biomolecules
A icle
Neu oimmune Response Media ed by Cy okines in Na u al
Sc apie a e Ch onic Dexame hasone T ea men
Isabel M. Guija o 1, Moisés Ga cés1, Pol And és-Beni o 2, Belén Ma ín1, Alicia O e o 1, Tomás Ba io 1,
Ma ga i a Ca mona 2, Isid o Fe e 2, Juan J. Badiola 1and Ma a Monzón1,*


Ci a ion: Guija o, I.M.; Ga cés, M.;
And és-Beni o, P.; Ma ín, B.; O e o,
A.; Ba io, T.; Ca mona, M.; Fe e , I.;
Badiola, J.J.; Monzón, M.
Neu oimmune Response Media ed
by Cy okines in Na u al Sc apie a e
Ch onic Dexame hasone T ea men .
Biomolecules 2021,11, 204.
h ps://doi.o g/10.3390/biom11020204
Recei ed: 14 Janua y 2021
Accep ed: 27 Janua y 2021
Published: 2 Feb ua y 2021
Publishe ’s No e: MDPI s ays neu al
wi h ega d o ju isdic ional claims in
published maps and ins i u ional a il-
ia ions.
Copy igh : © 2021 by he au ho s.
Licensee MDPI, Basel, Swi ze land.
This a icle is an open access a icle
dis ibu ed unde he e ms and
condi ions o he C ea i e Commons
A ibu ion (CC BY) license (h ps://
c ea i ecommons.o g/licenses/by/
4.0/).
1Resea ch Cen e o Encephalopa hies and T ansmissible Eme ging Diseases,
Ins i u e o Heal h Resea ch A agón (IIS), Uni e si y o Za agoza, C/Miguel Se e 155,
50013 Za agoza, Spain; [email p o ec ed] (I.M.G.); [email p o ec ed] (M.G.);
belenm@uniza .es (B.M.); [email p o ec ed] (A.O.); [email p o ec ed] (T.B.);
badiola@uniza .es (J.J.B.)
2Depa amen o de Pa ologíay Te apéu ica Expe imen al, Uni e sidad de Ba celona, 08907 Ba celona, Spain;
[email p o ec ed] (P.A.-B.); [email p o ec ed] (M.C.); [email p o ec ed] (I.F.)
*Co espondence: mmonzon@uniza .es; Tel.: +00-34-976-762944
Abs ac :
The ac ual ole o p ion p o ein-induced glial ac i a ion and subsequen cy okine sec e ion
du ing p ion diseases is s ill incomple ely unde s ood. The o e all aim o his s udy is o assess
he e ec o an an i-in lamma o y ea men wi h dexame hasone on di e en cy okines eleased
by neu oglial cells ha a e po en ially ela ed o neu oin lamma ion in na u al sc apie. This s udy
emphasizes he complex in e ac ions exis en among se e al pleio opic neu omodula o pep ides
and p o ides a global app oach o cla i y neu oin lamma o y p ocesses in p ion diseases. Addi ion-
ally, an impai men o communica ion be ween mic oglial and as oglial popula ions media ed by
cy okines, mainly IL-1, is sugges ed. The main no el y o his s udy is ha i is he i s one assessing
in si u neu oin lamma o y ac i i y in ela ion o ch onic an i-in lamma o y he apy, gaining ele ance
because i is based on a na u al model. The cy okine p o ile da a would sugges he ac i a ion o some
neu o oxici y-associa ed ou e. Consequen ly, a ge ing such a pa hway migh be a new app oach o
modi y he damaging e ec s o neu oin lamma ion.
Keywo ds: sc apie; cy okines; dexame hasone; neu oin lamma ion; p ion diseases
1. In oduc ion
Sc apie is conside ed he p o o ype o p ion diseases, which a e a g oup o neu ode-
gene a i e diso de s caused by he con e sion o a cellula p o ein in o a pa hological
iso o m called p ion.
Neu oin lamma ion is cu en ly a widely accep ed concep in neu odegene a ion,
pa icula ly in p ion diseases [
1
–
4
]. The neu oin lamma o y p ocess is de ined as he
p olonged ac i a ion o neu oglial cells wi h he co esponding p oduc ion o in lamma o y
cy okines [
5
]. Consequen ly, he e is a pa icula in e es in in es iga ing he oles o
he inna e and adap i e immune sys ems in se e al neu odegene a i e diso de s, wi h
neu oglia as a key elemen in he neu opa hological p ocess [6–8].
A ele an numbe o s udies ha e p oposed a c ucial ole o cy okines as neu oin-
lamma o y media o s in he cellula communica ion in hese p ion diseases [
3
,
9
–
14
]. The
de ec ion o hese cy okines was desc ibed coinciding wi h he onse o clinical signs
in bo h a mu ine model [
9
] and C eu z eld –Jakob disease (CJD) [
15
]. Mo eo e , a e-
cen s udy desc ibed he p esence o se e al genes implica ed in in lamma ion ha a e
up egula ed in ea ly phases o p ion in ec ion [
16
]. Ne e heless, al hough an al e ed
p o ile o in lamma o y in e media ies has been e idenced in some expe imen al mu ine
models [
9
,
14
,
15
,
17
–
21
], sca ce s udies ha e ocused on in si u issue exp ession o hese
p o eins [10,22], and none o hem on a na u al model.
Biomolecules 2021,11, 204. h ps://doi.o g/10.3390/biom11020204 h ps://www.mdpi.com/jou nal/biomolecules
Biomolecules 2021,11, 204 2 o 18
P e ious s udies de eloped in sc apie-a ec ed animals ha e led o conclusions abou
he glial ole in he neu odegene a i e p og ess ha was ex apola ed no only o o he
p ion bu also o he neu odegene a i e diso de s [
23
–
25
]. Mo e ecen ly, his same
in i o
model has been used o assess he changes o ac i a ion o glial cells associa ed wi h an i-
in lamma o y he apy [
26
]. This s udy cons i u ed a powe ul app oach o he in ol emen
o immune esponse in his neu odegene a i e disease, con i ming he occu ence o
neu oin lamma ion in neu odegene a ion. Speci ically, a po en ial ailu e o as ocy es
and a s imula ion o phagocy osis o p ion p o ein deposi s by mic oglia we e e idenced
a e dexame hasone (DEX) ea men . To examine he in e glial communica ion media ed
by cy okines in dep h cons i u es a main ool o ad ancemen o he knowledge o how
hese media o s a e eally in ol ed in neu oin lamma o y mechanisms con ibu ing o
neu odegene a ion [
27
–
30
]. I is indispensable o s udy he possible al e a ion o glial
c oss alk ha migh enhance ins ead o p e en neu onal damage. Thus, i could be a
c ucial a ge o he apeu ic app oaches in p ion diseases [31].
O e all, he ac ual ole o p ion p o ein-induced glial ac i a ion and subsequen cy-
okine exp ession du ing p ion diseases is s ill incomple ely unde s ood. Consequen ly, o
in es iga e he possible al e a ions o in si u cy okine exp ession in b ain samples om ani-
mals na u ally a ec ed by sc apie and DEX ea men would be eally help ul o de e mine
whe he hese p o eins could be signi ican ac o s in he p og ess o neu odegene a ion
in his g oup o diseases. Thus, he speci ic aim o his s udy is o assess he e ec o he
an i-in lamma o y ea men on di e en cy okines which could be po en ially ela ed o
neu oin lamma ion. Bo h immunohis ochemical and exp ession pa e ns o di e en p o-
and an i-in lamma o y cy okines in se e al b ain egions om ea ed and non- ea ed
sc apie-a ec ed sheep a e compa ed in his s udy as a i s s ep owa ds he ul ima e goal
ha is o de e mine whe he hese p o eins ep esen ele an a ge s in he immunopa ho-
genesis o neu odegene a ion.
2. Ma e ial and Me hods
All he ollowing expe imen al p ocedu es we e p e iously app o ed by he E hical
Commi ee o Uni e si y o Za agoza (Re e ence numbe : PI41/16, 03/10/2016). All e o s
we e made o minimize animal su e ing du ing he expe imen s and o educe he numbe
o animals used.
All he analyses we e pe o med on samples coming om animals included in a p e-
iously published s udy whe e, as ci ed abo e, he glial ac i a ion esponse in he p og ess
o na u al sc apie a e ch onic DEX ea men had been assessed [
26
]. All expe imen al
de ails we e p o ided in his p e ious manusc ip , bu b ie ly, a o al o 25 sheep (age
anging om 4 o 10 yea s and all excep o one o hem wi h he e ozygous alanine-
a ginine- glu amine and alanine-a ginine-his idine, ARQ/ARH, p esen ing homozygous
alanine-a ginine- glu amine, ARQ/ARQ geno ype) we e included in his s udy: 10 heal hy
con ol (o which 4 ea ed and 6 non- ea ed) and 15 clinical sc apie Rasa A agonesa ewes
(10 ea ed plus 5 non- ea ed). Heal hy con ols we e conside ed essen ial in o de o
speci ically obse e he e ec o ea men (daily in amuscula , IM 0.04 mg/kg dose un il
eu hanasia by endpoin c i e ia, 16 mon hs he longes ) in no mal condi ions in o ine
species. A e eu hanasia wi h in a enous pen oba bi al injec ion, nec opsy o each sheep
was pe o med and 80 samples we e subsequen ly collec ed and dis ibu ed o di e en
s udies. One hemi-sec ion om each sample was ixed by imme sion in 4% pa a o malde-
hyde o immunohis ochemical s udies and he o he hemi-sec ion was ozen a
−
80
◦
C
o molecula s udies (RT-qPCR).
2.1. Immunohis ochemical Techniques
Immunohis ochemis y (IHC) was ca ied ou in o de o assess in si u neu oin lamma-
o y p o ile associa ed wi h DEX ea men in all sheep. I was compa ed wi h non- ea ed
sheep g oup in ou encephalic a eas ( on al co ex: Fc, ce ebellum: Cb, obex: O and
medulla oblonga a: MO).
Biomolecules 2021,11, 204 3 o 18
P io 4
µ
m sec ioning, pa a in-embedding o ixed samples was de eloped. A e
speci ic p e- ea men s o an igen e ie al, speci ic immunohis ochemical p o ocols by
using speci ic p ima y an ibodies agains hose cy okines o hei ecep o s mainly s udied
in li e a u e ela ed ( o ou knowledge, IL-1
α
, IL-1R, IL-2R, IL-6, IL-10R, TNFR and IFN
γ
R)
we e applied. EnVision sys em (DAKO, Glos up, Denma k) and diaminobenzidine (DAB;
DAKO, Glos up, Denma k) we e used as he isualiza ion sys em and ch omogen, espec-
i ely. Hema oxylin coun e s aining and moun ing in DPX was inally pe o med on all
sec ions.
All slides we e analysed by wo independen obse e s sco ing he in ensi y o im-
munos aining om 0 (absence) o 4 (maximum p esence) by coun ing posi i e cells [
32
] in
5 mic oscopic ields in each b ain egion examined. Mo eo e , p o ided ha ce ebellum
has been p oposed as a pseudo e e ence egion o de ec neu oin lamma ion [
33
], close
a en ion was paid o di e en p o iles o cy okine dis ibu ion in his b ain egion. The
ocus was on Pu kinje cells based on p e ious esul s e e ing his neu onal ype as he
mos damaged while hey a e he mos p o ec ed neu ons in his a ea [
23
,
26
,
34
]. Table 1
summa izes all he p ima y an ibodies and he p o ocols applied as ollowing desc ibed.
Table 1. An ibodies used o immunohis ochemical echniques and e ie al me hod applied o each one.
An ibody An igen Type Dilu ion Re ie al Me hod Sou ce
IL-1 alpha IL-1 Polyclonal 1:100 Au ocla e 121 ◦C
(ci a e bu e 10%)
The moFishe
An i-IL-1RN IL-1R Polyclonal 1:100 Au ocla e 121 ◦C
(ci a e bu e 10%)
Sigma
IL-2R.1 IL-2R Monoclonal 1:1000 PTLink 96 ◦C The moFishe
8H12 IL-6 Monoclonal 1:40 Au ocla e 121 ◦C
(ci a e bu e 10%)
The moFishe
OTI1D10 IL-10R Monoclonal 1:250 PTLink 96 ◦C The moFishe
Be -H2 TNFR Monoclonal Ready o use PTLink 96 ◦C Dako
IFNGR1 IFNγR Polyclonal 1:200 Au ocla e 121 ◦C
(ci a e bu e 10%)
The moFishe
2.1.1. IL-1, IL-1R, IL-6 and IFNγR De ec ion
A p e- ea men consis ing o hyd a ed hea ing a 121
◦
C in ci a e bu e 10% o
20 min p eceded he endogenous pe oxidase blocking (DAKO, Glos up, Denma k) o
5 min and incuba ion o e nigh 4
◦
C wi h di e en p ima y an ibodies: polyclonal IL-1
α
(1:100; The moFishe Scien i ic, Wal ham, MA, USA), polyclonal IL-1RN (1:100, Sigma, S .
Louis, MO, USA), monoclonal 8H12 (1:40; The moFishe Scien i ic, Wal ham, MA, USA) o
polyclonal IFNGR1 (1:200, The moFishe Scien i ic, Wal ham, MA, USA).
2.1.2. IL-2R, IL-10R and TNFR De ec ion
A p e- ea men consis ing o hyd a ed hea ing a 96
◦
C in ci a e bu e 10% o
20 min p eceded he endogenous pe oxidase blocking (DAKO, Glos up, Denma k) o
5 min and incuba ion wi h di e en p ima y monoclonal an ibodies: IL-2R.1 (1:1000,
o e nigh 4
◦
C; The moFishe Scien i ic, Wal ham, MA, USA), OTI1D10 (1:250, o e nigh
4
◦
C; The moFishe Scien i ic, Wal ham, MA, USA) o Be -H2 ( eady o use, 30 min RT;
DAKO, Glos up, Denma k).
2.2. RT-qPCR
Ce ebellum and on al co ex ozen issues om ea ed and non- ea ed sc apie
animals we e included in he ollowing compa a i e molecula analysis o some in lam-
ma o y ma ke s.
Biomolecules 2021,11, 204 4 o 18
2.2.1. RNA Pu i ica ion
The pu i ica ion o RNA was pe o med ollowing he ins uc ions o he supplie
(RNeasy Lipid Tissue Mini ki , Qiagen, GmbH, Hilden, Ge many). RNA in eg i y and
28S/18S a ios we e de e mined wi h he Agilen Bioanalyze (Agilen Technologies Inc,
San a Cla a, CA, USA). Samples we e ea ed wi h DNase diges ion, and RNA concen-
a ion was e alua ed using a NanoD op Spec opho ome e (The moFishe Scien i ic,
Wal ham, MA, USA).
RNA samples wi h OD 260/280 a ios close o 5.0 we e selec ed o e e se ansc ip-
ion. Finally, a o al o 5 ea ed and 4 non- ea ed clinical sheep we e included in his
molecula analysis.
2.2.2. Re o ansc ip ion
Re o ansc ip ion o RNA in o cDNA was pe o med acco ding o he manu ac u e ’s
ins uc ions (High-Capaci y CDNA Re e se T ansc ip ion Ki , Applied Bio sys ems, Fos e
Ci y, CA, USA).
2.2.3. RT-qPCR
Gene exp ession o IL-1, IL-6, IL-10Ra, IL-10Rb and IFN
γ
in bo h clinical non- ea ed
and ea ed sheep was assessed. The pa ame e s o he eac ions we e 50
◦
C o 2 min,
95 ◦C o 10 min, and 40 cycles o 95 ◦C o 15 sec and 60 ◦C o 1 min.
Da a we e assessed using he
∆∆
C me hod, using Hypoxan hine Phospho ibosyl
ans e ase 1 (HRPT-1) and
β
-glucu onidase (GUS-
β
) as e e ence genes. Table 2shows
TaqMan p obes used o hese molecula s udies.
Table 2. Taqman p obes used o RT-qPCR analysis.
Gene Full Name Re e ence Sou ce
Gus-β β-glucu onidase ( e e ence gene)
Oa04828868_m1
The moFishe
HPRT-1 Hypoxan hine Phospho ibosyl ans e ase 1 ( e e ence gene)
Oa04825272_gH
The moFishe
IL-1αIn e leukin 1 alpha
Oa04658681_m1
The moFishe
IL-6 In e leukin 6
Oa04656315_m1
The moFishe
IL-10Ra In e leukin 10 ecep o alpha
Oa04822455_m1
The moFishe
IL-10Rb In e leukin 10 ecep o be a
Oa04894070_m1
The moFishe
IFNγIn e e on gamma
Oa04657364_m1
The moFishe
2.3. S a is ical Analysis
Fo IHC esul s, he no mali y o dis ibu ion was i s assessed by Kolmogo o -
Smi no es . The non-pa ame ic Mann–Whi ney U es was used o assess quan i a i e
di e ences be ween non- ea ed and DEX ea ed g oups.
Da a o RT-qPCR we e e alua ed by S uden ’s es a e assessing no mali y also by
Kolmogo o –Smi no es .
SPSS so wa e (SPSS S a is ics o Windows, Ve sion 17.0, Chicago, IL, USA) was used
o hese analyses and signi icance in all cases was aken a * p< 0.05. All g aphs we e
pe o med wi h G aphPad P ism 6.0 (San Diego, CA, USA). Da a p esen ed in igu es a e
exp essed as means and he s anda d e o o he mean (mean +/−SEM).
3. Resul s
3.1. Immunohis ochemis y
3.1.1. IL-1
DEX- ea ed con ols always displayed highe in ensi y o IL-1 s aining compa ed
o he un ea ed samples. Howe e , clinically ea ed animals showed lowe di e ences
compa ed o hei espec i e un ea ed g oup and we e e en e e sed (Figu e 1A).
Biomolecules 2021,11, 204 5 o 18
Biomolecules 2021, 11, x FOR PEER REVIEW 5 o 18
Figu e 1. IL-1 immunos aining. (A) No e he e iden highe in ensi y o immunos aining in me-
dulla oblonga a, MO om DEX ea ed con ol sheep compa ed o an un ea ed one. Scale ba s:
100 µm. (B) A signi ican e ec o DEX was obse ed in MO om ea ed con ols (* p < 0.05).
Howe e , in clinical s age, no signi ican changes we e de ec ed despi e o he ea men .
The Mann–Whi ney U es e ealed signi ican e ec s o DEX in ea ed con ols com-
pa ed o un ea ed samples, showing an inc ease o IL-1 immunos aining in MO (* p =
0.024). Meanwhile, no changes we e de ec ed in clinically ea ed sc apie sheep depending
on he ea men (Figu e 1B).
In Cb, immunos aining o IL-1 was widesp ead in all laye s (Figu e 2A).
Figu e 2. Mo phological indings in ce ebellum immunos ained wi h di e en p ima y an ibodies. (A) Immunos aining
o IL-1 was widesp ead in all laye s. (B) IL-1R immuno eac i i y was loca ed mainly su ounding Pu kinje cells, sugges -
Figu e 1.
IL-1 immunos aining. (
A
) No e he e iden highe in ensi y o immunos aining in medulla
oblonga a, MO om DEX ea ed con ol sheep compa ed o an un ea ed one. Scale ba s: 100
µ
m.
(
B
) A signi ican e ec o DEX was obse ed in MO om ea ed con ols (* p< 0.05). Howe e , in
clinical s age, no signi ican changes we e de ec ed despi e o he ea men .
The Mann–Whi ney U es e ealed signi ican e ec s o DEX in ea ed con ols
compa ed o un ea ed samples, showing an inc ease o IL-1 immunos aining in MO
(
*p= 0.024
). Meanwhile, no changes we e de ec ed in clinically ea ed sc apie sheep
depending on he ea men (Figu e 1B).
In Cb, immunos aining o IL-1 was widesp ead in all laye s (Figu e 2A).
3.1.2. IL-1R
In gene al, immunos aining was lowe in ea ed animals compa ed o non- ea ed
ones, excep o con ols in Fc, whe e he pa e ns we e exac ly he opposi e (highe ). These
di e ences e iden ly inc eased in O and Cb a clinical s age (Figu e 3A).
Despi e he ac ha he e we e no s a is ically signi ican changes obse ed o his
ma ke , a end o a educ ion in immunos aining in Cb om animals in he clinical s age
was de ec ed (# p= 0.082) (Figu e 3B).
IL-1R immuno eac i i y in Cb was nea ly exclusi ely loca ed in cells su ounding
Pu kinje cells, sugges ing mo phology consis en wi h speci ic as ocy es (Figu e 2B).
3.1.3. IL-2R
I is only ele an o poin ou ha Fc showed an exace ba ed inc ease o eac i i y
agains his cy okine, ega dless o ea men , and disease, in compa ison wi h he o he
b ain a eas.
No s a is ically signi ican di e ences we e ound ega ding ea men in bo h he
con ol and clinical g oups (Figu e 4A).

Biomolecules 2021,11, 204 6 o 18
Biomolecules 2021, 11, x FOR PEER REVIEW 5 o 18
Figu e 1. IL-1 immunos aining. (A) No e he e iden highe in ensi y o immunos aining in me-
dulla oblonga a, MO om DEX ea ed con ol sheep compa ed o an un ea ed one. Scale ba s:
100 µm. (B) A signi ican e ec o DEX was obse ed in MO om ea ed con ols (* p < 0.05).
Howe e , in clinical s age, no signi ican changes we e de ec ed despi e o he ea men .
The Mann–Whi ney U es e ealed signi ican e ec s o DEX in ea ed con ols com-
pa ed o un ea ed samples, showing an inc ease o IL-1 immunos aining in MO (* p =
0.024). Meanwhile, no changes we e de ec ed in clinically ea ed sc apie sheep depending
on he ea men (Figu e 1B).
In Cb, immunos aining o IL-1 was widesp ead in all laye s (Figu e 2A).
Figu e 2. Mo phological indings in ce ebellum immunos ained wi h di e en p ima y an ibodies. (A) Immunos aining
o IL-1 was widesp ead in all laye s. (B) IL-1R immuno eac i i y was loca ed mainly su ounding Pu kinje cells, sugges -
Figu e 2.
Mo phological indings in ce ebellum immunos ained wi h di e en p ima y an ibodies. (
A
) Immunos aining o
IL-1 was widesp ead in all laye s. (
B
) IL-1R immuno eac i i y was loca ed mainly su ounding Pu kinje cells, sugges ing
a mo phology consis en wi h as ocy es. (
C
) IL-2R was mainly ound in cy oplasm o Pu kinje cells and cells appea ing
as ocy es. (
D
) IL-6 immunos aining was mainly p esen in Pu kinje cells as in acy oplasmic s aining, as well as s ained cells
esembling glial cells in bo h g anula and molecula laye s. (
E
) IL-10R immunos aining appea ed spo in acy oplasmic in
Pu kinje cells as well as g anula cells and o he cellula ype wi h as ocy ic mo phology. (
F
) Pu kinje cells exp essed TNFR
in a e y lowe ex en han he es o ma ke s. (
G
) IFN
γ
R immunos aining pa e n was mainly localized in he cy oplasm
and dend i ic spines o Pu kinje cells. Scale ba s: 50 µm.
Biomolecules 2021, 11, x FOR PEER REVIEW 6 o 18
ing a mo phology consis en wi h as ocy es. (C) IL-2R was mainly ound in cy oplasm o Pu kinje cells and cells appea -
ing as ocy es. (D) IL-6 immunos aining was mainly p esen in Pu kinje cells as in acy oplasmic s aining, as well as
s ained cells esembling glial cells in bo h g anula and molecula laye s. (E) IL-10R immunos aining appea ed spo in a-
cy oplasmic in Pu kinje cells as well as g anula cells and o he cellula ype wi h as ocy ic mo phology. (F) Pu kinje cells
exp essed TNFR in a e y lowe ex en han he es o ma ke s. (G) IFNγR immunos aining pa e n was mainly localized
in he cy oplasm and dend i ic spines o Pu kinje cells. Scale ba s: 50 µm.
3.1.2. IL-1R
In gene al, immunos aining was lowe in ea ed animals compa ed o non- ea ed
ones, excep o con ols in Fc, whe e he pa e ns we e exac ly he opposi e (highe ).
These di e ences e iden ly inc eased in O and Cb a clinical s age (Figu e 3A).
Despi e he ac ha he e we e no s a is ically signi ican changes obse ed o his
ma ke , a end o a educ ion in immunos aining in Cb om animals in he clinical s age
was de ec ed (# p = 0.082) (Figu e 3B).
Figu e 3. IL-1R immunos aining. (A) Di e ences obse ed a e ea men in Cb a clinical s age
a e illus a ed. Scale ba s: 100 µm. (B) While a sub le educ ion o IL-1R in ea ed con ol g oup
was obse ed in he es o a eas examined, in Fc i inc eased a e DEX ea men . A clinical
s age, a dec ease o immunos aining in O and a end (# p = 0.082) o educ ion in Cb we e de-
ec ed a e ea men .
IL-1R immuno eac i i y in Cb was nea ly exclusi ely loca ed in cells su ounding
Pu kinje cells, sugges ing mo phology consis en wi h speci ic as ocy es (Figu e 2B).
3.1.3. IL-2R
I is only ele an o poin ou ha Fc showed an exace ba ed inc ease o eac i i y
agains his cy okine, ega dless o ea men , and disease, in compa ison wi h he o he
b ain a eas.
Figu e 3. Con .
Biomolecules 2021,11, 204 7 o 18
Biomolecules 2021, 11, x FOR PEER REVIEW 6 o 18
ing a mo phology consis en wi h as ocy es. (C) IL-2R was mainly ound in cy oplasm o Pu kinje cells and cells appea -
ing as ocy es. (D) IL-6 immunos aining was mainly p esen in Pu kinje cells as in acy oplasmic s aining, as well as
s ained cells esembling glial cells in bo h g anula and molecula laye s. (E) IL-10R immunos aining appea ed spo in a-
cy oplasmic in Pu kinje cells as well as g anula cells and o he cellula ype wi h as ocy ic mo phology. (F) Pu kinje cells
exp essed TNFR in a e y lowe ex en han he es o ma ke s. (G) IFNγR immunos aining pa e n was mainly localized
in he cy oplasm and dend i ic spines o Pu kinje cells. Scale ba s: 50 µm.
3.1.2. IL-1R
In gene al, immunos aining was lowe in ea ed animals compa ed o non- ea ed
ones, excep o con ols in Fc, whe e he pa e ns we e exac ly he opposi e (highe ).
These di e ences e iden ly inc eased in O and Cb a clinical s age (Figu e 3A).
Despi e he ac ha he e we e no s a is ically signi ican changes obse ed o his
ma ke , a end o a educ ion in immunos aining in Cb om animals in he clinical s age
was de ec ed (# p = 0.082) (Figu e 3B).
Figu e 3. IL-1R immunos aining. (A) Di e ences obse ed a e ea men in Cb a clinical s age
a e illus a ed. Scale ba s: 100 µm. (B) While a sub le educ ion o IL-1R in ea ed con ol g oup
was obse ed in he es o a eas examined, in Fc i inc eased a e DEX ea men . A clinical
s age, a dec ease o immunos aining in O and a end (# p = 0.082) o educ ion in Cb we e de-
ec ed a e ea men .
IL-1R immuno eac i i y in Cb was nea ly exclusi ely loca ed in cells su ounding
Pu kinje cells, sugges ing mo phology consis en wi h speci ic as ocy es (Figu e 2B).
3.1.3. IL-2R
I is only ele an o poin ou ha Fc showed an exace ba ed inc ease o eac i i y
agains his cy okine, ega dless o ea men , and disease, in compa ison wi h he o he
b ain a eas.
Figu e 3.
IL-1R immunos aining. (
A
) Di e ences obse ed a e ea men in Cb a clinical s age a e illus a ed. Scale ba s:
100
µ
m. (
B
) While a sub le educ ion o IL-1R in ea ed con ol g oup was obse ed in he es o a eas examined, in Fc i
inc eased a e DEX ea men . A clinical s age, a dec ease o immunos aining in O and a end (# p= 0.082) o educ ion in
Cb we e de ec ed a e ea men .
Biomolecules 2021, 11, x FOR PEER REVIEW 7 o 18
No s a is ically signi ican di e ences we e ound ega ding ea men in bo h he
con ol and clinical g oups (Figu e 4A).
Figu e 4. No signi ican di e ences we e ound ei he in con ol o in clinical g oups o immunos aining in ensi y o (A)
IL-2R (B) TNFR o (C) IFNγR. A highe in ensi y in obex, O and medulla oblonga a, MO a e ea men was e idenced in
con ols ha e e sed in DEX clinical animals.
In Cb, IL-2R was limi ed o he cy oplasm o Pu kinje cells and cells wi h an as ocy ic
appea ance (Figu e 2C).
3.1.4. IL-6
Immunos aining o IL-6 in DEX- ea ed con ols was highe in all b ain a eas com-
pa ed o un ea ed con ols, excep in O, whe e he labeling in ensi y was lowe . This e-
duc ion in O was mo e e iden a clinical s ages o disease (Figu e 5A).
The inc ease o immunos aining in ensi y eached signi icance in Fc o he con ol
g oup when animals we e DEX- ea ed (* p = 0.040). Meanwhile, he immuno eac i i y o
his cy okine showed a dec easing end in con ol animals (# p = 0.071) ha con e ed
in o signi ican in O in he clinical g oup (* p = 0.041) (Figu e 5B).
This neu omodula o pep ide was mainly p esen in Pu kinje cells as in acy oplas-
mic s aining (Figu e 2D). Mo eo e , he cy okine de ec ed was p esen in speci ic cells
esembling glial cells in bo h g anula and molecula laye s.
Figu e 4.
No signi ican di e ences we e ound ei he in con ol o in clinical g oups o immunos aining in ensi y o (
A
)
IL-2R (
B
) TNFR o (
C
) IFN
γ
R. A highe in ensi y in obex, O and medulla oblonga a, MO a e ea men was e idenced in
con ols ha e e sed in DEX clinical animals.
In Cb, IL-2R was limi ed o he cy oplasm o Pu kinje cells and cells wi h an as ocy ic
appea ance (Figu e 2C).
3.1.4. IL-6
Immunos aining o IL-6 in DEX- ea ed con ols was highe in all b ain a eas com-
pa ed o un ea ed con ols, excep in O, whe e he labeling in ensi y was lowe . This
educ ion in O was mo e e iden a clinical s ages o disease (Figu e 5A).
Biomolecules 2021,11, 204 8 o 18
Biomolecules 2021, 11, x FOR PEER REVIEW 8 o 18
Figu e 5.
IL-6 immunos aining. (
A
) Mic og aphs ep esen an inc ease o IL-6 immuno eac i i y in on al co ex, Fc in
ea ed con ols while sligh ly educed in bo h ea ed g oups in obex, O. Scale ba s: 100
µ
m. (
B
) DEX con ol g oup
p esen ed a signi ican inc ease in Fc (* p< 0.05) and a end o a dec ease in O (# p= 0.071). I was signi ican ly dec eased in
his a ea when clinical g oup was ea ed (* p< 0.05).
Biomolecules 2021,11, 204 9 o 18
The inc ease o immunos aining in ensi y eached signi icance in Fc o he con ol
g oup when animals we e DEX- ea ed (* p= 0.040). Meanwhile, he immuno eac i i y o
his cy okine showed a dec easing end in con ol animals (# p= 0.071) ha con e ed in o
signi ican in O in he clinical g oup (* p= 0.041) (Figu e 5B).
This neu omodula o pep ide was mainly p esen in Pu kinje cells as in acy oplas-
mic s aining (Figu e 2D). Mo eo e , he cy okine de ec ed was p esen in speci ic cells
esembling glial cells in bo h g anula and molecula laye s.
3.1.5. IL-10R
Immunos aining o his ma ke was he highes in all assessed samples in bo h he
con ol and clinical g oups, ega dless o ea men , bu especially in Fc, whe e he highes
sco e was eached ega dless o ea men o disease.
Along he same lines, al hough no signi ican , a end owa d a dec ease (# p= 0.074)
in Cb o clinical sheep a e ea men was obse ed (Figu e 6A). In gene al, s a is ically,
immunos aining o IL-10R did no e eal any changes be ween g oups a e ea men
(
p> 0.05
). Ne e heless, a e y ele an dec ease in O in ensi y in ea ed clinical animals
was e idenced o each a lowe in ensi y han un ea ed, while he s aining in ensi y was
highe in con ols (Figu e 6B).
Biomolecules 2021, 11, x FOR PEER REVIEW 9 o 18
Figu e 5. IL-6 immunos aining. (A) Mic og aphs ep esen an inc ease o IL-6 immuno eac i i y in on al co ex, Fc in
ea ed con ols while sligh ly educed in bo h ea ed g oups in obex, O. Scale ba s: 100 µm. (B) DEX con ol g oup p e-
sen ed a signi ican inc ease in Fc (*p < 0.05) and a end o a dec ease in O (# p = 0.071). I was signi ican ly dec eased in
his a ea when clinical g oup was ea ed (* p < 0.05).
3.1.5. IL-10R
Immunos aining o his ma ke was he highes in all assessed samples in bo h he
con ol and clinical g oups, ega dless o ea men , bu especially in Fc, whe e he highes
sco e was eached ega dless o ea men o disease.
Along he same lines, al hough no signi ican , a end owa d a dec ease (# p = 0.074)
in Cb o clinical sheep a e ea men was obse ed (Figu e 6A). In gene al, s a is ically,
immunos aining o IL-10R did no e eal any changes be ween g oups a e ea men (p
> 0.05). Ne e heless, a e y ele an dec ease in O in ensi y in ea ed clinical animals
was e idenced o each a lowe in ensi y han un ea ed, while he s aining in ensi y was
highe in con ols (Figu e 6B).
Figu e 6. IL-10R immunos aining. (A) Dec ease in in ensi y in Cb o ea ed clinical sheep is illus a ed. Scale ba s: 100 µm.
(B) No signi ican changes we e de ec ed a e ea men in con ol g oup, only a highe inc ease in O was ou s anding.
No signi ican changes we e ei he obse ed in clinical s age a e ea men , jus a end o lowe in ensi y in Cb o ea ed
clinical sheep (# p = 0.074).
This ma ke s ained Pu kinje neu ons consis en wi h a spo in acy oplasmic pa -
e n. Addi ionally, he s aining showed a high in ensi y in g anula cells and some o he s
wi h as ocy ic mo phology (Figu e 2E).
Figu e 6.
IL-10R immunos aining. (
A
) Dec ease in in ensi y in Cb o ea ed clinical sheep is illus a ed. Scale ba s: 100
µ
m.
(
B
) No signi ican changes we e de ec ed a e ea men in con ol g oup, only a highe inc ease in O was ou s anding. No
signi ican changes we e ei he obse ed in clinical s age a e ea men , jus a end o lowe in ensi y in Cb o ea ed
clinical sheep (# p= 0.074).
Biomolecules 2021,11, 204 16 o 18
22.
Cunningham, C.; Wilcockson, D.C.; Campion, S.; Lunnon, K.; Pe y, V.H. Cen al and Sys emic Endo oxin Challenges Exace ba e
he Local In lamma o y Response and Inc ease Neu onal Dea h du ing Ch onic Neu odegene a ion. J. Neu osci.
2005
,25,
9275–9284. [PubMed]
23.
He nandez, R.S.; Sa asa, R.; Toledano, A.; Badiola, J.J.; Monzon, M. Mo phological App oach o Assess he In ol emen o
As ocy es in P ion P opaga ion. Cell Tissue Res. 2014,358, 57–63. [C ossRe ] [PubMed]
24.
Monzon, M.; He nandez, R.S.; Ga ces, M.; Sa asa, R.; Badiola, J.J. Glial Al e a ions in Human P ion Diseases: A Co ela i e S udy
o As oglia, Reac i e Mic oglia, P o ein Deposi ion, and Neu opa hological Lesions. Medicine 2018,97, e0320. [C ossRe ]
25.
Ga ces, M.; Guija o, M.I.; Va gas, A.; Badiola, J.J.; Monzon, M. Neu oglial Pa e ns A e Sha ed by Ce ebella om P ion and
P ion-Like Diso de A ec ed Pa ien s. Mech. Ageing De . 2019,184, 111176. [C ossRe ]
26.
Guija o, I.M.; Ga ces, M.; And es-Beni o, P.; Ma in, B.; O e o, A.; Ba io, T.; Ca mona, M.; Fe e , I.; Badiola, J.J.; Monzon, M.
Assessmen o Glial Ac i a ion Response in he P og ess o Na u al Sc apie a e Ch onic Dexame hasone T ea men . In . J. Mol.
Sci. 2020,21, 3231. [C ossRe ]
27.
Mo ales, I.; Guzman-Ma inez, L.; Ce da-T oncoso, C.; Fa ias, G.A.; Maccioni, R.B. Neu oin lamma ion in he Pa hogenesis o
Alzheime ’s Disease. A Ra ional F amewo k o he Sea ch o No el The apeu ic App oaches. F on . Cell. Neu osci.
2014
,8,
112. [C ossRe ]
28.
DiSaba o, D.J.; Quan, N.; Godbou , J.P. Neu oin lamma ion: The De il Is in he De ails. J. Neu ochem.
2016
,139 (Suppl. S2),
136–153. [C ossRe ]
29. Ransoho , R.M. How Neu oin lamma ion Con ibu es o Neu odegene a ion. Science 2016,353, 777–783. [C ossRe ]
30.
Kempu aj, D.; Thanga el, R.; Na e u, P.A.; Sel akuma , G.P.; Saeed, D.; Zahoo , H.; Zahee , S.; Iye , S.S.; Zahee , A. Neu oin lam-
ma ion Induces Neu odegene a ion. J. Neu ol. Neu osu g. Spine 2016,1, 1003.
31.
Ga wood, C.J.; Poole , A.M.; A he on, J.; Hange , D.P.; Noble, W. As ocy es A e Impo an Media o s o Abe a-Induced
Neu o oxici y and Tau Phospho yla ion in P ima y Cul u e. Cell Dea h Dis. 2011,2, e167. [C ossRe ] [PubMed]
32.
Rome o-T e ejo, J.L.; Gomez-Villamandos, J.C.; Ped e a, M.; Blanco, A.; Bau is a, M.J.; Sanchez-Co don, P.J. Immunohis o-
chemical S udy o Mac ophage and Cy okine Dynamics in he Gu o Sc apie-In ec ed Mice. His ol. His opa hol.
2010
,25,
1025–1038. [PubMed]
33.
Lyoo, C.H.; Ikawa, M.; Liow, J.S.; Zoghbi, S.S.; Mo se, C.L.; Pike, V.W.; Fuji a, M.; Innis, R.B.; K eisl, W.C. Ce ebellum Can Se e
as a Pseudo-Re e ence Region in Alzheime Disease o De ec Neu oin lamma ion Measu ed wi h PET Radioligand Binding o
T ansloca o P o ein. J. Nucl. Med. 2015,56, 701–706. [C ossRe ] [PubMed]
34.
Sa asa, R.; Ma inez, A.; Monleon, E.; Bolea, R.; Va gas, A.; Badiola, J.J.; Monzon, M. In ol emen o As ocy es in T ansmissible
Spongi o m Encephalopa hies: A Con ocal Mic oscopy S udy. Cell Tissue Res. 2012,350, 127–134. [C ossRe ] [PubMed]
35.
Vincen i, J.E.; Mu phy, L.; G abe , K.; McColl, B.W.; Cancello i, E.; F eeman, T.C.; Manson, J.C. De ining he Mic oglia Response
du ing he Time Cou se o Ch onic Neu odegene a ion. J. Vi ol. 2015,90, 3003–3017. [C ossRe ] [PubMed]
36. Pe y, V.H. Mic oglia. Mic obiol. Spec . 2016,4. [C ossRe ]
37.
Obs , J.; Simon, E.; Mancuso, R.; Gomez-Nicola, D. The Role o Mic oglia in P ion Diseases: A Pa adigm o Func ional Di e si y.
F on . Aging Neu osci. 2017,9, 207. [C ossRe ]
38.
Walsh, D.T.; Be mouni, S.; Pe y, V.H. Absence o De ec able IL-1be a P oduc ion in Mu ine P ion Disease: A Model o Ch onic
Neu odegene a ion. J. Neu opa hol. Exp. Neu ol. 2001,60, 173–182. [C ossRe ]
39.
Pe y, V.H.; Cunningham, C.; Boche, D. A ypical In lamma ion in he Cen al Ne ous Sys em in P ion Disease. Cu . Opin.
Neu ol. 2002,15, 349–354. [C ossRe ]
40.
Boche, D.; Pe y, V.H.; Nicoll, J.A. Re iew: Ac i a ion Pa e ns o Mic oglia and Thei Iden i ica ion in he Human B ain.
Neu opa hol. Appl. Neu obiol. 2013,39, 3–18. [C ossRe ]
41.
Ve kh a sky, A.; Rod igues, J.J.; Pi o iunas, A.; Zo ec, R.; Semyano , A. As oglial A ophy in Alzheime ’s Disease. P lug. A ch.
2019,471, 1247–1261. [C ossRe ] [PubMed]
42.
Llo ens, F.; Lopez-Gonzalez, I.; Thune, K.; Ca mona, M.; Za a , S.; And eole i, O.; Ze , I.; Fe e , I. Sub ype and Regional-Speci ic
Neu oin lamma ion in Spo adic C eu z eld -Jakob Disease. F on . Aging Neu osci. 2014,6, 198. [C ossRe ] [PubMed]
43.
Fo s e , C.; Waschke, J.; Bu ek, M.; Lee s, J.; D enckhahn, D. Glucoco icoid E ec s on Mouse Mic o ascula Endo helial Ba ie
Pe meabili y A e B ain Speci ic. J. Physiol. 2006,573 P 2, 413–425. [C ossRe ]
44.
De Bossche , K.; Vanden Be ghe, W.; Haegeman, G. The In e play be ween he Glucoco icoid Recep o and Nuclea Fac o -kappaB
o Ac i a o P o ein-1: Molecula Mechanisms o Gene Rep ession. Endoc . Re . 2003,24, 488–522. [C ossRe ] [PubMed]
45.
Munck, A.; Guy e, P.M.; Holb ook, N.J. Physiological Func ions o Glucoco icoids in S ess and Thei Rela ion o Pha macological
Ac ions. Endoc . Re . 1984,5, 25–44. [C ossRe ] [PubMed]
46.
Gue ne, P.A.; Ca son, D.A.; Lo z, M. IL-6 p oduc ion by Human A icula Chond ocy es. Modula ion o I s Syn hesis by
Cy okines, G ow h Fac o s, and Ho mones In Vi o. J. Immunol. 1990,144, 499–505. [PubMed]
47.
Bowe s, S.L.; Bilbo, S.D.; Dhabha , F.S.; Nelson, R.J. S esso -Speci ic Al e a ions in Co icos e one and Immune Responses in
Mice. B ain Beha . Immun. 2008,22, 105–113. [C ossRe ] [PubMed]
48.
So ells, S.F.; Caso, J.R.; Munhoz, C.D.; Sapolsky, R.M. The S essed CNS: When Glucoco icoids Agg a a e In lamma ion. Neu on
2009,64, 33–39. [C ossRe ]

Biomolecules 2021,11, 204 17 o 18
49.
Hu, W.; Zhang, Y.; Wu, W.; Yin, Y.; Huang, D.; Wang, Y.; Li, W.; Li, W. Ch onic Glucoco icoids Exposu e Enhances Neu odegene -
a ion in he F on al Co ex and Hippocampus ia NLRP-1 In lammasome Ac i a ion in Male Mice. B ain Beha . Immun.
2016
,52,
58–70. [C ossRe ]
50.
Ki wan, J.R. Glucoco icoid Resis ance in Pa ien s wi h Rheuma oid A h i is. Scand. J. Rheuma ol.
2007
,36, 165–166. [C ossRe ]
51.
M ak, R.E.; Sheng, J.G.; G i in, W.S. Glial Cy okines in Alzheime ’s Disease: Re iew and Pa hogenic Implica ions. Hum. Pa hol.
1995,26, 816–823. [C ossRe ]
52.
Akiyama, H.; Ba ge , S.; Ba num, S.; B ad , B.; Baue , J.; Cole, G.M.; Coope , N.R.; Eikelenboom, P.; Emme ling, M.; Fiebich, B.L.;
e al. In lamma ion and Alzheime ’s Disease. Neu obiol. Aging 2000,21, 383–421. [C ossRe ]
53.
G i in, W.S.; M ak, R.E. In e leukin-1 in he Genesis and P og ession o and Risk o De elopmen o Neu onal Degene a ion in
Alzheime ’s Disease. J. Leukoc. Biol. 2002,72, 233–238. [PubMed]
54.
Sas e, M.; Klockge he , T.; Heneka, M.T. Con ibu ion o In lamma o y P ocesses o Alzheime ’s Disease: Molecula Mechanisms.
In . J. De . Neu osci. 2006,24, 167–176. [C ossRe ] [PubMed]
55.
Se ou, M.J.; DeCos e , M.A.; Bazan, N.G. In e leukin-1 Be a Ac i a es Exp ession o Cyclooxygenase-2 and Inducible Ni ic oxide
Syn hase in P ima y Hippocampal Neu onal Cul u e: Pla ele -Ac i a ing Fac o as a P e e en ial Media o o Cyclooxygenase-2
Exp ession. J. Neu osci. Res. 1999,58, 593–598. [C ossRe ]
56.
Zheng, Y.; Ho ii, A.; Apple on, I.; Da ling on, C.L.; Smi h, P.F. Damage o he Ves ibula Inne Ea Causes Long-Te m Changes in
Neu onal Ni ic Oxide Syn hase Exp ession in he Ra Hippocampus. Neu oscience 2001,105, 1–5. [C ossRe ]
57.
Dinkel, K.; MacPhe son, A.; Sapolsky, R.M. No el Glucoco icoid E ec s on Acu e In lamma ion in he CNS. J. Neu ochem.
2003
,
84, 705–716. [C ossRe ]
58.
Tamguney, G.; Giles, K.; Glidden, D.V.; Lessa d, P.; Wille, H.; T emblay, P.; G o h, D.F.; Yehiely, F.; Ko h, C.; Moo e, R.C.; e al.
Genes Con ibu ing o P ion Pa hogenesis. J. Gen. Vi ol. 2008,89 P 7, 1777–1788. [C ossRe ]
59.
Liddelow, S.A.; Gu enplan, K.A.; Cla ke, L.E.; Benne , F.C.; Bohlen, C.J.; Schi me , L.; Benne , M.L.; Munch, A.E.; Chung, W.S.;
Pe e son, T.C.; e al. Neu o oxic Reac i e As ocy es A e Induced by Ac i a ed Mic oglia. Na u e 2017,541, 481–487. [C ossRe ]
60.
B u ge , J.; Ka am, K.; Wo ge, S.; Regen, T.; Ma ini, F.; Hoppmann, N.; Klein, M.; Blank, T.; Yona, S.; Wol , Y.; e al. Gene ic
Cell Abla ion Re eals Clus e s o Local Sel -Renewing Mic oglia in he Mammalian Cen al Ne ous Sys em. Immuni y
2015
,43,
92–106. [C ossRe ]
61.
Ban, E.; Milon, G.; P udhomme, N.; Fillion, G.; Haou , F. Recep o s o In e leukin-1 (Alpha and Be a) in Mouse B ain: Mapping
and Neu onal Localiza ion in Hippocampus. Neu oscience 1991,43, 21–30. [PubMed]
62.
F ench, R.A.; VanHoy, R.W.; Chizzoni e, R.; Zacha y, J.F.; Dan ze , R.; Pa ne , P.; Blu he, R.M.; Kelley, K.W. Exp ession and Local-
iza ion o p80 and p68 In e leukin-1 Recep o P o eins in he B ain o Adul Mice. J. Neu oimmunol.
1999
,93,
194–202. [C ossRe ]
63.
Ra izza, T.; Vezzani, A. S a us Epilep icus Induces Time-Dependen Neu onal and As ocy ic Exp ession o In e leukin-1 Recep o
Type I in he Ra Limbic Sys em. Neu oscience 2006,137, 301–308. [C ossRe ] [PubMed]
64.
Spulbe , S.; Ba ai, T.; Schul zbe g, M. IL-1/IL-1 a Balance in he B ain Re isi ed—E idence om T ansgenic Mouse Models.
B ain Beha . Immun. 2009,23, 573–579. [C ossRe ] [PubMed]
65. Dina ello, C.A. Biologic Basis o In e leukin-1 in Disease. Blood 1996,87, 2095–2147. [C ossRe ]
66.
Al es, S.; Chu laud, G.; Aud ain, M.; Michaelsen-P eusse, K.; Fol, R.; Souche , B.; B audeau, J.; Ko e, M.; Kla zmann, D.;
Ca ie , N.
In e leukin-2 Imp o es Amyloid Pa hology, Synap ic Failu e and Memo y in Alzheime ’s Disease Mice. B ain
2017
,
140, 826–842. [C ossRe ]
67.
Saadoun, D.; Te ie , B.; Cacoub, P. In e leukin-25: Key Regula o o In lamma o y and Au oimmune Diseases. Cu . Pha m. Des.
2011,17, 3781–3785. [C ossRe ]
68.
Lube -Na od, J.; Roge s, J. Immune Sys em Associa ed An igens Exp essed by Cells o he Human Cen al Ne ous Sys em.
Neu osci. Le . 1988,94, 17–22. [C ossRe ]
69.
Sa de , M.; Abe, K.; Sai o, H.; Nishiyama, N. Compa a i e E ec o IL-2 and IL-6 on Mo phology o Cul u ed Hippocampal
Neu ons om Fe al Ra B ain. B ain Res. 1996,715, 9–16. [C ossRe ]
70.
Dansokho, C.; Ahmed, D.A.; Aid, S.; Toly-Ndou , C.; Chaigneau, T.; Calle, V.; Cagna d, N.; Holzenbe ge , M.;
Piaggio, E.
;
Aucou u ie , P.; e al. Regula o y T Cells Delay Disease P og ession in Alzheime -Like Pa hology. B ain
2016
,139 P 4,
1237–1251. [C ossRe ]
71.
Campbell, I.L.; Ab aham, C.R.; Masliah, E.; Kempe , P.; Inglis, J.D.; Olds one, M.B.; Mucke, L. Neu ologic Disease Induced in T ans-
genic Mice by Ce eb al O e exp ession o In e leukin 6. P oc. Na l. Acad. Sci. USA 1993,90, 10061–10065. [C ossRe ] [PubMed]
72.
Ha iz, F.B.; B own, D.R. A Model o he Mechanism o As ogliosis in P ion Disease. Mol. Cell. Neu osci.
2000
,16, 221–232.
[C ossRe ] [PubMed]
73. Gabay, C. In e leukin-6 and Ch onic In lamma ion. A h i is Res. The . 2006,8(Suppl. S2), S3. [C ossRe ] [PubMed]
74.
Xing, Z.; Gauldie, J.; Cox, G.; Baumann, H.; Jo dana, M.; Lei, X.F.; Achong, M.K. IL-6 Is an An iin lamma o y Cy okine Requi ed
o Con olling Local o Sys emic Acu e In lamma o y Responses. J. Clin. In es ig. 1998,101, 311–320. [C ossRe ]
75.
Yasukawa, H.; Ohishi, M.; Mo i, H.; Mu akami, M.; Chinen, T.; Aki, D.; Hanada, T.; Takeda, K.; Aki a, S.; Hoshijima, M.; e al. IL-6
Induces an An i-In lamma o y Response in he Absence o SOCS3 in Mac ophages. Na . Immunol. 2003,4, 551–556. [C ossRe ]
76.
Tilg, H.; Dina ello, C.A.; Mie , J.W. IL-6 and APPs: An i-In lamma o y and Immunosupp essi e Media o s. Immunol. Today
1997
,
18, 428–432. [C ossRe ]
Biomolecules 2021,11, 204 18 o 18
77.
Yamamo o, M.; Yoshizaki, K.; Kishimo o, T.; I o, H. IL-6 Is Requi ed o he De elopmen o Th1 Cell-Media ed Mu ine Coli is.
J. Immunol. 2000,164, 4878–4882. [C ossRe ]
78. Allan, S.M.; Ro hwell, N.J. Cy okines and Acu e Neu odegene a ion. Na . Re . Neu osci. 2001,2, 734–744. [C ossRe ]
79.
Mabbo , N.A.; Williams, A.; Fa quha , C.F.; Paspa akis, M.; Kollias, G.; B uce, M.E. Tumo Nec osis Fac o Alpha-De icien , Bu
no In e leukin-6-de icien , Mice Resis Pe iphe al In ec ion wi h Sc apie. J. Vi ol. 2000,74, 3338–3344. [C ossRe ]
80.
Baue , J.; S auss, S.; Volk, B.; Be ge , M. IL-6-media ed E en s in Alzheime ’s Disease Pa hology. Immunol. Today
1991
,12,
422. [C ossRe ]
81.
S auss, S.; Baue , J.; Gan e , U.; Jonas, U.; Be ge , M.; Volk, B. De ec ion o In e leukin-6 and Alpha 2-mac oglobulin Immuno eac-
i i y in Co ex and Hippocampus o Alzheime ’s Disease Pa ien s. Lab. In es ig. 1992,66, 223–230. [PubMed]
82.
Wood, J.A.; Wood, P.L.; Ryan, R.; G a -Rad o d, N.R.; Pilapil, C.; Robi aille, Y.; Qui ion, R. Cy okine Indices in Alzheime ’s
Tempo al Co ex: No Changes in Ma u e IL-1 Be a o IL-1RA bu Inc eases in he Associa ed Acu e Phase P o eins IL-6, Alpha
2-mac oglobulin and C- eac i e P o ein. B ain Res. 1993,629, 245–252. [C ossRe ]
83.
Coupe , K.N.; Bloun , D.G.; Riley, E.M. IL-10: The Mas e Regula o o Immuni y o In ec ion. J. Immunol.
2008
,180, 5771–5777.
[C ossRe ] [PubMed]
84.
Wang, P.; Wu, P.; Siegel, M.I.; Egan, R.W.; Billah, M.M. In e leukin (IL)-10 Inhibi s Nuclea Fac o Kappa B (NF Kappa B)
Ac i a ion in Human Monocy es. IL-10 and IL-4 Supp ess Cy okine Syn hesis by Di e en Mechanisms. J. Biol. Chem.
1995
,270,
9558–9563. [C ossRe ]
85.
Lobo-Sil a, D.; Ca iche, G.M.; Cas o, A.G.; Roque, S.; Sa ai a, M. Balancing he Immune Response in he B ain: IL-10 and I s
Regula ion. J. Neu oin lamm. 2016,13, 297. [C ossRe ]
86.
Chak aba y, P.; Li, A.; Ceballos-Diaz, C.; Eddy, J.A.; Funk, C.C.; Moo e, B.; DiNunno, N.; Rosa io, A.M.; C uz, P.E.;
Ve beeck, C.; e al.
IL-10 Al e s Immunop o eos asis in APP Mice, Inc easing Plaque Bu den and Wo sening Cogni i e Beha io .
Neu on 2015,85, 519–533. [C ossRe ]
87.
Guillo -Ses ie , M.V.; Do y, K.R.; Ga e, D.; Rod iguez, J., J .; Leung, B.P.; Rezai-Zadeh, K.; Town, T. Il10 De iciency Rebalances
Inna e Immuni y o Mi iga e Alzheime -Like Pa hology. Neu on 2015,85, 534–548. [C ossRe ]
88.
P inz, M.; Mon asio, F.; Klein, M.A.; Schwa z, P.; P ille , J.; Ode ma , B.; P e e , K.; Aguzzi, A. Lymph Nodal P ion Replica ion
and Neu oin asion in Mice De oid o Follicula Dend i ic Cells. P oc. Na l. Acad. Sci. USA 2002,99, 919–924. [C ossRe ]
89.
Mabbo , N.A.; McGo e n, G.; Je ey, M.; B uce, M.E. Tempo a y Blockade o he Tumo Nec osis Fac o Recep o Signaling
Pa hway Impedes he Sp ead o Sc apie o he B ain. J. Vi ol. 2002,76, 5131–5139. [C ossRe ]
90.
Zhao, M.; C ibbs, D.H.; Ande son, A.J.; Cummings, B.J.; Su, J.H.; Wasse man, A.J.; Co man, C.W. The Induc ion o he TNFalpha
Dea h Domain Signaling Pa hway in Alzheime ’s Disease B ain. Neu ochem. Res. 2003,28, 307–318. [C ossRe ]
91.
Cacabelos, R.; Al a ez, X.A.; F anco-Maside, A.; Fe nandez-No oa, L.; Caamano, J. Se um Tumo Nec osis Fac o (TNF) in
Alzheime ’s Disease and Mul i-In a c Demen ia. Me hods Find. Exp. Clin. Pha macol. 1994,16, 29–35. [PubMed]
92.
Sakudo, A.; Lee, D.C.; Saeki, K.; Ma sumo o, Y.; I oha a, S.; Onode a, T. Tumo Nec osis Fac o A enua es P ion P o ein-De icien
Neu onal Cell Dea h by Inc eases in An i-Apop o ic Bcl-2 Family P o eins. Biochem. Biophys. Res. Commun.
2003
,310, 725–729.
[C ossRe ] [PubMed]
93. Goe z, F.W.; Planas, J.V.; MacKenzie, S. Tumo Nec osis Fac o s. De . Comp. Immunol. 2004,28, 487–497. [C ossRe ]
94.
Sche bel, U.; Raghupa hi, R.; Nakamu a, M.; Saa man, K.E.; T ojanowski, J.Q.; Neugebaue , E.; Ma ino, M.W.; McIn osh, T.K.
Di e en ial Acu e and Ch onic Responses o Tumo Nec osis Fac o -De icien Mice o Expe imen al B ain Inju y. P oc. Na l. Acad.
Sci. USA 1999,96, 8721–8726. [C ossRe ] [PubMed]
95. Kim, S.U.; de Vellis, J. Mic oglia in Heal h and Disease. J. Neu osci. Res. 2005,81, 302–313. [C ossRe ]
96. Sa o, K. E ec s o Mic oglia on Neu ogenesis. Glia 2015,63, 1394–1405. [C ossRe ] [PubMed]
97.
Co bin, J.G.; Kelly, D.; Ra h, E.M.; Bae wald, K.D.; Suzuki, K.; Popko, B. Ta ge ed CNS Exp ession o In e e on-Gamma in
T ansgenic Mice Leads o Hypomyelina ion, Reac i e Gliosis, and Abno mal Ce ebella De elopmen . Mol. Cell. Neu osci.
1996
,7,
354–370. [C ossRe ]
98.
Hashioka, S.; Klege is, A.; Schwab, C.; McGee , P.L. In e e on-Gamma-Dependen Cy o oxic Ac i a ion o Human As ocy es
and As ocy oma Cells. Neu obiol. Aging 2009,30, 1924–1935. [C ossRe ]
99. Sa na, J.R.; Hawkes, R. Pa e ned Pu kinje Cell Dea h in he Ce ebellum. P og. Neu obiol. 2003,70, 473–507. [C ossRe ]
100.
Nelson, T.E.; Campbell, I.L.; G uol, D.L. Al e ed Physiology o Pu kinje Neu ons in Ce ebella Slices om T ansgenic Mice wi h
Ch onic Cen al Ne ous Sys em Exp ession o In e leukin-6. Neu oscience 1999,89, 127–136.
101.
Ragagnin, A.; Ezpele a, J.; Guillemain, A.; Boude -De aud, F.; Haebe le, A.M.; Demais, V.; Vidal, C.; Demu h, S.; Be ingue, V.;
Kelle mann, O.; e al. Ce ebella Compa men a ion o P ion Pa hogenesis. B ain Pa hol.
2017
,28, 240–263. [C ossRe ] [PubMed]
102.
Munck, A.; Na ay-Fejes-To h, A. The Ups and Downs o Glucoco icoid Physiology. Pe missi e and Supp essi e E ec s Re isi ed.
Mol. Cell. Endoc inol. 1992,90, C1–C4. [C ossRe ]
103.
Fila e o a, L.; Pod igina, T.; Bagae a, T.; Mo ozo a, O. Dual Ac ion o Glucoco icoid Ho mones on he Gas ic Mucosa: How he
Gas op o ec i e Ac ion Can Be T ans o med o he Ulce ogenic One. In lammopha macology
2009
,17, 15–22. [C ossRe ] [PubMed]