cells Dend i ic cells Epi helial cells
Lymphocy es (T cells, B cells, and NK cells) Lymphocy es (T cells and B cells)
Fib oblas s Epi helial cells
Epi helial cells
(A) M. ube culosis sp eads as an ae osol, and i s in ec s al eola mac ophages. In mos indi iduals, he in ec ion is main ained in a la en , subclinical s a e, which
is cha ac e ized by he o ma ion o g anulomas (le ). The ma u e g anulomas ha e a caseous, nec o ic co e, su ounded by in ec ed mac ophages and
lymphocy es. Upon eac i a ion, he g anulomas a e dis up ed, causing ca i ies in he lungs. The mycobac e ia escape om he g anulomas and a e dissemina ed
in cough d ople s, which acili a es he ansmission o he disease. (B) Upon in ec ion wi h M. ma inum, he g anulomas in he zeb a ish emb yo de elop wi hin a
ew days and mainly consis o in ec ed and unin ec ed mac ophages and ec ui ed neu ophils. (C) Depending on he bac e ial dose, he adul zeb a ish
M. ma inum in ec ion can lead o a la en o an ac i e, p og essi e disease. A la en in ec ion is cha ac e ized by he o ma ion o g anulomas in a ious o gans. As
he ea ly g anulomas ma u e, hei inne pa s become caseous and su ounded by a ib ous wall. The zeb a ish wi h a la en in ec ion emain asymp oma ic. A ( e)
ac i a ed, p og essi e mycobac e ial in ec ion is cha ac e ized by he dis up ion o he g anuloma ous s uc u es, apid eplica ion and dissemina ion o
mycobac e ia and p o ound issue damage. E en ually, a p og essi e mycobac e ial disease will lead o dea h in mos ish. The zeb a ish g anulomas we e isualized
wi h Ziehl–Neelsen s aining, mycobac e ia a e seen as pu ple ods. c u, colony- o ming uni .
FiGURe 1 | Con inued
3
Myllymäki e al.
Zeb a ish as a Tube culosis Model
F on ie s in Immunology | www. on ie sin.o g May 2016 | Volume 7 | A icle 196
blood-de i ed in ec ed and unin ec ed mac ophages, oamy
mac ophages, and epi helioid cells (3). The inne cell mass
is usually su ounded by a ing o leukocy es and ib oblas s
(4) (Figu e1A; Table 1). The localiza ion and con ol o he
bac e ia and he es ic ion o he immune esponse o a
de ined a ea a e gene ally ega ded as he main unc ions o
g anulomas (5).
Mycobac e ia can pe sis asymp oma ic wi hin he g anuloma
o decades. Howe e , a dys egula ion o he immune sys em can
eac i a e he mycobac e ia, which leads o disease p og ession
(seconda y TB) (2) (Figu e1A) P ima y TB mainly occu s in
child en, who a e a he highes isk o TB meningi is and a dis-
semina ed o m o he disease (19).
E o s o e adica e TB a e obs uc ed by he lack o unambigu-
ous diagnos ic ools, he leng hy an ibio ic ea men s equi ed
o cu a ion, he g owing p oblem o mul i-d ug- esis an
bac e ia, and he poo p o ec ion p o ided by he Bacillus
Calme e–Gué in (BCG), he only accine a ailable (1). As a
li e accine, BCG imposes a isk o a dissemina ed in ec ion in
immunocomp omised pa ien s (1, 20, 21). Thus, he e is a need
o de elop new e ec i e d ugs and accines agains TB. Fo his
pu pose, ele an animal models a e essen ial. The mos com-
monly used animal models in TB esea ch a e mice, guinea pigs,
and non-human p ima es (NHP), all o which ha e hei limi a-
ions ela ed o ei he space, cos s, e hical aspec s, o hei abili y
o eplica e he human disease pa hology (22, 23). Recen ly, he
zeb a ish–M. ma inum model has gained popula i y as a na u al
pa hogen–hos sys em ha closely ecapi ula es he pa hology o
human TB (Table1) (13, 23). The in ec ion model and i s applica-
ions a e discussed in mo e de ail below.
THe ZeBRAFiSH–MYCOBACTERIUM
MARINUM inFeCTiOn MODeL
Mycobac e ium ma inum, he causa i e agen o ish mycobac-
e iosis, is a close ela i e o M. ube culosis (24). M. ma inum
4
Myllymäki e al.
Zeb a ish as a Tube culosis Model
F on ie s in Immunology | www. on ie sin.o g May 2016 | Volume 7 | A icle 196
sp eads ia wa e , and i also occasionally in ec s humans, bu
he in ec ion is usually limi ed o he skin ( ish ank g anu-
loma) (11). Thus, M. ma inum is sa e o wo k wi h and has a
sho e eplica ion ime han M. ube culosis (9, 23). Simila o
human TB, ish mycobac e iosis displays an acu e and ch onic
o m and he subsequen ly o med g anulomas also esemble
he lesions caused by M. ube culosis [Figu e1 (13–15)]. Bo h
bac e ia a e able o su i e and eplica e wi hin mac ophages
(23, 25). In a labo a o y se ing, he zeb a ish is an ad an a-
geous choice as a hos o ganism o M. ma inum o se e al
easons: mul iple in ec ion echniques can be used o bo h
zeb a ish emb yos and adul s (26); o a e iew, see Re . (8,
27–29) (Figu es1B,C). The anspa ency o he emb yos allows
he use o sophis ica ed in i o eal- ime imaging echniques,
including mul iple leukocy e and mac ophage luo escen
epo e lines (30–32), and se e al echniques o gene ic
manipula ion (6, 33–35). Mo eo e , zeb a ish a e small in size,
and p oduce nume ous o sp ing, making hem also sui able
o la ge-scale sc eening s udies, including d ug sc eens (6).
Despi e he ana omical di e ences be ween ish and mammals,
he zeb a ish is a e eb a e model wi h an inna e and adap-
i e immuni y consis ing o he same p ima y componen s as
p esen in humans (6, 36, 37). As zeb a ish lack lymph nodes,
immune cells mainly de elop and pe o m hei unc ions in
he spleen, he kidney, and he hymus (6, 38, 39). Zeb a ish
emb yos ely solely on inna e immuni y. In he emb yos,
unc ional mac ophages and neu ophils eme ge 1 and 2days
pos - e iliza ion (dp ), espec i ely, while lymphocy es s a
de eloping a e 4dp and he adap i e immuni y becomes ully
unc ional a 4weeks pos - e iliza ion (wp ) (7). This acili-
a es s udying he unc ion o he inna e and adap i e immune
sys em, as well as di e en cell ypes, sepa a ely (8, 33).
HOST–MYCOBACTERIUM
inTeRACTiOnS–inSiGHTS inTO
eARLY inFeCTiOn e enTS FROM
ZeBRAFiSH LAR Ae
Zeb a ish la ae ha e been especially use ul in elucida ing he
ole o mac ophages and he s a egies he phagocy osed myco-
bac e ia use o supp ess phagosomal ma u a ion, apop osis, and
he an ibac e ial inna e immune esponse (8, 25, 40). Sca enge
ecep o s o di e en classes ha e been s udied in he con ex
o he phagocy osis o mycobac e ia. Fo example, Ma co binds
he glycolipid ehalose 6,69-dimycola e on he mycobac e ial
cell wall and a ec s he egula ion o he subsequen p oin lam-
ma o y esponse (41). CD36 also appea s o be in ol ed in
mycobac e ial con ol, hough i s ole and he egula ion o i s
exp ession seem qui e complica ed (42). Following phagocy o-
sis, Toll-like ecep o (TLR) signaling ia MyD88 is needed o
esis ance agains an ea ly mycobac e ial in ec ion (43, 44). Fo
example, he ac i a ion o he TLR signaling leads o he p o-
duc ion o an ibac e ial molecules by mac ophages, such as he
pe o in Mpeg1.2 (45). In he zeb a ish, he myd88−/− mu an s
ha e been a use ul ool in elucida ing he ole o TLR signaling
in mycobac e ial esis ance (46, 47).
Toll-like ecep o signaling is in ol ed in he ini ia ion o he
p oduc ion o eac i e oxygen and ni ogen species (ROS and
RNS, espec i ely). In he zeb a ish, ea ly s abiliza ion o he
ansc ip ion ac o hypoxia-inducible ac o 1α (Hi -1α) in neu-
ophils limi s bac e ial g ow h by inducing iNOS, which in u n
leads o inc eased p o ein ni osyla ion (48). La e in he cou se
o he in ec ion, his esponse is dec eased in he de eloping
g anulomas by a mechanism independen o he ea ly sec e o y
an igenic a ge 6 sys em 1/ egion o di e ence 1 (ESX-1/RD1)
i ulence locus (49).
As a mean o coun e ac he mycobac e ial e asion s a egies,
he hos can induce au ophagy, a p ocess which enables cells o
diges hei cy oplasmic con en s, including mic oo ganisms and
memb anous s uc u es, in lysosomes (50). This is dependen
on TLR signaling and Myd88, which a e linked ia he DNA
damage- egula ed au ophagy modula o DRAM1. DRAM1 is
needed o he o ma ion o au ophagosomes and o hei usion
wi h lysosomes, while a DRAM1 de iciency leads o de ec s in
main aining he mycobac e ia inside esicles in mac ophages
and in he con ol o mycobac e ial g ow h (51), Again, zeb a ish
la ae p o ide easible ools o obse ing he e en s o au ophagy
in i o and in eal- ime u ilizing bo h ligh and elec on mic os-
copy, including he GFP-Lc3- ansgenic line, o he isualiza ion
o au ophagosomal s uc u es (52).
ZeBRAFiSH LAR Ae MODeL
CHALLenGe OLD DOGMAS in
MYCOBACTeRiAL ReSiSTAnCe
A e a success ul in ec ion by mycobac e ia, g anulomas a e
seeded. The g anulomas in zeb a ish emb yos mainly consis o
agg ega ed mac ophages, in a- and ex acellula mycobac e ia,
oge he wi h ec ui ed neu ophils, and o m wi hin a ew days
pos -in ec ion (18, 53); o a e iew, see Re . (54). Despi e hei
a he p imi i e s uc u e, he emb yonic zeb a ish g anulomas
p o ide a physiological model o s udying cellula p ocesses
a ec ing mycobac e ial in ec ions, such as he gene a ion o
hypoxia and angiogenesis. Injec ion o mycobac e ia in o he cau-
dal ein esul s in he de elopmen o non-hypoxic g anulomas
in he ichly ascula ized a ea o he caudal hema opoie ic issue
(48). In con as , g anulomas esul ing om a unk in ec ion
eside in a spa sely ascula ized a ea and can become hypoxic
and induce ascula iza ion (55).
Impo an ly, s udies on ea ly g anulomas in zeb a ish la ae
ha e challenged some o he old dogmas. In gene al, g anulomas
ha e long been conside ed a p o ec ion mechanism elici ed by he
hos . Howe e , ecen e idence om zeb a ish emb yos sugges s
ha M. ma inum uses he mac ophages and g anulomas o i s own
expansion and dissemina ion (16, 17). The bac e ia can ec ui
new, unin ec ed mac ophages o he g anuloma si e using he
RD1 locus (16). The a i ing mac ophages phagocy ize in ec ed,
dead cells and hus con ibu e o he sp eading o he bac e ia.
The ec ui men o new mac ophages is enhanced by he bac e ia
by ESAT-6-media ed p oduc ion o ma ix me allop o einase 9
(17). Mycobac e ia also need he RD1 locus o escaping om he
phagosomes in o he cy oplasm o in ec ed mac ophages bo h in
5
Myllymäki e al.
Zeb a ish as a Tube culosis Model
F on ie s in Immunology | www. on ie sin.o g May 2016 | Volume 7 | A icle 196
human cells and in zeb a ish (51, 56). Addi ionally, mycobac e ia
use cell su ace lipids o mask pa hogen-associa ed molecula pa -
e ns, hence selec i ely in ec pe missi e mac ophages and a oid
mic obicidal ones (57). Thus, g anulomas p esen a comba zone
o he hos immune sys em and he bac e ial esponse, a he
han pu ely a p o ec ion mechanism o he hos o p e en he
sp ead o bac e ia (4).
In addi ion o basic esea ch, he zeb a ish la ae p o ide a
easible ool o ea ly-s age d ug de elopmen and la ge-scale
sc eens (33, 58, 59). Two majo , and ela ed, issues in imp o -
ing he TB d ug de elopmen a e he leng hiness o he cu a i e
ea men s and he inc easing eme gence o d ug- esis an
bac e ia (1). Disco e ies made in zeb a ish la ae e ealed ha
in acellula mycobac e ia use hei e lux pumps o acqui e a
ole ance agains he an ibio ics commonly used o ea human
TB, which allows he bac e ia o pe sis and eplica e in he
cy osol. M. ube culosis uses he same mechanism (60). Howe e ,
his ole ance was e e sed by an e lux pump inhibi o , such as
e apamil, which can he e o e educe he ole ance o an ibio ics
and he eby sho en ea men imes (61). These esul s p o e ha
he zeb a ish la al model can eplica e he unc ion o an i ube -
cula compounds (60). In addi ion, angiogenesis has been shown
o be impo an o g anuloma o ma ion. The e o e, a ge ing
ascula iza ion, o example, by inhibi ing ascula endo helial
g ow h ac o ecep o (VEGFR) signaling could p o ide a means
o a ge mycobac e ial in ec ions and inhibi mycobac e ial dis-
semina ion, esembling he s a egy used in cance he apies (55).
On he hos ’s side, cy okines and hei espec i e ecep o s
play an impo an ole in p o ec ion agains mycobac e ia. Fo
example, he signaling axis media ed by chemokine CXC-mo i e
con aining ecep o 3 (CXCR3) has been implica ed in mycobac-
e ial sp eading and could hus p o ide a he apeu ic a ge (62).
In addi ion, in bo h humans and zeb a ish la ae he leuko iene
A4 hyd olase (LTA4H) locus con ols p o- and an i-in lamma o y
media o s ha in u n con ol he exp ession o TNF (63). While
TNF is equi ed o he hos esponse agains mycobac e ia, i s
excess ende s he hos mo e suscep ible o an in ec ion, high-
ligh ing he impo ance o a balanced esponse (64). Thus, he
zeb a ish can elucida e he pa hways con olling he hos immune
esponses, and his in o ma ion can be u he applied o a ge -
ing hese pa hways wi h new d ugs and de eloping hos -di ec ed
he apies (8, 63–66). In addi ion, u he in es iga ion in o he
e asion s a egies ha mycobac e ia use o in e e e wi h he hos
de ense mechanisms can po en ially lead o he disco e y o no el
d ug a ge s o comba mycobac e ial diseases (49, 67).
MYCOBACTeRiAL LATenCY S.
ADAPTi e iMMUniTY–THe ADULT
ZeBRAFiSH AS A MODeL FOR TB
One o he main ad an ages o he zeb a ish–M. ma inum model
may lie in g anuloma o ma ion, which has no been easy o ep o-
duce in he adi ional model animals (22). In he adul zeb a ish,
howe e , he his ology o he ma u e g anulomas esembles hose
seen in human TB wi h hei caseous, nec o ic co e su ounded
by leukocy es and epi helial cells (4, 17). G anulomas a e ound
in a ious o gans such as he panc eas, gonads, spleen, and li e
se e al weeks pos -in ec ion (Figu e1C) (13–15).
The la ency o TB is ano he aspec ha has been di icul o
eplica e expe imen ally (22). As his is also challenging o s udy
in humans, ou knowledge o he equi ed immunological mecha-
nisms o he con ol o a mycobac e ial in ec ion a i s di e en
s ages emains limi ed (17). Howe e , his could be imp o ed
by s udying he zeb a ish model, as like humans, adul zeb a ish
de elop a la en , non-p og essi e disease wi h do man bac e ia
esiding wi hin well-s uc u ed g anulomas, and he ish emain
asymp oma ic (13, 14). Mo eo e , eac i a ion o he bac e ia
can occu spon aneously, o can be induced expe imen ally by
an immune de iciency, such as exposu e o γ-i adia ion. In ei he
case, eac i a ion will lead o he ac i e sp eading o he bac e ia
and he de elopmen o symp oms simila o an ac i e in ec ion
as well as high le hali y, much as in human TB (13).
Al hough he ea ly cy okine esponse media ed by he inna e
immuni y plays an impo an ole in de e mining whe he a
mycobac e ial in ec ion leads o an acu e o la en disease, he
adap i e immuni y is also equi ed o con ol he bac e ia. This
is seen in he adul ag−/− zeb a ish ha a e de oid o lympho-
cy es. The mu an ish a e unable o gene a e a la en s a e o
he in ec ion and a e, he e o e, hype suscep ible o M. ma inum
(13, 14, 68). So a , he signi icance o di e en lymphocy e
subse s has no been ex ensi ely s udied in he zeb a ish myco-
bac e ial in ec ion. Ne e heless, Th1 as well as Th2 cells seem
o be in ol ed in he e ec i e con ol o mycobac e ial in ec ions
(69, 70). In e es ingly, a no el Th2-like subse o cells capable
o inhibi ing he g ow h o M. ube culosis has been ound in
human TB pa ien s. This obse a ion challenges he old idea
ha only Th1 cells a e impo an o mycobac e ial con ol (71).
Gene ic di e ences in he mycobac e ial s ains also seem o
a ec hei i ulence, o example, s ains isola ed om in ec ed
humans mo e commonly causing an acu e disease, and isola es
om poikilo he mic species causing a ch onic in ec ion in he
zeb a ish (72).
While he zeb a ish la ae p o ide a easible ool o sc eening
o d ugs agains TB, he adul zeb a ish appea s o be a p omis-
ing model o ea ly accine de elopmen . The zeb a ish can be
pa ially p o ec ed agains mycobac e iosis by BCG (68, 73) o
a enua ed M. ma inum (74), sugges ing he use o adul zeb a ish
as a model o s udying he easibili y o conse ed mycobac e ial
an igens as accines. Fo example, he RD1 i ulence locus and
he ESX-1 sec e ion sys em, which a e absen om BCG, ha e
been shown o be impo an o i ulence in M. ube culosis. In
M. ma inum, RD1 is also is also equi ed o g anuloma o ma ion
in bo h la ae and adul ish (14, 49, 75, 76). Indeed, he an igens
in his egion do show some po en ial as a ge s o accines in he
zeb a ish as well as in o he models (68, 73, 77–79). The zeb a ish
can also p o ide a easible model o sea ching o he mos e ec-
i e an igen combina ions and o s udying di e en accina ion
s a egies. Fo ins ance, a DNA-based accine consis ing o h ee
mycobac e ial an igens (Ag85, ESAT-6, and CFP10), which has
also been s udied in o he TB models, con e s p o ec ion agains
mycobac e ia in zeb a ish (68, 80–82). Fu he mo e, he e ec o
BCG can be boos ed by a DNA accine (68, 73, 80–82). Adul
zeb a ish could he e o e be used o de eloping accines o

6
Myllymäki e al.
Zeb a ish as a Tube culosis Model
F on ie s in Immunology | www. on ie sin.o g May 2016 | Volume 7 | A icle 196
bo h eplacing and boos ing BCG, as well as o s udying he
immunological co ela es equi ed o p o ec ion.
DiSCUSSiOn
The lack o an animal model ha ecapi ula es he human disease
s ages and pa hology has in pa hampe ed he de elopmen
o new d ugs, accines, and diagnos ic ools agains TB (22).
E en hough he mammalian animal models mos ly used o
TB esea ch, namely, mice, abbi s, and guinea pigs, do de elop
g anuloma ous s uc u es, only p ima es a e a na u al hos o
M. ube culosis and show ue la ency and eac i a ion (83).
Howe e , he use o p ima es as labo a o y animals is di icul in
e ms o e hical and economic issues as well as space limi a ions.
When d ug and accine de elopmen is conside ed, a na u al
hos –pa hogen pai is likely o be a mo e eliable model. Mo eo e ,
choosing o wo k wi h (zeb a) ish migh ha e an addi ional
p ac ical ad an age: as mycobac e ial in ec ions a e able o cause
epidemics in ish a ms, aqua iums, and zeb a ish acili ies (9, 84),
accina ing ish agains M. ma inum is o po en ial economic and
ecological ele ance. This has been s udied o some ex en , o
example, in he s iped bass wi h he Ag85A DNA accine (85)
and in he Japanese lounde wi h BCG (86). Thus, he esul s
ob ained in human biomedical esea ch and e e ina y s udies
could po en ially augmen each o he .
The zeb a ish–M. ma inum in ec ion exhibi s essen ially he
same disease phases as hose seen in human TB, including la ency
and eac i a ion–ei he spon aneously o ollowing immuno-
supp ession (13, 14). This migh ha e impo an implica ions
as he di e en disease phases p obably also ep esen on he
one hand di e en s a egies o bac e ial adap a ion, and on he
o he hand, di e en s ages o he hos immune esponse (87).
Since hese aspec s can be eplica ed in he zeb a ish in ec ion,
he model should acili a e a mo e de ailed dissec ion o bo h he
e ec i e (and dele e ious) immune esponses and he bac e ial
coun e s a egies in each s age o he in ec ion. The zeb a ish
model can also be used o complemen he human pa ien da a in
he iden i ica ion o eliable bioma ke s o he diagnosis o he
di e en s ages o TB (88).
Besides bioma ke s, new d ugs and accines a e needed o
comba TB. Fo his, a be e knowledge conce ning he co ela es
o a p o ec i e immune esponse is essen ial (89). The HI i us
a acks CD4+ T cells, and a co-in ec ion ende s he pa ien s
highly suscep ible o TB. The e o e, i seems ha CD4+ T cells a e
impo an o he hos esponse (90). This does no mean, howe e ,
ha he s udy o o he cell ypes should be neglec ed, as hey oo
can e eal new immunological mechanisms (70, 71). Mo eo e ,
de iciencies in he IFN-γ signaling axis lead o hype suscep ibili y
owa d TB, and hus IFN-γ exp ession has been associa ed wi h
p o ec ion agains he disease (90). Howe e , despi e inducing
high le els o IFN-γ p oduc ion, a p omising accine candida e,
MVA85A, ailed ecen ly o enhance p o ec ion in an e icacy ial
(91). This sugges s ha i is unlikely ha a single immunological
ac o could p edic he cou se o a TB in ec ion (90, 92). Thus,
he e a e s ill gaps in ou knowledge o how an e ec i e hos
de ense agains TB is elici ed, and ele an animal models a e
needed o ill in he missing in o ma ion. Fu he mo e, once he
pic u e o p o ec ion mechanisms is mo e comple e, he animal
models can aid us in ansla ing his in o ma ion in o he bene i
o clinical medicine. We belie e he zeb a ish will be an impo an
playe in ul illing bo h o hese asks.
AUTHOR COnTRiBUTiOnS
All au ho s con ibu ed o planning and w i ing he manusc ip
and designing o he igu es.
ACKnOwLeDGMenTS
We hank D . Helen Coope o e ising he language o he
manusc ip and MSc Mi ja Niskanen o echnical help wi h
his ology and MSc Me i Uusi-Mäkelä o help wi h he igu e.
FUnDinG
This wo k was suppo ed by he Founda ion o he Finnish
An i-Tube culosis Associa ion (HM), he Tampe e Tube culosis
Founda ion (HM and MR), he Finnish Academy (MR) (g an
numbe on 277495), he Sig id Juselius Founda ion (MR), he
Jane and Aa os E kko Founda ion (MR), he Compe i i e S a e
Resea ch Financing o he Expe Responsibili y A ea o Tampe e
Uni e si y Hospi al (MR), and Compe i i e S a e Resea ch
Financing o he Expe Responsibili y a ea o Oulu Uni e si y
Hospi al (MR).
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