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Johannes S. Gach, Margaux Bouzin, Marcus P. Wong, Veronika Chromikova, Andrea Gorlani, Kuan-Ting Yu, Brijesh Sharma, Enrico Gratton, Donald N. Forthal
Human immunodeficiency virus type-1 (HIV-1) evades antibody-dependent phagocytosis
www.plospathogens.org
www.plospathogens.org
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0.6577 0 Td
(final)Tj
2.0012 0 Td
(concentration)Tj
5.8167 0 Td
(of)Tj
1.0148 0 Td
(0.01)Tj
1.8765 0 Td
(mg/mL)Tj
3.1918 0 Td
(DEAE-dextran)Tj
6.1965 0 Td
(and)Tj
1.7291 0 Td
(incubated)Tj
4.1782 0 Td
(for)Tj
-35.5404 -1.2982 Td
(48)Tj
1.1678 0 Td
(hours)Tj
2.5171 0 Td
(at)Tj
0.9524 0 Td
(37C.)Tj
2.4491 0 Td
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2.16 0 Td
(were)Tj
2.126 0 Td
(then)Tj
2.0183 0 Td
(washed)Tj
3.1747 0 Td
(with)Tj
2.0013 0 Td
(PBS,)Tj
2.0579 0 Td
(lysed,)Tj
2.4605 0 Td
(and)Tj
1.7291 0 Td
(finally)Tj
2.7042 0 Td
(developed)Tj
4.269 0 Td
(with)Tj
2.0012 0 Td
(luciferase)Tj
-33.7886 -1.2983 Td
(assay)Tj
2.262 0 Td
(reagent)Tj
3.186 0 Td
(\(Promega\))Tj
4.4787 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
(instructions.)Tj
5.2214 0 Td
(Luminescence)Tj
5.9527 0 Td
(in)Tj
1.0318 0 Td
(rela-)Tj
-35.0812 -1.3039 Td
(tive)Tj
1.6611 0 Td
(light)Tj
2.0238 0 Td
(units)Tj
2.2223 0 Td
(\(RLU\))Tj
2.8007 0 Td
(was)Tj
1.6951 0 Td
(measured)Tj
4.1102 0 Td
(using)Tj
2.3924 0 Td
(a)Tj
0.652 0 Td
(Synergy)Tj
3.4129 0 Td
(2)Tj
0.6973 0 Td
(microplate)Tj
4.524 0 Td
(luminometer)Tj
5.4765 0 Td
(\(BioTek\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 606.5574 cm
BT
/F12 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Virus)Tj
2.5417 0 Td
(capture)Tj
3.3827 0 Td
(assay)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 589.5496 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Ninety-six)Tj
4.3936 0 Td
(well)Tj
1.8199 0 Td
(plates)Tj
2.5001 0 Td
(\(Corning\))Tj
4.2689 0 Td
(were)Tj
2.1203 0 Td
(coated)Tj
2.8347 0 Td
(with)Tj
2.0012 0 Td
(250)Tj
1.6441 0 Td
(ng)Tj
1.2359 0 Td
(\(0.005)Tj
2.6872 0 Td
(mg/mL\))Tj
3.532 0 Td
(of)Tj
1.0148 0 Td
(rabbit)Tj
2.6022 0 Td
(anti-human)Tj
-32.6549 -1.2982 Td
(IgG)Tj
1.7404 0 Td
(gamma)Tj
3.1918 0 Td
(chain-specific)Tj
5.743 0 Td
(antibody)Tj
3.7643 0 Td
(\(Rockland\))Tj
4.7055 0 Td
(per)Tj
1.5307 0 Td
(well)Tj
1.8199 0 Td
(and)Tj
1.7291 0 Td
(incubated)Tj
4.1782 0 Td
(over)Tj
1.9786 0 Td
(night)Tj
2.3244 0 Td
(at)Tj
0.9524 0 Td
(4C.)Tj
-33.6583 -1.2983 Td
(Plates)Tj
2.5398 0 Td
(were)Tj
2.1259 0 Td
(then)Tj
2.0183 0 Td
(washed)Tj
3.1748 0 Td
(three)Tj
2.2677 0 Td
(times)Tj
2.3867 0 Td
(with)Tj
1.9956 0 Td
(PBS)Tj
1.8369 0 Td
(and)Tj
1.7291 0 Td
(blocked)Tj
3.3505 0 Td
(with)Tj
2.0013 0 Td
(5%)Tj
1.4513 0 Td
(non-fat)Tj
3.1918 0 Td
(dry)Tj
1.576 0 Td
(milk)Tj
2.0466 0 Td
(in)Tj
1.0375 0 Td
(PBS)Tj
1.8368 0 Td
(at)Tj
-36.5666 -1.3039 Td
(37C.)Tj
2.4491 0 Td
(After)Tj
2.2847 0 Td
(one)Tj
1.7008 0 Td
(hour,)Tj
2.381 0 Td
(plates)Tj
2.5002 0 Td
(were)Tj
2.1203 0 Td
(washed)Tj
3.1804 0 Td
(again)Tj
2.3698 0 Td
(three)Tj
2.262 0 Td
(times)Tj
2.3868 0 Td
(and)Tj
1.7348 0 Td
(incubated)Tj
4.1782 0 Td
(with)Tj
1.9956 0 Td
(100)Tj
1.6441 0 Td
(ng)Tj
1.2359 0 Td
(\(0.002)Tj
-34.4237 -1.2983 Td
(mg/mL\))Tj
3.5319 0 Td
(of)Tj
1.0148 0 Td
(HIV-1-specific)Tj
6.1681 0 Td
(antibodies)Tj
4.3597 0 Td
(for)Tj
1.389 0 Td
(one)Tj
1.7007 0 Td
(hour)Tj
2.1487 0 Td
(at)Tj
0.9524 0 Td
(37C.)Tj
2.4548 0 Td
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2.2847 0 Td
(three)Tj
2.2677 0 Td
(washes,)Tj
3.2315 0 Td
(HIV-1)Tj
2.8516 0 Td
(virions)Tj
-34.3556 -1.2982 Td
(were)Tj
2.1203 0 Td
(diluted)Tj
3.0387 0 Td
(in)Tj
1.0374 0 Td
(DMEM)Tj
3.3109 0 Td
(growth)Tj
3.0727 0 Td
(medium)Tj
3.6057 0 Td
(and)Tj
1.7291 0 Td
(transferred)Tj
4.6374 0 Td
(to)Tj
1.0262 0 Td
(wells)Tj
2.1826 0 Td
(\(10)Tj
1.5137 0 Td
(ng)Tj
1.2303 0 Td
(p24/well\).)Tj
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(4-hours)Tj
3.3391 0 Td
(incubation)Tj
4.5468 0 Td
(at)Tj
0.9524 0 Td
(37C,)Tj
2.4548 0 Td
(plates)Tj
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(were)Tj
2.1204 0 Td
(washed)Tj
3.1804 0 Td
(five)Tj
1.6554 0 Td
(times)Tj
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(with)Tj
1.9956 0 Td
(PBS.)Tj
2.0636 0 Td
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1.1055 0 Td
(a)Tj
0.6576 0 Td
(final)Tj
2.0013 0 Td
(step,)Tj
2.0466 0 Td
(the)Tj
1.4683 0 Td
(infec-)Tj
-34.4747 -1.2983 Td
(tivity)Tj
2.2563 0 Td
(of)Tj
1.0148 0 Td
(captured)Tj
3.7474 0 Td
(virions)Tj
2.9934 0 Td
(was)Tj
1.7007 0 Td
(determined)Tj
4.8529 0 Td
(using)Tj
2.3868 0 Td
(TZM-bl)Tj
3.4299 0 Td
(reporter)Tj
3.5036 0 Td
(cells)Tj
1.9218 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0536 0 Td
(above,)Tj
2.7666 0 Td
(or)Tj
-35.6426 -1.2983 Td
(the)Tj
1.4683 0 Td
(p24)Tj
1.6894 0 Td
(content)Tj
3.2655 0 Td
(captured)Tj
3.7474 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.4683 0 Td
(plate)Tj
2.1373 0 Td
(was)Tj
1.7008 0 Td
(directly)Tj
3.2315 0 Td
(measured)Tj
4.1102 0 Td
(with)Tj
1.9956 0 Td
(a)Tj
0.6519 0 Td
(p24)Tj
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(ELISA)Tj
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(assay.)Tj
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2.1203 0 Td
(analyzed)Tj
3.702 0 Td
(in)Tj
1.0374 0 Td
(triplicate,)Tj
3.9912 0 Td
(and)Tj
1.7291 0 Td
(assays)Tj
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(were)Tj
2.126 0 Td
(repeated)Tj
3.6396 0 Td
(three)Tj
2.2621 0 Td
(times.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 442.0346 cm
BT
/F12 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(HIV-1)Tj
2.9102 0 Td
(antigen)Tj
3.3496 0 Td
(depletion)Tj
4.1623 0 Td
(assay)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 425.0267 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 226.2614 423.3826 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(iGFP/JR-FL)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 264.1322 425.0267 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(virus)Tj
2.1996 0 Td
(preparations)Tj
5.2724 0 Td
(\(50)Tj
1.5137 0 Td
(ng)Tj
1.2302 0 Td
(p24/reaction\))Tj
5.6353 0 Td
(were)Tj
2.1203 0 Td
(incubated)Tj
4.1782 0 Td
(with)Tj
1.9956 0 Td
(10)Tj
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1.1735 0 Td
[()]TJ
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(M)Tj
1.6158 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 559.7858 423.3826 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(MN)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 412.044 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(gp41)Tj
2.1543 0 Td
(recombinant)Tj
5.3688 0 Td
(protein)Tj
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(at)Tj
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(37C.)Tj
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(one)Tj
1.7008 0 Td
(hour,)Tj
2.3811 0 Td
(virions)Tj
2.9934 0 Td
(were)Tj
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(pelleted)Tj
3.3222 0 Td
(\(60)Tj
1.5137 0 Td
(min)Tj
1.8538 0 Td
(at)Tj
0.9525 0 Td
(14,000)Tj
-33.2048 -1.2982 Td
(rpm)Tj
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(and)Tj
1.7291 0 Td
(4C\),)Tj
2.3187 0 Td
(and)Tj
1.7292 0 Td
(the)Tj
1.4683 0 Td
(supernatant)Tj
4.9833 0 Td
(was)Tj
1.7008 0 Td
(carefully)Tj
3.634 0 Td
(removed)Tj
3.7473 0 Td
(and)Tj
1.7292 0 Td
(analyzed)Tj
3.702 0 Td
(for)Tj
1.3889 0 Td
(unbound)Tj
3.9118 0 Td
(HIV-)Tj
-33.9701 -1.3039 Td
(1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 204.7748 384.3779 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(MN)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 219.1748 386.022 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(gp41)Tj
2.1543 0 Td
(protein.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 355.5212 cm
BT
/F12 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(ELISA)Tj
2.9197 0 Td
(for)Tj
1.4693 0 Td
(antigen)Tj
3.3496 0 Td
(depletion)Tj
4.167 0 Td
(assay)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 338.5133 cm
BT
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1 TL
-0.004 Tc
0 0 Td
(ELISA)Tj
2.8289 0 Td
(plates)Tj
2.5002 0 Td
(were)Tj
2.1203 0 Td
(coated)Tj
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(over)Tj
1.9729 0 Td
(night)Tj
2.33 0 Td
(at)Tj
0.9525 0 Td
(4C)Tj
1.7518 0 Td
(with)Tj
1.9956 0 Td
(0.005)Tj
2.347 0 Td
(mg/mL)Tj
3.1918 0 Td
(\(250)Tj
1.9899 0 Td
(ng)Tj
1.2303 0 Td
(per)Tj
1.5363 0 Td
(well\))Tj
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(of)Tj
1.0148 0 Td
(the)Tj
-32.7569 -1.2982 Td
(gp41-specific)Tj
5.4765 0 Td
(mouse)Tj
2.8573 0 Td
(monoclonal)Tj
5.0059 0 Td
(antibody)Tj
3.7644 0 Td
(D50.)Tj
2.1316 0 Td
(Samples)Tj
3.498 0 Td
(consisting)Tj
4.2689 0 Td
(of)Tj
1.0148 0 Td
(supernatant)Tj
4.9833 0 Td
(fluid)Tj
2.0749 0 Td
(from)Tj
-35.0756 -1.2983 Td
(gp41-decorated)Tj
6.4402 0 Td
(virus)Tj
2.1997 0 Td
(and)Tj
1.7291 0 Td
(a)Tj
0.6577 0 Td
(gp41)Tj
2.1543 0 Td
(standard)Tj
3.719 0 Td
(were)Tj
2.126 0 Td
(serially)Tj
3.0217 0 Td
(diluted)Tj
3.0387 0 Td
(1:3)Tj
1.3946 0 Td
(and)Tj
1.7348 0 Td
(added)Tj
2.6589 0 Td
(to)Tj
1.0262 0 Td
(the)Tj
1.4683 0 Td
(blocked)Tj
-33.3692 -1.3039 Td
(\(5%)Tj
1.7914 0 Td
(non-fat)Tj
3.1975 0 Td
(dry)Tj
1.5704 0 Td
(milk)Tj
2.0522 0 Td
(in)Tj
1.0319 0 Td
(PBS)Tj
1.8368 0 Td
(containing)Tj
4.5297 0 Td
(0.05%)Tj
2.6249 0 Td
(Tween-20\))Tj
4.5524 0 Td
(wells.)Tj
2.4094 0 Td
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3.498 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(gp41)Tj
2.1543 0 Td
(stan-)Tj
-34.4463 -1.2983 Td
(dard)Tj
2.0806 0 Td
(\(starting)Tj
3.6056 0 Td
(at)Tj
0.9468 0 Td
(0.003)Tj
2.3527 0 Td
(mg/mL\))Tj
3.532 0 Td
(were)Tj
2.1203 0 Td
(incubated)Tj
4.1782 0 Td
(for)Tj
1.3833 0 Td
(one)Tj
1.7008 0 Td
(hour)Tj
2.1543 0 Td
(at)Tj
0.9524 0 Td
(37C.)Tj
2.4548 0 Td
(Unbound)Tj
4.1216 0 Td
(proteins)Tj
3.5149 0 Td
(were)Tj
-35.0983 -1.2982 Td
(washed)Tj
3.1747 0 Td
(away)Tj
2.2281 0 Td
(and)Tj
1.7291 0 Td
(plates)Tj
2.5001 0 Td
(were)Tj
2.126 0 Td
(further)Tj
3.0274 0 Td
(incubated)Tj
4.1782 0 Td
(with)Tj
1.9956 0 Td
(the)Tj
1.4683 0 Td
(gp41-specific)Tj
5.4765 0 Td
(human)Tj
3.0784 0 Td
(monoclonal)Tj
-30.9824 -1.3039 Td
(antibody)Tj
3.7643 0 Td
(Z13e1)Tj
2.6703 0 Td
(\(0.001)Tj
2.6872 0 Td
(mg/mL\).)Tj
3.7587 0 Td
(After)Tj
2.2847 0 Td
(one)Tj
1.7008 0 Td
(hour,)Tj
2.3754 0 Td
(plates)Tj
2.5001 0 Td
(were)Tj
2.126 0 Td
(washed)Tj
3.1748 0 Td
(and)Tj
1.7348 0 Td
(incubated)Tj
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(with)Tj
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(an)Tj
-34.9509 -1.2983 Td
(HRP-labeled)Tj
5.3234 0 Td
(anti-human)Tj
4.9776 0 Td
(Fab-specific)Tj
5.0116 0 Td
(antibody.)Tj
3.9912 0 Td
(Plates)Tj
2.5398 0 Td
(were)Tj
2.1203 0 Td
(then)Tj
2.0182 0 Td
(incubated)Tj
4.1783 0 Td
(for)Tj
1.389 0 Td
(an)Tj
1.2018 0 Td
(additional)Tj
-32.7512 -1.2983 Td
(hour)Tj
2.1543 0 Td
(at)Tj
0.9524 0 Td
(37C,)Tj
2.4491 0 Td
(washed,)Tj
3.4072 0 Td
(developed)Tj
4.269 0 Td
(\(TMB\),)Tj
3.2088 0 Td
(stopped,)Tj
3.6 0 Td
(and)Tj
1.7291 0 Td
(read)Tj
1.9729 0 Td
(according)Tj
4.1896 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4683 0 Td
(manufacturer's)Tj
-30.4268 -1.3039 Td
(instructions)Tj
5.0002 0 Td
(\(Invitrogen\).)Tj
5.3461 0 Td
(Samples)Tj
3.498 0 Td
(were)Tj
2.1259 0 Td
(analyzed)Tj
3.7021 0 Td
(in)Tj
1.0318 0 Td
(triplicate,)Tj
3.9911 0 Td
(and)Tj
1.7348 0 Td
(experiments)Tj
5.1364 0 Td
(were)Tj
2.1203 0 Td
(per-)Tj
-33.6867 -1.2983 Td
(formed)Tj
3.1577 0 Td
(at)Tj
0.9525 0 Td
(least)Tj
1.9785 0 Td
(twice.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 178.0157 cm
BT
/F12 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(HIV-1)Tj
2.9102 0 Td
(antigen)Tj
3.3496 0 Td
(ELISA)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 161.0078 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(ELISA)Tj
2.8289 0 Td
(plates)Tj
2.5002 0 Td
(were)Tj
2.1203 0 Td
(coated)Tj
2.8346 0 Td
(with)Tj
1.9956 0 Td
(either)Tj
2.5341 0 Td
(D50)Tj
1.9049 0 Td
(antibody)Tj
3.7644 0 Td
(\(250)Tj
1.9899 0 Td
(ng/well\))Tj
3.5036 0 Td
(or)Tj
1.0941 0 Td
(NeutrAvidin)Tj
5.3291 0 Td
(protein)Tj
3.1522 0 Td
(\(250)Tj
-35.5519 -1.2982 Td
(ng/well\).)Tj
3.7303 0 Td
(The)Tj
1.7802 0 Td
(washed)Tj
3.1804 0 Td
(and)Tj
1.7291 0 Td
(blocked)Tj
3.3506 0 Td
(plates)Tj
2.5001 0 Td
(\(5%)Tj
1.7915 0 Td
(non-fat)Tj
3.1975 0 Td
(dry)Tj
1.5703 0 Td
(milk)Tj
2.0523 0 Td
(in)Tj
1.0318 0 Td
(PBS)Tj
1.8369 0 Td
(containing)Tj
4.5297 0 Td
(0.05%)Tj
-32.2807 -1.3039 Td
(Tween-20\))Tj
4.5524 0 Td
(were)Tj
2.1203 0 Td
(further)Tj
3.0273 0 Td
(incubated)Tj
4.1783 0 Td
(with)Tj
1.9956 0 Td
(0.001)Tj
2.3527 0 Td
(mg/mL)Tj
3.1918 0 Td
(of)Tj
1.0148 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 450.5952 133.3417 cm
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1.7008 0 Td
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(and)Tj
1.7291 0 Td
(reported)Tj
3.6623 0 Td
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(number)Tj
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(of)Tj
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(viral)Tj
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(particles)Tj
3.5603 0 Td
(based)Tj
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1.2756 0 Td
(a)Tj
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(standard)Tj
3.7247 0 Td
(curve)Tj
2.4208 0 Td
(generated)Tj
4.1328 0 Td
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1.1849 0 Td
(serial)Tj
2.3187 0 Td
(dilu-)Tj
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(extracted)Tj
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2.2054 0 Td
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(dilutions)Tj
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(and)Tj
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(tran-)Tj
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(as)Tj
1.0148 0 Td
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Q
q
1 j
1 J
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11.9999 0 0 11.9999 200.0125 563.9244 cm
BT
/F12 1 Tf
1 TL
-0.0033 Tc
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5.6315 0 Td
(of)Tj
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(aggregated)Tj
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(HIV-1)Tj
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Q
q
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1 J
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10 0 0 10 200.0125 546.9165 cm
BT
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1 TL
-0.004 Tc
0 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 226.2614 545.2724 cm
BT
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1 TL
-0.0053 Tc
0 0 Td
(iGFP/JR-FL)Tj
ET
Q
q
1 j
1 J
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BT
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1 TL
-0.004 Tc
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(virions)Tj
2.999 0 Td
(\(50)Tj
1.508 0 Td
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1 J
0 w
7.4999 0 0 6.6659 218.3811 506.2677 cm
BT
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1 TL
-0.0053 Tc
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(2)Tj
ET
Q
q
1 j
1 J
0 w
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BT
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1 TL
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(anti-human)Tj
4.9776 0 Td
(F\(ab'\))Tj
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Q
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1 J
0 w
7.4999 0 0 6.6659 297.5244 506.2677 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
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BT
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1 TL
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(antibody)Tj
3.7643 0 Td
(\(Jackson)Tj
3.6624 0 Td
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1.0148 0 Td
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(uptake)Tj
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(performed)Tj
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(described)Tj
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q
1 j
1 J
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11.9999 0 0 11.9999 200.0125 468.4535 cm
BT
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1 TL
-0.0033 Tc
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1.8803 0 Td
(surface-decorated)Tj
7.6583 0 Td
(HIV-1)Tj
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(phagocytosis)Tj
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(assay)Tj
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Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 451.4456 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 226.2614 449.8015 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(iGFP/JR-FL)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 264.1322 451.4456 cm
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1 TL
-0.004 Tc
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(virions)Tj
2.999 0 Td
(were)Tj
2.1203 0 Td
(pelleted)Tj
3.3278 0 Td
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(one)Tj
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(hour)Tj
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(14,000)Tj
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(CEM.NKr-)Tj
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(CCR5)Tj
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(target)Tj
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(cells)Tj
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(PBS)Tj
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(and)Tj
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(transferred)Tj
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1.0261 0 Td
(a)Tj
0.652 0 Td
(96-well)Tj
3.1238 0 Td
(flat-bottom)Tj
4.7735 0 Td
(plate)Tj
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7.4999 0 0 6.6659 565.2283 442.885 cm
BT
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1 TL
-0.0053 Tc
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(5)Tj
ET
Q
q
1 j
1 J
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1 TL
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(cells/well\))Tj
4.1895 0 Td
(and)Tj
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(500)Tj
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(p24)Tj
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(\(~)Tj
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(1.25)Tj
1.8765 0 Td
(x)Tj
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(10)Tj
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1 j
1 J
0 w
7.4999 0 0 6.6659 452.6362 429.9023 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(4)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 458.4188 425.4803 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(virions)Tj
2.9933 0 Td
(per)Tj
1.5364 0 Td
(target)Tj
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(cell\))Tj
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1.0148 0 Td
(0.1)Tj
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1.3833 0 Td
(2)Tj
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(hours)Tj
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(2500)Tj
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(10C.)Tj
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(virus-coated)Tj
5.159 0 Td
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(ice-cold)Tj
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(S1)Tj
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1 J
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11.9999 0 0 11.9999 200.0125 281.9905 cm
BT
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1 TL
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(ImageStream)Tj
5.7921 0 Td
(cytometry)Tj
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Q
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1 TL
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BT
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1 TL
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(iGFP/JR-FL)Tj
ET
Q
q
1 j
1 J
0 w
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BT
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1 TL
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(internalization)Tj
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(19)Tj
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1.1112 0 Td
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7.4999 0 0 6.6659 380.0692 243.4393 cm
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1 TL
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(6)Tj
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(Millipore\),)Tj
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(algorithms,)Tj
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(internalization)Tj
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(0.3)Tj
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1.0261 0 Td
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3.2541 0 Td
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6.0207 0 Td
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(mediate)Tj
3.4129 0 Td
(detectable)Tj
4.2236 0 Td
(internalization)Tj
6.0944 0 Td
(of)Tj
1.0148 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 373.7763 630.4251 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(iGFP/JR-FL)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 411.5905 632.0692 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(virions)Tj
2.999 0 Td
(in)Tj
1.0318 0 Td
(U937)Tj
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(cells)Tj
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(\()Tj
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(1B)Tj
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(monocytes)Tj
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(\()Tj
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(Fig)Tj
1.8198 0 Td
(1C)Tj
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q
1 j
1 J
0 w
7.4999 0 0 6.6659 446.6267 604.4031 cm
BT
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1 TL
-0.0053 Tc
0 0 Td
(JR-FL)Tj
ET
Q
q
1 j
1 J
0 w
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BT
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1 TL
-0.004 Tc
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(considered)Tj
4.6034 0 Td
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Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 461.9905 591.4204 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(iGFP/NL4-3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 499.1811 593.0645 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(,)Tj
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(a)Tj
0.652 0 Td
(neutralization)Tj
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(R4)Tj
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(1)Tj
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(U937)Tj
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(Fig)Tj
1.8198 0 Td
(1D)Tj
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(\).)Tj
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(\(PS\))Tj
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(21)Tj
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(in)Tj
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(all)Tj
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(phagocytic)Tj
4.5354 0 Td
(cell)Tj
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(\()Tj
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(Fig)Tj
1.8198 0 Td
(1)Tj
0 g
(\).)Tj
1.2585 0 Td
(A)Tj
0.9071 0 Td
(F\(ab'\))Tj
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Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 550.3748 539.433 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(2)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 556.1007 541.0771 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(ver-)Tj
-35.6085 -1.2982 Td
(sion)Tj
1.9048 0 Td
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1.0148 0 Td
(PGN635)Tj
3.6567 0 Td
(reduced)Tj
3.4412 0 Td
(the)Tj
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(phagocytic)Tj
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(score)Tj
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(by)Tj
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(88%)Tj
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(implicating)Tj
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(an)Tj
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(Fc-dependent)Tj
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(process)Tj
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(in)Tj
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(phagocytosis)Tj
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(the)Tj
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(antibody)Tj
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(further)Tj
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(that)Tj
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(antibodies)Tj
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(phagocytosis)Tj
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(HIV-1)Tj
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(our)Tj
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(findings)Tj
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1.0262 0 Td
(HIV-1)Tj
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(grown)Tj
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(in)Tj
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(PBMCs,)Tj
3.5036 0 Td
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(HIV-1)Tj
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Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 465.7889 474.4062 cm
BT
/F5 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(JRFL)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 483.0803 476.107 cm
BT
/F5 1 Tf
1 TL
-0.004 Tc
0 0 Td
(in)Tj
1.0318 0 Td
(an)Tj
1.2075 0 Td
(experimental)Tj
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2.9821 0 Td
(to)Tj
1.0261 0 Td
(that)Tj
1.7745 0 Td
(used)Tj
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(with)Tj
1.9956 0 Td
(the)Tj
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(GFP-expressing)Tj
6.5764 0 Td
(virions.)Tj
3.2201 0 Td
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(because)Tj
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(artifacts)Tj
3.3959 0 Td
(intro-)Tj
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1.1849 0 Td
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(virus)Tj
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(\(see)Tj
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(below\),)Tj
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1.3322 0 Td
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(the)Tj
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(virus)Tj
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(U937)Tj
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(lysates.)Tj
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(none)Tj
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(of)Tj
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(the)Tj
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(HIVIG)Tj
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(\(anti-PS\))Tj
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(hand,)Tj
2.4831 0 Td
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(ADP)Tj
2.1997 0 Td
(\()Tj
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(S3)Tj
1.5137 0 Td
(Fig)Tj
0 g
(\).)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 381.6 cm
BT
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1 TL
-0.0033 Tc
0 0 Td
(HIV-1-specific)Tj
6.3638 0 Td
(antibodies)Tj
4.5685 0 Td
(efficiently)Tj
4.389 0 Td
(capture)Tj
3.3827 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
8.9997 0 0 7.999 456.7181 379.6157 cm
BT
/F12 1 Tf
1 TL
-0.0044 Tc
0 0 Td
(iGFP/JR-FL)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 503.6598 381.6 cm
BT
/F12 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(and)Tj
1.7905 0 Td
(HIV-)Tj
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(1)Tj
ET
Q
q
1 j
1 J
0 w
8.9997 0 0 7.999 205.9653 365.6125 cm
BT
/F12 1 Tf
1 TL
-0.0044 Tc
0 0 Td
(iGFP/NL4-3)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 254.3811 367.5968 cm
BT
/F12 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(virions)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 350.5889 cm
BT
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1 TL
0.003 Tc
0 0 Td
(Although)Tj
4.0591 0 Td
(unlikely)Tj
3.4866 0 Td
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1.406 0 Td
(neutralizing)Tj
5.1137 0 Td
(antibodies,)Tj
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(the)Tj
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(lack)Tj
1.8482 0 Td
(of)Tj
1.0261 0 Td
(ADP)Tj
2.2223 0 Td
(could)Tj
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(be)Tj
1.1622 0 Td
(explained)Tj
4.1386 0 Td
(by)Tj
1.1962 0 Td
(insuffi-)Tj
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(cient)Tj
2.2166 0 Td
(capture)Tj
3.2655 0 Td
(of)Tj
1.0262 0 Td
(virus.)Tj
2.4718 0 Td
(Thus,)Tj
2.5058 0 Td
(we)Tj
1.3436 0 Td
(examined)Tj
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(the)Tj
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(antibodies)Tj
4.4277 0 Td
(for)Tj
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(their)Tj
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(ability)Tj
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(to)Tj
1.0431 0 Td
(capture)Tj
3.2655 0 Td
(HIV-)Tj
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(1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 204.8314 322.9228 cm
BT
/F5 1 Tf
1 TL
0.004 Tc
0 0 Td
(iGFP/JR-FL)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 243.3826 324.6236 cm
BT
/F5 1 Tf
1 TL
0.003 Tc
0 0 Td
(and)Tj
1.7517 0 Td
(HIV-1)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 287.5464 322.9228 cm
BT
/F5 1 Tf
1 TL
0.004 Tc
0 0 Td
(iGFP/NL4-3)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 325.4173 324.6236 cm
BT
/F5 1 Tf
1 TL
0.003 Tc
0 0 Td
(.)Tj
0.4535 0 Td
(For)Tj
1.6497 0 Td
(this)Tj
1.7008 0 Td
(approach,)Tj
4.2406 0 Td
(we)Tj
1.338 0 Td
(coated)Tj
2.8799 0 Td
(ELISA)Tj
2.863 0 Td
(plates)Tj
2.5399 0 Td
(with)Tj
2.0239 0 Td
(an)Tj
1.2189 0 Td
(anti-)Tj
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(human)Tj
3.1124 0 Td
(IgG)Tj
1.7631 0 Td
(antibody)Tj
3.8154 0 Td
(and)Tj
1.7518 0 Td
(stepwise)Tj
3.6112 0 Td
(added)Tj
2.6986 0 Td
(the)Tj
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(HIV-specific)Tj
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(antibodies)Tj
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(and)Tj
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(the)Tj
1.491 0 Td
(GFP-express-)Tj
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(ing)Tj
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(virus.)Tj
2.4718 0 Td
(We)Tj
1.6214 0 Td
(then)Tj
2.0466 0 Td
(either)Tj
2.5739 0 Td
(added)Tj
2.6928 0 Td
(TZM-bl)Tj
3.4752 0 Td
(cells)Tj
1.9502 0 Td
(to)Tj
1.0432 0 Td
(measure)Tj
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(the)Tj
1.491 0 Td
(quantity)Tj
3.6113 0 Td
(of)Tj
1.0318 0 Td
(infectious)Tj
4.201 0 Td
(virus)Tj
ET
Q
q
0.45 0.36 0.35 0.01 k
1 j
1 J
0 w
200.0001 270.5017 m
576 270.5017 l
576 270.0017 l
200.0001 270.0017 l
f*
442.2657 270.0017 m
442.2657 257.5018 l
441.7656 257.5018 l
441.7656 270.0017 l
f*
441.7656 257.5018 m
200.0001 257.5018 l
200.0001 258.0018 l
441.7656 258.0018 l
f*
575.9997 257.5018 m
442.2657 257.5018 l
442.2657 258.0018 l
575.9997 258.0018 l
f*
442.2657 257.5018 m
442.2657 220.002 l
441.7656 220.002 l
441.7656 257.5018 l
f*
441.7656 220.002 m
200.0001 220.002 l
200.0001 220.502 l
441.7656 220.502 l
f*
575.9997 245.0018 m
442.2657 245.0018 l
442.2657 245.5019 l
575.9997 245.5019 l
f*
575.9997 232.5019 m
442.2657 232.5019 l
442.2657 233.0019 l
575.9997 233.0019 l
f*
575.9997 220.002 m
442.2657 220.002 l
442.2657 220.502 l
575.9997 220.502 l
f*
442.2657 220.002 m
442.2657 207.502 l
441.7656 207.502 l
441.7656 220.002 l
f*
441.7656 207.502 m
200.0001 207.502 l
200.0001 208.002 l
441.7656 208.002 l
f*
575.9997 207.502 m
442.2657 207.502 l
442.2657 208.002 l
575.9997 208.002 l
f*
442.2657 207.502 m
442.2657 182.5021 l
441.7656 182.5021 l
441.7656 207.502 l
f*
441.7656 182.5021 m
200.0001 182.5021 l
200.0001 183.0022 l
441.7656 183.0022 l
f*
575.9997 195.0021 m
442.2657 195.0021 l
442.2657 195.5021 l
575.9997 195.5021 l
f*
575.9997 182.5021 m
442.2657 182.5021 l
442.2657 183.0022 l
575.9997 183.0022 l
f*
442.2657 182.5021 m
442.2657 170.0022 l
441.7656 170.0022 l
441.7656 182.5021 l
f*
441.7656 170.0022 m
200.0001 170.0022 l
200.0001 170.5022 l
441.7656 170.5022 l
f*
575.9997 170.0022 m
442.2657 170.0022 l
442.2657 170.5022 l
575.9997 170.5022 l
f*
442.2657 170.0022 m
442.2657 120.0025 l
441.7656 120.0025 l
441.7656 170.0022 l
f*
441.7656 120.0025 m
200.0001 120.0025 l
200.0001 120.5025 l
441.7656 120.5025 l
f*
575.9997 157.5023 m
442.2657 157.5023 l
442.2657 158.0023 l
575.9997 158.0023 l
f*
575.9997 145.0023 m
442.2657 145.0023 l
442.2657 145.5024 l
575.9997 145.5024 l
f*
575.9997 132.5024 m
442.2657 132.5024 l
442.2657 133.0024 l
575.9997 133.0024 l
f*
575.9997 120.0025 m
442.2657 120.0025 l
442.2657 120.5025 l
575.9997 120.5025 l
f*
442.2657 120.0025 m
442.2657 107.5025 l
441.7656 107.5025 l
441.7656 120.0025 l
f*
441.7656 107.5025 m
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