%PDF-1.6
%
1 0 obj
<>
endobj
2 0 obj
<>stream
Sietske Kooijman, Jolanda Brummelman, Cécile A. C. M. van Els, Fabio Marino, Albert J. R. Heck, Elly van Riet, Bernard Metz, Gideon F. A. Kersten, Jeroen L. A. Pennings, Hugo D. Meiring
Vaccine antigens modulate the innate response of monocytes to Al(OH)3
www.plosone.org
www.plosone.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 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]
endobj
16 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref001)>>
endobj
17 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref002)>>
endobj
18 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref002)>>
endobj
19 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref003)>>
endobj
20 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref004)>>
endobj
21 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref005)>>
endobj
22 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref007)>>
endobj
23 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref004)>>
endobj
24 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref008)>>
endobj
25 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref009)>>
endobj
26 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref010)>>
endobj
27 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref007)>>
endobj
28 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref011)>>
endobj
29 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref012)>>
endobj
30 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref013)>>
endobj
31 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref014)>>
endobj
32 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref015)>>
endobj
33 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref004)>>
endobj
34 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref014)>>
endobj
35 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref016)>>
endobj
36 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref007)>>
endobj
37 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref008)>>
endobj
38 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref011)>>
endobj
39 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref015)>>
endobj
40 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref015)>>
endobj
41 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref017)>>
endobj
42 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref018)>>
endobj
43 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref019)>>
endobj
44 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref020)>>
endobj
45 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref021)>>
endobj
46 0 obj
<>/Border[0 0 0]/A 47 0 R>>
endobj
47 0 obj
<>
endobj
15 0 obj
[48 0 R 49 0 R 50 0 R 51 0 R 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]
endobj
48 0 obj
<>stream
q
0.83 0.64 0.02 0 k
336.8693 706.337 m
341.5748 706.337 l
h
f*
273.7134 654.3496 m
278.4756 654.3496 l
h
f*
203.4142 615.3449 m
208.1764 615.3449 l
h
f*
212.5984 615.3449 m
217.3606 615.3449 l
h
f*
292.7055 576.3402 m
297.411 576.3402 l
h
f*
244.7433 563.3008 m
249.5055 563.3008 l
h
f*
254.6646 563.3008 m
259.4268 563.3008 l
h
f*
437.6693 563.3008 m
442.4315 563.3008 l
h
f*
446.9102 563.3008 m
451.6157 563.3008 l
h
f*
291.7984 550.3181 m
296.5606 550.3181 l
h
f*
300.9827 550.3181 m
310.5071 550.3181 l
h
f*
425.1969 550.3181 m
429.9591 550.3181 l
h
f*
434.3811 550.3181 m
443.9055 550.3181 l
h
f*
448.3843 550.3181 m
457.852 550.3181 l
h
f*
203.4142 537.3354 m
212.9386 537.3354 l
h
f*
439.937 537.3354 m
449.4614 537.3354 l
h
f*
554.9102 537.3354 m
564.4346 537.3354 l
h
f*
534.2173 524.2961 m
538.9795 524.2961 l
h
f*
543.4016 524.2961 m
552.926 524.2961 l
h
f*
557.4047 524.2961 m
566.8724 524.2961 l
h
f*
511.4268 511.3134 m
516.189 511.3134 l
h
f*
520.611 511.3134 m
525.3732 511.3134 l
h
f*
529.852 511.3134 m
539.3764 511.3134 l
h
f*
543.7984 511.3134 m
553.3228 511.3134 l
h
f*
241.5118 420.3213 m
251.2063 420.3213 l
h
f*
255.7984 420.3213 m
265.4929 420.3213 l
h
f*
270.0283 420.3213 m
279.7228 420.3213 l
h
f*
311.9244 290.3244 m
321.4488 290.3244 l
h
f*
325.9276 290.3244 m
335.3953 290.3244 l
h
f*
342.0283 264.3024 m
351.5528 264.3024 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(the)Tj
1.4683 0 Td
(toxoid)Tj
2.7949 0 Td
(than)Tj
2.024 0 Td
(the)Tj
1.474 0 Td
(antigen)Tj
3.2031 0 Td
(alone)Tj
2.3811 0 Td
([)Tj
0.83 0.64 0.02 0 k
(1)Tj
0 g
(].)Tj
1.5987 0 Td
(Since)Tj
2.3584 0 Td
(the)Tj
1.4684 0 Td
(discovery)Tj
4.0138 0 Td
(of)Tj
1.0148 0 Td
(their)Tj
2.1089 0 Td
(adjuvant)Tj
3.6964 0 Td
(activity,)Tj
3.3449 0 Td
(aluminum)Tj
-32.9497 -1.3039 Td
(salts)Tj
1.9218 0 Td
(have)Tj
2.058 0 Td
(been)Tj
2.1203 0 Td
(widely)Tj
2.8233 0 Td
(used)Tj
2.0636 0 Td
(as)Tj
1.0148 0 Td
(vaccine)Tj
3.1918 0 Td
(adjuvants)Tj
4.0648 0 Td
(in)Tj
1.0318 0 Td
(human)Tj
3.0784 0 Td
(vaccines.)Tj
3.7814 0 Td
(For)Tj
1.6271 0 Td
(a)Tj
0.652 0 Td
(long)Tj
1.9899 0 Td
(time)Tj
2.0296 0 Td
(the)Tj
1.4683 0 Td
(mech-)Tj
-34.9169 -1.2982 Td
(anism)Tj
2.6475 0 Td
(of)Tj
1.0148 0 Td
(action)Tj
2.6986 0 Td
(of)Tj
1.0148 0 Td
(aluminum)Tj
4.4106 0 Td
(adjuvants)Tj
4.0649 0 Td
(was)Tj
1.7008 0 Td
(largely)Tj
2.8573 0 Td
(unknown.)Tj
4.3086 0 Td
(Based)Tj
2.5455 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.4683 0 Td
(observation)Tj
4.8983 0 Td
(that)Tj
-34.9056 -1.2983 Td
(aluminum)Tj
4.4106 0 Td
(salt-adsorbed)Tj
5.5672 0 Td
(toxoids)Tj
3.1521 0 Td
(were)Tj
2.126 0 Td
(cleared)Tj
3.0671 0 Td
(more)Tj
2.3414 0 Td
(slowly)Tj
2.7212 0 Td
(from)Tj
2.2054 0 Td
(the)Tj
1.4683 0 Td
(injection)Tj
3.7587 0 Td
(site)Tj
1.5704 0 Td
(than)Tj
2.0296 0 Td
(non-)Tj
-34.418 -1.2983 Td
(adsorbed)Tj
3.8777 0 Td
(toxoids)Tj
3.1521 0 Td
([)Tj
0.83 0.64 0.02 0 k
(2)Tj
0 g
(],)Tj
1.6044 0 Td
(it)Tj
0.7824 0 Td
(was)Tj
1.7008 0 Td
(hypothesized)Tj
5.4651 0 Td
(that)Tj
1.7802 0 Td
(antigen-aluminum)Tj
7.7498 0 Td
(salts)Tj
1.9162 0 Td
(act)Tj
1.3777 0 Td
(as)Tj
1.0148 0 Td
(a)Tj
0.6519 0 Td
(depot)Tj
2.4945 0 Td
(at)Tj
0.9524 0 Td
(the)Tj
1.4684 0 Td
(site)Tj
-35.9884 -1.3039 Td
(of)Tj
1.0147 0 Td
(injection,)Tj
3.9855 0 Td
(causing)Tj
3.2485 0 Td
(a)Tj
0.652 0 Td
(slow)Tj
2.0182 0 Td
(antigen)Tj
3.2032 0 Td
(release.)Tj
3.1294 0 Td
(However,)Tj
4.0762 0 Td
(in)Tj
1.0375 0 Td
(guinea)Tj
2.8856 0 Td
(pig)Tj
1.4684 0 Td
(experiments,)Tj
5.3631 0 Td
(the)Tj
1.474 0 Td
(immune)Tj
-33.5563 -1.2982 Td
(response)Tj
3.7303 0 Td
(was)Tj
1.7008 0 Td
(not)Tj
1.576 0 Td
(compromised)Tj
5.76 0 Td
(when)Tj
2.4038 0 Td
(the)Tj
1.474 0 Td
(salt)Tj
1.559 0 Td
(deposit)Tj
3.1181 0 Td
(was)Tj
1.7008 0 Td
(removed)Tj
3.7474 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.4684 0 Td
(injection)Tj
3.7643 0 Td
(site)Tj
-34.2082 -1.2983 Td
([)Tj
0.83 0.64 0.02 0 k
(2)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.2585 0 Td
(3)Tj
0 g
(],)Tj
1.2643 0 Td
(indicating)Tj
4.2632 0 Td
(that)Tj
1.7802 0 Td
(this)Tj
1.6724 0 Td
(is)Tj
0.8504 0 Td
(at)Tj
0.9524 0 Td
(least)Tj
1.9843 0 Td
(not)Tj
1.5704 0 Td
(the)Tj
1.474 0 Td
(only)Tj
1.9729 0 Td
(mechanism)Tj
4.8472 0 Td
(by)Tj
1.1792 0 Td
(which)Tj
2.6532 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 508.4787 614.7212 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 514.2047 616.422 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(affects)Tj
2.7382 0 Td
(the)Tj
-34.1572 -1.3039 Td
(immune)Tj
3.6227 0 Td
(response)Tj
3.736 0 Td
(towards)Tj
3.4016 0 Td
(antigens.)Tj
3.7927 0 Td
(Other)Tj
2.5852 0 Td
(modes)Tj
2.863 0 Td
(of)Tj
1.0148 0 Td
(action)Tj
2.6985 0 Td
(suggested)Tj
4.0706 0 Td
(for)Tj
1.3889 0 Td
(the)Tj
1.4684 0 Td
(adjuvant)Tj
3.7019 0 Td
(effect)Tj
2.3641 0 Td
(of)Tj
-36.7084 -1.2982 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 231.2503 588.7559 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 236.9763 590.4 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(include)Tj
3.1804 0 Td
(early)Tj
2.1487 0 Td
(innate)Tj
2.7325 0 Td
(mechanisms)Tj
5.2044 0 Td
(such)Tj
2.0579 0 Td
(as)Tj
1.0092 0 Td
(differentiation)Tj
5.9754 0 Td
(of)Tj
1.0091 0 Td
(monocytes)Tj
4.5581 0 Td
(to)Tj
1.0261 0 Td
(antigen-pre-)Tj
-32.5981 -1.2982 Td
(senting)Tj
3.1294 0 Td
(dendritic)Tj
3.8777 0 Td
(cells)Tj
1.9219 0 Td
([)Tj
0.83 0.64 0.02 0 k
(4)Tj
0 g
(],)Tj
1.5987 0 Td
(triggering)Tj
4.1612 0 Td
(and)Tj
1.7292 0 Td
(recognition)Tj
4.8472 0 Td
(of)Tj
1.0148 0 Td
(Danger)Tj
3.2031 0 Td
(Associated)Tj
4.5071 0 Td
(Molecular)Tj
4.2916 0 Td
(Patterns)Tj
-34.2819 -1.3039 Td
(\(DAMPs\))Tj
4.1328 0 Td
([)Tj
0.83 0.64 0.02 0 k
(5)Tj
0 g
()Tj
0.83 0.64 0.02 0 k
(7)Tj
0 g
(])Tj
2.3697 0 Td
(promoting)Tj
4.5184 0 Td
(T-helper)Tj
3.6907 0 Td
(\(Th\))Tj
2.0409 0 Td
(2)Tj
0.6974 0 Td
(differentiation)Tj
5.9754 0 Td
([)Tj
0.83 0.64 0.02 0 k
(4)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.2642 0 Td
(8)Tj
0 g
(],)Tj
1.2586 0 Td
(recruitment)Tj
4.9946 0 Td
(of)Tj
1.0148 0 Td
(immune)Tj
3.6226 0 Td
(cells)Tj
-35.5801 -1.2983 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(site)Tj
1.5703 0 Td
(of)Tj
1.0147 0 Td
(injection)Tj
3.7588 0 Td
([)Tj
0.83 0.64 0.02 0 k
(9)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.2585 0 Td
(10)Tj
0 g
(],)Tj
1.7405 0 Td
(inflammasome)Tj
6.1852 0 Td
(activation)Tj
4.1555 0 Td
([)Tj
0.83 0.64 0.02 0 k
(7)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.2586 0 Td
(11)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.4003 0 Td
(12)Tj
0 g
(],)Tj
1.7348 0 Td
(complement)Tj
5.2441 0 Td
(activation)Tj
-31.8158 -1.2983 Td
([)Tj
0.83 0.64 0.02 0 k
(13)Tj
0 g
(],)Tj
2.0749 0 Td
(increasing)Tj
4.3313 0 Td
(antigen)Tj
3.2031 0 Td
(presentation)Tj
5.2214 0 Td
(via)Tj
1.3719 0 Td
(HLA)Tj
2.211 0 Td
(class)Tj
2.0466 0 Td
(I)Tj
0.5613 0 Td
(and)Tj
1.7291 0 Td
(II)Tj
0.9014 0 Td
([)Tj
0.83 0.64 0.02 0 k
(14)Tj
0 g
(],)Tj
2.075 0 Td
(enhanced)Tj
4.0762 0 Td
(phagocytosis)Tj
5.3461 0 Td
([)Tj
0.83 0.64 0.02 0 k
(15)Tj
0 g
(].)Tj
-33.9531 -1.3039 Td
(Most)Tj
2.279 0 Td
(of)Tj
1.0147 0 Td
(the)Tj
1.4741 0 Td
(previous)Tj
3.6453 0 Td
(studies)Tj
2.9877 0 Td
(analyzed)Tj
3.702 0 Td
(the)Tj
1.4683 0 Td
(effect)Tj
2.3641 0 Td
(of)Tj
1.0148 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 442.7716 523.7291 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 448.4976 525.3732 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(with)Tj
1.9955 0 Td
(either)Tj
2.5342 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 525.033 523.7291 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 530.8157 525.3732 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
([)Tj
0.83 0.64 0.02 0 k
(4)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.2585 0 Td
(14)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.4003 0 Td
(16)Tj
0 g
(])Tj
-35.7389 -1.2982 Td
(alone)Tj
2.3755 0 Td
(or)Tj
1.0998 0 Td
(in)Tj
1.0375 0 Td
(combination)Tj
5.3461 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6576 0 Td
(model)Tj
2.7439 0 Td
(antigen)Tj
3.2031 0 Td
(like)Tj
1.6555 0 Td
(ovalbumin)Tj
4.5297 0 Td
(or)Tj
1.0998 0 Td
(alpha)Tj
2.3811 0 Td
(casein)Tj
2.6759 0 Td
([)Tj
0.83 0.64 0.02 0 k
(7)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.2586 0 Td
(8)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
0.9241 0 Td
(11)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.3945 0 Td
(15)Tj
0 g
(].)Tj
-34.3783 -1.2983 Td
(Since)Tj
2.3585 0 Td
(antigens)Tj
3.5659 0 Td
(may)Tj
1.9276 0 Td
(have)Tj
2.0579 0 Td
(a)Tj
0.6576 0 Td
(distinct)Tj
3.2088 0 Td
(effect)Tj
2.3641 0 Td
(on)Tj
1.2756 0 Td
(innate)Tj
2.7439 0 Td
(immune)Tj
3.6227 0 Td
(cells,)Tj
2.1486 0 Td
(the)Tj
1.4684 0 Td
(question)Tj
3.6623 0 Td
(arises)Tj
2.4377 0 Td
(to)Tj
1.0204 0 Td
(what)Tj
-34.52 -1.2982 Td
(extent)Tj
2.6759 0 Td
(particular)Tj
4.1102 0 Td
(antigens)Tj
3.566 0 Td
(skew)Tj
2.1883 0 Td
(the)Tj
1.4684 0 Td
(innate)Tj
2.7382 0 Td
(effects)Tj
2.7269 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(adjuvant.)Tj
0.005 Tc
-20.7608 -1.304 Td
(DTaP)Tj
2.5965 0 Td
(vaccine)Tj
3.2598 0 Td
(was)Tj
1.7235 0 Td
(implemented)Tj
5.6352 0 Td
(for)Tj
1.4173 0 Td
(active)Tj
2.5738 0 Td
(immunization)Tj
6.0435 0 Td
(against)Tj
3.0897 0 Td
(diphtheria,)Tj
4.7055 0 Td
(tetanus)Tj
3.1747 0 Td
(and)Tj
-35.4158 -1.2982 Td
(pertussis)Tj
3.7871 0 Td
(in)Tj
1.0545 0 Td
(infants)Tj
3.0274 0 Td
(and)Tj
1.7574 0 Td
(children.)Tj
3.8608 0 Td
(Besides)Tj
3.2258 0 Td
(diphtheria)Tj
4.4731 0 Td
(toxoid)Tj
2.8459 0 Td
(and)Tj
1.7575 0 Td
(tetanus)Tj
3.1748 0 Td
(toxoid,)Tj
3.0784 0 Td
(DTaP)Tj
2.5964 0 Td
(com-)Tj
-34.6391 -1.2983 Td
(prises)Tj
/F13 1 Tf
2.5739 0 Td
[()-2()11()20()4()8()11()15()4()4()]TJ
4.2406 0 Td
[( )4()2()-19()7(\n)-4()10()10()-4()]TJ
/F9 1 Tf
(-derived)Tj
7.0299 0 Td
(filamentous)Tj
5.0399 0 Td
(hemagglutinin,)Tj
6.412 0 Td
(pertussis)Tj
3.787 0 Td
(toxoid)Tj
2.846 0 Td
(and)Tj
1.7574 0 Td
(pertactin)Tj
-33.6867 -1.3039 Td
(P.69)Tj
1.99 0 Td
(antigens.)Tj
3.8721 0 Td
(DTaP)Tj
2.5965 0 Td
(vaccination)Tj
4.9266 0 Td
(typically)Tj
3.6396 0 Td
(induces)Tj
3.3619 0 Td
(a)Tj
0.6633 0 Td
(Th2-biased)Tj
4.7962 0 Td
(and)Tj
1.7574 0 Td
(regulatory)Tj
4.4107 0 Td
(T)Tj
0.8391 0 Td
(cell)Tj
-32.8534 -1.2983 Td
(response)Tj
3.8041 0 Td
([)Tj
0.83 0.64 0.02 0 k
(15)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.7745 0 Td
(17)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.423 0 Td
(18)Tj
0 g
(])Tj
1.5363 0 Td
(not)Tj
1.5988 0 Td
(optimally)Tj
4.0932 0 Td
(conferring)Tj
4.524 0 Td
(long-term)Tj
4.3314 0 Td
(protective)Tj
4.2972 0 Td
(immunity)Tj
4.2917 0 Td
(to)Tj
1.0431 0 Td
(pertussis.)Tj
-32.7173 -1.2982 Td
(For)Tj
1.6498 0 Td
(vaccine)Tj
3.2598 0 Td
(development)Tj
5.5105 0 Td
(it)Tj
0.7994 0 Td
(is)Tj
0.8674 0 Td
(important)Tj
4.3653 0 Td
(to)Tj
1.0432 0 Td
(understand)Tj
4.8755 0 Td
(whether)Tj
3.5376 0 Td
(antigens)Tj
3.6397 0 Td
(affect)Tj
2.4321 0 Td
(the)Tj
1.4966 0 Td
(innate)Tj
-33.4769 -1.304 Td
(phase)Tj
2.5285 0 Td
(of)Tj
1.0318 0 Td
(the)Tj
1.4967 0 Td
(immune)Tj
3.6794 0 Td
(response)Tj
3.804 0 Td
(or)Tj
1.1169 0 Td
(the)Tj
1.4967 0 Td
(instructions)Tj
5.108 0 Td
(of)Tj
1.0318 0 Td
(the)Tj
1.4967 0 Td
(adaptive)Tj
3.651 0 Td
(response)Tj
3.804 0 Td
(of)Tj
1.0318 0 Td
(the)Tj
1.4967 0 Td
(adjuvant,)Tj
-32.774 -1.2982 Td
(or)Tj
1.1169 0 Td
(both.)Tj
-0.004 Tc
0.0794 -1.2983 Td
(In)Tj
1.1111 0 Td
(this)Tj
1.6725 0 Td
(study,)Tj
2.6192 0 Td
(we)Tj
1.3266 0 Td
(compared)Tj
4.2406 0 Td
(the)Tj
1.474 0 Td
(innate)Tj
2.7382 0 Td
(immune)Tj
3.6283 0 Td
(responses)Tj
4.0932 0 Td
(induced)Tj
3.464 0 Td
(by)Tj
1.1792 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 518.7401 367.7102 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 524.4661 369.411 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(alone)Tj
/F13 1 Tf
2.381 0 Td
[(\r)3()2()5()10(\n)-4()]TJ
/F9 1 Tf
-34.8261 -1.2982 Td
(that)Tj
1.7745 0 Td
(of)Tj
1.0148 0 Td
(a)Tj
0.6576 0 Td
(licensed)Tj
3.4412 0 Td
(combination)Tj
5.3462 0 Td
(DTaP)Tj
2.5625 0 Td
(vaccine)Tj
3.1917 0 Td
(containing)Tj
4.5298 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 456.4346 354.7275 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 460.0062 356.4283 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
0.4422 0 Td
(using)Tj
2.3924 0 Td
(cytokine)Tj
3.6396 0 Td
(analysis,)Tj
-32.4734 -1.3039 Td
(transcriptomics)Tj
6.4857 0 Td
(and)Tj
1.7348 0 Td
(proteomics,)Tj
4.9379 0 Td
(to)Tj
1.0261 0 Td
(determine)Tj
4.3257 0 Td
(unique,)Tj
3.2428 0 Td
(shared)Tj
2.8686 0 Td
(and)Tj
1.729 0 Td
(potential)Tj
3.7588 0 Td
(synergistic)Tj
4.4617 0 Td
(or)Tj
-34.5711 -1.2982 Td
(antagonistic)Tj
5.074 0 Td
(effects)Tj
2.7213 0 Td
(of)Tj
1.0148 0 Td
(adjuvant)Tj
3.702 0 Td
(and)Tj
1.7291 0 Td
(antigen)Tj
3.2088 0 Td
(components.)Tj
5.3971 0 Td
(Primary)Tj
3.4979 0 Td
(human)Tj
3.0784 0 Td
(monocytes)Tj
4.5637 0 Td
(were)Tj
-33.9871 -1.2983 Td
(used)Tj
2.058 0 Td
(as)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(main)Tj
2.2904 0 Td
(innate)Tj
2.7382 0 Td
(cell)Tj
1.5648 0 Td
(platform)Tj
3.702 0 Td
(since)Tj
2.2393 0 Td
(these)Tj
2.2564 0 Td
(are)Tj
1.4457 0 Td
(prominent)Tj
4.524 0 Td
(mononuclear)Tj
5.5785 0 Td
(phagocytes)Tj
4.6318 0 Td
(in)Tj
-35.5122 -1.3039 Td
(the)Tj
1.4684 0 Td
(blood)Tj
2.5171 0 Td
(and)Tj
1.7292 0 Td
(play,)Tj
2.0976 0 Td
(when)Tj
2.4037 0 Td
(activated,)Tj
4.0082 0 Td
(an)Tj
1.2019 0 Td
(important)Tj
4.2803 0 Td
(role)Tj
1.7688 0 Td
(in)Tj
1.0318 0 Td
(bridging)Tj
3.6396 0 Td
(the)Tj
1.474 0 Td
(innate)Tj
2.7382 0 Td
(and)Tj
1.7291 0 Td
(adaptive)Tj
-32.0879 -1.2983 Td
(immune)Tj
3.6227 0 Td
(response)Tj
3.736 0 Td
(in)Tj
1.0375 0 Td
(tissue)Tj
2.4548 0 Td
([)Tj
0.83 0.64 0.02 0 k
(19)Tj
0 g
(,)Tj
0.83 0.64 0.02 0 k
1.7405 0 Td
(20)Tj
0 g
(].)Tj
1.7348 0 Td
(Besides)Tj
3.1634 0 Td
(this,)Tj
1.8992 0 Td
(their)Tj
2.109 0 Td
(known)Tj
3.0047 0 Td
(differentiation)Tj
5.9753 0 Td
(into)Tj
1.8425 0 Td
(monocyte-)Tj
-32.3204 -1.2982 Td
(derived)Tj
3.2145 0 Td
(dendritic)Tj
3.8778 0 Td
(cells)Tj
1.9162 0 Td
(\(MoDCs\))Tj
4.0592 0 Td
(monocyte)Tj
4.2066 0 Td
(can)Tj
1.6214 0 Td
(also)Tj
1.7688 0 Td
(enforce)Tj
3.2088 0 Td
(their)Tj
2.1089 0 Td
(antigen)Tj
3.2031 0 Td
(presenting)Tj
4.4447 0 Td
(role)Tj
1.7688 0 Td
(to)Tj
1.0261 0 Td
(T)Tj
-36.4249 -1.304 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
(various)Tj
3.1408 0 Td
(stimuli)Tj
3.0047 0 Td
([)Tj
0.83 0.64 0.02 0 k
(21)Tj
0 g
(].)Tj
2.0806 0 Td
(This)Tj
1.9842 0 Td
(systems-based)Tj
5.9017 0 Td
(approach)Tj
3.9514 0 Td
(results)Tj
2.8119 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.652 0 Td
(comprehen-)Tj
-32.2807 -1.2982 Td
(sive)Tj
1.7235 0 Td
(insight)Tj
2.9593 0 Td
(in)Tj
1.0319 0 Td
(the)Tj
1.4683 0 Td
(molecular)Tj
4.2236 0 Td
(pathways)Tj
3.9288 0 Td
(involved)Tj
3.651 0 Td
(in)Tj
1.0318 0 Td
(innate)Tj
2.7439 0 Td
(immune)Tj
3.6226 0 Td
(activation)Tj
4.1555 0 Td
(of)Tj
1.0148 0 Td
(monocytes)Tj
-31.555 -1.2983 Td
(upon)Tj
2.3188 0 Td
(stimulation)Tj
4.7962 0 Td
(with)Tj
1.9955 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 322.3559 237.7133 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 325.9275 239.4141 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(-based)Tj
2.8232 0 Td
(vaccines.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 195.9874 cm
BT
/F2 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Materials)Tj
4.5071 0 Td
(and)Tj
1.989 0 Td
(methods)Tj
/F14 1 Tf
-6.4961 -1.4976 Td
(Ethics)Tj
2.7779 0 Td
(statement)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 161.0078 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(This)Tj
1.9842 0 Td
(study)Tj
2.3924 0 Td
(was)Tj
1.6951 0 Td
(conducted)Tj
4.4277 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(principles)Tj
4.1612 0 Td
(expressed)Tj
4.0989 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.474 0 Td
(Declaration)Tj
4.9096 0 Td
(of)Tj
1.0148 0 Td
(Helsinki.)Tj
-33.8738 -1.2982 Td
(Written)Tj
3.3902 0 Td
(informed)Tj
3.9741 0 Td
(consent)Tj
3.3279 0 Td
(was)Tj
1.7007 0 Td
(obtained)Tj
3.7361 0 Td
(from)Tj
2.2053 0 Td
(all)Tj
1.1452 0 Td
(blood)Tj
2.5115 0 Td
(donors)Tj
3.0444 0 Td
(before)Tj
2.7382 0 Td
(collection)Tj
4.1159 0 Td
(and)Tj
1.7291 0 Td
(use)Tj
1.5307 0 Td
(of)Tj
-35.1493 -1.3039 Td
(their)Tj
2.1089 0 Td
(samples.)Tj
3.6057 0 Td
(All)Tj
1.3946 0 Td
(blood)Tj
2.5171 0 Td
(samples)Tj
3.3846 0 Td
(were)Tj
2.1203 0 Td
(processed)Tj
4.1329 0 Td
(anonymously.)Tj
5.8563 0 Td
(All)Tj
1.3947 0 Td
(blood)Tj
2.5171 0 Td
(donations,)Tj
4.4447 0 Td
(provided)Tj
-33.4769 -1.2983 Td
(by)Tj
1.1792 0 Td
(the)Tj
1.4683 0 Td
(Dutch)Tj
2.7382 0 Td
(National)Tj
3.6907 0 Td
(Institute)Tj
3.583 0 Td
(for)Tj
1.3833 0 Td
(Public)Tj
2.7496 0 Td
(Health)Tj
2.9197 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(Environment)Tj
5.5786 0 Td
(\(RIVM,)Tj
3.3335 0 Td
(Bilthoven;)Tj
-31.8215 -1.2983 Td
(The)Tj
1.7801 0 Td
(Netherlands\),)Tj
5.6863 0 Td
(were)Tj
2.1203 0 Td
(specifically)Tj
4.5751 0 Td
(given)Tj
2.3754 0 Td
(for)Tj
1.3889 0 Td
(primary)Tj
3.4526 0 Td
(cell)Tj
1.5591 0 Td
(isolation,)Tj
3.8834 0 Td
(a)Tj
0.652 0 Td
(research)Tj
3.5603 0 Td
(goal)Tj
1.8708 0 Td
(explicitly)Tj
-32.9043 -1.3039 Td
(approved)Tj
3.9741 0 Td
(by)Tj
1.1792 0 Td
(the)Tj
1.4683 0 Td
(accredited)Tj
4.3427 0 Td
(Medical)Tj
3.4356 0 Td
(Research)Tj
3.804 0 Td
(Ethics)Tj
2.6646 0 Td
(Committee)Tj
4.7451 0 Td
(\(MREC\),)Tj
3.8721 0 Td
(METC,)Tj
3.1805 0 Td
(Noord-Hol-)Tj
-32.6662 -1.2982 Td
(land)Tj
1.9729 0 Td
(in)Tj
1.0374 0 Td
(The)Tj
1.7858 0 Td
(Netherlands.)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 107.1496 23.6976 re
W* n
q
107.0929 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 350.1354 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Antigens)Tj
4.0394 0 Td
(modulate)Tj
4.259 0 Td
(the)Tj
1.5875 0 Td
(adjuvant-indu)Tj
5.889 0 Td
(ced)Tj
1.8 0 Td
(innate)Tj
2.8914 0 Td
(immune)Tj
3.7205 0 Td
(response)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*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
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
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01978)Tj
4.089 0 Td
(85)Tj
0 g
2.0906 0 Td
(May)Tj
2.0906 0 Td
(29,)Tj
1.5803 0 Td
(2018)Tj
35.2349 0 Td
(2)Tj
0.7654 0 Td
(/)Tj
0.496 0 Td
(22)Tj
/F11 1 Tf
-0.025 Tc
-66.402 84.0904 Td
(`author)Tj
2.8204 0 Td
(contributi)Tj
3.515 0 Td
(ons')Tj
1.7859 0 Td
(section.)Tj
3.1039 0 Td
(This)Tj
1.7788 0 Td
(work)Tj
2.0622 0 Td
(was)Tj
1.7008 0 Td
(also)Tj
-16.767 -1.3748 Td
(partly)Tj
2.2889 0 Td
(supported)Tj
3.9969 0 Td
(by)Tj
1.1126 0 Td
(the)Tj
1.3748 0 Td
(Protein)Tj
2.6434 0 Td
(s@Work,)Tj
3.6496 0 Td
(a)Tj
0.6449 0 Td
(628)Tj
-15.7111 -1.3749 Td
(program)Tj
3.4299 0 Td
(of)Tj
0.9496 0 Td
(the)Tj
1.3677 0 Td
(Netherlands)Tj
4.6702 0 Td
(Proteomics)Tj
4.4646 0 Td
(Centre)Tj
-14.882 -1.3748 Td
(financed)Tj
3.3519 0 Td
(by)Tj
1.1127 0 Td
(the)Tj
1.3748 0 Td
(Netherlan)Tj
3.5504 0 Td
(ds)Tj
1.1197 0 Td
(Organisa)Tj
3.2953 0 Td
(tion)Tj
1.6157 0 Td
(for)Tj
-15.4205 -1.3748 Td
(Scientific)Tj
3.5716 0 Td
(Research)Tj
3.6993 0 Td
(\(NWO\))Tj
2.8134 0 Td
(as)Tj
1.063 0 Td
(part)Tj
1.6795 0 Td
(of)Tj
0.9496 0 Td
(the)Tj
1.3678 0 Td
(National)Tj
-15.1442 -1.3748 Td
(Roadmap)Tj
3.8268 0 Td
(Large-s)Tj
2.8204 0 Td
(cale)Tj
1.6796 0 Td
(Research)Tj
3.6992 0 Td
(Facilities)Tj
3.352 0 Td
(of)Tj
0.9496 0 Td
(the)Tj
-16.3276 -1.3748 Td
(Netherland)Tj
4.0252 0 Td
(s)Tj
0.6449 0 Td
(\(project)Tj
3.0685 0 Td
(number)Tj
3.1252 0 Td
(184.032.201\))Tj
5.0174 0 Td
(.)Tj
-15.8812 -1.3748 Td
(Proteins@W)Tj
4.6488 0 Td
(ork)Tj
1.4244 0 Td
(funded)Tj
2.7922 0 Td
(the)Tj
1.3677 0 Td
(salary)Tj
2.4095 0 Td
(of)Tj
0.9496 0 Td
(FM)Tj
1.389 0 Td
(but)Tj
1.4244 0 Td
(did)Tj
1.3606 0 Td
(not)Tj
-17.7662 -1.3748 Td
(have)Tj
1.9559 0 Td
(a)Tj
0.6449 0 Td
(role)Tj
1.6157 0 Td
(in)Tj
0.8859 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
-17.171 -1.3748 Td
(analysis,)Tj
3.4086 0 Td
(decision)Tj
3.2953 0 Td
(to)Tj
0.9497 0 Td
(publish)Tj
2.9197 0 Td
(or)Tj
0.9992 0 Td
(preparation)Tj
4.4433 0 Td
(of)Tj
0.9496 0 Td
(the)Tj
-16.9654 -1.3749 Td
(manuscript.)Tj
4.6417 0 Td
(The)Tj
1.5874 0 Td
(specific)Tj
3.019 0 Td
(role)Tj
1.6157 0 Td
(of)Tj
0.9496 0 Td
(this)Tj
1.5591 0 Td
(author)Tj
2.6292 0 Td
(is)Tj
-16.0017 -1.3748 Td
(articulated)Tj
4.0535 0 Td
(in)Tj
0.8929 0 Td
(the)Tj
1.3678 0 Td
(`author)Tj
2.8134 0 Td
(contributions')Tj
5.3008 0 Td
(section.)Tj
/F10 1 Tf
-14.4284 -2.126 Td
(Competing)Tj
4.3795 0 Td
(interests)Tj
3.3307 0 Td
(:)Tj
/F11 1 Tf
0.4819 0 Td
(The)Tj
1.5875 0 Td
(authors)Tj
3.0401 0 Td
(have)Tj
1.963 0 Td
(the)Tj
-14.7827 -1.3748 Td
(following)Tj
3.5858 0 Td
(competing)Tj
4.1599 0 Td
(interest)Tj
2.9693 0 Td
(to)Tj
0.9496 0 Td
(declare:)Tj
3.104 0 Td
(BM,)Tj
1.722 0 Td
(GFAK,)Tj
-16.4906 -1.3748 Td
(HDM)Tj
2.1472 0 Td
(and)Tj
1.5874 0 Td
(SK)Tj
1.2756 0 Td
(are)Tj
1.3678 0 Td
(affiliated)Tj
3.3023 0 Td
(with)Tj
1.7788 0 Td
(the)Tj
1.3748 0 Td
(commerci)Tj
3.7418 0 Td
(al)Tj
-16.5757 -1.3748 Td
(company,)Tj
3.8834 0 Td
(Intravacc)Tj
3.3946 0 Td
(.)Tj
0.4464 0 Td
(There)Tj
2.3174 0 Td
(are)Tj
1.3677 0 Td
(no)Tj
1.1693 0 Td
(patents,)Tj
-12.5788 -1.3748 Td
(products)Tj
3.522 0 Td
(in)Tj
0.8859 0 Td
(developm)Tj
3.6283 0 Td
(ent)Tj
1.3678 0 Td
(or)Tj
1.0063 0 Td
(markete)Tj
2.9976 0 Td
(d)Tj
0.6945 0 Td
(products)Tj
3.5221 0 Td
(to)Tj
-17.6245 -1.3748 Td
(declare.)Tj
3.1039 0 Td
(This)Tj
1.7788 0 Td
(affiliation)Tj
3.5504 0 Td
(does)Tj
2.0126 0 Td
(not)Tj
1.4173 0 Td
(alter)Tj
1.8213 0 Td
(authors')Tj
-13.6843 -1.3749 Td
(adherence)Tj
4.0535 0 Td
(to)Tj
0.9496 0 Td
(data)Tj
1.793 0 Td
(sharing)Tj
2.9835 0 Td
(policies)Tj
3.0118 0 Td
(of)Tj
0.9496 0 Td
(PLOS)Tj
2.3315 0 Td
(ONE.)Tj
-16.0725 -1.3748 Td
(The)Tj
1.5874 0 Td
(authors)Tj
3.0472 0 Td
(declare)Tj
2.8772 0 Td
(no)Tj
1.1693 0 Td
(competing)Tj
4.1599 0 Td
(financial)Tj
ET
endstream
endobj
49 0 obj
<>stream
endstream
endobj
50 0 obj
<>stream
endstream
endobj
51 0 obj
<>stream
endstream
endobj
52 0 obj
<>stream
endstream
endobj
53 0 obj
<>stream
endstream
endobj
54 0 obj
<>stream
endstream
endobj
55 0 obj
<>stream
endstream
endobj
56 0 obj
<>stream
endstream
endobj
57 0 obj
<>stream
endstream
endobj
58 0 obj
<>stream
endstream
endobj
59 0 obj
<>stream
endstream
endobj
60 0 obj
<>stream
endstream
endobj
61 0 obj
<>stream
endstream
endobj
62 0 obj
<>stream
endstream
endobj
63 0 obj
<>stream
endstream
endobj
64 0 obj
<>stream
endstream
endobj
65 0 obj
<>stream
endstream
endobj
66 0 obj
<>stream
endstream
endobj
67 0 obj
<>stream
endstream
endobj
68 0 obj
<>stream
endstream
endobj
69 0 obj
<>stream
endstream
endobj
70 0 obj
<>stream
BT
0 55.8429 Td
(interest.)Tj
ET
Q
endstream
endobj
71 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 75 0 R/Contents 76 0 R/TrimBox[0 0 612 792]>>
endobj
75 0 obj
[77 0 R 78 0 R 79 0 R 80 0 R]
endobj
77 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref016)>>
endobj
78 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.s001)>>
endobj
79 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref016)>>
endobj
80 0 obj
<>/Border[0 0 0]/A 81 0 R>>
endobj
81 0 obj
<>
endobj
76 0 obj
[82 0 R 83 0 R 84 0 R 85 0 R 86 0 R 87 0 R]
endobj
82 0 obj
<>stream
q
0.83 0.64 0.02 0 k
246.8976 293.9528 m
256.422 293.9528 l
h
f*
265.4362 293.9528 m
289.7575 293.9528 l
h
f*
470.8346 172.9134 m
480.3591 172.9134 l
h
f*
0 g
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 706.0535 cm
BT
/F14 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Materials)Tj
4.1527 0 Td
(used)Tj
2.1355 0 Td
(for)Tj
1.4693 0 Td
(cell)Tj
1.6299 0 Td
(stimulation)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 689.1023 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Aluminum)Tj
4.6601 0 Td
(hydroxide)Tj
4.2916 0 Td
(\(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 324.17 687.4015 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 327.7417 689.1023 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\))Tj
0.5612 0 Td
(Alhydrogel)Tj
4.6715 0 Td
(2%,)Tj
1.6781 0 Td
(Brenntag)Tj
3.8891 0 Td
(\(Frederikssund;)Tj
6.5196 0 Td
(Denmark\))Tj
4.3937 0 Td
(was)Tj
-34.486 -1.3039 Td
(used)Tj
2.058 0 Td
(as)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(adjuvant.)Tj
3.9288 0 Td
(The)Tj
1.7801 0 Td
(registered)Tj
4.1555 0 Td
(DTaP)Tj
2.5625 0 Td
(vaccine)Tj
3.1918 0 Td
(Infanrix)Tj
3.4809 0 Td
(was)Tj
1.7008 0 Td
(obtained)Tj
3.736 0 Td
(from)Tj
2.2054 0 Td
(GlaxoSmithK-)Tj
-31.2829 -1.2982 Td
(line)Tj
1.7065 0 Td
(\(Brentford;)Tj
4.6998 0 Td
(Middlesex;)Tj
4.5807 0 Td
(UK\).)Tj
2.1939 0 Td
(The)Tj
1.7802 0 Td
(vaccine)Tj
3.1974 0 Td
(contains)Tj
3.6057 0 Td
(combined)Tj
4.2463 0 Td
(antigens,)Tj
/F13 1 Tf
3.7927 0 Td
[()]TJ
/F9 1 Tf
(.)Tj
/F13 1 Tf
[()]TJ
/F9 1 Tf
(.)Tj
1.3266 0 Td
(a)Tj
0.6576 0 Td
(minimum)Tj
4.286 0 Td
(of)Tj
-36.0734 -1.2983 Td
(30)Tj
1.1679 0 Td
(international)Tj
5.4141 0 Td
(units)Tj
2.2224 0 Td
(\(I.U.\))Tj
2.4264 0 Td
(diphtheria)Tj
4.3822 0 Td
(toxoid,)Tj
3.0161 0 Td
(a)Tj
0.6576 0 Td
(minimum)Tj
4.286 0 Td
(of)Tj
1.0148 0 Td
(40)Tj
1.1735 0 Td
(I.U.)Tj
1.7461 0 Td
(tetanus)Tj
3.1125 0 Td
(toxoid,)Tj
3.016 0 Td
(25)Tj
/F15 1 Tf
1.1735 0 Td
[()]TJ
/F9 1 Tf
(g)Tj
1.1906 0 Td
(fila-)Tj
-35.9997 -1.3039 Td
(mentous)Tj
3.702 0 Td
(hemagglutinin)Tj
6.0605 0 Td
(\(FHA\),)Tj
3.1066 0 Td
(25)Tj
/F15 1 Tf
1.1679 0 Td
[()]TJ
/F9 1 Tf
(g)Tj
1.1962 0 Td
(pertussis)Tj
3.7077 0 Td
(toxoid,)Tj
3.016 0 Td
(and)Tj
1.7292 0 Td
(8)Tj
/F15 1 Tf
0.6973 0 Td
[()]TJ
/F9 1 Tf
(g)Tj
1.1905 0 Td
(pertactin)Tj
3.8041 0 Td
(P.69)Tj
1.9559 0 Td
(\(PRN\),)Tj
3.0387 0 Td
(all)Tj
-34.3726 -1.2983 Td
(absorbed)Tj
3.8551 0 Td
(to)Tj
1.0261 0 Td
(0.5)Tj
1.4004 0 Td
(mg)Tj
1.5023 0 Td
(of)Tj
1.0148 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 319.2377 622.4314 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 325.0204 624.0755 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(per)Tj
1.5306 0 Td
(dose.)Tj
2.2621 0 Td
(LPS)Tj
1.7971 0 Td
(from)Tj
/F13 1 Tf
2.2054 0 Td
[()]TJ
/F9 1 Tf
(.)Tj
/F13 1 Tf
[()7()10()12()]TJ
/F9 1 Tf
2.33 0 Td
(K12)Tj
1.8482 0 Td
(\(Invivogen;)Tj
4.7962 0 Td
(San)Tj
1.6781 0 Td
(Diego;)Tj
2.8403 0 Td
(Califor-)Tj
-33.7887 -1.2982 Td
(nia;)Tj
1.6951 0 Td
(USA\))Tj
2.4548 0 Td
(was)Tj
1.7008 0 Td
(used)Tj
2.058 0 Td
(as)Tj
1.0147 0 Td
(positive)Tj
3.3166 0 Td
(control.)Tj
3.3447 0 Td
(Non-adsorbed)Tj
/F13 1 Tf
6.0208 0 Td
[()-2()11()20()4()8()11()15()4()4()]TJ
4.1555 0 Td
[( )4()2()-19()7(\n)-4()10()10()-4()]TJ
/F9 1 Tf
3.532 0 Td
(antigens)Tj
3.5659 0 Td
(PRN)Tj
2.1374 0 Td
(and)Tj
-34.9963 -1.3039 Td
(Pertussis)Tj
3.7474 0 Td
(toxin)Tj
2.3074 0 Td
(\(PTx\))Tj
2.5342 0 Td
(were)Tj
2.1259 0 Td
(obtained)Tj
3.7303 0 Td
(from)Tj
2.2053 0 Td
(Biotrend)Tj
3.7418 0 Td
(\(Cologne;)Tj
4.1498 0 Td
(Germany\))Tj
4.3257 0 Td
(and)Tj
1.7291 0 Td
(FHA)Tj
2.1997 0 Td
(was)Tj
1.7008 0 Td
(pur-)Tj
-34.4974 -1.2983 Td
(chased)Tj
2.914 0 Td
(from)Tj
2.2054 0 Td
(Sigma)Tj
2.6815 0 Td
(Aldrich)Tj
3.2712 0 Td
(\(Darmstadt;)Tj
5.0625 0 Td
(Germany\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 559.0488 cm
BT
/F14 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Monocyte)Tj
4.3559 0 Td
(isolation)Tj
3.8551 0 Td
(and)Tj
1.7906 0 Td
(stimulation)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 542.0409 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Blood)Tj
2.5851 0 Td
(from)Tj
2.2054 0 Td
(5)Tj
0.6973 0 Td
(healthy)Tj
3.1351 0 Td
(adult)Tj
2.2507 0 Td
(donors)Tj
3.0444 0 Td
(was)Tj
1.6951 0 Td
(used)Tj
2.0636 0 Td
(for)Tj
1.3833 0 Td
(Peripheral)Tj
4.3653 0 Td
(blood)Tj
2.5115 0 Td
(mononuclear)Tj
5.5785 0 Td
(cell)Tj
1.5591 0 Td
(\(PBMC\))Tj
-33.0744 -1.2982 Td
(isolation)Tj
3.6566 0 Td
(and)Tj
1.7291 0 Td
(subsequent)Tj
4.7055 0 Td
(monocyte)Tj
4.2009 0 Td
(isolation.)Tj
3.8835 0 Td
(PBMCs)Tj
3.2711 0 Td
(were)Tj
2.126 0 Td
(obtained)Tj
3.7304 0 Td
(by)Tj
1.1849 0 Td
(density)Tj
3.101 0 Td
(gradient)Tj
3.5547 0 Td
(cen-)Tj
-35.1437 -1.3039 Td
(trifugation)Tj
4.49 0 Td
(on)Tj
1.2756 0 Td
(Lymphoprep)Tj
5.4141 0 Td
(\(Nycomed;)Tj
4.6772 0 Td
(Zurich;)Tj
3.1634 0 Td
(Switzerland\))Tj
5.2497 0 Td
(at)Tj
0.9525 0 Td
(1,000)Tj
/F13 1 Tf
[()]TJ
/F9 1 Tf
(g)Tj
3.2825 0 Td
(for)Tj
1.3833 0 Td
(30)Tj
1.1735 0 Td
(minutes.)Tj
3.6907 0 Td
(Subse-)Tj
-34.7525 -1.2983 Td
(quently,)Tj
3.4525 0 Td
(monocytes)Tj
4.5581 0 Td
(were)Tj
2.126 0 Td
(isolated)Tj
3.2825 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.474 0 Td
(obtained)Tj
3.7304 0 Td
(PBMC)Tj
2.914 0 Td
(fraction)Tj
3.3562 0 Td
(using)Tj
2.3924 0 Td
(anti-CD14)Tj
4.4674 0 Td
(MACS)Tj
-33.9588 -1.2983 Td
(beads)Tj
2.4717 0 Td
(in)Tj
1.0375 0 Td
(combination)Tj
5.3461 0 Td
(with)Tj
1.9956 0 Td
(MACS)Tj
2.9367 0 Td
(\(Miltenyi)Tj
3.9571 0 Td
(Biotech;)Tj
3.4753 0 Td
(Bergisch)Tj
3.6396 0 Td
(Gladbach;)Tj
4.2576 0 Td
(Germany\).)Tj
4.5525 0 Td
(A)Tj
0.907 0 Td
(purity)Tj
-34.5767 -1.3039 Td
(check)Tj
2.5114 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.474 0 Td
(monocytes)Tj
4.5581 0 Td
(was)Tj
1.7008 0 Td
(performed)Tj
4.4674 0 Td
(by)Tj
1.1848 0 Td
(flow)Tj
1.9446 0 Td
(cytometric)Tj
4.5071 0 Td
(analysis)Tj
3.3278 0 Td
(of)Tj
1.0148 0 Td
(CD14)Tj
2.5682 0 Td
(cell)Tj
1.559 0 Td
(surface)Tj
-31.8328 -1.2982 Td
(expression)Tj
4.4617 0 Td
(and)Tj
1.7347 0 Td
(only)Tj
1.9729 0 Td
(if)Tj
0.7767 0 Td
(the)Tj
1.4684 0 Td
(purity)Tj
2.6589 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(monocyte)Tj
4.2009 0 Td
(population)Tj
4.5921 0 Td
(was)Tj
/F16 1 Tf
0 Tc
1.7008 0 Td
()Tj
/F9 1 Tf
-0.004 Tc
(95%)Tj
2.6985 0 Td
(the)Tj
1.4684 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(used)Tj
2.0636 0 Td
(for)Tj
-36.3229 -1.2983 Td
(proteome)Tj
4.0875 0 Td
(and)Tj
1.7291 0 Td
(transcriptome)Tj
5.862 0 Td
(analysis.)Tj
-10.4824 -1.2983 Td
(The)Tj
1.7858 0 Td
(isolated)Tj
3.2825 0 Td
(monocytes,)Tj
4.7905 0 Td
(600,000)Tj
3.2995 0 Td
(cells/well)Tj
3.8494 0 Td
(were)Tj
2.1203 0 Td
(cultured)Tj
3.566 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.6519 0 Td
(24-wells)Tj
3.4866 0 Td
(plate)Tj
2.1373 0 Td
(\(Corning;)Tj
-31.2035 -1.3039 Td
(Corning;)Tj
3.8154 0 Td
(New)Tj
2.0636 0 Td
(York;)Tj
2.4548 0 Td
(USA\))Tj
2.4547 0 Td
(in)Tj
1.0318 0 Td
(1.5)Tj
1.4004 0 Td
(ml)Tj
1.2869 0 Td
(of)Tj
1.0148 0 Td
(RPMI)Tj
2.6305 0 Td
(\(Gibco/Thermo)Tj
6.5707 0 Td
(Fisher;)Tj
2.9253 0 Td
(Waltham;)Tj
4.1669 0 Td
(Massachu-)Tj
-31.8158 -1.2983 Td
(setts;)Tj
2.1883 0 Td
(USA\))Tj
2.4548 0 Td
(containing)Tj
4.5297 0 Td
(10%)Tj
1.9276 0 Td
(Fetal)Tj
2.1486 0 Td
(Calf)Tj
1.8482 0 Td
(Serum)Tj
2.8403 0 Td
(\(FSC\))Tj
2.5681 0 Td
(\(Hyclone\),)Tj
4.4958 0 Td
(100)Tj
1.6441 0 Td
(units/ml)Tj
3.6226 0 Td
(of)Tj
1.0148 0 Td
(penicillin)Tj
-31.2829 -1.2982 Td
(\(Gibco\))Tj
3.3165 0 Td
(100)Tj
1.6497 0 Td
(units/ml)Tj
3.617 0 Td
(streptomycin)Tj
5.5105 0 Td
(\(Gibco\))Tj
3.3222 0 Td
(and)Tj
1.7291 0 Td
(2.92)Tj
1.8709 0 Td
(mg/ml)Tj
2.9026 0 Td
(L-glutamin)Tj
4.7169 0 Td
(\(Gibco\))Tj
3.3165 0 Td
(\(culture)Tj
-31.9519 -1.3039 Td
(medium\).)Tj
4.1669 0 Td
(Monocytes)Tj
4.6374 0 Td
(were)Tj
2.126 0 Td
(either)Tj
2.5285 0 Td
(left)Tj
1.4796 0 Td
(unstimulated)Tj
5.4992 0 Td
(or)Tj
1.0942 0 Td
(were)Tj
2.126 0 Td
(stimulated)Tj
4.422 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6576 0 Td
(final)Tj
2.0012 0 Td
(concentra-)Tj
-32.7342 -1.2983 Td
(tion)Tj
1.8425 0 Td
(of)Tj
1.0148 0 Td
(0.1)Tj
/F15 1 Tf
1.3946 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5852 0 Td
(LPS,)Tj
2.0182 0 Td
(10)Tj
/F15 1 Tf
1.1736 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5851 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 357.3921 371.3952 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 363.1748 373.0393 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(or)Tj
1.0998 0 Td
(DTaP)Tj
2.5568 0 Td
(containing)Tj
4.5298 0 Td
(a)Tj
0.6519 0 Td
(final)Tj
2.0069 0 Td
(concentration)Tj
5.8167 0 Td
(of)Tj
1.0148 0 Td
(10)Tj
/F15 1 Tf
1.1735 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
-35.1663 -1.2982 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 231.2503 358.3559 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 237.033 360.0566 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(per)Tj
1.5306 0 Td
(stimulation)Tj
4.7962 0 Td
(condition)Tj
4.1102 0 Td
(per)Tj
1.5364 0 Td
(donor.)Tj
2.9027 0 Td
(After)Tj
2.2847 0 Td
(24)Tj
1.1735 0 Td
(and)Tj
1.7291 0 Td
(48)Tj
1.1736 0 Td
(hours)Tj
2.5115 0 Td
(of)Tj
1.0148 0 Td
(stimulation,)Tj
5.0229 0 Td
(culture)Tj
-33.4882 -1.3039 Td
(supernatants)Tj
5.3404 0 Td
(were)Tj
2.1259 0 Td
(collected)Tj
3.7361 0 Td
(for)Tj
1.3889 0 Td
(cytokine)Tj
3.6397 0 Td
(assays)Tj
2.6249 0 Td
(and)Tj
1.7291 0 Td
(monocytes)Tj
4.5637 0 Td
(were)Tj
2.1203 0 Td
(harvested)Tj
4.0422 0 Td
(for)Tj
1.389 0 Td
(targeted)Tj
-32.7002 -1.2982 Td
(transcriptome)Tj
5.8619 0 Td
(and)Tj
1.7292 0 Td
(whole)Tj
2.6078 0 Td
(proteome)Tj
4.0876 0 Td
(analysis.)Tj
3.5489 0 Td
(For)Tj
1.6271 0 Td
(both)Tj
2.0636 0 Td
(proteomics)Tj
4.7111 0 Td
(and)Tj
1.7348 0 Td
(gene)Tj
2.0807 0 Td
(expression)Tj
4.4617 0 Td
(analy-)Tj
-34.5144 -1.2983 Td
(sis,)Tj
1.4343 0 Td
(the)Tj
1.4684 0 Td
(material)Tj
3.5148 0 Td
(of)Tj
1.0148 0 Td
(three)Tj
2.2621 0 Td
(individual)Tj
4.2916 0 Td
(donors)Tj
3.0387 0 Td
(was)Tj
1.7008 0 Td
(available.)Tj
3.8891 0 Td
(LPS)Tj
1.7971 0 Td
(was)Tj
1.6951 0 Td
(used)Tj
2.0637 0 Td
(as)Tj
1.0148 0 Td
(a)Tj
0.6519 0 Td
(positive)Tj
3.3165 0 Td
(control)Tj
3.1238 0 Td
(for)Tj
-36.2775 -1.3039 Td
(the)Tj
1.4684 0 Td
(cell)Tj
1.559 0 Td
(culture)Tj
3.0386 0 Td
(and)Tj
1.7292 0 Td
(only)Tj
1.9785 0 Td
(when)Tj
2.4038 0 Td
(LPS)Tj
1.7915 0 Td
(performed)Tj
4.4673 0 Td
(as)Tj
1.0148 0 Td
(expected,)Tj
3.9515 0 Td
(by)Tj
1.1792 0 Td
(inducing)Tj
3.7984 0 Td
(CD80,)Tj
2.795 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
-32.19 -1.2983 Td
(previously)Tj
4.3482 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(])Tj
1.8539 0 Td
(\()Tj
0.83 0.64 0.02 0 k
(S1)Tj
1.508 0 Td
(Fig)Tj
0 g
(\),)Tj
2.0466 0 Td
(the)Tj
1.4684 0 Td
(samples)Tj
3.3845 0 Td
(were)Tj
2.1203 0 Td
(used.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 269.0078 cm
BT
/F14 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Culture)Tj
3.4441 0 Td
(and)Tj
1.7905 0 Td
(stimulation)Tj
5.0646 0 Td
(of)Tj
1.0678 0 Td
(THP-1)Tj
3.0756 0 Td
(cells)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 252 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(A)Tj
0.9014 0 Td
(human)Tj
3.0784 0 Td
(monocytic)Tj
4.4673 0 Td
(cell)Tj
1.5648 0 Td
(line)Tj
1.7007 0 Td
(THP-1)Tj
2.9877 0 Td
(\(ATCC;)Tj
3.4073 0 Td
(Teddington;)Tj
5.1703 0 Td
(Middlesex;)Tj
4.5808 0 Td
(U.K.\),)Tj
2.6418 0 Td
(was)Tj
1.7008 0 Td
(used)Tj
2.058 0 Td
(to)Tj
1.0261 0 Td
(verify)Tj
-35.2854 -1.2982 Td
(pathways)Tj
3.9287 0 Td
(or)Tj
1.0999 0 Td
(leads)Tj
2.211 0 Td
(identified)Tj
4.0592 0 Td
(in)Tj
1.0375 0 Td
(primary)Tj
3.4525 0 Td
(monocytes,)Tj
/F13 1 Tf
4.7905 0 Td
[()]TJ
/F9 1 Tf
(.)Tj
/F13 1 Tf
[()]TJ
/F9 1 Tf
(.)Tj
1.3267 0 Td
(Inflammasome)Tj
6.2645 0 Td
(activation)Tj
4.1499 0 Td
(and)Tj
1.7348 0 Td
(IL-10)Tj
-34.0552 -1.3039 Td
(secretion.)Tj
1.1962 -1.2983 Td
(THP-1)Tj
2.9877 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.1203 0 Td
(cultured)Tj
3.566 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0204 0 Td
(the)Tj
1.4741 0 Td
(supplier's)Tj
4.0251 0 Td
(protocol.)Tj
3.8211 0 Td
(To)Tj
1.3379 0 Td
(investigate)Tj
4.4618 0 Td
(the)Tj
1.474 0 Td
(IL-1)Tj
/F17 1 Tf
()Tj
/F9 1 Tf
-33.596 -1.2983 Td
(secretion)Tj
3.8324 0 Td
(induced)Tj
3.4582 0 Td
(by)Tj
1.1735 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 315.8929 198.3118 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 321.6188 200.0125 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(or)Tj
1.0884 0 Td
(DTaP,)Tj
2.778 0 Td
(the)Tj
1.4627 0 Td
(cells)Tj
1.9105 0 Td
(were)Tj
2.1146 0 Td
(primed)Tj
3.1351 0 Td
(with)Tj
1.9899 0 Td
(300)Tj
1.6384 0 Td
(ng/ml)Tj
2.6249 0 Td
(phorbol)Tj
3.3959 0 Td
(12-myr-)Tj
-34.2989 -1.3039 Td
(istate)Tj
2.3017 0 Td
(\(PMA\))Tj
3.0331 0 Td
(\(Sigma-Aldrich;)Tj
6.65 0 Td
(Darmstadt,)Tj
4.7224 0 Td
(Germany\))Tj
4.3256 0 Td
(for)Tj
1.389 0 Td
(24)Tj
1.1735 0 Td
(hours.)Tj
2.7383 0 Td
(After)Tj
2.2847 0 Td
(24)Tj
1.1735 0 Td
(hours)Tj
2.5115 0 Td
(of)Tj
1.0148 0 Td
(priming,)Tj
-33.3181 -1.2982 Td
(the)Tj
1.4683 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(placed)Tj
2.795 0 Td
(in)Tj
1.0318 0 Td
(culture)Tj
3.0386 0 Td
(medium)Tj
3.6056 0 Td
(without)Tj
3.3336 0 Td
(PMA)Tj
2.3527 0 Td
(for)Tj
1.389 0 Td
(24)Tj
1.1678 0 Td
(hours)Tj
2.5172 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(].)Tj
2.0749 0 Td
(The)Tj
1.7858 0 Td
(medium)Tj
3.6057 0 Td
(was)Tj
-34.2082 -1.2983 Td
(refreshed)Tj
3.9345 0 Td
(again)Tj
2.3697 0 Td
(and)Tj
1.7291 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(either)Tj
2.5341 0 Td
(left)Tj
1.474 0 Td
(unstimulated)Tj
5.5048 0 Td
(or)Tj
1.0942 0 Td
(stimulated)Tj
4.4277 0 Td
(with)Tj
1.9956 0 Td
(50)Tj
/F15 1 Tf
1.1678 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5909 0 Td
(DTaP)Tj
2.5568 0 Td
(or)Tj
-35.4214 -1.2983 Td
(the)Tj
1.4683 0 Td
(same)Tj
2.2564 0 Td
(concentration)Tj
5.8223 0 Td
(of)Tj
1.0148 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 336.8692 146.3244 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 342.5952 148.0251 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\(based)Tj
2.8176 0 Td
(on)Tj
1.2756 0 Td
(a)Tj
0.6519 0 Td
(dose)Tj
2.0409 0 Td
(response)Tj
3.7304 0 Td
(curve)Tj
2.4208 0 Td
(in)Tj
1.0375 0 Td
(THP-1)Tj
2.982 0 Td
(cells\),)Tj
2.4888 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
-34.7354 -1.3039 Td
(presence)Tj
3.7133 0 Td
(or)Tj
1.0942 0 Td
(absence)Tj
3.3449 0 Td
(of)Tj
1.0148 0 Td
(25)Tj
/F15 1 Tf
1.1678 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5909 0 Td
(of)Tj
1.0148 0 Td
(an)Tj
1.2018 0 Td
(inflammasome)Tj
6.1851 0 Td
(blocker)Tj
3.1975 0 Td
(\(Glybenclamide\))Tj
6.9335 0 Td
(\(Invivogen;)Tj
-31.4586 -1.2982 Td
(San)Tj
1.6781 0 Td
(Diego;)Tj
2.8403 0 Td
(California;)Tj
4.473 0 Td
(USA\))Tj
2.4548 0 Td
(for)Tj
1.389 0 Td
(48)Tj
1.1735 0 Td
(hours.)Tj
-12.8125 -1.2983 Td
(For)Tj
1.6271 0 Td
(determination)Tj
5.9583 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(IL-10-inducing)Tj
6.3268 0 Td
(component,)Tj
5.0343 0 Td
(THP-1)Tj
2.9877 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.1203 0 Td
(stimulated)Tj
4.4221 0 Td
(with)Tj
2.0012 0 Td
(50)Tj
-36.079 -1.3039 Td
(or)Tj
1.0941 0 Td
(100)Tj
/F15 1 Tf
1.6498 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5852 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 284.5984 94.337 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 290.3244 95.9811 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(alone)Tj
2.381 0 Td
(or)Tj
1.0999 0 Td
(the)Tj
1.4683 0 Td
(same)Tj
2.2564 0 Td
(concentrations)Tj
6.1795 0 Td
(of)Tj
1.0148 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 465.5622 94.337 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 471.3448 95.9811 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(in)Tj
1.0374 0 Td
(DTaP)Tj
2.5569 0 Td
(or)Tj
1.0998 0 Td
(with)Tj
1.9956 0 Td
(PTx)Tj
-33.8227 -1.2982 Td
(\(0.165)Tj
/F15 1 Tf
2.6872 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5852 0 Td
(to)Tj
1.0261 0 Td
(5.28)Tj
/F15 1 Tf
1.8709 0 Td
[()]TJ
/F9 1 Tf
(g/ml\),)Tj
3.1578 0 Td
(FHA)Tj
2.1996 0 Td
(\(0.165)Tj
/F15 1 Tf
2.6873 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5851 0 Td
(to)Tj
1.0262 0 Td
(5.28)Tj
/F15 1 Tf
1.8708 0 Td
[()]TJ
/F9 1 Tf
(g/ml\),)Tj
3.1521 0 Td
(PRN)Tj
2.1373 0 Td
(\(0.05)Tj
/F15 1 Tf
2.2166 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5852 0 Td
(to)Tj
1.0261 0 Td
(1.6)Tj
/F15 1 Tf
1.3947 0 Td
[()]TJ
/F9 1 Tf
(g/ml\))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 107.1496 23.6976 re
W* n
q
107.0929 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 350.1354 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Antigens)Tj
4.0394 0 Td
(modulate)Tj
4.259 0 Td
(the)Tj
1.5875 0 Td
(adjuvant-indu)Tj
5.889 0 Td
(ced)Tj
1.8 0 Td
(innate)Tj
2.8914 0 Td
(immune)Tj
3.7205 0 Td
(response)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*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
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
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01978)Tj
4.089 0 Td
(85)Tj
0 g
2.0906 0 Td
(May)Tj
2.0906 0 Td
(29,)Tj
1.5803 0 Td
(2018)Tj
35.2349 0 Td
(3)Tj
0.7654 0 Td
(/)Tj
ET
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
BT
66.402 0 Td
(22)Tj
ET
Q
endstream
endobj
88 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 89 0 R/Contents 90 0 R/TrimBox[0 0 612 792]>>
endobj
89 0 obj
[91 0 R 92 0 R 93 0 R]
endobj
91 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref016)>>
endobj
92 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.s002)>>
endobj
93 0 obj
<>/Border[0 0 0]/A 94 0 R>>
endobj
94 0 obj
<>
endobj
90 0 obj
[95 0 R 96 0 R 97 0 R 98 0 R 99 0 R]
endobj
95 0 obj
<>stream
q
0.83 0.64 0.02 0 k
278.022 465.6189 m
287.5465 465.6189 l
h
f*
356.1449 361.6441 m
390.1039 361.6441 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(alone)Tj
2.3754 0 Td
(all)Tj
1.1508 0 Td
(in)Tj
1.0318 0 Td
(a)Tj
0.6577 0 Td
(twofold)Tj
3.2768 0 Td
(dilution)Tj
3.4072 0 Td
(series.)Tj
2.6532 0 Td
(The)Tj
1.7858 0 Td
(concentration)Tj
5.8167 0 Td
(range)Tj
2.4661 0 Td
(used)Tj
2.058 0 Td
(was)Tj
1.7007 0 Td
(based)Tj
2.4718 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.4684 0 Td
(concen-)Tj
-33.596 -1.3039 Td
(trations)Tj
3.3051 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(individual)Tj
4.2859 0 Td
(antigens)Tj
3.566 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(complete)Tj
3.8834 0 Td
(vaccine.)Tj
3.4186 0 Td
(Synergy)Tj
3.4129 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.6519 0 Td
(complete)Tj
3.8778 0 Td
(vaccine)Tj
3.1975 0 Td
(can-)Tj
-35.6256 -1.2982 Td
(not)Tj
1.5703 0 Td
(be)Tj
1.1509 0 Td
(excluded;)Tj
4.0138 0 Td
(therefore)Tj
3.8551 0 Td
(the)Tj
1.4684 0 Td
(concentration)Tj
5.8166 0 Td
(range)Tj
2.4661 0 Td
(started)Tj
2.931 0 Td
(just)Tj
1.6668 0 Td
(below)Tj
2.5852 0 Td
(the)Tj
1.4683 0 Td
(lowest)Tj
2.7439 0 Td
(vaccine)Tj
3.1918 0 Td
(dose)Tj
-34.9282 -1.2983 Td
(used)Tj
2.0579 0 Td
(in)Tj
1.0375 0 Td
(THP-1)Tj
2.982 0 Td
(cell)Tj
1.559 0 Td
(stimulations.)Tj
5.3802 0 Td
(The)Tj
1.7858 0 Td
(DTaP)Tj
2.5568 0 Td
(stimulations)Tj
5.159 0 Td
(contained)Tj
4.1953 0 Td
(0.33)Tj
1.8765 0 Td
(or)Tj
1.0942 0 Td
(0.66)Tj
/F15 1 Tf
1.8765 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5852 0 Td
(of)Tj
1.0148 0 Td
(PTx)Tj
-35.1607 -1.2983 Td
(or)Tj
1.0941 0 Td
(0.33)Tj
1.8765 0 Td
(or)Tj
1.0942 0 Td
(0.66)Tj
/F15 1 Tf
1.8765 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5852 0 Td
(of)Tj
1.0148 0 Td
(FHA)Tj
2.1997 0 Td
(or)Tj
1.0998 0 Td
(0.2)Tj
1.3947 0 Td
(or)Tj
1.0998 0 Td
(0.4)Tj
/F15 1 Tf
1.3946 0 Td
[()]TJ
/F9 1 Tf
(g/ml)Tj
2.5909 0 Td
(of)Tj
1.0148 0 Td
(PRN,)Tj
2.3584 0 Td
(respectively.)Tj
5.1136 0 Td
(Cells)Tj
2.16 0 Td
(were)Tj
2.1203 0 Td
(stimulated)Tj
-32.0879 -1.3039 Td
(for)Tj
1.3832 0 Td
(48)Tj
1.1736 0 Td
(hours.)Tj
2.7382 0 Td
(Supernatants)Tj
5.4595 0 Td
(were)Tj
2.1203 0 Td
(analyzed)Tj
3.7021 0 Td
(for)Tj
1.3889 0 Td
(the)Tj
1.4684 0 Td
(presence)Tj
3.7133 0 Td
(of)Tj
1.0148 0 Td
(IL-10)Tj
2.3981 0 Td
(by)Tj
1.1849 0 Td
(ELISA.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 613.7007 cm
BT
/F14 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(mRNA)Tj
3.1512 0 Td
(expression)Tj
4.6394 0 Td
(analysis)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 596.6929 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(mRNA)Tj
3.0784 0 Td
(Isolation)Tj
3.7303 0 Td
(from)Tj
2.2054 0 Td
(monocytes)Tj
4.5637 0 Td
(\(from)Tj
2.5455 0 Td
(three)Tj
2.2677 0 Td
(different)Tj
3.651 0 Td
(donors\))Tj
3.3846 0 Td
(was)Tj
1.7007 0 Td
(performed)Tj
4.4674 0 Td
(using)Tj
2.3924 0 Td
(the)Tj
-33.9871 -1.2982 Td
(RNeasy)Tj
3.2484 0 Td
(mini)Tj
2.126 0 Td
(kit)Tj
1.2813 0 Td
(\(Qiagen;)Tj
3.668 0 Td
(Venlo;)Tj
2.8686 0 Td
(The)Tj
1.7802 0 Td
(Netherlands\),)Tj
5.6805 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0262 0 Td
(the)Tj
1.4683 0 Td
(manufacturer's)Tj
6.2645 0 Td
(animal)Tj
-33.6016 -1.2983 Td
(cell)Tj
1.559 0 Td
(spin)Tj
1.9105 0 Td
(protocol.)Tj
3.8268 0 Td
(RNA)Tj
2.262 0 Td
(purity)Tj
2.6532 0 Td
(and)Tj
1.7348 0 Td
(concentration)Tj
5.8167 0 Td
(was)Tj
1.7008 0 Td
(determined)Tj
4.8472 0 Td
(using)Tj
2.3924 0 Td
(spectrophotometric)Tj
-28.7034 -1.3039 Td
(analysis)Tj
3.3278 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(260-nm)Tj
3.3619 0 Td
(and)Tj
1.7348 0 Td
(280-nm)Tj
3.3619 0 Td
(absorbance,)Tj
4.9662 0 Td
(on)Tj
1.2699 0 Td
(the)Tj
1.4741 0 Td
(NanoDrop)Tj
4.575 0 Td
(2000)Tj
2.126 0 Td
(\(Thermo)Tj
3.8154 0 Td
(Fisher;)Tj
2.9254 0 Td
(Wal-)Tj
-35.4215 -1.2983 Td
(tham)Tj
2.296 0 Td
(MA;)Tj
2.0239 0 Td
(USA\).)Tj
2.6816 0 Td
(cDNA)Tj
2.8006 0 Td
(synthesis)Tj
3.8211 0 Td
(of)Tj
1.0148 0 Td
(12)Tj
1.1735 0 Td
(ng)Tj
1.2359 0 Td
(of)Tj
1.0148 0 Td
(RNA)Tj
2.262 0 Td
(was)Tj
1.6951 0 Td
(performed)Tj
4.4731 0 Td
(using)Tj
2.3867 0 Td
(the)Tj
1.4684 0 Td
(RT)Tj
1.457 0 Td
(cDNA)Tj
2.7949 0 Td
(synthe-)Tj
-34.5994 -1.2982 Td
(sis)Tj
1.2075 0 Td
(kit)Tj
1.2813 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.474 0 Td
(RT)Tj
1.4513 0 Td
(preAMP)Tj
3.668 0 Td
(pathway)Tj
3.5717 0 Td
(primer)Tj
2.9876 0 Td
(mix)Tj
1.7745 0 Td
(innate)Tj
2.7383 0 Td
(and)Tj
1.7291 0 Td
(adaptive)Tj
3.5773 0 Td
(immunity)Tj
4.2236 0 Td
(\(both)Tj
2.3981 0 Td
(from)Tj
-33.8114 -1.3039 Td
(Qiagen;)Tj
3.3278 0 Td
(Venlo;)Tj
2.8686 0 Td
(The)Tj
1.7802 0 Td
(Netherlands\);)Tj
5.6863 0 Td
(cDNA)Tj
2.7949 0 Td
(was)Tj
1.7008 0 Td
(frozen)Tj
2.7722 0 Td
(at)Tj
0.9525 0 Td
(-20C.)Tj
2.8006 0 Td
(qPCR)Tj
2.5738 0 Td
(measurements)Tj
6.0321 0 Td
(were)Tj
2.1203 0 Td
(per-)Tj
-35.4101 -1.2983 Td
(formed)Tj
3.1577 0 Td
(using)Tj
2.3868 0 Td
(the)Tj
1.4683 0 Td
(Roche)Tj
2.7269 0 Td
(Light)Tj
2.3131 0 Td
(Cycler)Tj
2.8063 0 Td
(96)Tj
1.1678 0 Td
(\(Roche;)Tj
3.2882 0 Td
(Basel;)Tj
2.4945 0 Td
(Switzerland\))Tj
5.2497 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4684 0 Td
(innate)Tj
2.7382 0 Td
(and)Tj
1.7348 0 Td
(adap-)Tj
-34.7298 -1.2983 Td
(tive)Tj
1.661 0 Td
(immune)Tj
3.6284 0 Td
(response)Tj
3.7303 0 Td
(RT)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 302.57 497.1401 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 308.296 492.7181 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(profiler)Tj
3.2144 0 Td
(arrays)Tj
2.6476 0 Td
(\(Qiagen;)Tj
3.668 0 Td
(Venlo;)Tj
2.8629 0 Td
(The)Tj
1.7859 0 Td
(Netherlands\),)Tj
5.6806 0 Td
(comprising)Tj
4.7678 0 Td
(89)Tj
-35.4554 -1.3039 Td
(functional)Tj
4.3086 0 Td
(genes)Tj
2.4435 0 Td
(and)Tj
1.7291 0 Td
(7)Tj
0.6916 0 Td
(controls.)Tj
3.7133 0 Td
(A)Tj
0.9014 0 Td
(melt)Tj
2.0069 0 Td
(curve)Tj
2.4208 0 Td
(determination)Tj
5.9584 0 Td
(was)Tj
1.6951 0 Td
(included)Tj
3.7133 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4684 0 Td
(measurement)Tj
-32.0879 -1.2982 Td
(for)Tj
1.3833 0 Td
(quality)Tj
2.9593 0 Td
(control)Tj
3.1181 0 Td
([)Tj
0.83 0.64 0.02 0 k
(16)Tj
0 g
(].)Tj
-6.2645 -1.2983 Td
(Gene)Tj
2.3357 0 Td
(expression)Tj
4.4617 0 Td
(of)Tj
1.0148 0 Td
(each)Tj
2.0295 0 Td
(donor)Tj
2.6816 0 Td
(was)Tj
1.7007 0 Td
(normalized)Tj
4.7792 0 Td
(to)Tj
1.0205 0 Td
(the)Tj
1.474 0 Td
(three)Tj
2.262 0 Td
(most)Tj
2.2054 0 Td
(stable)Tj
2.4888 0 Td
(housekeeping)Tj
5.7089 0 Td
(genes)Tj
-35.359 -1.3039 Td
(ACTB,)Tj
2.982 0 Td
(HPRT1)Tj
3.2542 0 Td
(and)Tj
1.7291 0 Td
(RPLP0.)Tj
3.203 0 Td
(After)Tj
2.2847 0 Td
(normalization,)Tj
6.1058 0 Td
(the)Tj
1.4683 0 Td
(fold-change)Tj
4.9606 0 Td
(was)Tj
1.7008 0 Td
(determined,)Tj
5.0797 0 Td
(meaning)Tj
-32.7682 -1.2983 Td
(normalized)Tj
4.7792 0 Td
(gene)Tj
2.0806 0 Td
(expression)Tj
4.4616 0 Td
(\(2)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 321.3921 432.17 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
/F17 1 Tf
( )Tj
/F9 1 Tf
(Ct)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 335.622 427.6913 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\))Tj
0.5612 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(test)Tj
1.5987 0 Td
(sample)Tj
3.0218 0 Td
(divided)Tj
3.2144 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4683 0 Td
(normalized)Tj
4.7792 0 Td
(gene)Tj
2.0806 0 Td
(expres-)Tj
-33.9757 -1.2982 Td
(sion)Tj
1.9049 0 Td
(\(2)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 227.1685 419.1307 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(-)Tj
/F17 1 Tf
( )Tj
/F9 1 Tf
(Ct)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 241.3984 414.7086 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\))Tj
0.5612 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(control)Tj
3.1238 0 Td
(sample.)Tj
3.2428 0 Td
(Fold)Tj
2.0296 0 Td
(change)Tj
3.0444 0 Td
(values)Tj
2.6645 0 Td
(greater)Tj
3.0104 0 Td
(than)Tj
2.0296 0 Td
(one)Tj
1.6951 0 Td
(indicates)Tj
3.7757 0 Td
(an)Tj
1.2019 0 Td
(up-regula-)Tj
-33.0233 -1.2982 Td
(tion)Tj
1.8425 0 Td
(of)Tj
1.0148 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 233.2913 406.148 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 239.074 401.7259 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(or)Tj
1.0998 0 Td
(more.)Tj
2.5625 0 Td
(Fold-change)Tj
5.2044 0 Td
(values)Tj
2.6645 0 Td
(less)Tj
1.6158 0 Td
(than)Tj
2.0239 0 Td
(one)Tj
1.7008 0 Td
(indicate)Tj
3.4128 0 Td
(down-regulation.)Tj
7.149 0 Td
(Genes)Tj
2.6929 0 Td
(were)Tj
-34.0325 -1.3039 Td
(considered)Tj
4.5977 0 Td
(regulated)Tj
3.9401 0 Td
(when)Tj
2.4038 0 Td
(they)Tj
1.9219 0 Td
(differed)Tj
3.3392 0 Td
(a)Tj
0.6519 0 Td
(factor)Tj
2.5455 0 Td
(2)Tj
0.6917 0 Td
(or)Tj
1.0998 0 Td
(more)Tj
2.3414 0 Td
(from)Tj
2.2054 0 Td
(the)Tj
1.4683 0 Td
(control)Tj
3.1238 0 Td
(in)Tj
1.0374 0 Td
(two)Tj
1.7065 0 Td
(out)Tj
1.5533 0 Td
(of)Tj
-34.6277 -1.2982 Td
(three)Tj
2.2621 0 Td
(donors,)Tj
3.271 0 Td
(based)Tj
2.4718 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.4683 0 Td
(SD)Tj
1.4344 0 Td
(between)Tj
3.5262 0 Td
(technical)Tj
3.8098 0 Td
(replicates)Tj
3.9911 0 Td
(of)Tj
1.0148 0 Td
(0.13)Tj
/F13 1 Tf
[()]TJ
/F9 1 Tf
(Ct)Tj
3.2939 0 Td
(values,)Tj
2.897 0 Td
(corresponding)Tj
6.0547 0 Td
(to)Tj
-36.7707 -1.2983 Td
(a)Tj
0.652 0 Td
(coefficient)Tj
4.3766 0 Td
(of)Tj
1.0148 0 Td
(variation)Tj
3.804 0 Td
(\(CV\))Tj
2.2564 0 Td
(of)Tj
1.0148 0 Td
(9.4%)Tj
2.1543 0 Td
(\()Tj
0.83 0.64 0.02 0 k
(S1)Tj
1.5137 0 Td
(Table)Tj
0 g
(\).)Tj
3.0047 0 Td
(This)Tj
1.9899 0 Td
(means)Tj
2.8063 0 Td
(that)Tj
1.7745 0 Td
(a)Tj
0.6576 0 Td
(two-fold)Tj
3.6283 0 Td
(change)Tj
3.0387 0 Td
(in)Tj
1.0375 0 Td
(gene)Tj
-34.7241 -1.3039 Td
(expression)Tj
4.4616 0 Td
(more)Tj
2.3414 0 Td
(than)Tj
2.0296 0 Td
(three)Tj
2.2677 0 Td
(times)Tj
2.3868 0 Td
(exceeds)Tj
3.2712 0 Td
(the)Tj
1.4683 0 Td
(CV)Tj
1.576 0 Td
(and)Tj
1.7292 0 Td
(is)Tj
0.8504 0 Td
(a)Tj
0.6519 0 Td
(meaningful)Tj
4.7905 0 Td
(difference.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 320.9952 cm
BT
/F14 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Protein)Tj
3.359 0 Td
(isolation,)Tj
4.1009 0 Td
(digestion)Tj
4.063 0 Td
(and)Tj
1.7905 0 Td
(labeling)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 303.9874 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(To)Tj
1.3379 0 Td
(isolate)Tj
2.7553 0 Td
(the)Tj
1.4683 0 Td
(proteins)Tj
3.5149 0 Td
(from)Tj
2.2054 0 Td
(the)Tj
1.4683 0 Td
(monocytes)Tj
4.5638 0 Td
(\(from)Tj
2.5455 0 Td
(3)Tj
0.6973 0 Td
(three)Tj
2.2677 0 Td
(different)Tj
3.651 0 Td
(donors\))Tj
3.3845 0 Td
(the)Tj
1.4684 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.1203 0 Td
(incu-)Tj
-35.3704 -1.2982 Td
(bated)Tj
2.4094 0 Td
(with)Tj
1.9956 0 Td
(500)Tj
/F15 1 Tf
1.6497 0 Td
[()]TJ
/F9 1 Tf
(l)Tj
0.9695 0 Td
(of)Tj
1.0148 0 Td
(4)Tj
0.6973 0 Td
(M)Tj
1.1111 0 Td
(guanidine)Tj
/F16 1 Tf
0 Tc
()Tj
/F9 1 Tf
-0.004 Tc
(HCl)Tj
6.1795 0 Td
(in)Tj
1.0375 0 Td
(phosphate)Tj
4.3313 0 Td
(buffer,)Tj
2.8517 0 Td
(pH)Tj
1.5023 0 Td
(7.5,)Tj
1.6271 0 Td
(at)Tj
0.9524 0 Td
(4C)Tj
1.7462 0 Td
(for)Tj
1.3889 0 Td
(two)Tj
1.7065 0 Td
(hours.)Tj
-33.1708 -1.2983 Td
(During)Tj
3.135 0 Td
(incubation,)Tj
4.7736 0 Td
(the)Tj
1.4683 0 Td
(cells)Tj
1.9219 0 Td
(were)Tj
2.1203 0 Td
(subjected)Tj
3.9685 0 Td
(to)Tj
1.0261 0 Td
(a)Tj
0.6576 0 Td
(freeze-thaw)Tj
4.8586 0 Td
(step.)Tj
2.0409 0 Td
(After)Tj
2.2847 0 Td
(the)Tj
1.474 0 Td
(cell)Tj
1.5591 0 Td
(lysis,)Tj
2.1316 0 Td
(50)Tj
/F15 1 Tf
1.1735 0 Td
[()]TJ
/F9 1 Tf
(l)Tj
0.9752 0 Td
(of)Tj
-35.5689 -1.3039 Td
(the)Tj
1.4683 0 Td
(lysate)Tj
2.4378 0 Td
(of)Tj
1.0148 0 Td
(each)Tj
2.0296 0 Td
(sample)Tj
3.0217 0 Td
(was)Tj
1.6951 0 Td
(used)Tj
2.0636 0 Td
(to)Tj
1.0261 0 Td
(determine)Tj
4.32 0 Td
(the)Tj
1.474 0 Td
(protein)Tj
3.1521 0 Td
(concentration)Tj
5.8167 0 Td
(using)Tj
2.3924 0 Td
(the)Tj
1.4683 0 Td
(BCA)Tj
2.1487 0 Td
(pro-)Tj
-35.5292 -1.2983 Td
(tein)Tj
1.7574 0 Td
(assay)Tj
2.2621 0 Td
(\(Pierce)Tj
3.0273 0 Td
(Biotechnology;)Tj
6.2022 0 Td
(Waltham;)Tj
4.1726 0 Td
(Massachusetts;)Tj
6.1454 0 Td
(USA\),)Tj
2.6816 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(manufac-)Tj
-32.9327 -1.2982 Td
(turer's)Tj
2.7892 0 Td
(protocol.)Tj
3.8211 0 Td
(The)Tj
1.7801 0 Td
(remaining)Tj
4.3654 0 Td
(lysed)Tj
2.2337 0 Td
(cells)Tj
1.9218 0 Td
(were)Tj
2.1203 0 Td
(stored)Tj
2.7099 0 Td
(at)Tj
0.9525 0 Td
(-80C.)Tj
-21.4978 -1.3039 Td
(To)Tj
1.3379 0 Td
(reduce)Tj
2.9197 0 Td
(the)Tj
1.4683 0 Td
(guanidine)Tj
/F16 1 Tf
0 Tc
()Tj
/F9 1 Tf
-0.004 Tc
(HCl)Tj
6.1795 0 Td
(concentration,)Tj
6.0434 0 Td
(protein)Tj
3.1521 0 Td
(samples)Tj
3.3846 0 Td
(were)Tj
2.1203 0 Td
(diluted)Tj
3.0387 0 Td
(four)Tj
1.9162 0 Td
(times)Tj
-32.7569 -1.2983 Td
(with)Tj
1.9955 0 Td
(100)Tj
1.6498 0 Td
(mM)Tj
1.9275 0 Td
(phosphate)Tj
4.337 0 Td
(buffer)Tj
2.6249 0 Td
(pH)Tj
1.5023 0 Td
(7.5.)Tj
1.6214 0 Td
(Subsequently,)Tj
5.7487 0 Td
(the)Tj
1.4683 0 Td
(proteins)Tj
3.5149 0 Td
(were)Tj
2.126 0 Td
(digested)Tj
3.5093 0 Td
(with)Tj
2.0012 0 Td
(Lys-C)Tj
-34.0268 -1.2983 Td
(\(Roche\))Tj
3.4015 0 Td
(in)Tj
1.0375 0 Td
(an)Tj
1.2019 0 Td
(enzyme-to-substrate)Tj
8.3905 0 Td
(ratio)Tj
2.0976 0 Td
(of)Tj
1.0148 0 Td
(1:10)Tj
1.8708 0 Td
(\(w/w\))Tj
2.6022 0 Td
(at)Tj
0.9525 0 Td
(37C.)Tj
2.4491 0 Td
(After)Tj
2.2847 0 Td
(4)Tj
0.6973 0 Td
(hours,)Tj
2.7383 0 Td
(fresh)Tj
2.194 0 Td
(Lys-C)Tj
2.5851 0 Td
(was)Tj
-35.5178 -1.3039 Td
(added)Tj
2.6588 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.652 0 Td
(1:10)Tj
1.8708 0 Td
(\(w/w\))Tj
2.6022 0 Td
(enzyme-to-substrate)Tj
8.3905 0 Td
(ratio)Tj
2.0976 0 Td
(for)Tj
1.3833 0 Td
(an)Tj
1.2019 0 Td
(overnight)Tj
4.0875 0 Td
(incubation.)Tj
-24.7859 -1.2983 Td
(Normalization)Tj
6.0717 0 Td
(on)Tj
1.2756 0 Td
(protein)Tj
3.1521 0 Td
(content)Tj
3.2712 0 Td
(was)Tj
1.6951 0 Td
(performed)Tj
4.473 0 Td
(on)Tj
1.2756 0 Td
(aliquots)Tj
3.3676 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(digested)Tj
3.5149 0 Td
(protein)Tj
3.1521 0 Td
(sam-)Tj
-34.9282 -1.2982 Td
(ples)Tj
1.7688 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.4683 0 Td
(6)Tj
0.6974 0 Td
(conditions)Tj
4.473 0 Td
(\(medium,)Tj
4.1726 0 Td
(Al\(OH\))Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 379.1055 159.307 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 384.8881 161.0078 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(and)Tj
1.7291 0 Td
(DTaP)Tj
2.5568 0 Td
(all)Tj
1.1509 0 Td
(at)Tj
0.9524 0 Td
(24)Tj
1.1679 0 Td
(and)Tj
1.7291 0 Td
(48)Tj
1.1735 0 Td
(hours\))Tj
2.8573 0 Td
(per)Tj
1.5307 0 Td
(individual)Tj
-33.3351 -1.2982 Td
(donor.)Tj
2.9083 0 Td
(The)Tj
1.7802 0 Td
(samples)Tj
3.3845 0 Td
(were)Tj
2.1203 0 Td
(labeled)Tj
3.0274 0 Td
(per)Tj
1.5307 0 Td
(condition)Tj
4.1158 0 Td
(on)Tj
1.2756 0 Td
(solid)Tj
2.1316 0 Td
(phase)Tj
2.4831 0 Td
(extraction)Tj
4.2577 0 Td
(\(SPE\))Tj
2.5001 0 Td
(columns)Tj
-31.5153 -1.3039 Td
(\(Waters;)Tj
3.6397 0 Td
(Milford;)Tj
3.5433 0 Td
(MA;)Tj
2.0239 0 Td
(USA\))Tj
2.4548 0 Td
(using)Tj
2.3867 0 Td
(tandem)Tj
3.2769 0 Td
(mass)Tj
2.1939 0 Td
(tag)Tj
1.4173 0 Td
(labeling-6plex)Tj
5.8336 0 Td
(\(TMT\(6\),)Tj
4.0592 0 Td
(Thermo)Tj
3.4809 0 Td
(Fisher\).)Tj
-34.3102 -1.2983 Td
(The)Tj
1.7802 0 Td
(SPE)Tj
1.8255 0 Td
(columns)Tj
3.651 0 Td
(were)Tj
2.1259 0 Td
(equilibrated)Tj
5.0173 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0534 0 Td
(by)Tj
1.1849 0 Td
(the)Tj
1.4683 0 Td
(manufacturer.)Tj
5.9131 0 Td
(Columns)Tj
3.8948 0 Td
(were)Tj
2.1203 0 Td
(washed)Tj
-34.0495 -1.2983 Td
(with)Tj
1.9956 0 Td
(100)Tj
1.6497 0 Td
(mM)Tj
1.9276 0 Td
(phosphate)Tj
4.337 0 Td
(buffer)Tj
2.6248 0 Td
(pH)Tj
1.5024 0 Td
(7.5.)Tj
-12.8409 -1.3039 Td
(The)Tj
1.7858 0 Td
(digested)Tj
3.515 0 Td
(samples)Tj
3.3789 0 Td
(were)Tj
2.1259 0 Td
(loaded)Tj
2.8857 0 Td
(onto)Tj
2.0806 0 Td
(individual)Tj
4.2858 0 Td
(SPE)Tj
1.8255 0 Td
(columns)Tj
3.651 0 Td
(per)Tj
1.5364 0 Td
(condition,)Tj
4.337 0 Td
(using)Tj
2.3924 0 Td
(a)Tj
0.652 0 Td
(vac-)Tj
-35.6482 -1.2982 Td
(uum)Tj
2.092 0 Td
(manifold)Tj
3.8607 0 Td
(\(Waters\),)Tj
3.9798 0 Td
(after)Tj
2.0353 0 Td
(which)Tj
2.6475 0 Td
(samples)Tj
3.3846 0 Td
(were)Tj
2.1202 0 Td
(washed)Tj
3.1804 0 Td
(with)Tj
1.9956 0 Td
(100)Tj
1.6498 0 Td
(mM)Tj
1.9275 0 Td
(phosphate)Tj
4.337 0 Td
(buffer)Tj
2.6249 0 Td
(pH)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 107.1496 23.6976 re
W* n
q
107.0929 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 350.1354 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Antigens)Tj
4.0394 0 Td
(modulate)Tj
4.259 0 Td
(the)Tj
1.5875 0 Td
(adjuvant-indu)Tj
5.889 0 Td
(ced)Tj
1.8 0 Td
(innate)Tj
2.8914 0 Td
(immune)Tj
3.7205 0 Td
(response)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*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
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
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01978)Tj
4.089 0 Td
(85)Tj
0 g
2.0906 0 Td
(May)Tj
2.0906 0 Td
(29,)Tj
1.5803 0 Td
(2018)Tj
35.2349 0 Td
(4)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
96 0 obj
<>stream
endstream
endobj
97 0 obj
<>stream
endstream
endobj
98 0 obj
<>stream
endstream
endobj
99 0 obj
<>stream
BT
66.402 0 Td
(22)Tj
ET
Q
endstream
endobj
100 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 102 0 R/Contents 103 0 R/TrimBox[0 0 612 792]>>
endobj
102 0 obj
[104 0 R 105 0 R 106 0 R]
endobj
104 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref022)>>
endobj
105 0 obj
<>/Border[0 0 0]/Dest(Rpone.0197885.ref023)>>
endobj
106 0 obj
<>/Border[0 0 0]/A 107 0 R>>
endobj
107 0 obj
<>
endobj
103 0 obj
[108 0 R 109 0 R 110 0 R 111 0 R 112 0 R]
endobj
108 0 obj
<>stream
q
0.83 0.64 0.02 0 k
418.6772 530.6457 m
428.2016 530.6457 l
h
f*
393.6189 289.9276 m
403.1433 289.9276 l
h
f*
0 g
1 j
1 J
0 w
10 0 0 10 200.0125 707.4141 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(7.5.)Tj
1.6214 0 Td
(The)Tj
1.7801 0 Td
(labeling)Tj
3.3619 0 Td
(reagent)Tj
3.1804 0 Td
(\(TMT\(6\)\))Tj
4.1726 0 Td
(was)Tj
1.7008 0 Td
(reconstituted)Tj
5.4935 0 Td
(in)Tj
1.0375 0 Td
(acetonitrile)Tj
4.7281 0 Td
(\(AcN\))Tj
2.7439 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0205 0 Td
(the)Tj
-35.0303 -1.3039 Td
(supplier's)Tj
4.0251 0 Td
(protocol)Tj
3.6 0 Td
(\(0.8)Tj
1.7348 0 Td
(mg)Tj
1.508 0 Td
(per)Tj
1.5307 0 Td
(individual)Tj
4.286 0 Td
(label)Tj
2.0749 0 Td
(in)Tj
1.0375 0 Td
(41)Tj
/F15 1 Tf
1.1735 0 Td
[()]TJ
/F9 1 Tf
(l)Tj
0.9695 0 Td
(AcN\),)Tj
2.6305 0 Td
(after)Tj
2.0353 0 Td
(which)Tj
2.6532 0 Td
(the)Tj
1.4683 0 Td
(AcN)Tj
2.0636 0 Td
(concentra-)Tj
-32.7909 -1.2982 Td
(tion)Tj
1.8311 0 Td
(was)Tj
1.6895 0 Td
(reduced)Tj
3.4355 0 Td
(to)Tj
1.0148 0 Td
(a)Tj
0.6463 0 Td
(maximum)Tj
4.3653 0 Td
(of)Tj
1.0035 0 Td
(2.5%)Tj
2.143 0 Td
(\(v/v\))Tj
2.1316 0 Td
(with)Tj
1.9899 0 Td
(100)Tj
1.6384 0 Td
(mM)Tj
1.9219 0 Td
(phosphate)Tj
4.3257 0 Td
(buffer)Tj
2.6135 0 Td
(pH)Tj
1.4967 0 Td
(7.5.)Tj
1.6157 0 Td
(The)Tj
1.7688 0 Td
(indi-)Tj
-35.6312 -1.2983 Td
(vidual)Tj
2.6758 0 Td
(TMT\(6\))Tj
3.4923 0 Td
(labels)Tj
2.4378 0 Td
(were)Tj
2.1203 0 Td
(loaded)Tj
2.8856 0 Td
(onto)Tj
2.0806 0 Td
(the)Tj
1.4684 0 Td
(6)Tj
0.6973 0 Td
(individual)Tj
4.2916 0 Td
(SPE)Tj
1.8199 0 Td
(columns)Tj
3.6566 0 Td
(leaving)Tj
3.0558 0 Td
(0.5)Tj
1.4003 0 Td
(ml)Tj
1.2869 0 Td
(reagent)Tj
-33.3692 -1.2983 Td
(on)Tj
1.2755 0 Td
(top)Tj
1.5421 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(column)Tj
3.2938 0 Td
(for)Tj
1.3833 0 Td
(a)Tj
0.6577 0 Td
(30)Tj
1.1678 0 Td
(minute)Tj
3.1068 0 Td
(incubation.)Tj
4.7735 0 Td
(After)Tj
2.2847 0 Td
(30)Tj
1.1736 0 Td
(minutes,)Tj
3.6906 0 Td
(fresh)Tj
2.1884 0 Td
(label)Tj
2.0806 0 Td
(was)Tj
1.6951 0 Td
(added)Tj
2.6645 0 Td
(for)Tj
-35.4611 -1.3039 Td
(another)Tj
3.3278 0 Td
(30)Tj
1.1735 0 Td
(minutes)Tj
3.464 0 Td
(of)Tj
1.0148 0 Td
(incubation)Tj
4.5524 0 Td
(subsequently,)Tj
5.6295 0 Td
(the)Tj
1.4684 0 Td
(columns)Tj
3.6566 0 Td
(were)Tj
2.1203 0 Td
(washed)Tj
3.1805 0 Td
(with)Tj
1.9956 0 Td
(water)Tj
2.4321 0 Td
(contain-)Tj
-34.0155 -1.2982 Td
(ing)Tj
1.5023 0 Td
(0.5%)Tj
2.1487 0 Td
(formic)Tj
2.8969 0 Td
(acid)Tj
1.8766 0 Td
(\(FA\).)Tj
2.3357 0 Td
(The)Tj
1.7801 0 Td
(individual)Tj
4.2917 0 Td
(stimulation)Tj
4.7905 0 Td
(conditions)Tj
4.4787 0 Td
(per)Tj
1.5307 0 Td
(donor)Tj
2.6816 0 Td
(were)Tj
2.1259 0 Td
(eluted)Tj
2.6646 0 Td
(from)Tj
-35.104 -1.2983 Td
(the)Tj
1.4683 0 Td
(column)Tj
3.2938 0 Td
(with)Tj
1.9956 0 Td
(90%)Tj
1.9275 0 Td
(Acetonitrile)Tj
4.9777 0 Td
(\(AcN\))Tj
2.7439 0 Td
(containing)Tj
4.5297 0 Td
(0.5%)Tj
2.1543 0 Td
(formic)Tj
2.897 0 Td
(acid)Tj
1.8709 0 Td
(\(FA\),)Tj
2.3357 0 Td
(were)Tj
2.126 0 Td
(pooled)Tj
2.948 0 Td
(and)Tj
-35.2684 -1.3039 Td
(then)Tj
2.0182 0 Td
(dried)Tj
2.3357 0 Td
(by)Tj
1.1849 0 Td
(centrifugation)Tj
5.8847 0 Td
(under)Tj
2.6192 0 Td
(reduced)Tj
3.4469 0 Td
(pressure)Tj
3.5773 0 Td
(and)Tj
1.7291 0 Td
(reconstituted)Tj
5.4935 0 Td
(in)Tj
1.0375 0 Td
(Trifluoroacetic)Tj
6.1852 0 Td
(acid)Tj
-35.5122 -1.2983 Td
(\(0.1%TFA\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 561.7133 cm
BT
/F14 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Peptide)Tj
3.411 0 Td
(fractionation)Tj
5.7119 0 Td
(by)Tj
1.2141 0 Td
(SCX)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 544.7055 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(To)Tj
1.3379 0 Td
(purify)Tj
2.6475 0 Td
(and)Tj
1.7292 0 Td
(fractionate)Tj
4.5127 0 Td
(the)Tj
1.4683 0 Td
(labeled)Tj
3.0274 0 Td
(monocyte-derived)Tj
7.5515 0 Td
(digested)Tj
3.5149 0 Td
(protein)Tj
3.1521 0 Td
(samples,)Tj
3.6056 0 Td
(Strong)Tj
2.8857 0 Td
(Cat-)Tj
-35.4328 -1.2982 Td
(ion)Tj
1.542 0 Td
(eXchange)Tj
4.1159 0 Td
(\(SCX\))Tj
2.6929 0 Td
(was)Tj
1.7007 0 Td
(used)Tj
2.058 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
(previously)Tj
4.3483 0 Td
([)Tj
0.83 0.64 0.02 0 k
(22)Tj
0 g
(].)Tj
2.0806 0 Td
(The)Tj
1.7802 0 Td
(system)Tj
2.9367 0 Td
(comprised)Tj
4.439 0 Td
(an)Tj
1.2019 0 Td
(in-house)Tj
-33.9645 -1.3039 Td
(made)Tj
2.4207 0 Td
(Hypercarb)Tj
4.4957 0 Td
(trapping)Tj
3.634 0 Td
(column)Tj
3.2939 0 Td
(\(200)Tj
/F15 1 Tf
1.9899 0 Td
[()]TJ
/F9 1 Tf
(m)Tj
1.5477 0 Td
(I.D.)Tj
1.7461 0 Td
(x)Tj
0.686 0 Td
(5)Tj
0.6916 0 Td
(mm)Tj
1.8596 0 Td
(length,)Tj
2.9536 0 Td
(7)Tj
/F15 1 Tf
0.6974 0 Td
[()]TJ
/F9 1 Tf
(m)Tj
1.5477 0 Td
(particle)Tj
3.2031 0 Td
(size\))Tj
2.0239 0 Td
(and)Tj
1.7291 0 Td
(an)Tj
1.2019 0 Td
(in-)Tj
-35.7219 -1.2983 Td
(house)Tj
2.5681 0 Td
(made)Tj
2.4208 0 Td
(SCX)Tj
2.0126 0 Td
(column)Tj
3.2938 0 Td
(\(200)Tj
/F15 1 Tf
1.9843 0 Td
[()]TJ
/F9 1 Tf
(m)Tj
1.5477 0 Td
(I.D.)Tj
1.7461 0 Td
(x)Tj
0.686 0 Td
(11)Tj
1.1735 0 Td
(cm)Tj
1.4627 0 Td
(length)Tj
2.7269 0 Td
(PolySULFOETHYL)Tj
8.1297 0 Td
(Aspartamide,)Tj
5.5842 0 Td
(5)Tj
/F15 1 Tf
0.6973 0 Td
[()]TJ
/F9 1 Tf
(m,)Tj
-36.0337 -1.2983 Td
(PolyLC\).)Tj
3.753 0 Td
(Elution)Tj
3.1861 0 Td
(was)Tj
1.6951 0 Td
(12)Tj
1.1736 0 Td
(min)Tj
1.8538 0 Td
(at)Tj
0.9524 0 Td
(100%)Tj
2.4038 0 Td
(solvent)Tj
3.0614 0 Td
(A)Tj
0.9014 0 Td
(\(water)Tj
2.7723 0 Td
(+)Tj
0.7993 0 Td
(0.5%)Tj
2.1544 0 Td
(HOAc\))Tj
3.1804 0 Td
(followed)Tj
3.651 0 Td
(by)Tj
1.1849 0 Td
(a)Tj
0.652 0 Td
(16.5)Tj
1.8708 0 Td
(min-)Tj
-35.2457 -1.3039 Td
(ute)Tj
1.4683 0 Td
(linear)Tj
2.5115 0 Td
(gradient)Tj
3.5546 0 Td
(to)Tj
1.0261 0 Td
(100%)Tj
2.4038 0 Td
(solvent)Tj
3.0614 0 Td
(B)Tj
0.7994 0 Td
(\(250)Tj
1.9842 0 Td
(mM)Tj
1.9332 0 Td
(KCl)Tj
1.8028 0 Td
(+)Tj
0.7994 0 Td
(35%)Tj
1.9275 0 Td
(AcN+)Tj
2.6476 0 Td
(0.5%)Tj
2.1543 0 Td
(HOAc)Tj
2.8403 0 Td
(in)Tj
1.0375 0 Td
(water\))Tj
2.7666 0 Td
(and)Tj
1.7348 0 Td
(a)Tj
-36.4533 -1.2982 Td
(second)Tj
3.0103 0 Td
(linear)Tj
2.5115 0 Td
(gradient)Tj
3.5546 0 Td
(of)Tj
1.0148 0 Td
(16.5)Tj
1.8709 0 Td
(min)Tj
1.8595 0 Td
(to)Tj
1.0205 0 Td
(100%)Tj
2.4037 0 Td
(solvent)Tj
3.0614 0 Td
(C)Tj
0.8844 0 Td
(\(500)Tj
1.9843 0 Td
(mM)Tj
1.9332 0 Td
(KCl)Tj
1.8028 0 Td
(+)Tj
0.7994 0 Td
(35%)Tj
1.9275 0 Td
(AcN+)Tj
2.6476 0 Td
(0.5%)Tj
2.1543 0 Td
(HOAc)Tj
-34.4407 -1.2983 Td
(in)Tj
1.0318 0 Td
(water\).)Tj
2.9933 0 Td
(Twenty-six)Tj
4.6772 0 Td
(SCX)Tj
2.0069 0 Td
(fractions)Tj
3.719 0 Td
(were)Tj
2.1146 0 Td
(obtained)Tj
3.7304 0 Td
(and)Tj
1.7235 0 Td
(of)Tj
1.0148 0 Td
(each)Tj
2.0239 0 Td
(4)Tj
/F15 1 Tf
0.6916 0 Td
[()]TJ
/F9 1 Tf
(L)Tj
1.2586 0 Td
(was)Tj
1.6951 0 Td
(subjected)Tj
3.9685 0 Td
(to)Tj
1.0205 0 Td
(nanoscale)Tj
-33.6697 -1.3039 Td
(LC-MS)Tj
3.1407 0 Td
(analysis.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 411.9874 cm
BT
/F14 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(LC-MS/MS)Tj
4.8567 0 Td
(analysis)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 394.9795 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Peptide)Tj
3.2371 0 Td
(separation)Tj
4.3823 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4684 0 Td
(individual)Tj
4.2859 0 Td
(SCX)Tj
2.0126 0 Td
(fractions)Tj
3.719 0 Td
(was)Tj
1.7008 0 Td
(performed)Tj
4.4674 0 Td
(on)Tj
1.2756 0 Td
(a)Tj
0.6519 0 Td
(Proxeon)Tj
3.6057 0 Td
(Easy-nLC)Tj
-31.8215 -1.2982 Td
(1000)Tj
2.1203 0 Td
(system)Tj
2.9366 0 Td
(\(Thermo)Tj
3.8211 0 Td
(Scientific;)Tj
4.1272 0 Td
(San)Tj
1.6781 0 Td
(Jose;)Tj
2.0693 0 Td
(CA;)Tj
1.7915 0 Td
(USA\).)Tj
2.6815 0 Td
(Peptides)Tj
3.6 0 Td
(were)Tj
2.1203 0 Td
(trapped)Tj
3.3109 0 Td
(on)Tj
1.2755 0 Td
(a)Tj
0.652 0 Td
(double-frit-)Tj
-32.1843 -1.2983 Td
(ted)Tj
1.4683 0 Td
(trapping)Tj
3.6396 0 Td
(column)Tj
3.2939 0 Td
(Reprosil)Tj
3.5319 0 Td
(\(Dr.)Tj
1.8936 0 Td
(Maisch;)Tj
3.3505 0 Td
(Ammerbuch;)Tj
5.5502 0 Td
(Germany\))Tj
4.3256 0 Td
(C18;)Tj
2.0636 0 Td
(df)Tj
1.0375 0 Td
(=)Tj
0.7994 0 Td
(3)Tj
/F15 1 Tf
0.6916 0 Td
[()]TJ
/F9 1 Tf
(m,)Tj
1.7745 0 Td
(2)Tj
0.6973 0 Td
(cm)Tj
-34.1175 -1.3039 Td
(length)Tj
/F15 1 Tf
2.7269 0 Td
[()]TJ
/F9 1 Tf
0.7937 0 Td
(100)Tj
/F15 1 Tf
1.644 0 Td
[()]TJ
/F9 1 Tf
(m)Tj
1.5477 0 Td
(I.D.,)Tj
1.9729 0 Td
(made)Tj
2.4208 0 Td
(in-house)Tj
3.7361 0 Td
(and)Tj
1.7291 0 Td
(separated)Tj
4.0138 0 Td
(on)Tj
1.2756 0 Td
(an)Tj
1.2019 0 Td
(in-house-packed)Tj
6.9051 0 Td
(analytical)Tj
4.0082 0 Td
(column)Tj
-33.9758 -1.2983 Td
(Poroshell)Tj
3.9684 0 Td
(\(Agilent;)Tj
3.7191 0 Td
(Waldbron;)Tj
4.5524 0 Td
(Germany\))Tj
4.3256 0 Td
(120)Tj
1.6498 0 Td
(EC-C18;)Tj
3.6396 0 Td
(df)Tj
1.0375 0 Td
(=)Tj
0.7994 0 Td
(2.7)Tj
/F15 1 Tf
1.3946 0 Td
[()]TJ
/F9 1 Tf
(m,)Tj
1.7745 0 Td
(50)Tj
1.1678 0 Td
(cm)Tj
1.4684 0 Td
(length)Tj
/F15 1 Tf
2.7269 0 Td
[()]TJ
/F9 1 Tf
0.788 0 Td
(50)Tj
/F15 1 Tf
1.1736 0 Td
[()]TJ
/F9 1 Tf
(m)Tj
1.5477 0 Td
(I.)Tj
-35.7333 -1.2982 Td
(D.\),)Tj
1.7404 0 Td
(at)Tj
0.9524 0 Td
(a)Tj
0.652 0 Td
(column)Tj
3.2882 0 Td
(temperature)Tj
5.125 0 Td
(of)Tj
1.0091 0 Td
(40C.)Tj
2.4491 0 Td
(Solvent)Tj
3.1748 0 Td
(A)Tj
0.9014 0 Td
(was)Tj
1.6951 0 Td
(MilliQ)Tj
2.88 0 Td
(water)Tj
2.4321 0 Td
(containing)Tj
4.5241 0 Td
(0.1%)Tj
2.1486 0 Td
(FA)Tj
1.4287 0 Td
(and)Tj
1.7291 0 Td
(sol-)Tj
-36.1301 -1.2983 Td
(vent)Tj
1.9445 0 Td
(B)Tj
0.7994 0 Td
(was)Tj
1.7007 0 Td
(0.1%)Tj
2.1544 0 Td
(FA)Tj
1.4286 0 Td
(in)Tj
1.0375 0 Td
(AcN)Tj
2.0636 0 Td
(\(Biosolve\).)Tj
4.473 0 Td
(The)Tj
1.7802 0 Td
(peptides)Tj
3.5546 0 Td
(were)Tj
2.126 0 Td
(separated)Tj
4.0081 0 Td
(in)Tj
1.0375 0 Td
(133)Tj
1.6441 0 Td
(minutes)Tj
3.4696 0 Td
(\(10)Tj
1.508 0 Td
(min-)Tj
-34.7298 -1.3039 Td
(utes)Tj
1.8311 0 Td
(at)Tj
0.9525 0 Td
(2%)Tj
1.4513 0 Td
(B,)Tj
1.0261 0 Td
(from)Tj
2.2054 0 Td
(2%)Tj
1.4513 0 Td
(to)Tj
1.0261 0 Td
(30%)Tj
1.9276 0 Td
(B)Tj
0.7993 0 Td
(in)Tj
1.0375 0 Td
(118)Tj
1.6441 0 Td
(minutes)Tj
3.4696 0 Td
(and)Tj
1.7291 0 Td
(5)Tj
0.6973 0 Td
(minutes)Tj
3.4639 0 Td
(at)Tj
0.9525 0 Td
(70%)Tj
1.9275 0 Td
(B\))Tj
1.1395 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.652 0 Td
(non-linear)Tj
-30.4212 -1.2983 Td
(gradient)Tj
3.5546 0 Td
(optimized)Tj
4.2633 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0478 0 Td
(by)Tj
1.1849 0 Td
(Morus)Tj
/F13 1 Tf
2.88 0 Td
[()8()]TJ
0.907 0 Td
[()11()]TJ
/F9 1 Tf
(.)Tj
1.1679 0 Td
([)Tj
0.83 0.64 0.02 0 k
(23)Tj
0 g
(].)Tj
2.0806 0 Td
(After)Tj
2.2847 0 Td
(133)Tj
1.6441 0 Td
(minutes,)Tj
3.6907 0 Td
(the)Tj
1.4683 0 Td
(system)Tj
2.9424 0 Td
(was)Tj
1.6951 0 Td
(kept)Tj
1.9559 0 Td
(at)Tj
-36.7821 -1.2982 Td
(5%)Tj
1.4513 0 Td
(B)Tj
0.7993 0 Td
(for)Tj
1.389 0 Td
(15)Tj
1.1679 0 Td
(minutes)Tj
3.4696 0 Td
(to)Tj
1.0204 0 Td
(equilibrate)Tj
4.4957 0 Td
(the)Tj
1.4684 0 Td
(column)Tj
3.2938 0 Td
(for)Tj
1.3833 0 Td
(the)Tj
1.474 0 Td
(next)Tj
1.9559 0 Td
(injection.)Tj
3.9855 0 Td
(The)Tj
1.7802 0 Td
(column)Tj
3.2938 0 Td
(effluent)Tj
3.2655 0 Td
(was)Tj
-35.6936 -1.304 Td
(electro-sprayed)Tj
6.3495 0 Td
(directly)Tj
3.2372 0 Td
(into)Tj
1.8368 0 Td
(the)Tj
1.474 0 Td
(MS)Tj
1.5874 0 Td
(using)Tj
2.3868 0 Td
(a)Tj
0.6576 0 Td
(gold-coated)Tj
4.9266 0 Td
(fused)Tj
2.347 0 Td
(silica)Tj
2.2224 0 Td
(tapered)Tj
3.2145 0 Td
(tip)Tj
1.3039 0 Td
(of)Tj
1.0148 0 Td
(5)Tj
/F15 1 Tf
0.6973 0 Td
[()]TJ
/F9 1 Tf
(m,)Tj
1.7688 0 Td
(at)Tj
0.9524 0 Td
(a)Tj
-35.977 -1.2982 Td
(spray)Tj
2.3584 0 Td
(voltage)Tj
3.05 0 Td
(of)Tj
1.0148 0 Td
(1.8)Tj
1.3947 0 Td
(kV.)Tj
-6.6217 -1.2983 Td
(Mass)Tj
2.2733 0 Td
(spectrometric)Tj
5.7203 0 Td
(data)Tj
1.9162 0 Td
(were)Tj
2.1203 0 Td
(acquired)Tj
3.702 0 Td
(on)Tj
1.2756 0 Td
(a)Tj
0.6577 0 Td
(Tribrid-Orbitrap)Tj
7.0072 0 Td
(Fusion)Tj
2.9593 0 Td
(\(Thermo)Tj
3.8154 0 Td
(Fisher)Tj
2.6986 0 Td
(Sci-)Tj
-35.3421 -1.3039 Td
(entific;)Tj
2.9593 0 Td
(San)Tj
1.6781 0 Td
(Jose;)Tj
2.0693 0 Td
(CA;)Tj
1.7915 0 Td
(USA\).)Tj
2.6815 0 Td
(The)Tj
1.7802 0 Td
(full)Tj
1.5307 0 Td
(scan)Tj
1.9842 0 Td
(\(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 381.8834 230.4566 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 385.3984 225.9779 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\))Tj
0.5612 0 Td
(spectra)Tj
3.0501 0 Td
(were)Tj
2.1203 0 Td
(acquired)Tj
3.702 0 Td
(with)Tj
2.0012 0 Td
(a)Tj
0.652 0 Td
(scan)Tj
1.9842 0 Td
(mass)Tj
2.1997 0 Td
(range)Tj
-34.8091 -1.2982 Td
(of)Tj
/F13 1 Tf
1.0148 0 Td
[()]TJ
/F9 1 Tf
1.7178 0 Td
(3501500)Tj
4.0648 0 Td
(at)Tj
0.9525 0 Td
(120,000)Tj
3.2995 0 Td
(resolution)Tj
4.2803 0 Td
(\(FWHM\))Tj
4.0534 0 Td
(with)Tj
1.9955 0 Td
(an)Tj
1.2019 0 Td
(Orbitrap)Tj
3.7361 0 Td
(readout.)Tj
3.5319 0 Td
(For)Tj
1.6271 0 Td
(the)Tj
1.4683 0 Td
(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 543.1748 217.4173 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 548.9007 212.9952 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(the)Tj
-34.8885 -1.2982 Td
(maximum)Tj
4.3709 0 Td
(injection)Tj
3.7588 0 Td
(time)Tj
2.0296 0 Td
(was)Tj
1.7007 0 Td
(50)Tj
1.1736 0 Td
(ms)Tj
1.4003 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(automatic)Tj
4.2293 0 Td
(gain)Tj
1.9332 0 Td
(control)Tj
3.1238 0 Td
(\(AGC\))Tj
2.965 0 Td
(was)Tj
1.6951 0 Td
(set)Tj
1.3039 0 Td
(to)Tj
1.0262 0 Td
(200,000.)Tj
-33.9078 -1.3039 Td
(Top)Tj
1.8538 0 Td
(speed)Tj
2.4718 0 Td
(mode)Tj
2.5001 0 Td
(was)Tj
1.7008 0 Td
(chosen)Tj
3.0104 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6519 0 Td
(duration)Tj
3.7021 0 Td
(of)Tj
1.0147 0 Td
(3)Tj
0.6974 0 Td
(s)Tj
0.5782 0 Td
(where)Tj
2.6532 0 Td
(precursor)Tj
4.0932 0 Td
(ions)Tj
1.9049 0 Td
(with)Tj
1.9956 0 Td
(an)Tj
1.2019 0 Td
(intensity)Tj
/F18 1 Tf
0 Tc
-32.0256 -1.2983 Td
(>)Tj
/F9 1 Tf
-0.004 Tc
(5,000)Tj
3.118 0 Td
(were)Tj
2.1203 0 Td
(selected)Tj
3.3506 0 Td
(for)Tj
1.3833 0 Td
(fragmentation)Tj
5.9243 0 Td
(\(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 376.1007 178.4692 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 379.6157 173.9905 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(\).)Tj
0.788 0 Td
(For)Tj
1.6271 0 Td
(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 417.4299 178.4692 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 423.2125 173.9905 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(charge)Tj
2.8629 0 Td
(states)Tj
2.3981 0 Td
(between)Tj
3.5263 0 Td
(1)Tj
0.6973 0 Td
(and)Tj
1.7291 0 Td
(7)Tj
0.6974 0 Td
(were)Tj
-34.2309 -1.2982 Td
(selected.)Tj
3.5716 0 Td
(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 249.3921 165.4299 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 255.1748 161.0078 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(was)Tj
1.7007 0 Td
(performed)Tj
4.4674 0 Td
(using)Tj
2.3868 0 Td
(Collision-Induced)Tj
7.5061 0 Td
(Dissociation)Tj
5.1987 0 Td
(\(CID\))Tj
2.6362 0 Td
(in)Tj
1.0374 0 Td
(the)Tj
1.4684 0 Td
(linear)Tj
2.5115 0 Td
(ion)Tj
1.542 0 Td
(trap)Tj
-35.9713 -1.2982 Td
(\(LTQ\))Tj
2.7949 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6576 0 Td
(normalized)Tj
4.7734 0 Td
(collision)Tj
3.5943 0 Td
(energy)Tj
2.9084 0 Td
(of)Tj
1.0148 0 Td
(35%.)Tj
2.1543 0 Td
(In)Tj
1.1112 0 Td
(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 423.7228 152.4472 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 429.4488 148.0251 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(the)Tj
1.474 0 Td
(AGC)Tj
2.279 0 Td
(was)Tj
1.7008 0 Td
(set)Tj
1.3039 0 Td
(to)Tj
1.0261 0 Td
(10,000)Tj
2.8233 0 Td
(and)Tj
1.7292 0 Td
(the)Tj
-35.2797 -1.3039 Td
(maximum)Tj
4.371 0 Td
(injection)Tj
3.7587 0 Td
(time)Tj
2.0296 0 Td
(was)Tj
1.7008 0 Td
(35)Tj
1.1735 0 Td
(ms.)Tj
1.6214 0 Td
(Synchronous-Precurso)Tj
9.1501 0 Td
(r-Selection)Tj
4.5524 0 Td
(\(SPS\))Tj
2.4151 0 Td
(was)Tj
1.6951 0 Td
(used)Tj
2.0636 0 Td
(to)Tj
-34.5313 -1.2982 Td
(include)Tj
3.1804 0 Td
(up)Tj
1.2699 0 Td
(to)Tj
1.0205 0 Td
(10)Tj
1.1735 0 Td
(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 280.1196 126.4251 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 285.9023 122.0031 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(fragment)Tj
3.8664 0 Td
(ions)Tj
1.9049 0 Td
(in)Tj
1.0374 0 Td
(MS)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 367.6535 126.4251 cm
BT
/F9 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 371.2251 122.0031 cm
BT
/F9 1 Tf
1 TL
-0.004 Tc
0 0 Td
(.)Tj
0.4422 0 Td
(These)Tj
2.5681 0 Td
(fragment)Tj
3.8721 0 Td
(ions)Tj
1.8992 0 Td
(were)Tj
2.126 0 Td
(further)Tj
3.0274 0 Td
(fragmented)Tj
4.8188 0 Td
(by)Tj
-35.8749 -1.2982 Td
(Higher)Tj
3.0387 0 Td
(energy)Tj
2.9083 0 Td
(Collision)Tj
3.8325 0 Td
(Dissociation)Tj
5.1987 0 Td
(\(HCD\))Tj
3.0669 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6577 0 Td
(normalized)Tj
4.7791 0 Td
(collision)Tj
3.5887 0 Td
(energy)Tj
2.9083 0 Td
(of)Tj
1.0148 0 Td
(50%.)Tj
2.1543 0 Td
(The)Tj
-35.1436 -1.3039 Td
(TMT)Tj
2.3357 0 Td
(reporter)Tj
3.5036 0 Td
(ions)Tj
1.8992 0 Td
(were)Tj
2.126 0 Td
(analyzed)Tj
3.702 0 Td
(in)Tj
1.0318 0 Td
(the)Tj
1.474 0 Td
(Orbitrap)Tj
3.7303 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6576 0 Td
(maximum)Tj
4.371 0 Td
(injection)Tj
3.7587 0 Td
(time)Tj
2.0296 0 Td
(of)Tj
1.0148 0 Td
(120)Tj
1.6498 0 Td
(ms,)Tj
-35.2797 -1.2983 Td
(and)Tj
1.7291 0 Td
(an)Tj
1.2019 0 Td
(AGC)Tj
2.2847 0 Td
(of)Tj
1.0148 0 Td
(100,000.)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 107.1496 23.6976 re
W* n
q
107.0929 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 350.1354 745.7952 cm
BT
/F0 1 Tf
1 TL
-0.005 Tc
0 0 Td
(Antigens)Tj
4.0394 0 Td
(modulate)Tj
4.259 0 Td
(the)Tj
1.5875 0 Td
(adjuvant-indu)Tj
5.889 0 Td
(ced)Tj
1.8 0 Td
(innate)Tj
2.8914 0 Td
(immune)Tj
3.7205 0 Td
(response)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*
0.83 0.64 0.02 0 k
81.5811 34.9228 m
237.8835 34.9228 l
h
f*
0 g
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
(ONE)Tj
2.3669 0 Td
(|)Tj
0.83 0.64 0.02 0 k
0.4748 0 Td
(https://doi.or)Tj
5.4142 0 Td
(g/10.137)Tj
3.8198 0 Td
(1/journal.po)Tj
5.1236 0 Td
(ne.01978)Tj
4.089 0 Td
(85)Tj
0 g
2.0906 0 Td
(May)Tj
2.0906 0 Td
(29,)Tj
1.5803 0 Td
(2018)Tj
35.2349 0 Td
(5)Tj
0.7654 0 Td
(/)Tj
ET
endstream
endobj
109 0 obj
<>stream
endstream
endobj
110 0 obj
<>stream
endstream
endobj
111 0 obj
<>stream
endstream
endobj
112 0 obj
<>stream
BT
66.402 0 Td
(22)Tj
ET
Q
endstream
endobj
113 0 obj
<>/XObject<>>>/CropBox[0 0 612 792]/MediaBox[0 0 612 792]/Parent 13 0 R/Annots 115 0 R/Contents 116 0 R/TrimBox[0 0 612 792]>>
endobj
114 0 obj
<>
endobj
117 0 obj
<>
endobj
118 0 obj
<>stream
%!PS-AdobeFont-1.0
11 dict begin
/FontName /PI_chars_3 def
/FontType 1 def
/FontBBox {-11 -345 1373 718} readonly def
/FontMatrix [0.001 0 0 0.001 0 0] readonly def
/PaintType 0 def
/FontInfo 8 dict dup begin
/FullName (PI_chars_3) readonly def
/BaseFontName (PI_chars_3) def
end def
/Encoding 256 array
0 1 255 {1 index exch /.notdef put} for
dup 33 /exclam put
dup 34 /quotedbl put
dup 35 /numbersign put
dup 36 /dollar put
dup 37 /percent put
dup 38 /ampersand put
dup 39 /quoteright put
dup 40 /parenleft put
dup 41 /parenright put
dup 42 /asterisk put
dup 43 /plus put
dup 44 /comma put
dup 45 /hyphen put
dup 46 /period put
dup 47 /slash put
dup 49 /one put
dup 50 /two put
dup 51 /three put
dup 52 /four put
dup 53 /five put
dup 54 /six put
dup 55 /seven put
dup 56 /eight put
dup 58 /colon put
dup 60 /less put
dup 61 /equal put
dup 62 /greater put
dup 63 /question put
dup 64 /at put
dup 65 /A put
dup 66 /B put
dup 67 /C put
dup 68 /D put
dup 69 /E put
dup 70 /F put
dup 71 /G put
dup 72 /H put
dup 73 /I put
dup 74 /J put
dup 75 /K put
dup 76 /L put
dup 77 /M put
dup 78 /N put
dup 79 /O put
dup 80 /P put
dup 81 /Q put
dup 82 /R put
dup 83 /S put
dup 84 /T put
dup 85 /U put
dup 86 /V put
dup 87 /W put
dup 88 /X put
dup 89 /Y put
dup 90 /Z put
dup 91 /bracketleft put
dup 92 /backslash put
dup 93 /bracketright put
dup 94 /asciicircum put
dup 97 /a put
dup 98 /b put
dup 99 /c put
dup 100 /d put
dup 101 /e put
dup 102 /f put
dup 103 /g put
dup 104 /h put
dup 105 /i put
dup 106 /j put
dup 107 /k put
dup 108 /l put
dup 109 /m put
dup 110 /n put
dup 111 /o put
dup 112 /p put
dup 113 /q put
dup 114 /r put
dup 115 /s put
dup 116 /t put
dup 117 /u put
dup 118 /v put
dup 119 /w put
dup 120 /x put
dup 121 /y put
dup 122 /z put
dup 123 /braceleft put
dup 124 /bar put
dup 125 /braceright put
dup 126 /asciitilde put
readonly def
currentdict end
currentfile eexec
M#1H5 J6o\&rtS(#v\0\V7Fe༙OYx5-cY9^%Y6
(efIOldN"wȰ;)UƽoFg/rvV$JS8_oc@
o9-eGJp}[Do]mN=4*{ܯq=eU^t6¤/{zM4aܪ
Q:д'B-r^ܦ6y?rRnu S\U{Oλrs(Βƚ
#d29%s'ƈT1F8&\0Xd&&Zug'VPd]:>u=Q,?#(CV4'..|ӱ5 h
sȀg)fɘ`UXK^>] q\l#5pE"JO}rquáQBr/E7
QW5\GSlu{7ZH(PSf/O;̟w{ɴ}-K hA8!b!U>`#u?Vlnǝ19V+mg'QmA}S Y$í8n uJ ڒx3>[x0Z]
ͩw4M&1+pkC,sr"%q&OjbH{:5$65*S`cr;%s4Oӆe>T+6~pBOD!$c]"~r#nh6$38i˞[~Z}}rHmte9(1½=j{<\ ;? RA?c{Rf1'VԞ_-Lwo]`.l|zjf"![G1BYro˓9[-+z g!}RN+pX!M~HcFh@g%J6B)6!Lq>TX\ݪGF
PiQݓX~Fs;Ölo4/(C?+,ɖxۼ:xU^)
fd,`*>-:}=q$:\Xzcw1Nf(
ZkEeJXfэi@zXc\{a5qKg\5ݩv\Qe%1>OY̗$Y#P[)."^+{"bP]Bt&zeN6
іijDE% 0;̅
~~lfB>u'i4znym%tfԟ^נBTDS6-ghiWk/(r!!˶fc>趕%U