%PDF-1.6
%
1 0 obj
<>
endobj
2 0 obj
<>stream
Cinthia C. Stempin, Jorge D. Rojas Marquez, Yamile Ana, Fabio M. Cerban
GRAIL and Otubain-1 are Related to T Cell Hyporesponsiveness during Trypanosoma cruzi Infection
www.plosntds.org
www.plosntds.org
endstream
endobj
3 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 14 0 R/Contents 15 0 R/TrimBox[0 0 612 792]>>
endobj
14 0 obj
[16 0 R 17 0 R 18 0 R 19 0 R 20 0 R 21 0 R 22 0 R 23 0 R 24 0 R 25 0 R 26 0 R 27 0 R 28 0 R 29 0 R 30 0 R 31 0 R 32 0 R]
endobj
16 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref001)>>
endobj
17 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref002)>>
endobj
18 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref003)>>
endobj
19 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref004)>>
endobj
20 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref005)>>
endobj
21 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref006)>>
endobj
22 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref007)>>
endobj
23 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref008)>>
endobj
24 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref009)>>
endobj
25 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref010)>>
endobj
26 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref008)>>
endobj
27 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref011)>>
endobj
28 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref011)>>
endobj
29 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref012)>>
endobj
30 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref011)>>
endobj
31 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref012)>>
endobj
32 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref013)>>
endobj
15 0 obj
[33 0 R 34 0 R 35 0 R 36 0 R 37 0 R 38 0 R 39 0 R 40 0 R 41 0 R 42 0 R 43 0 R 44 0 R 45 0 R 46 0 R 47 0 R]
endobj
33 0 obj
<>stream
q
0.955 g
0 w
199.9998 641.23 m
575.9997 641.23 l
575.9997 415.2305 l
199.9998 415.2305 l
f*
0.45 0.36 0.35 0.01 k
575.9997 666.2299 m
199.9998 666.2299 l
199.9998 666.7299 l
575.9997 666.7299 l
f*
0.83 0.64 0.02 0 k
455.1307 289.9276 m
459.8929 289.9276 l
h
f*
462.1606 289.9276 m
466.9228 289.9276 l
h
f*
533.4803 237.9402 m
538.2425 237.9402 l
h
f*
433.5307 198.9354 m
438.2929 198.9354 l
h
f*
440.5606 198.9354 m
445.3228 198.9354 l
h
f*
447.5339 198.9354 m
452.2961 198.9354 l
h
f*
454.5638 198.9354 m
459.326 198.9354 l
h
f*
461.5937 198.9354 m
466.3559 198.9354 l
h
f*
306.4819 172.9134 m
311.2441 172.9134 l
h
f*
313.5118 172.9134 m
323.0362 172.9134 l
h
f*
464.8252 159.9307 m
469.5874 159.9307 l
h
f*
350.1921 133.9087 m
359.6598 133.9087 l
h
f*
515.622 107.9433 m
525.1465 107.9433 l
h
f*
527.4142 107.9433 m
536.9386 107.9433 l
h
f*
257.1591 81.9213 m
266.6835 81.9213 l
h
f*
268.9512 81.9213 m
278.4189 81.9213 l
h
f*
280.6866 81.9213 m
290.211 81.9213 l
h
f*
0 g
1 j
1 J
9.5 0 0 9.5 200.0125 707.7543 cm
BT
/F0 1 Tf
1 TL
-0.0042 Tc
0 0 Td
(maintained)Tj
5.0486 0 Td
(the)Tj
1.5934 0 Td
(brake)Tj
2.6914 0 Td
(on)Tj
1.3188 0 Td
(T)Tj
0.8295 0 Td
(cell)Tj
1.7008 0 Td
(proliferation.)Tj
5.6454 0 Td
(Our)Tj
1.8798 0 Td
(findings)Tj
3.6164 0 Td
(reveal)Tj
2.9063 0 Td
(a)Tj
0.7698 0 Td
(novel)Tj
2.578 0 Td
(association)Tj
5.162 0 Td
(between)Tj
-35.7402 -1.474 Td
(increased)Tj
4.4996 0 Td
(GRAIL)Tj
3.2046 0 Td
(expression)Tj
4.9949 0 Td
(and)Tj
1.8679 0 Td
(impaired)Tj
4.0043 0 Td
(CD4)Tj
2.2021 0 Td
(T)Tj
0.8295 0 Td
(cell)Tj
1.6948 0 Td
(proliferation)Tj
5.3709 0 Td
(during)Tj
/F6 1 Tf
2.9599 0 Td
(Trypanosoma)Tj
-31.6285 -1.474 Td
(cruzi)Tj
/F0 1 Tf
2.3035 0 Td
(infection.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 209.9905 623.2251 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Author)Tj
3.137 0 Td
(Summary)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 209.9905 605.2535 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Chagas)Tj
3.1067 0 Td
(disease)Tj
3.016 0 Td
(is)Tj
0.8504 0 Td
(caused)Tj
2.914 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4683 0 Td
(protozoan)Tj
4.3257 0 Td
(parasite)Tj
/F13 1 Tf
3.3222 0 Td
[()75()-20()-13()4()16()6()7()-3()11( )6()]TJ
5.6182 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(and)Tj
1.7348 0 Td
(is)Tj
0.8447 0 Td
(endemic)Tj
3.6567 0 Td
(in)Tj
-34.2026 -1.3039 Td
(Central)Tj
3.2087 0 Td
(and)Tj
1.7292 0 Td
(South)Tj
2.5568 0 Td
(America,)Tj
3.8664 0 Td
(where)Tj
2.6532 0 Td
(it)Tj
0.7824 0 Td
(affects)Tj
2.7383 0 Td
(about)Tj
2.4944 0 Td
(10)Tj
1.1679 0 Td
(million)Tj
3.1238 0 Td
(people.)Tj
3.0784 0 Td
(In)Tj
1.1111 0 Td
(addition,)Tj
3.8211 0 Td
(migra-)Tj
-32.3317 -1.2982 Td
(tion)Tj
1.8425 0 Td
(has)Tj
1.542 0 Td
(led)Tj
1.4173 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(disease)Tj
3.016 0 Td
(being)Tj
2.4321 0 Td
(established)Tj
4.5978 0 Td
(in)Tj
1.0375 0 Td
(non-endemic)Tj
5.6069 0 Td
(countries.)Tj
4.1725 0 Td
(Infection)Tj
3.8665 0 Td
(involves)Tj
-32.0256 -1.2983 Td
(an)Tj
1.2018 0 Td
(acute)Tj
2.3301 0 Td
(stage)Tj
2.1997 0 Td
(that)Tj
1.7801 0 Td
(evolves)Tj
3.0898 0 Td
(to)Tj
1.0261 0 Td
(a)Tj
0.652 0 Td
(chronic)Tj
3.2881 0 Td
(stage)Tj
2.2054 0 Td
(where)Tj
2.6475 0 Td
(infected)Tj
3.4186 0 Td
(individuals)Tj
4.6488 0 Td
(may)Tj
1.9332 0 Td
(or)Tj
1.0941 0 Td
(may)Tj
1.9276 0 Td
(not)Tj
-33.4429 -1.3039 Td
(show)Tj
2.3017 0 Td
(clinical)Tj
3.0727 0 Td
(symptoms)Tj
4.3597 0 Td
(or)Tj
1.0998 0 Td
(suffer)Tj
2.4775 0 Td
(progressive)Tj
4.7451 0 Td
(heart)Tj
2.2734 0 Td
(disease.)Tj
3.2428 0 Td
(The)Tj
1.7858 0 Td
(relevance)Tj
3.9685 0 Td
(of)Tj
1.0148 0 Td
(T)Tj
0.8277 0 Td
(cells)Tj
1.9219 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
-34.1289 -1.2983 Td
(control)Tj
3.118 0 Td
(of)Tj
/F13 1 Tf
1.0148 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0318 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(infection)Tj
3.7871 0 Td
(has)Tj
1.542 0 Td
(been)Tj
2.1203 0 Td
(demonstrated)Tj
5.7543 0 Td
(in)Tj
1.0318 0 Td
(human)Tj
3.0784 0 Td
(infection)Tj
3.7871 0 Td
(and)Tj
1.7291 0 Td
(in)Tj
1.0318 0 Td
(experimen-)Tj
-31.1865 -1.2982 Td
(tal)Tj
1.2018 0 Td
(models.)Tj
3.3279 0 Td
(However,)Tj
4.0762 0 Td
(the)Tj
/F13 1 Tf
1.4683 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0375 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(parasite)Tj
3.3278 0 Td
(employs)Tj
3.549 0 Td
(different)Tj
3.6567 0 Td
(strategies)Tj
3.9231 0 Td
(to)Tj
1.0261 0 Td
(downregulate)Tj
5.6749 0 Td
(the)Tj
-34.4293 -1.304 Td
(T)Tj
0.8333 0 Td
(cell)Tj
1.5591 0 Td
(function.)Tj
3.8551 0 Td
(These)Tj
2.5625 0 Td
(mechanisms)Tj
5.21 0 Td
(can)Tj
1.6214 0 Td
(act)Tj
1.3777 0 Td
(at)Tj
0.9524 0 Td
(the)Tj
1.4683 0 Td
(initial)Tj
2.5512 0 Td
(time)Tj
2.0296 0 Td
(of)Tj
1.0148 0 Td
(T)Tj
0.8277 0 Td
(cell)Tj
1.559 0 Td
(activation,)Tj
4.3824 0 Td
(leading)Tj
3.1294 0 Td
(to)Tj
-34.9339 -1.2982 Td
(a)Tj
0.6576 0 Td
(state)Tj
2.0296 0 Td
(of)Tj
1.0148 0 Td
(anergy)Tj
2.9196 0 Td
(where)Tj
2.6533 0 Td
(lymphocytes)Tj
5.2497 0 Td
(do)Tj
1.2586 0 Td
(not)Tj
1.5703 0 Td
(respond.)Tj
3.7021 0 Td
(However,)Tj
4.0762 0 Td
(the)Tj
1.474 0 Td
(molecular)Tj
4.2179 0 Td
(components)Tj
-30.8237 -1.2983 Td
(that)Tj
1.7801 0 Td
(regulate)Tj
3.4072 0 Td
(this)Tj
1.6781 0 Td
(process)Tj
3.1805 0 Td
(during)Tj
/F13 1 Tf
2.931 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0318 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(infection)Tj
3.7927 0 Td
(are)Tj
1.4513 0 Td
(not)Tj
1.5704 0 Td
(well)Tj
1.8198 0 Td
(understood.)Tj
5.0457 0 Td
(Our)Tj
1.8708 0 Td
(findings)Tj
-31.7194 -1.2983 Td
(demonstrate)Tj
5.2327 0 Td
(for)Tj
1.3889 0 Td
(the)Tj
1.4684 0 Td
(first)Tj
1.8028 0 Td
(time)Tj
2.0296 0 Td
(that)Tj
1.7801 0 Td
(this)Tj
1.6725 0 Td
(T)Tj
0.8334 0 Td
(cell)Tj
1.559 0 Td
(hyporesponsiveness)Tj
8.1581 0 Td
(could)Tj
2.4491 0 Td
(be)Tj
1.1508 0 Td
(linked)Tj
2.7269 0 Td
(to)Tj
1.0262 0 Td
(an)Tj
-33.2785 -1.3039 Td
(increased)Tj
4.0024 0 Td
(expression)Tj
4.4617 0 Td
(of)Tj
1.0148 0 Td
(GRAIL.)Tj
3.3449 0 Td
(We)Tj
1.6101 0 Td
(propose)Tj
3.4242 0 Td
(that)Tj
1.7802 0 Td
(GRAIL)Tj
3.118 0 Td
(expression)Tj
4.4674 0 Td
(induced)Tj
3.4639 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4683 0 Td
(para-)Tj
-33.3408 -1.2982 Td
(site)Tj
1.5703 0 Td
(could)Tj
2.4548 0 Td
(be)Tj
1.1509 0 Td
(maintained)Tj
4.7848 0 Td
(by)Tj
1.1849 0 Td
(increased)Tj
3.9968 0 Td
(expression)Tj
4.4674 0 Td
(of)Tj
1.0148 0 Td
(inhibitory)Tj
4.2349 0 Td
(molecules,)Tj
4.4221 0 Td
(which)Tj
2.6532 0 Td
(blocked)Tj
-31.9349 -1.2983 Td
(mTOR)Tj
3.0217 0 Td
(activation)Tj
4.1499 0 Td
(and)Tj
1.7291 0 Td
(IL-2)Tj
1.9275 0 Td
(secretion.)Tj
4.0706 0 Td
(GRAIL)Tj
3.118 0 Td
(could)Tj
2.4492 0 Td
(then)Tj
2.0182 0 Td
(play)Tj
1.8765 0 Td
(a)Tj
0.652 0 Td
(key)Tj
1.5931 0 Td
(role)Tj
1.7688 0 Td
(in)Tj
1.0374 0 Td
(downregulating)Tj
-29.412 -1.3039 Td
(T)Tj
0.8333 0 Td
(cell)Tj
1.5591 0 Td
(functions)Tj
3.9911 0 Td
(by)Tj
1.1792 0 Td
(allowing)Tj
3.617 0 Td
(the)Tj
1.4684 0 Td
(parasites)Tj
3.6906 0 Td
(to)Tj
1.0262 0 Td
(establish)Tj
3.6453 0 Td
(the)Tj
1.4683 0 Td
(chronic)Tj
3.2882 0 Td
(disease.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 372.9826 cm
BT
/F3 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Introduction)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 355.9748 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Chagas)Tj
3.101 0 Td
(disease,)Tj
3.2485 0 Td
(caused)Tj
2.914 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4683 0 Td
(intracellular)Tj
5.057 0 Td
(protozoa)Tj
/F13 1 Tf
3.7757 0 Td
[()75()-20()-13()4()16()6()7()-3()11( )6()]TJ
5.6183 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
(,)Tj
2.3867 0 Td
(is)Tj
0.8447 0 Td
(one)Tj
1.7008 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(major)Tj
-33.783 -1.2982 Td
(human)Tj
3.0784 0 Td
(health)Tj
2.6758 0 Td
(problems)Tj
3.9742 0 Td
(in)Tj
1.0318 0 Td
(Latin)Tj
2.3074 0 Td
(America.)Tj
3.8721 0 Td
(It)Tj
0.856 0 Td
(evolves)Tj
3.0898 0 Td
(from)Tj
2.2053 0 Td
(an)Tj
1.2019 0 Td
(acute)Tj
2.3301 0 Td
(to)Tj
1.0261 0 Td
(a)Tj
0.652 0 Td
(chronic)Tj
3.2881 0 Td
(phase,)Tj
2.7099 0 Td
(where)Tj
-34.2989 -1.2983 Td
(subjects)Tj
3.3788 0 Td
(may)Tj
1.9276 0 Td
(be)Tj
1.1508 0 Td
(clinically)Tj
3.7758 0 Td
(asymptomatic)Tj
5.8563 0 Td
(or)Tj
1.0942 0 Td
(show)Tj
2.3017 0 Td
(progressive)Tj
4.7451 0 Td
(heart)Tj
2.2734 0 Td
(disease)Tj
3.0161 0 Td
(and)Tj
1.7291 0 Td
(leads)Tj
2.2167 0 Td
(to)Tj
1.0204 0 Td
(an)Tj
1.2076 0 Td
(end-)Tj
-35.6936 -1.2982 Td
(stage)Tj
2.1996 0 Td
(dilated)Tj
2.948 0 Td
(cardiomyopathy)Tj
6.7635 0 Td
(in)Tj
1.0318 0 Td
(2030%)Tj
3.4015 0 Td
(of)Tj
1.0148 0 Td
(infected)Tj
3.4186 0 Td
(individuals.)Tj
4.8755 0 Td
(It)Tj
0.8561 0 Td
(is)Tj
0.8504 0 Td
(estimated)Tj
4.0705 0 Td
(that)Tj
1.7801 0 Td
(approxi-)Tj
-33.2104 -1.304 Td
(mately)Tj
2.8969 0 Td
(4)Tj
0.6917 0 Td
(million)Tj
3.1237 0 Td
(chagasic)Tj
3.5547 0 Td
(individuals)Tj
4.6488 0 Td
(have)Tj
2.0636 0 Td
(developed)Tj
4.2689 0 Td
(heart)Tj
2.2734 0 Td
(disease,)Tj
3.2428 0 Td
(making)Tj
3.2542 0 Td
(Chagas)Tj
3.101 0 Td
(disease)Tj
3.0218 0 Td
(the)Tj
-36.1415 -1.2982 Td
(most)Tj
2.2053 0 Td
(frequent)Tj
3.6056 0 Td
(cause)Tj
2.3925 0 Td
(of)Tj
1.0148 0 Td
(infectious)Tj
4.1328 0 Td
(cardiomyopathy)Tj
6.7635 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.474 0 Td
(world)Tj
2.5511 0 Td
([)Tj
0.83 0.64 0.02 0 k
(1)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(2)Tj
0 g
(].)Tj
-23.9752 -1.2983 Td
(The)Tj
1.7858 0 Td
(immune)Tj
3.6226 0 Td
(control)Tj
3.1238 0 Td
(of)Tj
/F13 1 Tf
1.0148 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0375 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.1599 0 Td
(is)Tj
0.8448 0 Td
(complex,)Tj
3.8494 0 Td
(requiring)Tj
3.9685 0 Td
(the)Tj
1.4683 0 Td
(generation)Tj
4.5071 0 Td
(of)Tj
1.0148 0 Td
(a)Tj
0.6576 0 Td
(substantial)Tj
4.5014 0 Td
(anti-)Tj
-34.7525 -1.3039 Td
(body)Tj
2.2166 0 Td
(response)Tj
3.7304 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.474 0 Td
(activation)Tj
4.1499 0 Td
(of)Tj
1.0148 0 Td
(both)Tj
2.058 0 Td
(CD4)Tj
2.0976 0 Td
(and)Tj
1.7291 0 Td
(CD8)Tj
2.0976 0 Td
(T)Tj
0.8277 0 Td
(cell)Tj
1.5591 0 Td
(responses.)Tj
4.32 0 Td
(Even)Tj
2.2166 0 Td
(in)Tj
1.0375 0 Td
(cases)Tj
2.2224 0 Td
(in)Tj
-34.4804 -1.2983 Td
(which)Tj
2.6475 0 Td
(these)Tj
2.2564 0 Td
(responses)Tj
4.0932 0 Td
(are)Tj
1.4513 0 Td
(sufficiently)Tj
4.6091 0 Td
(stimulated)Tj
4.4277 0 Td
(to)Tj
1.0204 0 Td
(be)Tj
1.1509 0 Td
(able)Tj
1.8312 0 Td
(to)Tj
1.0261 0 Td
(control)Tj
3.1238 0 Td
(the)Tj
1.4683 0 Td
(acute)Tj
2.3244 0 Td
(infection,)Tj
/F13 1 Tf
4.0195 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
-35.4498 -1.2982 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.1599 0 Td
(is)Tj
0.8448 0 Td
(not)Tj
1.576 0 Td
(completely)Tj
4.5808 0 Td
(eradicated,)Tj
4.5751 0 Td
(but)Tj
1.5533 0 Td
(instead)Tj
3.0784 0 Td
(persists)Tj
3.1805 0 Td
(in)Tj
1.0375 0 Td
(infected)Tj
3.4185 0 Td
(hosts)Tj
2.2791 0 Td
(for)Tj
1.3833 0 Td
(decades)Tj
3.3392 0 Td
([)Tj
0.83 0.64 0.02 0 k
(3)Tj
0 g
(].)Tj
/F13 1 Tf
-31.8102 -1.304 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0374 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(employs)Tj
3.549 0 Td
(a)Tj
0.6576 0 Td
(variety)Tj
2.9197 0 Td
(of)Tj
1.0148 0 Td
(strategies)Tj
3.9231 0 Td
(to)Tj
1.0261 0 Td
(evade)Tj
2.4832 0 Td
(the)Tj
1.4683 0 Td
(immune)Tj
3.6283 0 Td
(system)Tj
2.9367 0 Td
(and)Tj
1.7348 0 Td
(remain)Tj
3.0841 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
-33.8511 -1.2982 Td
(infected)Tj
3.4185 0 Td
(host.)Tj
2.1373 0 Td
(The)Tj
1.7858 0 Td
(main)Tj
2.2904 0 Td
(method)Tj
3.3222 0 Td
(involves)Tj
3.4809 0 Td
(the)Tj
1.4684 0 Td
(inhibition)Tj
4.2292 0 Td
(of)Tj
1.0148 0 Td
(specific)Tj
3.1918 0 Td
(T-cell)Tj
2.5172 0 Td
(responses,)Tj
4.3199 0 Td
(and)Tj
1.7292 0 Td
(conse-)Tj
-34.9056 -1.2983 Td
(quently,)Tj
3.4525 0 Td
(can)Tj
1.6214 0 Td
(favor)Tj
2.2791 0 Td
(the)Tj
1.4683 0 Td
(establishment)Tj
5.7373 0 Td
(of)Tj
1.0148 0 Td
(chronic)Tj
3.2882 0 Td
(infections)Tj
4.1499 0 Td
([)Tj
0.83 0.64 0.02 0 k
(4)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(5)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(6)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(7)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(8)Tj
0 g
(].)Tj
4.405 0 Td
(Related)Tj
3.1974 0 Td
(to)Tj
1.0262 0 Td
(this,)Tj
1.8992 0 Td
(a)Tj
0.6519 0 Td
(number)Tj
-34.1912 -1.3039 Td
(of)Tj
1.0147 0 Td
(both)Tj
2.058 0 Td
(host-dependent)Tj
6.5026 0 Td
(and)Tj
1.7291 0 Td
(parasite-induced)Tj
6.9279 0 Td
(mechanisms)Tj
5.2044 0 Td
(have)Tj
2.0579 0 Td
(been)Tj
2.126 0 Td
(previously)Tj
4.3483 0 Td
(shown)Tj
2.8459 0 Td
(to)Tj
-34.8148 -1.2983 Td
(affect)Tj
2.3754 0 Td
(immune)Tj
3.6226 0 Td
(regulation)Tj
4.3087 0 Td
([)Tj
0.83 0.64 0.02 0 k
(9)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(10)Tj
0 g
(].)Tj
2.7779 0 Td
(Moreover,)Tj
4.3993 0 Td
(T)Tj
0.8277 0 Td
(cells)Tj
1.9219 0 Td
(from)Tj
2.2054 0 Td
(infected)Tj
3.4242 0 Td
(hosts)Tj
2.2734 0 Td
(are)Tj
1.4456 0 Td
(largely)Tj
2.863 0 Td
(unrespon-)Tj
-32.4451 -1.2982 Td
(sive)Tj
1.7234 0 Td
(to)Tj
1.0261 0 Td
(antigens)Tj
3.566 0 Td
(and)Tj
1.7291 0 Td
(mitogens,)Tj
4.1386 0 Td
(resulting)Tj
3.7304 0 Td
(in)Tj
1.0318 0 Td
(reduced)Tj
3.4469 0 Td
(IL-2)Tj
1.9218 0 Td
(synthesis)Tj
3.8268 0 Td
([)Tj
0.83 0.64 0.02 0 k
(8)Tj
0 g
(].)Tj
-24.9447 -1.2983 Td
(IL-2)Tj
1.9275 0 Td
(production)Tj
4.7168 0 Td
(initiates)Tj
3.3846 0 Td
(proliferation,)Tj
5.4878 0 Td
(effector)Tj
3.2428 0 Td
(functions,)Tj
4.218 0 Td
(and)Tj
1.7291 0 Td
(clonal)Tj
2.6248 0 Td
(expansion)Tj
4.2917 0 Td
(via)Tj
1.3776 0 Td
(IL-2)Tj
-34.1969 -1.3039 Td
(receptor)Tj
3.5546 0 Td
(\(IL-2R\)-mediated)Tj
7.2963 0 Td
(signaling)Tj
3.8268 0 Td
([)Tj
0.83 0.64 0.02 0 k
(11)Tj
0 g
(].)Tj
2.0749 0 Td
(In)Tj
1.1055 0 Td
(the)Tj
1.474 0 Td
(absence)Tj
3.3392 0 Td
(of)Tj
1.0148 0 Td
(a)Tj
0.652 0 Td
(robust)Tj
2.7949 0 Td
(activation)Tj
4.1499 0 Td
(initiated)Tj
3.5546 0 Td
(by)Tj
-34.8375 -1.2983 Td
(TCR)Tj
2.1146 0 Td
(and)Tj
1.7291 0 Td
(CD28)Tj
2.5738 0 Td
(signaling,)Tj
4.0479 0 Td
(CD4)Tj
2.092 0 Td
(T)Tj
0.8333 0 Td
(cells)Tj
1.9219 0 Td
(fail)Tj
1.4513 0 Td
(to)Tj
1.0262 0 Td
(proliferate)Tj
4.371 0 Td
(or)Tj
1.0941 0 Td
(to)Tj
1.0262 0 Td
(produce)Tj
3.5206 0 Td
(IL-2)Tj
1.9219 0 Td
(and)Tj
1.7291 0 Td
(enter)Tj
2.2847 0 Td
(a)Tj
0.6576 0 Td
(state)Tj
2.0353 0 Td
(of)Tj
-36.4306 -1.2982 Td
(unresponsiveness)Tj
7.2226 0 Td
(following)Tj
3.9798 0 Td
(immunogenic)Tj
5.8393 0 Td
(stimulation,)Tj
5.0173 0 Td
(referred)Tj
3.4243 0 Td
(to)Tj
1.0204 0 Td
(as)Tj
1.0205 0 Td
(anergy)Tj
3.6963 0 Td
([)Tj
0.83 0.64 0.02 0 k
(11)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(12)Tj
0 g
(].)Tj
3.2542 0 Td
(In)Tj
1.1112 0 Td
(the)Tj
-35.5859 -1.304 Td
(case)Tj
1.8595 0 Td
(of)Tj
1.0148 0 Td
(CD4)Tj
2.0976 0 Td
(T)Tj
0.8277 0 Td
(cells,)Tj
2.1486 0 Td
(the)Tj
1.4684 0 Td
(development)Tj
5.4085 0 Td
(of)Tj
1.0148 0 Td
(anergy)Tj
2.9196 0 Td
(depends)Tj
3.549 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.4683 0 Td
(alteration)Tj
4.0478 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(expression)Tj
4.4617 0 Td
(of)Tj
-36.0507 -1.2982 Td
(several)Tj
2.931 0 Td
(genes)Tj
2.4434 0 Td
([)Tj
0.83 0.64 0.02 0 k
(11)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(12)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(13)Tj
0 g
(].)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 183.2882 23.6976 re
W* n
q
183.2315 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 399.0047 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(T)Tj
0.822 0 Td
(Cell)Tj
1.9063 0 Td
(Hyporespon)Tj
5.2938 0 Td
(siveness)Tj
4.0464 0 Td
(during)Tj
/F6 1 Tf
2.941 0 Td
(T)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.1055 0 Td
(cruzi)Tj
/F0 1 Tf
2.289 0 Td
(Infection)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(Neglected)Tj
4.6417 0 Td
(Tropical)Tj
3.7135 0 Td
(Disease)Tj
3.5433 0 Td
(s)Tj
0.7158 0 Td
(|)Tj
0.4748 0 Td
(DOI:10.137)Tj
5.0386 0 Td
(1/journal.pn)Tj
5.1236 0 Td
(td.00053)Tj
3.8198 0 Td
(07)Tj
2.0905 0 Td
(January)Tj
3.7134 0 Td
(23,)Tj
1.5875 0 Td
(2017)Tj
27.8222 0 Td
(2)Tj
0.7654 0 Td
(/)Tj
0.496 0 Td
(16)Tj
/F10 1 Tf
-0.025 Tc
-66.402 84.0904 Td
(Investigac)Tj
3.7559 0 Td
(iones)Tj
2.2039 0 Td
(Cient)Tj
0 Tc
1.8142 0.0212 Td
()Tj
-0.025 Tc
0.2481 -0.0212 Td
(ficas)Tj
1.9346 0 Td
(y)Tj
0.6378 0 Td
(Te)Tj
0 Tc
0.5386 0.0283 Td
()Tj
-0.025 Tc
0.3614 -0.0283 Td
(cnicas)Tj
2.5725 0 Td
(\(CONICET\))Tj
-14.067 -1.3748 Td
(PIP)Tj
1.5236 0 Td
(0620)Tj
2.1189 0 Td
(CCS;)Tj
2.0197 0 Td
(Secretar)Tj
3.0756 0 Td
()Tj
0 Tc
-0.0496 0.0212 Td
()Tj
-0.025 Tc
0.248 -0.0212 Td
(a)Tj
0.6378 0 Td
(de)Tj
1.1197 0 Td
(Ciencia)Tj
2.863 0 Td
(y)Tj
0.6449 0 Td
(Tecnolog)Tj
0 Tc
3.352 0.0212 Td
()Tj
-0.025 Tc
0.241 -0.0212 Td
(a)Tj
-17.7946 -1.3749 Td
(Universida)Tj
3.9118 0 Td
(d)Tj
0.6945 0 Td
(Nacional)Tj
3.4087 0 Td
(de)Tj
1.1126 0 Td
(Co)Tj
0 Tc
0.6094 0.0284 Td
()Tj
-0.025 Tc
0.3898 -0.0284 Td
(rdoba)Tj
2.3669 0 Td
(\(SECyT-UNC)Tj
4.7339 0 Td
(\))Tj
-17.2276 -1.3748 Td
(Res.)Tj
1.8779 0 Td
(103/15)Tj
2.8418 0 Td
(FMC;)Tj
2.1401 0 Td
(Secretar)Tj
0 Tc
3.0331 0.0213 Td
()Tj
-0.025 Tc
0.241 -0.0213 Td
(a)Tj
0.6449 0 Td
(de)Tj
1.1126 0 Td
(Ciencia)Tj
2.8701 0 Td
(y)Tj
-14.7615 -1.3748 Td
(Tecnolog)Tj
0 Tc
3.3519 0.0213 Td
()Tj
-0.025 Tc
0.241 -0.0213 Td
(a)Tj
0.6449 0 Td
(Universida)Tj
3.9118 0 Td
(d)Tj
0.6945 0 Td
(Nacional)Tj
3.4087 0 Td
(de)Tj
1.1126 0 Td
(Co)Tj
0 Tc
0.6095 0.0284 Td
()Tj
-0.025 Tc
0.3897 -0.0284 Td
(rdoba)Tj
-14.3646 -1.3748 Td
(\(SECyT-UNC)Tj
4.7338 0 Td
(\))Tj
0.5245 0 Td
(Res.203/14)Tj
4.5 0 Td
(CCS;)Tj
2.0197 0 Td
(Ministerio)Tj
3.9048 0 Td
(de)Tj
-15.6828 -1.3748 Td
(Ciencia)Tj
2.8701 0 Td
(y)Tj
0.6378 0 Td
(Tecnolog)Tj
0 Tc
3.3519 0.0213 Td
()Tj
-0.025 Tc
0.2481 -0.0213 Td
(a)Tj
0.6378 0 Td
(de)Tj
1.1197 0 Td
(la)Tj
0.8291 0 Td
(Provincia)Tj
3.6496 0 Td
(de)Tj
1.1197 0 Td
(Co)Tj
0 Tc
0.6095 0.0283 Td
()Tj
-0.025 Tc
0.3898 -0.0283 Td
(rdoba)Tj
-15.4631 -1.3748 Td
(\(Mincyt-CB)Tj
4.1811 0 Td
(A\))Tj
1.0559 0 Td
(Res)Tj
1.6512 0 Td
(113/11)Tj
2.8417 0 Td
(FMC)Tj
1.9205 0 Td
(and)Tj
1.5874 0 Td
(Fundacion)Tj
-13.2378 -1.3748 Td
(Bunge)Tj
2.5866 0 Td
(y)Tj
0.6449 0 Td
(Born)Tj
1.9984 0 Td
(2010)Tj
2.1118 0 Td
(CCS.)Tj
2.0197 0 Td
(The)Tj
1.5875 0 Td
(funders)Tj
3.0472 0 Td
(had)Tj
1.5874 0 Td
(no)Tj
1.1693 0 Td
(role)Tj
-16.7528 -1.3748 Td
(in)Tj
0.8858 0 Td
(the)Tj
1.3677 0 Td
(study)Tj
2.2677 0 Td
(design,)Tj
2.8985 0 Td
(data)Tj
1.7929 0 Td
(collection)Tj
3.7418 0 Td
(and)Tj
1.5874 0 Td
(analysis,)Tj
-14.5418 -1.3749 Td
(decision)Tj
3.2882 0 Td
(to)Tj
0.9496 0 Td
(publish,)Tj
3.1465 0 Td
(or)Tj
0.9992 0 Td
(preparation)Tj
4.4433 0 Td
(of)Tj
0.9496 0 Td
(the)Tj
-13.7764 -1.3748 Td
(manuscript.)Tj
/F9 1 Tf
0 -2.126 Td
(Competing)Tj
4.3795 0 Td
(Interests)Tj
3.3307 0 Td
(:)Tj
/F10 1 Tf
0.4819 0 Td
(The)Tj
1.5875 0 Td
(authors)Tj
3.0401 0 Td
(have)Tj
1.963 0 Td
(declared)Tj
-14.7827 -1.3748 Td
(that)Tj
1.6228 0 Td
(no)Tj
1.1693 0 Td
(competing)Tj
4.1599 0 Td
(interests)Tj
ET
endstream
endobj
34 0 obj
<>stream
endstream
endobj
35 0 obj
<>stream
endstream
endobj
36 0 obj
<>stream
endstream
endobj
37 0 obj
<>stream
endstream
endobj
38 0 obj
<>stream
endstream
endobj
39 0 obj
<>stream
endstream
endobj
40 0 obj
<>stream
endstream
endobj
41 0 obj
<>stream
endstream
endobj
42 0 obj
<>stream
endstream
endobj
43 0 obj
<>stream
endstream
endobj
44 0 obj
<>stream
endstream
endobj
45 0 obj
<>stream
endstream
endobj
46 0 obj
<>stream
endstream
endobj
47 0 obj
<>stream
BT
10.3394 65.4666 Td
(exist.)Tj
ET
Q
endstream
endobj
48 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 50 0 R/Contents 51 0 R/TrimBox[0 0 612 792]>>
endobj
50 0 obj
[52 0 R 53 0 R 54 0 R 55 0 R 56 0 R 57 0 R 58 0 R 59 0 R 60 0 R 61 0 R 62 0 R 63 0 R 64 0 R 65 0 R 66 0 R 67 0 R 68 0 R 69 0 R 70 0 R 71 0 R 72 0 R 73 0 R 74 0 R 75 0 R 76 0 R 77 0 R 78 0 R]
endobj
52 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref014)>>
endobj
53 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref015)>>
endobj
54 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref016)>>
endobj
55 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref017)>>
endobj
56 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref018)>>
endobj
57 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref019)>>
endobj
58 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref017)>>
endobj
59 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref020)>>
endobj
60 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref017)>>
endobj
61 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref021)>>
endobj
62 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref017)>>
endobj
63 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref020)>>
endobj
64 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref022)>>
endobj
65 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref023)>>
endobj
66 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref024)>>
endobj
67 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref025)>>
endobj
68 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref026)>>
endobj
69 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref027)>>
endobj
70 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref019)>>
endobj
71 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref019)>>
endobj
72 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref017)>>
endobj
73 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref020)>>
endobj
74 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref020)>>
endobj
75 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref028)>>
endobj
76 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref029)>>
endobj
77 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref030)>>
endobj
78 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref031)>>
endobj
51 0 obj
[79 0 R 80 0 R 81 0 R 82 0 R 83 0 R 84 0 R 85 0 R 86 0 R 87 0 R 88 0 R 89 0 R 90 0 R 91 0 R 92 0 R 93 0 R 94 0 R 95 0 R 96 0 R 97 0 R 98 0 R 99 0 R 100 0 R]
endobj
79 0 obj
<>stream
q
0.83 0.64 0.02 0 k
393.6756 693.2976 m
403.2 693.2976 l
h
f*
405.4677 693.2976 m
414.9921 693.2976 l
h
f*
350.7024 667.3323 m
360.1701 667.3323 l
h
f*
362.4378 667.3323 m
371.9622 667.3323 l
h
f*
374.2299 667.3323 m
383.7543 667.3323 l
h
f*
438.1795 641.3102 m
447.6472 641.3102 l
h
f*
269.5181 615.3449 m
279.0425 615.3449 l
h
f*
281.2535 615.3449 m
290.778 615.3449 l
h
f*
506.3811 615.3449 m
515.9055 615.3449 l
h
f*
518.1732 615.3449 m
527.6976 615.3449 l
h
f*
293.3291 602.3055 m
302.8535 602.3055 l
h
f*
265.2661 563.3008 m
274.7906 563.3008 l
h
f*
305.9717 537.3354 m
315.4961 537.3354 l
h
f*
317.7638 537.3354 m
327.2882 537.3354 l
h
f*
329.4992 537.3354 m
339.0236 537.3354 l
h
f*
341.2913 537.3354 m
350.8157 537.3354 l
h
f*
432.3402 524.2961 m
441.8646 524.2961 l
h
f*
317.8772 498.3307 m
327.4016 498.3307 l
h
f*
542.0976 485.348 m
551.622 485.348 l
h
f*
271.6724 420.3213 m
281.1969 420.3213 l
h
f*
372.1323 407.3386 m
381.6567 407.3386 l
h
f*
383.9244 407.3386 m
393.4488 407.3386 l
h
f*
409.7764 394.2992 m
419.3008 394.2992 l
h
f*
442.885 381.3165 m
452.4094 381.3165 l
h
f*
454.6772 381.3165 m
464.2016 381.3165 l
h
f*
466.4126 381.3165 m
475.937 381.3165 l
h
f*
478.2047 381.3165 m
487.7291 381.3165 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 211.9748 707.4141 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Post-translational)Tj
7.2906 0 Td
(modification)Tj
5.3745 0 Td
(of)Tj
1.0148 0 Td
(proteins)Tj
3.5149 0 Td
(via)Tj
1.3776 0 Td
(ubiquitination)Tj
5.9924 0 Td
(also)Tj
1.7688 0 Td
(plays)Tj
2.2337 0 Td
(an)Tj
1.2019 0 Td
(essential)Tj
3.583 0 Td
(role)Tj
1.7688 0 Td
(in)Tj
-36.3172 -1.3039 Td
(the)Tj
1.4626 0 Td
(regulatory)Tj
4.32 0 Td
(mechanism)Tj
4.8415 0 Td
(of)Tj
1.0091 0 Td
(CD4)Tj
2.092 0 Td
(T)Tj
0.8334 0 Td
(cell)Tj
1.5534 0 Td
(anergy)Tj
2.914 0 Td
([)Tj
0.83 0.64 0.02 0 k
(14)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(15)Tj
0 g
(].)Tj
3.2541 0 Td
(GRAIL,)Tj
3.3392 0 Td
(also)Tj
1.7631 0 Td
(known)Tj
3.0047 0 Td
(as)Tj
1.0092 0 Td
(ring)Tj
1.8708 0 Td
(finger)Tj
2.5795 0 Td
(pro-)Tj
-35.8466 -1.2982 Td
(tein-128)Tj
3.5319 0 Td
(\(RNF-128\),)Tj
4.7905 0 Td
(has)Tj
1.542 0 Td
(been)Tj
2.1203 0 Td
(identified)Tj
4.0592 0 Td
(as)Tj
1.0148 0 Td
(a)Tj
0.6577 0 Td
(novel)Tj
2.3981 0 Td
(E3)Tj
1.2642 0 Td
(ubiquitin-protein)Tj
7.2226 0 Td
(ligase)Tj
2.4151 0 Td
(that)Tj
1.7802 0 Td
(induces)Tj
-32.7966 -1.2983 Td
(and)Tj
1.7291 0 Td
(maintains)Tj
4.2009 0 Td
(anergy)Tj
2.914 0 Td
(in)Tj
1.0374 0 Td
(CD4)Tj
2.092 0 Td
(T)Tj
0.8334 0 Td
(cells)Tj
1.9219 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(18)Tj
0 g
(].)Tj
4.4277 0 Td
(It)Tj
0.8617 0 Td
(has)Tj
1.542 0 Td
(been)Tj
2.1203 0 Td
(shown)Tj
2.846 0 Td
(that)Tj
1.7801 0 Td
(GRAIL)Tj
3.1181 0 Td
(expression)Tj
-31.4246 -1.2983 Td
(could)Tj
2.4491 0 Td
(be)Tj
1.1508 0 Td
(correlated)Tj
4.2463 0 Td
(with)Tj
2.0012 0 Td
(the)Tj
1.4684 0 Td
(inhibition)Tj
4.2236 0 Td
(of)Tj
1.0148 0 Td
(CD4)Tj
2.0976 0 Td
(T)Tj
0.8277 0 Td
(cell)Tj
1.559 0 Td
(proliferation)Tj
5.2668 0 Td
(and)Tj
1.7348 0 Td
(antigen-induced)Tj
6.7974 0 Td
(IL-2)Tj
-34.8375 -1.3039 Td
(transcription)Tj
5.4311 0 Td
(by)Tj
1.1849 0 Td
(disrupting)Tj
4.371 0 Td
(the)Tj
1.4683 0 Td
(T)Tj
0.8334 0 Td
(cell)Tj
1.559 0 Td
(stimulatory)Tj
4.8019 0 Td
(signaling)Tj
3.8267 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
2.075 0 Td
(In)Tj
1.1055 0 Td
(support)Tj
3.3222 0 Td
(of)Tj
1.0148 0 Td
(this)Tj
1.6724 0 Td
(observation,)Tj
-32.6662 -1.2982 Td
(T)Tj
0.8277 0 Td
(cells)Tj
1.9218 0 Td
(from)Tj
2.2054 0 Td
(GRAIL)Tj
3.1181 0 Td
(knock-out)Tj
4.3766 0 Td
(mice)Tj
2.1543 0 Td
(were)Tj
2.126 0 Td
(shown)Tj
2.846 0 Td
(to)Tj
1.0261 0 Td
(be)Tj
1.1509 0 Td
(defective)Tj
3.7473 0 Td
(in)Tj
1.0375 0 Td
(anergy)Tj
2.914 0 Td
(induction)Tj
4.1386 0 Td
(both)Tj
/F13 1 Tf
2.0579 0 Td
[(\r)12()]TJ
-35.6482 -1.2983 Td
[()-13(\r)14()7()20()]TJ
/F8 1 Tf
2.0465 0 Td
(and)Tj
/F13 1 Tf
1.7292 0 Td
[(\r)12()]TJ
1.0148 0 Td
[()-13(\r)15()3()]TJ
/F8 1 Tf
1.8198 0 Td
([)Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(20)Tj
0 g
(].)Tj
3.2541 0 Td
(In)Tj
1.1055 0 Td
(particular,)Tj
4.337 0 Td
(GRAIL)Tj
3.1181 0 Td
(\(-/-\))Tj
1.9332 0 Td
(T)Tj
0.8334 0 Td
(cells)Tj
1.9162 0 Td
(hyperproliferated)Tj
7.1943 0 Td
([)Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(21)Tj
0 g
(])Tj
3.0274 0 Td
(and)Tj
1.7291 0 Td
(pro-)Tj
-35.0586 -1.3039 Td
(duced)Tj
2.6475 0 Td
(more)Tj
2.3414 0 Td
(cytokines)Tj
4.0025 0 Td
([)Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(])Tj
1.8482 0 Td
(compared)Tj
4.2462 0 Td
(with)Tj
1.9956 0 Td
(wild)Tj
1.9446 0 Td
(type)Tj
1.9162 0 Td
(\(WT\))Tj
2.4774 0 Td
(cells)Tj
1.9219 0 Td
(in)Tj
1.0318 0 Td
(response)Tj
3.7361 0 Td
(to)Tj
1.0261 0 Td
(TCR)Tj
2.1146 0 Td
(stimula-)Tj
-33.2501 -1.2983 Td
(tion)Tj
1.8424 0 Td
(alone)Tj
/F13 1 Tf
2.3755 0 Td
[(\r)12()]TJ
1.0148 0 Td
[()-13(\r)14()7()20()]TJ
/F8 1 Tf
2.0466 0 Td
(or)Tj
1.0998 0 Td
(with)Tj
1.9956 0 Td
(concomitant)Tj
5.2951 0 Td
(anti-CD28)Tj
4.4673 0 Td
(costimulation.)Tj
5.9528 0 Td
(It)Tj
0.856 0 Td
(has)Tj
1.5477 0 Td
(also)Tj
1.7632 0 Td
(been)Tj
2.1259 0 Td
(recently)Tj
-32.3827 -1.2983 Td
(demonstrated)Tj
5.7599 0 Td
(that)Tj
1.7801 0 Td
(GRAIL,)Tj
3.3449 0 Td
(by)Tj
1.1792 0 Td
(mediating)Tj
4.269 0 Td
(TCR-CD3)Tj
4.3426 0 Td
(degradation,)Tj
5.2497 0 Td
(regulated)Tj
3.9402 0 Td
(naive)Tj
2.347 0 Td
(T)Tj
0.8334 0 Td
(cell)Tj
1.5591 0 Td
(toler-)Tj
-34.6051 -1.3039 Td
(ance)Tj
2.0465 0 Td
(induction)Tj
4.1386 0 Td
([)Tj
0.83 0.64 0.02 0 k
(20)Tj
0 g
(].)Tj
2.0749 0 Td
(Furthermore,)Tj
5.6126 0 Td
(several)Tj
2.9367 0 Td
(investigations)Tj
5.7203 0 Td
(have)Tj
2.0636 0 Td
(shown)Tj
2.8459 0 Td
(how)Tj
1.9389 0 Td
(GRAIL)Tj
3.1181 0 Td
(interacts)Tj
-32.4961 -1.2982 Td
(with)Tj
1.9955 0 Td
(T-cells,)Tj
3.1068 0 Td
(antigen)Tj
3.2088 0 Td
(presenting)Tj
4.4447 0 Td
(cell)Tj
1.559 0 Td
(\(APC\))Tj
2.8063 0 Td
(receptors)Tj
3.9174 0 Td
(and)Tj
1.7292 0 Td
(cytoskeletal)Tj
4.8302 0 Td
(proteins,)Tj
3.7417 0 Td
(thereby)Tj
3.2258 0 Td
(pro-)Tj
-34.5654 -1.2983 Td
(moting)Tj
3.1237 0 Td
(their)Tj
2.109 0 Td
(degradation)Tj
5.0229 0 Td
([)Tj
0.83 0.64 0.02 0 k
(22)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(23)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(24)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(25)Tj
0 g
(].)Tj
-9.0594 -1.3039 Td
(GRAIL)Tj
3.118 0 Td
(expression)Tj
4.4561 0 Td
(is)Tj
0.8447 0 Td
(regulated)Tj
3.9345 0 Td
(by)Tj
1.1735 0 Td
(Otubain-1)Tj
4.371 0 Td
(\(Otub-1\))Tj
3.7984 0 Td
([)Tj
0.83 0.64 0.02 0 k
(26)Tj
0 g
(],)Tj
2.0749 0 Td
(which)Tj
2.6419 0 Td
(is)Tj
0.8447 0 Td
(a)Tj
0.6463 0 Td
(member)Tj
3.5773 0 Td
(of)Tj
1.0148 0 Td
(deubiqui-)Tj
-33.6923 -1.2983 Td
(tinating)Tj
3.3448 0 Td
(enzymes)Tj
3.6623 0 Td
(with)Tj
2.0013 0 Td
(the)Tj
1.4683 0 Td
(capability)Tj
4.0705 0 Td
(to)Tj
1.0262 0 Td
(cleave)Tj
2.6248 0 Td
(proteins)Tj
3.515 0 Td
(at)Tj
0.9524 0 Td
(the)Tj
1.4684 0 Td
(ubiquitin-protein)Tj
7.2169 0 Td
(bond)Tj
2.3131 0 Td
(by)Tj
1.1792 0 Td
(using)Tj
-34.8432 -1.2982 Td
(its)Tj
1.1452 0 Td
(cysteine)Tj
3.4185 0 Td
(protease)Tj
3.5546 0 Td
(domain)Tj
3.3279 0 Td
([)Tj
0.83 0.64 0.02 0 k
(27)Tj
0 g
(].)Tj
2.0749 0 Td
(It)Tj
0.8617 0 Td
(has)Tj
1.5421 0 Td
(been)Tj
2.1203 0 Td
(shown)Tj
2.8459 0 Td
(that)Tj
1.7802 0 Td
(Otub-1)Tj
3.1238 0 Td
(is)Tj
0.8503 0 Td
(expressed)Tj
4.0932 0 Td
(and)Tj
1.7292 0 Td
(GRAIL)Tj
-32.4678 -1.2983 Td
(degraded)Tj
3.9174 0 Td
(when)Tj
2.4038 0 Td
(naive)Tj
2.3527 0 Td
(CD4)Tj
2.092 0 Td
(T)Tj
0.8333 0 Td
(cells)Tj
1.9162 0 Td
(are)Tj
1.4514 0 Td
(productively)Tj
5.2384 0 Td
(activated)Tj
3.7814 0 Td
(to)Tj
1.0261 0 Td
(undergo)Tj
3.5886 0 Td
(proliferation)Tj
5.2668 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
-33.8681 -1.3039 Td
(Moreover,)Tj
4.3936 0 Td
(the)Tj
1.4683 0 Td
(loss)Tj
1.6952 0 Td
(of)Tj
1.0148 0 Td
(GRAIL)Tj
3.1237 0 Td
(was)Tj
1.6951 0 Td
(mechanistically)Tj
6.3949 0 Td
(controlled)Tj
4.32 0 Td
(through)Tj
3.4469 0 Td
(a)Tj
0.652 0 Td
(pathway)Tj
3.5716 0 Td
(involving)Tj
-31.7761 -1.2983 Td
(CD28)Tj
2.5681 0 Td
(co-stimulation,)Tj
6.3042 0 Td
(IL-2)Tj
1.9219 0 Td
(production)Tj
4.7168 0 Td
(and)Tj
1.7291 0 Td
(IL-2R)Tj
2.5455 0 Td
(signaling,)Tj
4.0479 0 Td
(and)Tj
1.7291 0 Td
(ultimately,)Tj
4.4617 0 Td
(mTOR-dependent)Tj
-30.0243 -1.2983 Td
(translation)Tj
4.5353 0 Td
(of)Tj
1.0148 0 Td
(select)Tj
2.3925 0 Td
(mRNAs.)Tj
3.668 0 Td
(Blocking)Tj
3.753 0 Td
(the)Tj
1.474 0 Td
(mTOR)Tj
3.0161 0 Td
(by)Tj
1.1848 0 Td
(using)Tj
2.3868 0 Td
(CTLA-4-Ig,)Tj
4.9266 0 Td
(anti-IL-2)Tj
3.8211 0 Td
(or)Tj
1.0998 0 Td
(rapamycin)Tj
-33.2728 -1.3039 Td
(prevented)Tj
4.2065 0 Td
(Otub-1)Tj
3.1295 0 Td
(protein)Tj
3.1521 0 Td
(expression)Tj
4.4617 0 Td
(and)Tj
1.7348 0 Td
(maintained)Tj
4.7848 0 Td
(GRAIL)Tj
3.1181 0 Td
(expression,)Tj
4.6941 0 Td
(which)Tj
2.6476 0 Td
(inhibited)Tj
3.8551 0 Td
(T)Tj
-35.7843 -1.2982 Td
(cell)Tj
1.559 0 Td
(proliferation)Tj
5.2667 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(].)Tj
2.075 0 Td
(Although)Tj
4.0025 0 Td
(the)Tj
1.474 0 Td
(function)Tj
3.6283 0 Td
(of)Tj
1.0148 0 Td
(GRAIL)Tj
3.1181 0 Td
(in)Tj
1.0375 0 Td
(CD4)Tj
2.0919 0 Td
(T)Tj
0.8334 0 Td
(cells)Tj
1.9219 0 Td
(has)Tj
1.542 0 Td
(been)Tj
2.1203 0 Td
(studied)Tj
3.1521 0 Td
(ex-)Tj
-34.8375 -1.2983 Td
(tensively)Tj
3.6963 0 Td
(for)Tj
1.389 0 Td
(the)Tj
1.4683 0 Td
(development)Tj
5.4085 0 Td
(of)Tj
1.0148 0 Td
(tolerance)Tj
3.8947 0 Td
([)Tj
0.83 0.64 0.02 0 k
(17)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(20)Tj
0 g
(],)Tj
3.2542 0 Td
(with)Tj
1.9956 0 Td
(its)Tj
1.1508 0 Td
(participation)Tj
5.3688 0 Td
(having)Tj
2.9197 0 Td
(been)Tj
2.1203 0 Td
(demon-)Tj
-33.681 -1.3039 Td
(strated)Tj
2.9309 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4684 0 Td
(development)Tj
5.4141 0 Td
(of)Tj
1.0148 0 Td
(autoimmune)Tj
5.3915 0 Td
(diseases)Tj
3.3788 0 Td
([)Tj
0.83 0.64 0.02 0 k
(20)Tj
0 g
(],)Tj
2.075 0 Td
(only)Tj
1.9785 0 Td
(recently)Tj
3.4073 0 Td
(has)Tj
1.542 0 Td
(its)Tj
1.1509 0 Td
(role)Tj
1.7631 0 Td
(been)Tj
-32.5528 -1.2983 Td
(studied)Tj
3.152 0 Td
(during)Tj
2.9254 0 Td
(T)Tj
0.8334 0 Td
(cell)Tj
1.559 0 Td
(dysfunction)Tj
4.9719 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4684 0 Td
(course)Tj
2.8346 0 Td
(of)Tj
1.0148 0 Td
(infections)Tj
4.1499 0 Td
([)Tj
0.83 0.64 0.02 0 k
(28)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(29)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(30)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
(31)Tj
0 g
(].)Tj
5.6069 0 Td
(Thus,)Tj
2.4774 0 Td
(the)Tj
1.4684 0 Td
(aim)Tj
1.7404 0 Td
(of)Tj
-35.24 -1.2983 Td
(this)Tj
1.6724 0 Td
(work)Tj
2.279 0 Td
(was)Tj
1.7008 0 Td
(to)Tj
1.0261 0 Td
(search)Tj
2.7609 0 Td
(for)Tj
1.3833 0 Td
(a)Tj
0.652 0 Td
(novel)Tj
2.4038 0 Td
(link)Tj
1.7745 0 Td
(between)Tj
3.5319 0 Td
(GRAIL)Tj
3.1181 0 Td
(and)Tj
1.7291 0 Td
(CD4)Tj
2.0976 0 Td
(T)Tj
0.8277 0 Td
(cell)Tj
1.5648 0 Td
(unresponsiveness)Tj
7.2226 0 Td
(in)Tj
-35.7446 -1.2982 Td
(the)Tj
1.4683 0 Td
(context)Tj
3.1861 0 Td
(of)Tj
1.0148 0 Td
(abnormalities)Tj
5.7033 0 Td
(of)Tj
1.0148 0 Td
(T)Tj
0.8333 0 Td
(cell)Tj
1.5591 0 Td
(proliferation)Tj
5.2667 0 Td
(observed)Tj
3.7928 0 Td
(during)Tj
/F13 1 Tf
2.931 0 Td
[()75()-20()-13()4()16()6()7()-3()11( )6()]TJ
5.6182 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(infec-)Tj
-34.5484 -1.3039 Td
(tion.)Tj
2.0635 0 Td
(Our)Tj
1.8652 0 Td
(results)Tj
2.812 0 Td
(provide)Tj
3.2882 0 Td
(evidence)Tj
3.7133 0 Td
(demonstrating)Tj
6.0888 0 Td
(that)Tj
1.7801 0 Td
(CD4)Tj
2.092 0 Td
(T)Tj
0.8334 0 Td
(cells)Tj
1.9219 0 Td
(from)Tj
/F13 1 Tf
2.2053 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0318 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(infected)Tj
3.4242 0 Td
(mice)Tj
-35.2797 -1.2983 Td
(exhibited)Tj
3.9231 0 Td
(an)Tj
1.2075 0 Td
(increase)Tj
3.4696 0 Td
(in)Tj
1.0375 0 Td
(GRAIL)Tj
3.1181 0 Td
(expression)Tj
4.4673 0 Td
(during)Tj
2.9254 0 Td
(the)Tj
1.4683 0 Td
(acute)Tj
2.3301 0 Td
(phase)Tj
2.4831 0 Td
(of)Tj
1.0148 0 Td
(infection,)Tj
4.0138 0 Td
(which)Tj
2.6532 0 Td
(was)Tj
1.7008 0 Td
(cor-)Tj
-35.8126 -1.2983 Td
(related)Tj
2.9423 0 Td
(with)Tj
1.9956 0 Td
(defects)Tj
2.9706 0 Td
(in)Tj
1.0319 0 Td
(proliferation)Tj
5.2667 0 Td
(and)Tj
1.7291 0 Td
(immune)Tj
3.6283 0 Td
(responsiveness.)Tj
6.3723 0 Td
(In)Tj
1.1112 0 Td
(addition,)Tj
3.821 0 Td
(we)Tj
1.3266 0 Td
(showed)Tj
3.2542 0 Td
(that)Tj
-35.4498 -1.3039 Td
(high)Tj
1.9955 0 Td
(expression)Tj
4.4617 0 Td
(of)Tj
1.0035 0 Td
(CTLA-4)Tj
3.5319 0 Td
(and)Tj
1.7235 0 Td
(low)Tj
1.6497 0 Td
(levels)Tj
2.3641 0 Td
(of)Tj
1.0091 0 Td
(IL-2)Tj
1.9162 0 Td
(prevented)Tj
4.201 0 Td
(mTOR)Tj
3.0103 0 Td
(activation)Tj
4.1443 0 Td
(and)Tj
1.7177 0 Td
(Otub-1)Tj
3.1181 0 Td
(pro-)Tj
-35.8466 -1.2982 Td
(tein)Tj
1.7517 0 Td
(expression,)Tj
4.6885 0 Td
(and)Tj
1.7235 0 Td
(maintained)Tj
4.7848 0 Td
(GRAIL)Tj
3.1124 0 Td
(expression,)Tj
4.6885 0 Td
(which)Tj
2.6419 0 Td
(inhibited)Tj
3.8494 0 Td
(T)Tj
0.8277 0 Td
(cell)Tj
1.5591 0 Td
(proliferation)Tj
5.261 0 Td
(during)Tj
-34.8885 -1.2983 Td
(the)Tj
1.4683 0 Td
(acute)Tj
2.3244 0 Td
(phase)Tj
2.4888 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(infection.)Tj
4.0138 0 Td
(Our)Tj
1.8652 0 Td
(results)Tj
2.812 0 Td
(therefore)Tj
3.8551 0 Td
(indicate)Tj
3.4072 0 Td
(that)Tj
1.7801 0 Td
(GRAIL)Tj
3.1181 0 Td
(is)Tj
0.8504 0 Td
(an)Tj
1.2019 0 Td
(important)Tj
-31.6684 -1.3039 Td
(player)Tj
2.6645 0 Td
(in)Tj
1.0375 0 Td
(CD4)Tj
2.0919 0 Td
(T)Tj
0.8334 0 Td
(cell)Tj
1.559 0 Td
(anergy)Tj
2.9197 0 Td
(during)Tj
2.9253 0 Td
(the)Tj
1.4684 0 Td
(acute)Tj
2.33 0 Td
(phase)Tj
2.4832 0 Td
(of)Tj
/F13 1 Tf
1.0148 0 Td
[()75()-20()-13()4()16()6()7()-3()11( )6()]TJ
5.6182 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(infection.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 239.1874 cm
BT
/F3 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Methods)Tj
/F0 1 Tf
0 -1.4976 Td
(Ethics)Tj
2.9149 0 Td
(statement)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 204.2078 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(All)Tj
1.3946 0 Td
(the)Tj
1.4683 0 Td
(animal)Tj
2.9707 0 Td
(experiments)Tj
5.142 0 Td
(were)Tj
2.1203 0 Td
(examined)Tj
4.1272 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4684 0 Td
(Institutional)Tj
5.1647 0 Td
(Experimentation)Tj
6.9788 0 Td
(Animal)Tj
-32.0199 -1.2982 Td
(Committee,)Tj
4.9662 0 Td
(from)Tj
2.2054 0 Td
(Facultad)Tj
3.6396 0 Td
(de)Tj
1.1679 0 Td
(Ciencias)Tj
3.6113 0 Td
(Qu)Tj
0 Tc
1.2132 0.051 Td
()Tj
-0.004 Tc
0.3288 -0.051 Td
(micas,)Tj
2.7496 0 Td
(Univesidad)Tj
4.7679 0 Td
(Nacional)Tj
3.8154 0 Td
(de)Tj
1.1735 0 Td
(Co)Tj
0 Tc
0.7257 0.051 Td
()Tj
-0.004 Tc
0.4422 -0.051 Td
(rdoba,)Tj
2.7893 0 Td
(which)Tj
-33.596 -1.3039 Td
(approved)Tj
3.9741 0 Td
(the)Tj
1.4683 0 Td
(experimental)Tj
5.4595 0 Td
(procedures)Tj
4.6715 0 Td
(\(authorization)Tj
5.9584 0 Td
(no.)Tj
1.5023 0 Td
(2016209\).)Tj
4.6318 0 Td
(This)Tj
1.9899 0 Td
(committee)Tj
4.5014 0 Td
(follows)Tj
-34.1572 -1.2983 Td
(the)Tj
1.4683 0 Td
(guidelines)Tj
4.2803 0 Td
(for)Tj
1.3833 0 Td
(animal)Tj
2.9707 0 Td
(care)Tj
1.8708 0 Td
(of)Tj
1.0148 0 Td
(Guide)Tj
3.0728 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(care)Tj
1.8765 0 Td
(and)Tj
1.7292 0 Td
(use)Tj
1.5307 0 Td
(of)Tj
1.0148 0 Td
(experime)Tj
/F14 1 Tf
(n)Tj
/F8 1 Tf
(tal)Tj
5.4708 0 Td
(animals)Tj
3.719 0 Td
(\(Cana-)Tj
-33.8964 -1.2983 Td
(dian)Tj
1.9955 0 Td
(Council)Tj
3.4016 0 Td
(on)Tj
1.2756 0 Td
(Animal)Tj
3.2201 0 Td
(Care,)Tj
2.3414 0 Td
(1993\))Tj
2.4605 0 Td
(and)Tj
1.7291 0 Td
(of)Tj
1.0148 0 Td
(Institutional)Tj
5.5559 0 Td
(Animal)Tj
3.2201 0 Td
(Care)Tj
2.109 0 Td
(and)Tj
1.7347 0 Td
(Use)Tj
1.7405 0 Td
(Comittee)Tj
-31.7988 -1.3039 Td
(Guidebook)Tj
5.0909 0 Td
(\(ARENA/OLAW)Tj
7.1206 0 Td
(IACUC)Tj
3.3052 0 Td
(Guidebook,)Tj
4.9266 0 Td
(Nacional)Tj
3.8154 0 Td
(Institutes)Tj
3.9401 0 Td
(of)Tj
1.0148 0 Td
(Health,)Tj
3.1465 0 Td
(2002\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 112.9889 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Mice)Tj
2.3102 0 Td
(and)Tj
/F6 1 Tf
1.8614 0 Td
(T)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.1103 0 Td
(cruzi)Tj
/F0 1 Tf
2.3055 0 Td
(infection)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 95.9811 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(BALB/c)Tj
3.3561 0 Td
(mice)Tj
2.1544 0 Td
(obtained)Tj
3.7303 0 Td
(from)Tj
2.2054 0 Td
(the)Tj
1.4683 0 Td
(Comisio)Tj
0 Tc
2.948 0.0566 Td
()Tj
-0.004 Tc
0.4479 -0.0566 Td
(n)Tj
0.7653 0 Td
(Nacional)Tj
3.8155 0 Td
(de)Tj
1.1735 0 Td
(Energ)Tj
0 Tc
2.313 0.0566 Td
()Tj
-0.004 Tc
0.3345 -0.0566 Td
(a)Tj
0.652 0 Td
(Ato)Tj
0 Tc
1.0431 0.0566 Td
()Tj
-0.004 Tc
0.4479 -0.0566 Td
(mica)Tj
2.16 0 Td
(\(CNEA;)Tj
3.4412 0 Td
(Buenos)Tj
-32.4564 -1.2982 Td
(Aires,)Tj
2.5511 0 Td
(Argentina\))Tj
4.6035 0 Td
(were)Tj
2.1203 0 Td
(inbred)Tj
2.8629 0 Td
(and)Tj
1.7348 0 Td
(housed)Tj
3.0898 0 Td
(according)Tj
4.1895 0 Td
(to)Tj
1.0262 0 Td
(institutional)Tj
5.091 0 Td
(guidelines.)Tj
4.5014 0 Td
(BALB/c)Tj
3.3562 0 Td
(mice,)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 183.2882 23.6976 re
W* n
q
183.2315 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 399.0047 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(T)Tj
0.822 0 Td
(Cell)Tj
1.9063 0 Td
(Hyporespon)Tj
5.2938 0 Td
(siveness)Tj
4.0464 0 Td
(during)Tj
/F6 1 Tf
2.941 0 Td
(T)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.1055 0 Td
(cruzi)Tj
/F0 1 Tf
2.289 0 Td
(Infection)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(Neglected)Tj
4.6417 0 Td
(Tropical)Tj
3.7135 0 Td
(Disease)Tj
3.5433 0 Td
(s)Tj
0.7158 0 Td
(|)Tj
0.4748 0 Td
(DOI:10.137)Tj
5.0386 0 Td
(1/journal.pn)Tj
5.1236 0 Td
(td.00053)Tj
3.8198 0 Td
(07)Tj
2.0905 0 Td
(January)Tj
3.7134 0 Td
(23,)Tj
1.5875 0 Td
(2017)Tj
27.8222 0 Td
(3)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
80 0 obj
<>stream
endstream
endobj
81 0 obj
<>stream
endstream
endobj
82 0 obj
<>stream
endstream
endobj
83 0 obj
<>stream
endstream
endobj
84 0 obj
<>stream
endstream
endobj
85 0 obj
<>stream
endstream
endobj
86 0 obj
<>stream
endstream
endobj
87 0 obj
<>stream
endstream
endobj
88 0 obj
<>stream
endstream
endobj
89 0 obj
<>stream
endstream
endobj
90 0 obj
<>stream
endstream
endobj
91 0 obj
<>stream
endstream
endobj
92 0 obj
<>stream
endstream
endobj
93 0 obj
<>stream
endstream
endobj
94 0 obj
<>stream
endstream
endobj
95 0 obj
<>stream
endstream
endobj
96 0 obj
<>stream
endstream
endobj
97 0 obj
<>stream
endstream
endobj
98 0 obj
<>stream
endstream
endobj
99 0 obj
<>stream
endstream
endobj
100 0 obj
<>stream
BT
66.402 0 Td
(16)Tj
ET
Q
endstream
endobj
101 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 103 0 R/Contents 104 0 R/TrimBox[0 0 612 792]>>
endobj
103 0 obj
[105 0 R]
endobj
105 0 obj
<>/Border[0 0 0]/Dest(Rpntd.0005307.ref032)>>
endobj
104 0 obj
[106 0 R 107 0 R 108 0 R 109 0 R 110 0 R 111 0 R]
endobj
106 0 obj
<>stream
q
0.83 0.64 0.02 0 k
283.8047 680.315 m
293.2724 680.315 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(when)Tj
2.4037 0 Td
(68)Tj
1.6895 0 Td
(weeks)Tj
2.6078 0 Td
(old,)Tj
1.7235 0 Td
(were)Tj
2.1259 0 Td
(intraperitoneally)Tj
6.8768 0 Td
(infected)Tj
3.4186 0 Td
(with)Tj
1.9956 0 Td
(1x10)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 447.3637 711.8362 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(6)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 453.1464 707.4141 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(blood-derived)Tj
/F13 1 Tf
5.8563 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0375 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.16 0 Td
(trypo-)Tj
-34.367 -1.3039 Td
(mastigote)Tj
4.0932 0 Td
(forms)Tj
2.5682 0 Td
(from)Tj
2.2053 0 Td
(Tulahue)Tj
0 Tc
2.897 0.0567 Td
()Tj
-0.004 Tc
0.4025 -0.0567 Td
(n)Tj
0.7654 0 Td
(strain,)Tj
2.7269 0 Td
(which)Tj
2.6476 0 Td
(was)Tj
1.7007 0 Td
(maintained)Tj
4.7905 0 Td
(through)Tj
3.4412 0 Td
(intraperitoneal)Tj
6.1795 0 Td
(inocu-)Tj
-34.418 -1.2982 Td
(lation)Tj
2.5172 0 Td
(every)Tj
2.3527 0 Td
(11)Tj
1.1679 0 Td
(days)Tj
2.0012 0 Td
([)Tj
0.83 0.64 0.02 0 k
(32)Tj
0 g
(].)Tj
-6.8428 -1.2983 Td
(Female)Tj
3.0955 0 Td
(BALB/c)Tj
3.3561 0 Td
(mice)Tj
2.1544 0 Td
(were)Tj
2.1203 0 Td
(infected)Tj
3.4242 0 Td
(intraperitoneally)Tj
6.8768 0 Td
(with)Tj
1.9956 0 Td
(500)Tj
1.6496 0 Td
(blood-derived)Tj
/F13 1 Tf
5.8564 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
1.0375 0 Td
[(\n)1()-9(\r)]TJ
/F8 1 Tf
2.1599 0 Td
(trypo-)Tj
-34.9225 -1.2983 Td
(mastigote)Tj
4.0932 0 Td
(forms)Tj
2.5682 0 Td
(diluted)Tj
3.0387 0 Td
(in)Tj
1.0318 0 Td
(saline)Tj
2.5002 0 Td
(solution.)Tj
3.7077 0 Td
(After)Tj
2.2903 0 Td
(different)Tj
3.651 0 Td
(days)Tj
2.0013 0 Td
(post)Tj
1.8991 0 Td
(infection)Tj
3.7927 0 Td
(\(p.i.\),)Tj
2.3641 0 Td
(these)Tj
2.2507 0 Td
(mice)Tj
-35.189 -1.3039 Td
(were)Tj
2.1203 0 Td
(sacrificed)Tj
4.0082 0 Td
(by)Tj
1.1792 0 Td
(CO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 287.2062 640.7433 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 292.9889 642.3874 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(asphyxiation)Tj
5.3007 0 Td
(and)Tj
1.7291 0 Td
(spleens)Tj
3.1011 0 Td
(were)Tj
2.1203 0 Td
(extracted.)Tj
4.1102 0 Td
(Non-infected)Tj
5.5672 0 Td
(animals)Tj
3.3279 0 Td
(were)Tj
-34.554 -1.2982 Td
(processed)Tj
4.1329 0 Td
(in)Tj
1.0374 0 Td
(parallel.)Tj
3.3619 0 Td
(Trypomastigotes)Tj
6.9164 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(Tulahuen)Tj
4.0649 0 Td
(and)Tj
1.7291 0 Td
(Y)Tj
0.8504 0 Td
(strains)Tj
2.8629 0 Td
(were)Tj
2.1203 0 Td
(obtained)Tj
3.7361 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
-35.5064 -1.2983 Td
(extracellular)Tj
5.1306 0 Td
(medium)Tj
3.6057 0 Td
(of)Tj
1.0147 0 Td
(infected)Tj
3.4186 0 Td
(monolayers)Tj
4.8925 0 Td
(of)Tj
1.0148 0 Td
(Vero)Tj
2.2167 0 Td
(or)Tj
1.0998 0 Td
(NIH3T3)Tj
3.6227 0 Td
(cells,)Tj
2.1486 0 Td
(respectively,)Tj
5.108 0 Td
(and)Tj
1.7348 0 Td
(were)Tj
-35.0075 -1.3039 Td
(collected)Tj
3.736 0 Td
(by)Tj
1.1849 0 Td
(centrifugation)Tj
5.8846 0 Td
(at)Tj
0.9524 0 Td
(4400)Tj
2.126 0 Td
(rpm)Tj
1.9275 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1736 0 Td
(min)Tj
1.8538 0 Td
(and)Tj
1.7348 0 Td
(resuspended)Tj
5.2327 0 Td
(in)Tj
1.0318 0 Td
(RPMI)Tj
2.6362 0 Td
(medium)Tj
3.6057 0 Td
(con-)Tj
-34.4633 -1.2983 Td
(taining)Tj
3.05 0 Td
(10%)Tj
1.9276 0 Td
(FCS.)Tj
2.1089 0 Td
(Parasites)Tj
3.7303 0 Td
(were)Tj
2.1203 0 Td
(counted)Tj
3.4809 0 Td
(using)Tj
2.3868 0 Td
(a)Tj
0.652 0 Td
(Neubauer)Tj
4.1725 0 Td
(chamber)Tj
3.7247 0 Td
(and)Tj
1.7348 0 Td
(used)Tj
2.058 0 Td
(for)Tj
/F13 1 Tf
1.3889 0 Td
[(\r)12()]TJ
1.0148 0 Td
[()-13(\r)14()7()20()]TJ
/F8 1 Tf
-33.5505 -1.2982 Td
(infection)Tj
3.7927 0 Td
(experiments)Tj
5.1363 0 Td
(as)Tj
1.0147 0 Td
(described)Tj
4.0536 0 Td
(below.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 548.7874 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Isolation)Tj
3.8929 0 Td
(of)Tj
1.0488 0 Td
(splenocytes)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 531.7795 cm
BT
/F8 1 Tf
1 TL
-0.01 Tc
0 0 Td
(Spleen)Tj
2.8176 0 Td
(cells)Tj
1.8878 0 Td
(were)Tj
2.1033 0 Td
(obtained)Tj
3.685 0 Td
(from)Tj
2.1827 0 Td
(control)Tj
3.0784 0 Td
(or)Tj
1.0828 0 Td
(infected)Tj
3.3732 0 Td
(animals)Tj
3.2882 0 Td
(by)Tj
1.1736 0 Td
(homogenizing)Tj
5.9073 0 Td
(the)Tj
1.4513 0 Td
(organs)Tj
2.8687 0 Td
(in)Tj
1.0261 0 Td
(a)Tj
-35.926 -1.2982 Td
(cell)Tj
1.5363 0 Td
(strainer.)Tj
3.4639 0 Td
(Then,)Tj
2.5285 0 Td
(the)Tj
1.4514 0 Td
(spleen)Tj
2.7042 0 Td
(cell)Tj
1.5364 0 Td
(suspensions)Tj
4.9379 0 Td
(were)Tj
2.0976 0 Td
(centrifuged)Tj
4.7168 0 Td
(\(1500)Tj
2.4321 0 Td
(rpm,)Tj
2.1317 0 Td
(5)Tj
0.6916 0 Td
(min,)Tj
2.0523 0 Td
(4C\))Tj
2.0693 0 Td
(and)Tj
1.7121 0 Td
(the)Tj
-36.0621 -1.2983 Td
(pellets)Tj
2.6928 0 Td
(treated)Tj
2.9537 0 Td
(with)Tj
1.9729 0 Td
(RBC)Tj
2.0693 0 Td
(lysis)Tj
1.8822 0 Td
(buffer)Tj
2.5852 0 Td
(\(GIBCO\).)Tj
4.1272 0 Td
(These)Tj
2.5342 0 Td
(cells)Tj
1.8878 0 Td
(were)Tj
2.1033 0 Td
(subsequently)Tj
5.3348 0 Td
(resuspended)Tj
5.1647 0 Td
(in)Tj
-35.3081 -1.3039 Td
(complete)Tj
3.8324 0 Td
(RPMI)Tj
2.6078 0 Td
(medium)Tj
3.566 0 Td
(containing)Tj
4.473 0 Td
(10%)Tj
1.9106 0 Td
(fetal)Tj
1.8765 0 Td
(bovine)Tj
2.8913 0 Td
(serum)Tj
2.6929 0 Td
(\(FBS,)Tj
2.3414 0 Td
(PAA)Tj
2.126 0 Td
(laboratories\),)Tj
5.4311 0 Td
(L-gluta-)Tj
-33.749 -1.2983 Td
(mine)Tj
2.2563 0 Td
(\(2)Tj
1.0205 0 Td
(mM,)Tj
2.143 0 Td
(GIBCO\))Tj
3.5659 0 Td
(and)Tj
1.7121 0 Td
(gentamicin)Tj
4.6545 0 Td
(\(40)Tj
1.4967 0 Td
(g/ml\),)Tj
2.6078 0 Td
(and)Tj
1.7122 0 Td
(the)Tj
1.4513 0 Td
(isolated)Tj
3.2371 0 Td
(cell)Tj
1.5364 0 Td
(suspensions)Tj
4.9379 0 Td
(were)Tj
2.0976 0 Td
(passed)Tj
-34.4293 -1.2982 Td
(through)Tj
3.4015 0 Td
(a)Tj
0.6463 0 Td
(50-mm)Tj
3.1351 0 Td
(nylon)Tj
2.4945 0 Td
(mesh)Tj
2.3244 0 Td
(\(BD)Tj
1.8595 0 Td
(Falcon\))Tj
3.2088 0 Td
(for)Tj
1.3663 0 Td
(cell)Tj
1.5363 0 Td
(culture,)Tj
3.2145 0 Td
(flow)Tj
1.9219 0 Td
(cytometry)Tj
4.2179 0 Td
(or)Tj
1.0828 0 Td
(cell)Tj
1.5364 0 Td
(isolation.)Tj
-31.9462 -1.2983 Td
(Finally,)Tj
3.118 0 Td
(the)Tj
1.457 0 Td
(T)Tj
0.8221 0 Td
(cells)Tj
1.8935 0 Td
(were)Tj
2.0976 0 Td
(isolated)Tj
3.2372 0 Td
(from)Tj
2.1826 0 Td
(spleens)Tj
3.0558 0 Td
(using)Tj
2.3584 0 Td
(a)Tj
0.6519 0 Td
(CD4+)Tj
2.6476 0 Td
(T)Tj
0.8277 0 Td
(cell)Tj
1.5364 0 Td
(isolation)Tj
3.5999 0 Td
(kit,)Tj
1.4854 0 Td
(according)Tj
4.1329 0 Td
(to)Tj
1.0148 0 Td
(the)Tj
-36.1188 -1.3039 Td
(manufacturer's)Tj
6.1794 0 Td
(instructions)Tj
4.9266 0 Td
(\(Miltenyi)Tj
3.9061 0 Td
(Biotec\),)Tj
3.2372 0 Td
(with)Tj
1.9729 0 Td
(the)Tj
1.4513 0 Td
(average)Tj
3.1861 0 Td
(purity)Tj
2.6192 0 Td
(found)Tj
2.5852 0 Td
(to)Tj
1.0148 0 Td
(be)Tj
1.1395 0 Td
(9598%.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 412.2141 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(HEK-293)Tj
4.2331 0 Td
(cell)Tj
1.696 0 Td
(line)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 395.2062 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(HEK-293)Tj
4.0081 0 Td
(cells)Tj
1.9219 0 Td
(\(ATCC\))Tj
3.5206 0 Td
(were)Tj
2.1203 0 Td
(maintained)Tj
4.7905 0 Td
(in)Tj
1.0318 0 Td
(Dulbecco's)Tj
4.5921 0 Td
(modified)Tj
3.8494 0 Td
(Eagle's)Tj
2.9311 0 Td
(medium)Tj
3.6056 0 Td
(\(Gibco\))Tj
3.3222 0 Td
(sup-)Tj
-35.6936 -1.2982 Td
(plemented)Tj
4.4446 0 Td
(with)Tj
2.0013 0 Td
(10%)Tj
1.9275 0 Td
(fetal)Tj
1.9106 0 Td
(bovine)Tj
2.9253 0 Td
(serum)Tj
2.7269 0 Td
(\(FBS,)Tj
2.3641 0 Td
(PAA)Tj
2.1487 0 Td
(laboratories\),)Tj
5.5161 0 Td
(L-glutamine)Tj
5.1364 0 Td
(\(2)Tj
1.0375 0 Td
(mM,)Tj
-32.139 -1.3039 Td
(GIBCO\))Tj
3.6056 0 Td
(and)Tj
1.7291 0 Td
(gentamicin)Tj
4.7168 0 Td
(\(40)Tj
1.5081 0 Td
(g/ml\),)Tj
2.6475 0 Td
(and)Tj
1.7291 0 Td
(these)Tj
2.2564 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(grown)Tj
2.7949 0 Td
(at)Tj
0.9524 0 Td
(37C)Tj
2.2281 0 Td
(under)Tj
2.6192 0 Td
(5%)Tj
1.4513 0 Td
(CO)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 536.9385 367.5401 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 540.5102 369.1842 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 340.611 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Fluorescence-act)Tj
7.5827 0 Td
(ivated)Tj
2.8583 0 Td
(cell-scanning)Tj
5.9717 0 Td
(\(FACS\))Tj
3.5291 0 Td
(analysis)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 323.6031 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(To)Tj
1.3379 0 Td
(examine)Tj
3.6 0 Td
(PD-1,)Tj
2.5681 0 Td
(CTLA-4)Tj
3.5433 0 Td
(and)Tj
1.7292 0 Td
(GRAIL)Tj
3.118 0 Td
(expression,)Tj
4.6942 0 Td
(splenocytes)Tj
4.7848 0 Td
(or)Tj
1.0942 0 Td
(CD4)Tj
2.0976 0 Td
(T)Tj
0.8277 0 Td
(cells)Tj
1.9219 0 Td
(from)Tj
2.2054 0 Td
(control)Tj
-33.5223 -1.2982 Td
(and)Tj
1.7291 0 Td
(infected)Tj
3.4185 0 Td
(animals)Tj
3.3336 0 Td
(or)Tj
/F13 1 Tf
1.0941 0 Td
[(\r)12()]TJ
1.0148 0 Td
[()-13(\r)14()7()20()]TJ
/F8 1 Tf
2.0466 0 Td
(infected)Tj
3.4243 0 Td
(CD4)Tj
2.0919 0 Td
(T)Tj
0.8334 0 Td
(cells)Tj
1.9162 0 Td
(were)Tj
2.126 0 Td
(washed)Tj
3.1747 0 Td
(with)Tj
2.0013 0 Td
(saline)Tj
2.5001 0 Td
(solution)Tj
3.481 0 Td
(2%)Tj
-34.1856 -1.3039 Td
(FBS)Tj
1.8028 0 Td
(and)Tj
1.7291 0 Td
(incubated)Tj
4.1782 0 Td
(with)Tj
1.9956 0 Td
(anti-mouse)Tj
4.7565 0 Td
(CD32/CD16)Tj
5.2554 0 Td
(antibody)Tj
3.7644 0 Td
(for)Tj
1.3833 0 Td
(20)Tj
1.1735 0 Td
(minutes)Tj
3.464 0 Td
(at)Tj
0.9524 0 Td
(4C)Tj
1.7518 0 Td
(to)Tj
1.0261 0 Td
(block)Tj
2.3981 0 Td
(Fc)Tj
-35.6312 -1.2983 Td
(receptors.)Tj
4.1385 0 Td
(Then,)Tj
2.5568 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(incubated)Tj
4.1783 0 Td
(with)Tj
2.0012 0 Td
(APC)Tj
2.126 0 Td
(labeled)Tj
3.0274 0 Td
(anti-CD4,)Tj
4.2179 0 Td
(PercP)Tj
2.5568 0 Td
(labeled)Tj
3.0274 0 Td
(anti-CD3)Tj
3.9912 0 Td
(\(BD)Tj
-35.8637 -1.2983 Td
(Pharmingen\),)Tj
5.7486 0 Td
(and)Tj
1.7291 0 Td
(with)Tj
2.0012 0 Td
(PE-labeled)Tj
4.5071 0 Td
(anti-PD-1)Tj
4.2349 0 Td
(or)Tj
1.0999 0 Td
(anti-CTLA-4)Tj
5.4425 0 Td
(\(BD)Tj
1.8765 0 Td
(Pharmingen\))Tj
5.5218 0 Td
(for)Tj
1.389 0 Td
(20)Tj
1.1735 0 Td
(min)Tj
1.8539 0 Td
(at)Tj
-36.578 -1.3039 Td
(4C.)Tj
1.1962 -1.2982 Td
(For)Tj
1.627 0 Td
(the)Tj
1.4684 0 Td
(assessment)Tj
4.6148 0 Td
(of)Tj
1.0147 0 Td
(intracellular)Tj
5.057 0 Td
(GRAIL)Tj
3.1181 0 Td
(expression,)Tj
4.6942 0 Td
(cells)Tj
1.9162 0 Td
(were)Tj
2.1259 0 Td
(first)Tj
1.8029 0 Td
(stained)Tj
3.084 0 Td
(with)Tj
1.9956 0 Td
(FITC-)Tj
-33.715 -1.2983 Td
(CD3)Tj
2.0919 0 Td
(and)Tj
1.7348 0 Td
(APC-CD4)Tj
4.3483 0 Td
(antibodies,)Tj
4.5864 0 Td
(and)Tj
1.7292 0 Td
(then)Tj
2.0182 0 Td
(fixed)Tj
2.194 0 Td
(and)Tj
1.7291 0 Td
(permeabilized)Tj
5.8621 0 Td
(with)Tj
1.9955 0 Td
(Citofix/Citoperm)Tj
7.1773 0 Td
(\(BD)Tj
-35.4668 -1.2983 Td
(Biosciences,\))Tj
5.3687 0 Td
(for)Tj
1.389 0 Td
(30)Tj
1.1679 0 Td
(min)Tj
1.8595 0 Td
(followed)Tj
3.6453 0 Td
(by)Tj
1.1849 0 Td
(reacting)Tj
3.4526 0 Td
(with)Tj
1.9955 0 Td
(rabbit)Tj
2.6022 0 Td
(anti-GRAIL)Tj
5.0173 0 Td
(primary)Tj
3.4583 0 Td
(Ab)Tj
1.4116 0 Td
(\(Abcam\))Tj
3.7644 0 Td
(for)Tj
-36.3172 -1.3039 Td
(45)Tj
1.1678 0 Td
(min.)Tj
2.0806 0 Td
(After)Tj
2.2904 0 Td
(washing,)Tj
3.7304 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(stained)Tj
3.0784 0 Td
(for)Tj
1.3889 0 Td
(20)Tj
1.1736 0 Td
(min)Tj
1.8538 0 Td
(with)Tj
1.9956 0 Td
(PEanti-rabbit)Tj
6.1511 0 Td
(IgG)Tj
1.7405 0 Td
(\(Biolegend\).)Tj
-30.6933 -1.2983 Td
(Finally,)Tj
3.1691 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.1203 0 Td
(washed)Tj
3.1805 0 Td
(twice)Tj
2.3131 0 Td
(with)Tj
2.0012 0 Td
(saline)Tj
2.5001 0 Td
(solution)Tj
3.481 0 Td
(of)Tj
1.0148 0 Td
(2%)Tj
1.4513 0 Td
(FBS,)Tj
2.0296 0 Td
(and)Tj
1.7291 0 Td
(stored)Tj
2.7099 0 Td
(at)Tj
0.9524 0 Td
(4C)Tj
1.7518 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4684 0 Td
(dark)Tj
-34.8319 -1.2982 Td
(until)Tj
2.1089 0 Td
(analysis)Tj
3.3222 0 Td
(using)Tj
2.3924 0 Td
(a)Tj
0.652 0 Td
(FACS)Tj
2.5738 0 Td
(flow)Tj
1.9446 0 Td
(cytometer)Tj
4.2406 0 Td
(\(FACS)Tj
2.9083 0 Td
(Canto)Tj
2.6702 0 Td
(II,)Tj
1.1282 0 Td
(BD)Tj
1.5364 0 Td
(Biosciences\).)Tj
5.3688 0 Td
(The)Tj
1.7858 0 Td
(results)Tj
2.8063 0 Td
(were)Tj
-35.4385 -1.3039 Td
(processed)Tj
4.1328 0 Td
(using)Tj
2.3925 0 Td
(Flow)Tj
2.1826 0 Td
(Jo)Tj
1.0602 0 Td
(software)Tj
3.5829 0 Td
(\(version)Tj
3.4923 0 Td
(7.6.2\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 139.011 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Cytokine)Tj
4.063 0 Td
(determination)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 122.0031 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Cytokines)Tj
4.2406 0 Td
(were)Tj
2.1203 0 Td
(measured)Tj
4.1102 0 Td
(in)Tj
1.0318 0 Td
(culture)Tj
3.0387 0 Td
(supernatants)Tj
5.3461 0 Td
(using)Tj
2.3868 0 Td
(a)Tj
0.6576 0 Td
(capture)Tj
3.2145 0 Td
(enzyme-linked)Tj
6.1624 0 Td
(immunosor-)Tj
-32.309 -1.2982 Td
(bent)Tj
1.9955 0 Td
(assay)Tj
2.2677 0 Td
(\(ELISA\).)Tj
3.7304 0 Td
(IFN-)Tj
/F15 1 Tf
(\r)Tj
/F8 1 Tf
2.6305 0 Td
(\(eBioscience\))Tj
5.5446 0 Td
(and)Tj
1.7348 0 Td
(IL-2)Tj
1.9218 0 Td
(\(Biolegend\))Tj
4.8869 0 Td
(were)Tj
2.126 0 Td
(used)Tj
2.0579 0 Td
(as)Tj
1.0148 0 Td
(paired)Tj
2.761 0 Td
(monoclonal)Tj
-32.6719 -1.3039 Td
(antibodies)Tj
4.3596 0 Td
(in)Tj
1.0375 0 Td
(combination)Tj
5.3461 0 Td
(with)Tj
1.9956 0 Td
(recombinant)Tj
5.3688 0 Td
(cytokine)Tj
3.6396 0 Td
(standards.)Tj
4.3086 0 Td
(All)Tj
1.4004 0 Td
(assays)Tj
2.6248 0 Td
(were)Tj
2.1203 0 Td
(performed)Tj
-32.2013 -1.2983 Td
(according)Tj
4.1839 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.474 0 Td
(manufacturer's)Tj
6.2589 0 Td
(guidelines.)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 183.2882 23.6976 re
W* n
q
183.2315 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 399.0047 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(T)Tj
0.822 0 Td
(Cell)Tj
1.9063 0 Td
(Hyporespon)Tj
5.2938 0 Td
(siveness)Tj
4.0464 0 Td
(during)Tj
/F6 1 Tf
2.941 0 Td
(T)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.1055 0 Td
(cruzi)Tj
/F0 1 Tf
2.289 0 Td
(Infection)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(Neglected)Tj
4.6417 0 Td
(Tropical)Tj
3.7135 0 Td
(Disease)Tj
3.5433 0 Td
(s)Tj
0.7158 0 Td
(|)Tj
0.4748 0 Td
(DOI:10.137)Tj
5.0386 0 Td
(1/journal.pn)Tj
5.1236 0 Td
(td.00053)Tj
3.8198 0 Td
(07)Tj
2.0905 0 Td
(January)Tj
3.7134 0 Td
(23,)Tj
1.5875 0 Td
(2017)Tj
27.8222 0 Td
(4)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
107 0 obj
<>stream
endstream
endobj
108 0 obj
<>stream
endstream
endobj
109 0 obj
<>stream
endstream
endobj
110 0 obj
<>stream
endstream
endobj
111 0 obj
<>stream
BT
66.402 0 Td
(16)Tj
ET
Q
endstream
endobj
112 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Contents 114 0 R/TrimBox[0 0 612 792]>>
endobj
114 0 obj
[115 0 R 116 0 R 117 0 R 118 0 R 119 0 R 120 0 R]
endobj
115 0 obj
<>stream
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 706.0535 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Immunoblot)Tj
5.3905 0 Td
(analysis)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 689.1023 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(CD4)Tj
2.0919 0 Td
(T)Tj
0.8334 0 Td
(cells)Tj
1.9219 0 Td
(from)Tj
2.2053 0 Td
(control)Tj
3.1181 0 Td
(and)Tj
1.7348 0 Td
(infected)Tj
3.4185 0 Td
(animals)Tj
3.3279 0 Td
(were)Tj
2.126 0 Td
(washed)Tj
3.1748 0 Td
(and)Tj
1.7347 0 Td
(lysed)Tj
2.2281 0 Td
(for)Tj
1.3889 0 Td
(30)Tj
1.1736 0 Td
(min)Tj
1.8538 0 Td
(at)Tj
0.9524 0 Td
(4C)Tj
1.7518 0 Td
(in)Tj
-35.0359 -1.3039 Td
(RIPA)Tj
2.4207 0 Td
(buffer)Tj
2.6306 0 Td
([1%)Tj
1.7914 0 Td
(Triton)Tj
2.8233 0 Td
(X-100)Tj
2.6476 0 Td
(\(v/v\),)Tj
2.3641 0 Td
(0.5%)Tj
2.1543 0 Td
(sodium)Tj
3.2314 0 Td
(deoxicolate)Tj
4.7452 0 Td
(\(p/v\),)Tj
2.4265 0 Td
(0.1%)Tj
2.1543 0 Td
(sodium)Tj
3.2258 0 Td
(dodecyl)Tj
3.3335 0 Td
(sul-)Tj
-35.9487 -1.2982 Td
(fate)Tj
1.661 0 Td
(\(SDS\)])Tj
2.931 0 Td
(containing)Tj
4.5241 0 Td
(a)Tj
0.6576 0 Td
(protease)Tj
3.5547 0 Td
(inhibitor)Tj
3.7814 0 Td
(cocktail)Tj
3.3165 0 Td
(\(Roche\),)Tj
3.6283 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(cell)Tj
1.559 0 Td
(debris)Tj
2.6816 0 Td
(was)Tj
1.6951 0 Td
(spun)Tj
-33.1878 -1.2983 Td
(down)Tj
2.4888 0 Td
(at)Tj
0.9524 0 Td
(13,000)Tj
2.8233 0 Td
(g)Tj
0.6803 0 Td
(for)Tj
1.389 0 Td
(15)Tj
1.1735 0 Td
(min.)Tj
2.0806 0 Td
(Precipitates)Tj
4.8586 0 Td
(were)Tj
2.1259 0 Td
(removed,)Tj
3.9742 0 Td
(and)Tj
1.7291 0 Td
(aliquots)Tj
3.3675 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(cell)Tj
1.559 0 Td
(lysates)Tj
2.8006 0 Td
(were)Tj
-34.486 -1.3039 Td
(diluted)Tj
3.0387 0 Td
(in)Tj
1.0318 0 Td
(SDS)Tj
1.9105 0 Td
(sample)Tj
3.0217 0 Td
(buffer,)Tj
2.8517 0 Td
(boiled)Tj
2.6985 0 Td
(at)Tj
0.9525 0 Td
(100C)Tj
2.6985 0 Td
(for)Tj
1.389 0 Td
(3)Tj
0.6973 0 Td
(min,)Tj
2.0806 0 Td
(spun)Tj
2.1714 0 Td
(down,)Tj
2.7155 0 Td
(and)Tj
1.7292 0 Td
(applied)Tj
3.1521 0 Td
(to)Tj
1.0261 0 Td
(precast)Tj
-33.1651 -1.2983 Td
(10%)Tj
1.9275 0 Td
(acrylamide)Tj
4.6204 0 Td
(Tris-glycine)Tj
5.0117 0 Td
(gels)Tj
1.7177 0 Td
(at)Tj
0.9468 0 Td
(40)Tj
/F16 1 Tf
1.1735 0 Td
[()]TJ
/F8 1 Tf
(g)Tj
1.1906 0 Td
(protein/lane)Tj
5.1363 0 Td
(and)Tj
1.7292 0 Td
(run)Tj
1.6667 0 Td
(at)Tj
0.9525 0 Td
(150)Tj
1.6497 0 Td
(V)Tj
0.9128 0 Td
(for)Tj
1.3889 0 Td
(1)Tj
0.6917 0 Td
(h.)Tj
0.9751 0 Td
(Samples)Tj
3.4979 0 Td
(were)Tj
-35.189 -1.2982 Td
(transferred)Tj
4.6374 0 Td
(to)Tj
1.0261 0 Td
(nitrocellulose)Tj
5.6183 0 Td
(membranes)Tj
4.9266 0 Td
(\(BioRad\))Tj
3.838 0 Td
(at)Tj
0.9525 0 Td
(100)Tj
1.6441 0 Td
(V)Tj
0.9184 0 Td
(for)Tj
1.3833 0 Td
(1)Tj
0.6973 0 Td
(h,)Tj
0.9694 0 Td
(and)Tj
1.7348 0 Td
(these)Tj
2.2507 0 Td
(membranes)Tj
4.9266 0 Td
(were)Tj
-35.5235 -1.304 Td
(probed)Tj
3.0727 0 Td
(using)Tj
2.3867 0 Td
(rabbit)Tj
2.6079 0 Td
(anti-mouse)Tj
4.7565 0 Td
(GRAIL)Tj
3.1181 0 Td
(\(Santa)Tj
2.7496 0 Td
(Cruz)Tj
2.1997 0 Td
(Biotechnology\),)Tj
6.5366 0 Td
(anti-mouse)Tj
4.7622 0 Td
(Otub-1)Tj
3.1237 0 Td
(or)Tj
-35.3137 -1.2982 Td
(anti-mouse)Tj
4.7565 0 Td
(p-4EBP1)Tj
3.7473 0 Td
(\(Cell)Tj
2.1373 0 Td
(Signaling)Tj
3.9402 0 Td
(Technology\))Tj
5.2724 0 Td
(followed)Tj
3.651 0 Td
(by)Tj
1.1792 0 Td
(peroxidase)Tj
4.5184 0 Td
(conjugated)Tj
4.6318 0 Td
(anti-rab-)Tj
-33.8341 -1.2983 Td
(bit)Tj
1.2925 0 Td
(antibody)Tj
3.7644 0 Td
(\(Sigma)Tj
3.0217 0 Td
(Chemical)Tj
4.0252 0 Td
(Co.\),)Tj
2.177 0 Td
(before)Tj
2.7439 0 Td
(being)Tj
2.4321 0 Td
(visualized)Tj
4.1386 0 Td
(using)Tj
2.3924 0 Td
(enhanced)Tj
4.0762 0 Td
(chemilumines-)Tj
-30.064 -1.2983 Td
(cence)Tj
2.4604 0 Td
(\(Pierce\))Tj
3.3675 0 Td
(for)Tj
1.389 0 Td
(detection.)Tj
4.1612 0 Td
(The)Tj
1.7802 0 Td
(protein)Tj
3.1578 0 Td
(loading)Tj
3.2144 0 Td
(was)Tj
1.7008 0 Td
(evaluated)Tj
3.9855 0 Td
(by)Tj
1.1849 0 Td
(actin)Tj
2.1883 0 Td
(expression.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 528.548 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Quantitative)Tj
5.4992 0 Td
(real-time)Tj
4.0583 0 Td
(PCR)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 511.5401 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(RNA)Tj
2.262 0 Td
(was)Tj
1.6951 0 Td
(extracted)Tj
3.8891 0 Td
(from)Tj
2.2053 0 Td
(splenocytes)Tj
4.7849 0 Td
(from)Tj
2.2053 0 Td
(infected)Tj
3.4186 0 Td
(or)Tj
1.0998 0 Td
(control)Tj
3.1238 0 Td
(animals)Tj
3.3278 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4684 0 Td
(Trizol)Tj
2.6362 0 Td
(reagent)Tj
-33.3012 -1.2982 Td
(\(Invitrogen\))Tj
5.125 0 Td
(and)Tj
1.7291 0 Td
(reverse-transcribed)Tj
7.9313 0 Td
(into)Tj
1.8425 0 Td
(cDNA)Tj
2.8006 0 Td
(by)Tj
1.1792 0 Td
(using)Tj
2.3924 0 Td
(Revert)Tj
2.812 0 Td
(Aid)Tj
1.7007 0 Td
(First)Tj
2.041 0 Td
(Strand)Tj
2.88 0 Td
(cDNA)Tj
2.8006 0 Td
(Syn-)Tj
-35.2344 -1.2983 Td
(thesis)Tj
2.4547 0 Td
(\(Fermentas\).)Tj
5.3348 0 Td
(Transcripts)Tj
4.7849 0 Td
(were)Tj
2.1203 0 Td
(quantified)Tj
4.3143 0 Td
(by)Tj
1.1792 0 Td
(real-time)Tj
3.8551 0 Td
(quantitative)Tj
4.9776 0 Td
(PCR)Tj
2.0636 0 Td
(on)Tj
1.2756 0 Td
(an)Tj
1.2018 0 Td
(ABI)Tj
-33.5619 -1.3039 Td
(Prism)Tj
2.5965 0 Td
(7500)Tj
2.1203 0 Td
(sequence)Tj
3.8607 0 Td
(detector)Tj
3.4923 0 Td
(\(Applied)Tj
3.7417 0 Td
(Biosystems\))Tj
4.9946 0 Td
(with)Tj
1.9956 0 Td
(predesigned)Tj
5.0796 0 Td
(TaqMan)Tj
3.6454 0 Td
(gene)Tj
2.0806 0 Td
(expres-)Tj
-33.6073 -1.2983 Td
(sion)Tj
1.9048 0 Td
(assays)Tj
2.6249 0 Td
(and)Tj
1.7291 0 Td
(reagents)Tj
3.5433 0 Td
(\(Applied)Tj
3.7474 0 Td
(Biosystems\),)Tj
5.2157 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(manufacturers)Tj
6.4403 0 Td
(instructions.)Tj
-31.8895 -1.2982 Td
(Probes)Tj
2.948 0 Td
(with)Tj
1.9955 0 Td
(the)Tj
1.4684 0 Td
(following)Tj
3.9798 0 Td
(Applied)Tj
3.4072 0 Td
(Biosystems)Tj
4.6488 0 Td
(assay)Tj
2.2677 0 Td
(identification)Tj
5.5842 0 Td
(numbers)Tj
3.7814 0 Td
(were)Tj
2.1203 0 Td
(used:)Tj
2.2847 0 Td
(Cblb,)Tj
-34.486 -1.2983 Td
(Mn01343092.m1;)Tj
7.2112 0 Td
(Rnf128,)Tj
3.3279 0 Td
(Mn00480990.m1;)Tj
7.217 0 Td
(Rn18s,)Tj
2.9253 0 Td
(Mn03928990.g1.)Tj
6.8598 0 Td
(For)Tj
1.6214 0 Td
(each)Tj
2.0296 0 Td
(sample,)Tj
3.2485 0 Td
(mRNA)Tj
-34.4407 -1.3039 Td
(abundance)Tj
4.592 0 Td
(was)Tj
1.7008 0 Td
(normalized)Tj
4.7792 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(amount)Tj
3.3562 0 Td
(of)Tj
1.0148 0 Td
(18S)Tj
1.6441 0 Td
(RNA)Tj
2.262 0 Td
(and)Tj
1.7291 0 Td
(expressed)Tj
4.0932 0 Td
(as)Tj
1.0148 0 Td
(arbitrary)Tj
3.7304 0 Td
(units.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 390.0472 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Cell)Tj
1.9134 0 Td
(proliferation)Tj
5.3669 0 Td
(assay)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 373.0393 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(CD4)Tj
2.0919 0 Td
(T)Tj
0.8334 0 Td
(cell)Tj
1.559 0 Td
(proliferation)Tj
5.2668 0 Td
(was)Tj
1.7007 0 Td
(measured)Tj
4.1046 0 Td
(using)Tj
2.3924 0 Td
(the)Tj
1.4684 0 Td
(cell)Tj
1.559 0 Td
(division)Tj
3.4186 0 Td
(tracking)Tj
3.5262 0 Td
(dye)Tj
1.6271 0 Td
(carboxyfluorescein)Tj
-29.5481 -1.2982 Td
(diacetate)Tj
3.7473 0 Td
(succinimidyl)Tj
5.3575 0 Td
(ester)Tj
2.0976 0 Td
(\(CFSE\))Tj
3.1351 0 Td
(\(Molecular)Tj
4.6318 0 Td
(Probes,)Tj
3.1748 0 Td
(Eugene,)Tj
3.4015 0 Td
(OR\).)Tj
2.1543 0 Td
(Spleen)Tj
2.8573 0 Td
(CD4)Tj
2.092 0 Td
(T)Tj
0.8334 0 Td
(cells)Tj
1.9162 0 Td
(iso-)Tj
-35.3988 -1.3039 Td
(lated)Tj
2.1486 0 Td
(from)Tj
2.2053 0 Td
(infected)Tj
3.4186 0 Td
(or)Tj
1.0998 0 Td
(control)Tj
3.1238 0 Td
(animals)Tj
3.3278 0 Td
(were)Tj
2.126 0 Td
(stained)Tj
3.0784 0 Td
(with)Tj
1.9956 0 Td
(CFSE)Tj
2.4548 0 Td
(dye)Tj
1.6271 0 Td
(at)Tj
0.9524 0 Td
(5)Tj
/F16 1 Tf
0.6973 0 Td
[()]TJ
/F8 1 Tf
(M)Tj
1.6214 0 Td
(concentrations.)Tj
-29.8769 -1.2983 Td
(Cells)Tj
2.1599 0 Td
(were)Tj
2.1203 0 Td
(incubated)Tj
4.1783 0 Td
(at)Tj
0.9524 0 Td
(37C)Tj
2.228 0 Td
(for)Tj
1.3833 0 Td
(10)Tj
1.1736 0 Td
(min,)Tj
2.0806 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(the)Tj
1.4683 0 Td
(reaction)Tj
3.4979 0 Td
(was)Tj
1.6951 0 Td
(stopped)Tj
3.3732 0 Td
(by)Tj
1.1792 0 Td
(adding)Tj
2.9934 0 Td
(10)Tj
1.1679 0 Td
(ml)Tj
1.2869 0 Td
(of)Tj
-36.6857 -1.2983 Td
(RPMI)Tj
2.6305 0 Td
(medium)Tj
3.6056 0 Td
(containing)Tj
4.5298 0 Td
(10%)Tj
1.9275 0 Td
(FBS.)Tj
2.0296 0 Td
(After)Tj
2.2847 0 Td
(washing,)Tj
3.7304 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1259 0 Td
(resuspended)Tj
5.2328 0 Td
(in)Tj
1.0318 0 Td
(warm)Tj
2.5285 0 Td
(RPMI)Tj
-33.579 -1.3039 Td
(complete)Tj
3.8777 0 Td
(medium)Tj
3.6057 0 Td
(before)Tj
2.7382 0 Td
(being)Tj
2.4321 0 Td
(plated)Tj
2.6702 0 Td
(in)Tj
1.0319 0 Td
(anti-CD3/CD28)Tj
6.6783 0 Td
(Abs)Tj
1.7688 0 Td
(\(1)Tj
/F16 1 Tf
1.0375 0 Td
[()]TJ
/F8 1 Tf
(g/mL)Tj
2.88 0 Td
(of)Tj
1.0148 0 Td
(each)Tj
2.0296 0 Td
(Abs\)-coated)Tj
-31.7648 -1.2982 Td
(plates.)Tj
2.7269 0 Td
(After)Tj
2.2847 0 Td
(72)Tj
1.1678 0 Td
(h)Tj
0.7484 0 Td
(of)Tj
1.0148 0 Td
(incubation,)Tj
4.7735 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(stained)Tj
3.084 0 Td
(with)Tj
1.9956 0 Td
(PerCP/Cy5.5-CD4)Tj
7.6478 0 Td
(Ab)Tj
1.4117 0 Td
(and)Tj
1.7291 0 Td
(acquired)Tj
3.702 0 Td
(for)Tj
-36.3285 -1.2983 Td
(FACS)Tj
2.5681 0 Td
(analysis.)Tj
3.5546 0 Td
(Unstimulated)Tj
5.709 0 Td
(CFSE-labeled)Tj
5.6125 0 Td
(cells)Tj
1.9219 0 Td
(served)Tj
2.778 0 Td
(as)Tj
1.0148 0 Td
(a)Tj
0.6576 0 Td
(non-dividing)Tj
5.4992 0 Td
(control.)Tj
3.3448 0 Td
(Data)Tj
2.126 0 Td
(analy-)Tj
-34.7865 -1.2983 Td
(sis)Tj
1.2075 0 Td
(was)Tj
1.7008 0 Td
(performed)Tj
4.4674 0 Td
(using)Tj
2.3924 0 Td
(a)Tj
0.6519 0 Td
(FACS)Tj
2.5682 0 Td
(flow)Tj
1.9446 0 Td
(cytometer)Tj
4.2406 0 Td
(\(FACS)Tj
2.914 0 Td
(Canto)Tj
2.6702 0 Td
(II,)Tj
1.1225 0 Td
(BD)Tj
1.5364 0 Td
(Biosciences\))Tj
5.1477 0 Td
(with)Tj
1.9955 0 Td
(FlowJo)Tj
-34.5597 -1.3039 Td
(software)Tj
3.5829 0 Td
(\(version)Tj
3.498 0 Td
(7.6.2\),)Tj
2.6588 0 Td
(by)Tj
1.1849 0 Td
(setting)Tj
2.88 0 Td
(a)Tj
0.6576 0 Td
(gate)Tj
1.8369 0 Td
(on)Tj
1.2755 0 Td
(the)Tj
1.4684 0 Td
(live)Tj
1.6157 0 Td
(cells)Tj
1.9162 0 Td
(to)Tj
1.0262 0 Td
(side-scatter)Tj
4.7621 0 Td
(versus)Tj
2.7213 0 Td
(forward-scatter)Tj
-31.0845 -1.2983 Td
(dot)Tj
1.5533 0 Td
(plots)Tj
2.1487 0 Td
(and)Tj
1.7291 0 Td
(determining)Tj
5.1817 0 Td
(the)Tj
1.4683 0 Td
(expression)Tj
4.4674 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(CFSE.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 212.5417 cm
BT
/F6 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(In)Tj
1.0441 0 Td
(vitro)Tj
/F0 1 Tf
2.0882 0 Td
(stimulation)Tj
4.937 0 Td
(of)Tj
1.0488 0 Td
(recombinant)Tj
5.6552 0 Td
(mouse)Tj
3.1937 0 Td
(IL-2)Tj
1.9229 0 Td
(\(rmIL-2\))Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 195.5338 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(CD4+)Tj
2.6758 0 Td
(T)Tj
0.8277 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(isolated)Tj
3.2882 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.4684 0 Td
(spleen)Tj
2.7382 0 Td
(of)Tj
1.0148 0 Td
(control)Tj
3.1238 0 Td
(or)Tj
1.0998 0 Td
(infected)Tj
3.4186 0 Td
(animals)Tj
3.3335 0 Td
(and)Tj
1.7291 0 Td
(then)Tj
2.0183 0 Td
(cultured)Tj
3.5659 0 Td
(in)Tj
-36.5496 -1.3039 Td
(complete)Tj
3.8777 0 Td
(RPMI)Tj
2.6306 0 Td
(with)Tj
2.0012 0 Td
(or)Tj
1.0998 0 Td
(without)Tj
3.3279 0 Td
(rmIL-2)Tj
3.1124 0 Td
(\(R&D)Tj
2.6249 0 Td
(Systems\))Tj
3.753 0 Td
(at)Tj
0.9525 0 Td
(a)Tj
0.6576 0 Td
(concentration)Tj
5.8167 0 Td
(of)Tj
1.0147 0 Td
(20)Tj
1.1736 0 Td
(ng/mL)Tj
2.9196 0 Td
(for)Tj
1.3833 0 Td
(3)Tj
-36.3455 -1.2982 Td
(days)Tj
1.9955 0 Td
(for)Tj
1.389 0 Td
(the)Tj
1.4683 0 Td
(cell)Tj
1.5591 0 Td
(proliferation)Tj
5.2667 0 Td
(assays,)Tj
2.8517 0 Td
(or)Tj
1.0941 0 Td
(for)Tj
1.389 0 Td
(1)Tj
0.6973 0 Td
(day)Tj
1.6328 0 Td
(for)Tj
1.3889 0 Td
(the)Tj
1.4684 0 Td
(assessment)Tj
4.6091 0 Td
(of)Tj
1.0148 0 Td
(GRAIL)Tj
3.1237 0 Td
(intracellular)Tj
-30.9484 -1.2983 Td
(expression.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 125.9716 cm
BT
/F6 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(In)Tj
1.0441 0 Td
(vitro)Tj
/F0 1 Tf
2.0882 0 Td
(blockade)Tj
4.1669 0 Td
(of)Tj
1.0488 0 Td
(CTLA-4)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 109.0204 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Splenocytes)Tj
4.8982 0 Td
(were)Tj
2.1203 0 Td
(isolated)Tj
3.2882 0 Td
(at)Tj
0.9524 0 Td
(21)Tj
1.1679 0 Td
(days)Tj
2.0012 0 Td
(p.i.)Tj
1.4513 0 Td
(and)Tj
1.7292 0 Td
(then)Tj
2.0182 0 Td
(cultured)Tj
3.5716 0 Td
(in)Tj
1.0318 0 Td
(complete)Tj
3.8778 0 Td
(RPMI)Tj
2.6362 0 Td
(medium)Tj
3.6057 0 Td
(in)Tj
1.0374 0 Td
(the)Tj
-35.3874 -1.3039 Td
(presence)Tj
3.7133 0 Td
(of)Tj
1.0148 0 Td
(CTLA-4)Tj
3.5433 0 Td
(blocking)Tj
3.6793 0 Td
(antibody)Tj
3.7644 0 Td
(or)Tj
1.0942 0 Td
(control)Tj
3.1238 0 Td
(antibody)Tj
3.7643 0 Td
(\(10)Tj
/F16 1 Tf
1.5137 0 Td
[()]TJ
/F8 1 Tf
(g/ml,)Tj
2.812 0 Td
(eBioscience\).)Tj
5.4311 0 Td
(Then,)Tj
-33.4542 -1.2982 Td
(intracellular)Tj
5.0569 0 Td
(GRAIL)Tj
3.1181 0 Td
(expression)Tj
4.4674 0 Td
(was)Tj
1.7008 0 Td
(evaluated)Tj
3.9855 0 Td
(48)Tj
1.1735 0 Td
(h)Tj
0.7483 0 Td
(later)Tj
1.9899 0 Td
(by)Tj
1.1849 0 Td
(flow)Tj
1.9446 0 Td
(cytometry.)Tj
ET
Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 183.2882 23.6976 re
W* n
q
183.2315 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
Q
Q
q
0 0 612 792 re
W* n
1 j
1 J
0 w
7.9999 0 0 7.9999 399.0047 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(T)Tj
0.822 0 Td
(Cell)Tj
1.9063 0 Td
(Hyporespon)Tj
5.2938 0 Td
(siveness)Tj
4.0464 0 Td
(during)Tj
/F6 1 Tf
2.941 0 Td
(T)Tj
/F0 1 Tf
(.)Tj
/F6 1 Tf
1.1055 0 Td
(cruzi)Tj
/F0 1 Tf
2.289 0 Td
(Infection)Tj
ET
Q
q
1 j
1 J
0 w
36 48.0002 m
576 48.0002 l
576 47.5002 l
36 47.5002 l
f*
7.9999 0 0 7.9999 36 36 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(PLOS)Tj
2.8559 0 Td
(Neglected)Tj
4.6417 0 Td
(Tropical)Tj
3.7135 0 Td
(Disease)Tj
3.5433 0 Td
(s)Tj
0.7158 0 Td
(|)Tj
0.4748 0 Td
(DOI:10.137)Tj
5.0386 0 Td
(1/journal.pn)Tj
5.1236 0 Td
(td.00053)Tj
3.8198 0 Td
(07)Tj
2.0905 0 Td
(January)Tj
3.7134 0 Td
(23,)Tj
1.5875 0 Td
(2017)Tj
27.8222 0 Td
(5)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
116 0 obj
<>stream
endstream
endobj
117 0 obj
<>stream
endstream
endobj
118 0 obj
<>stream
endstream
endobj
119 0 obj
<>stream
endstream
endobj
120 0 obj
<>stream
BT
66.402 0 Td
(16)Tj
ET
Q
endstream
endobj
121 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 127 0 R/Contents 128 0 R/TrimBox[0 0 612 792]>>
endobj
126 0 obj
<>stream
JFIF C
$, !$4.763.22:ASF:=N>22HbINVX]^]8EfmeZlS[]Y C**Y;2;YYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYYY y"
} !1AQa"q2#BR$3br
%&'()*456789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz
w !1AQaq"2B #3Rbr
$4%&'()*56789:CDEFGHIJSTUVWXYZcdefghijstuvwxyz ? J(+6_t0v1EfYxI[{Kf!kN((((((((((((((((((((((((((((((((((((((((((vu-yun@()##404(?4L}" ߉(ب$[>S'uٰפZžݶ@<2}
4i)hQE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE 6GXѝFI:gu̻PzSZӼ7>'=kkKd,~J*³@NIm{W*M
K:S
yĭ<
˔ݟ~_c2,;U-P5gE(((((((((((((((((((((((((((((((((((((((((W "Σ \Z+,?Ūe>%*LLiV2[ڞ3 Uݝ7;k_k^fé 5ϓ Uqn\ Nn>
.41iGD%-%- QE QE QE QE QE QE QE QE QE QE QE QE QE RQ@E% QE QE QI@E% QE QE QI@E% QE QE QE QE QE QE QE QE QE QE QE QE QE QE QE RQ@E% RQ@EPEPE RQ@E%- QE QE QE QE QE QE QE QE QE QE QE QE QE QE QI@E RQ@E% RREPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPQEGq2[$b{I5/?Tlx՟k:^ywRS }k]s_g.zd^>Y`fpqN*%(x^%sEnR&m+?+?,pq>{|.@$Ik7_\#>ܥ1~"QKE( ( ( ))i(l-%pF&5Xτt_@7QcAqYĝq}*i-l*
rǭJ7$H !8:WH"208>V^Y.i>7B֝ދMZHßL5{k_o Ss?EuZDla(()jKy&8Ha'-PS#%]ѺȧEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPEPX+ gQ -[[ZPXX I-<.ǣ*6mKJ[8eI }_S'@2Ldן|;XU
N/E6 P:"T~5-ԋSz#RR0( ( ( ( ( ( ( ( ( ( ( ( (
ז4t[a1456[Ճ(e!Ey$udJs-gVY!HI-{0=E`t#"oEwE'y'
J*qﲺp?yKm>ծnBX4bm mJo&)d 5)7.<_5{m"qENZ^#
fYlȸVrO9IjZ//NXZ".'pGw]DKi2NAxעC-ݱXxBc!'6`ǟ4q{EPmIc