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Luisina Martorelli, Sergio Garbaccio, Daniel A. Vilte, Adriana A. Albanese, María P. Mejías, Marina S. Palermo, Elsa C. Mercado, Cristina E. Ibarra, Angel A. Cataldi
Immune Response in Calves Vaccinated with Type Three Secretion System Antigens and Shiga Toxin 2B Subunit of Escherichia coli O157:H7
www.plosone.org
www.plosone.org
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1.6724 0 Td
(antigen)Tj
3.2031 0 Td
(on)Tj
1.2756 0 Td
(the)Tj
1.474 0 Td
(response)Tj
3.7304 0 Td
(to)Tj
1.0261 0 Td
(IntiminC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 518.57 692.7307 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(280)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 531.4393 694.3748 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(and)Tj
1.7291 0 Td
(EspB,)Tj
-34.8715 -1.2982 Td
(as)Tj
1.0148 0 Td
(well)Tj
1.8198 0 Td
(as)Tj
1.0148 0 Td
(to)Tj
1.0261 0 Td
(evaluate)Tj
3.4583 0 Td
(the)Tj
1.474 0 Td
(ability)Tj
2.6929 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(antibodies)Tj
4.3597 0 Td
(generated)Tj
4.1385 0 Td
(to)Tj
1.0262 0 Td
(inhibit)Tj
2.9026 0 Td
(virulence)Tj
3.9061 0 Td
(traits)Tj
2.2506 0 Td
(of)Tj
/F13 1 Tf
1.0148 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
0.9922 0 Td
[()7( )10(\n)12()]TJ
/F8 1 Tf
-35.5745 -1.2983 Td
(0157:H7)Tj
/F13 1 Tf
3.5886 0 Td
[()12()]TJ
1.0148 0 Td
[()-13()15()3( )]TJ
/F8 1 Tf
(.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 642.1039 cm
BT
/F3 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Materials)Tj
4.5071 0 Td
(and)Tj
1.989 0 Td
(Methods)Tj
/F0 1 Tf
-6.4961 -1.4976 Td
(Animals)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 607.1244 cm
BT
/F8 1 Tf
1 TL
-0.014 Tc
0 0 Td
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3.6623 0 Td
(four-month-old)Tj
6.4573 0 Td
(conventionally)Tj
5.9867 0 Td
(reared)Tj
2.7156 0 Td
(Holstein)Tj
3.5546 0 Td
(Friesian)Tj
3.3449 0 Td
(male)Tj
2.1033 0 Td
(calves)Tj
2.5001 0 Td
(were)Tj
2.0863 0 Td
(obtained)Tj
-32.4111 -1.2982 Td
(from)Tj
2.1656 0 Td
(a)Tj
0.6406 0 Td
(dairy)Tj
2.228 0 Td
(farm)Tj
2.092 0 Td
(in)Tj
1.0148 0 Td
(Buenos)Tj
3.1068 0 Td
(Aires)Tj
2.279 0 Td
(Province,)Tj
3.9118 0 Td
(Argentina)Tj
4.1726 0 Td
(and)Tj
1.6951 0 Td
(housed)Tj
3.0387 0 Td
(at)Tj
0.9297 0 Td
(the)Tj
1.44 0 Td
(Instituto)Tj
3.5773 0 Td
(Nacional)Tj
3.7361 0 Td
(de)Tj
-36.0281 -1.3039 Td
(Tecnolog)Tj
0 Tc
3.5886 0.051 Td
()Tj
-0.014 Tc
0.3175 -0.051 Td
(a)Tj
0.6463 0 Td
(Agropecuaria)Tj
5.5275 0 Td
(\(INTA\))Tj
3.2088 0 Td
(research)Tj
3.4752 0 Td
(centre.)Tj
2.8687 0 Td
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3.5093 0 Td
(were)Tj
2.0806 0 Td
(selected)Tj
3.2711 0 Td
(on)Tj
1.2529 0 Td
(the)Tj
1.44 0 Td
(basis)Tj
2.0976 0 Td
(of)Tj
0.9978 0 Td
(absence)Tj
-34.2819 -1.2983 Td
(of)Tj
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0.9921 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
0.9751 0 Td
[()7( )10(\n)12()]TJ
/F8 1 Tf
1.5137 0 Td
(O157:H7)Tj
3.7927 0 Td
(by)Tj
1.1622 0 Td
(enrichment)Tj
4.7678 0 Td
(of)Tj
0.9922 0 Td
(rectoanal)Tj
3.8154 0 Td
(mucosal)Tj
3.4696 0 Td
(swabs)Tj
2.5171 0 Td
(followed)Tj
3.566 0 Td
(by)Tj
1.1622 0 Td
(immunomagnetic)Tj
7.2453 0 Td
(sep-)Tj
-35.9714 -1.2983 Td
(aration)Tj
3.0103 0 Td
(following)Tj
3.8891 0 Td
(manufacturer`s)Tj
6.1342 0 Td
(instructions)Tj
4.8812 0 Td
(\(Dynabeads)Tj
4.8869 0 Td
(anti-)Tj
/F13 1 Tf
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
[()7( )10(\n)12()]TJ
/F8 1 Tf
4.1215 0 Td
(O157,)Tj
2.5682 0 Td
(Invitrogen)Tj
4.3427 0 Td
(Dynal)Tj
-33.8341 -1.3039 Td
(AS,)Tj
1.576 0 Td
(Oslo,)Tj
2.2564 0 Td
(Norway\),)Tj
3.8891 0 Td
(and)Tj
1.7008 0 Td
(low)Tj
1.627 0 Td
(levels)Tj
2.3188 0 Td
(of)Tj
0.9921 0 Td
(serum)Tj
2.6759 0 Td
(specific)Tj
3.1067 0 Td
(antibodies)Tj
4.2633 0 Td
(against)Tj
2.9593 0 Td
(IntiminC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 510.4062 540.4535 cm
BT
/F8 1 Tf
1 TL
-0.0186 Tc
0 0 Td
(280)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 522.9921 542.0976 cm
BT
/F8 1 Tf
1 TL
-0.014 Tc
0 0 Td
(and)Tj
1.6951 0 Td
(EspB.)Tj
-0.004 Tc
-32.7966 -1.2982 Td
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2.8063 0 Td
(were)Tj
2.1203 0 Td
(fed)Tj
1.4627 0 Td
(alfalfa)Tj
2.5851 0 Td
(pellets,)Tj
2.9651 0 Td
(with)Tj
1.9955 0 Td
(free)Tj
1.7292 0 Td
(access)Tj
2.6418 0 Td
(to)Tj
1.0262 0 Td
(hay)Tj
1.6384 0 Td
(and)Tj
1.7291 0 Td
(water)Tj
2.4321 0 Td
(and)Tj
1.7291 0 Td
(treated)Tj
2.9991 0 Td
(prophylactically)Tj
-31.0562 -1.2983 Td
(upon)Tj
2.3187 0 Td
(arrival)Tj
2.8006 0 Td
(with)Tj
1.9956 0 Td
(1%)Tj
1.4513 0 Td
(ivermectin)Tj
4.5184 0 Td
(to)Tj
1.0262 0 Td
(control)Tj
3.1237 0 Td
(intestinal)Tj
3.9118 0 Td
(nematodes.)Tj
4.7849 0 Td
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1.4003 0 Td
(animal)Tj
2.9707 0 Td
(experiments)Tj
5.1362 0 Td
(were)Tj
-35.4384 -1.3039 Td
(performed)Tj
4.4674 0 Td
(with)Tj
1.9956 0 Td
(the)Tj
1.474 0 Td
(approval)Tj
3.702 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(Institutional)Tj
5.159 0 Td
(Animal)Tj
3.2202 0 Td
(Care)Tj
2.1146 0 Td
(and)Tj
1.7291 0 Td
(Use)Tj
1.7405 0 Td
(of)Tj
1.0148 0 Td
(Experimentation)Tj
-29.1003 -1.2983 Td
(Animals)Tj
3.5773 0 Td
(Committee)Tj
4.7452 0 Td
(\(CICUAE\))Tj
4.5524 0 Td
(of)Tj
1.0148 0 Td
(the)Tj
1.4683 0 Td
(National)Tj
3.6964 0 Td
(Institution)Tj
4.4787 0 Td
(of)Tj
1.0148 0 Td
(Agricultural)Tj
5.0853 0 Td
(Reserach.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 463.8614 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Production)Tj
4.9417 0 Td
(of)Tj
1.0441 0 Td
(recombinant)Tj
5.6599 0 Td
(E.)Tj
1.1622 0 Td
(coli)Tj
1.6914 0 Td
(O157:H7)Tj
4.1858 0 Td
(proteins)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 446.8535 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(The)Tj
1.7801 0 Td
(coding)Tj
2.9593 0 Td
(sequence)Tj
3.8608 0 Td
(of)Tj
1.0148 0 Td
(EspB)Tj
2.245 0 Td
(and)Tj
1.7292 0 Td
(IntiminC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 373.4362 445.1527 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
0 0 Td
(280)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 386.3055 446.8535 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.474 0 Td
(bovine)Tj
/F13 1 Tf
2.9253 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
0.9922 0 Td
[()7( )10(\n)12()]TJ
/F8 1 Tf
1.559 0 Td
(O157:H7)Tj
3.8664 0 Td
(strain)Tj
2.5002 0 Td
(146N)Tj
-34.1515 -1.3039 Td
(were)Tj
2.1203 0 Td
(cloned)Tj
2.897 0 Td
(in)Tj
1.0374 0 Td
(pRSET-A)Tj
4.0479 0 Td
(vector)Tj
2.6986 0 Td
(\(Invitrogen)Tj
4.7848 0 Td
(Corp.,)Tj
2.7268 0 Td
(Carlsbad,)Tj
3.9912 0 Td
(CA\))Tj
1.9105 0 Td
(and)Tj
1.7291 0 Td
(expressed)Tj
4.0932 0 Td
(in)Tj
/F13 1 Tf
1.0375 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F13 1 Tf
0.9921 0 Td
[()7( )10(\n)12()]TJ
/F8 1 Tf
-34.0664 -1.2982 Td
(BL21)Tj
2.2904 0 Td
(\(D3\)/pLysS,)Tj
5.0116 0 Td
(as)Tj
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(described)Tj
4.0535 0 Td
(previously)Tj
4.3483 0 Td
([)Tj
0.83 0.64 0.02 0 k
(29)Tj
0 g
(].)Tj
2.0749 0 Td
(Briefly,)Tj
3.0784 0 Td
(the)Tj
1.4683 0 Td
(amino)Tj
2.8006 0 Td
(terminal-His-tagged)Tj
8.3338 0 Td
(pro-)Tj
-34.4746 -1.2983 Td
(teins)Tj
2.1146 0 Td
(were)Tj
2.126 0 Td
(purified)Tj
3.4072 0 Td
(from)Tj
2.2053 0 Td
(the)Tj
1.474 0 Td
(lysates)Tj
2.795 0 Td
(by)Tj
1.1849 0 Td
(affinity)Tj
3.0614 0 Td
(chromatography)Tj
6.8823 0 Td
(on)Tj
1.2756 0 Td
(Nickel-agarose)Tj
6.1568 0 Td
(columns)Tj
-32.6831 -1.2983 Td
(\(ProBond)Tj
4.1612 0 Td
(nickel-chelating)Tj
6.6104 0 Td
(resin;)Tj
2.4151 0 Td
(Invitrogen)Tj
4.4447 0 Td
(Corp.\),)Tj
3.0669 0 Td
(eluted)Tj
2.6702 0 Td
(under)Tj
2.6192 0 Td
(denaturing)Tj
4.6318 0 Td
(conditions)Tj
4.4731 0 Td
(and)Tj
-35.0926 -1.3039 Td
(dialyzed)Tj
3.5149 0 Td
(against)Tj
3.0274 0 Td
(PBS)Tj
1.8312 0 Td
(at)Tj
0.9524 0 Td
(pH7.4.)Tj
-8.1297 -1.2982 Td
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1.7858 0 Td
(B)Tj
0.7994 0 Td
(subunit)Tj
3.2598 0 Td
(of)Tj
1.0148 0 Td
(Shiga)Tj
2.3867 0 Td
(toxin)Tj
2.3074 0 Td
(2)Tj
0.6917 0 Td
(was)Tj
1.7008 0 Td
(cloned)Tj
2.8969 0 Td
(upstream)Tj
3.9741 0 Td
(to)Tj
1.0261 0 Td
(the)Tj
1.4684 0 Td
(Brucella)Tj
3.4696 0 Td
(Lumazine)Tj
4.1952 0 Td
(Synthase)Tj
-32.1729 -1.2983 Td
(\(BLS\))Tj
2.4944 0 Td
(gene)Tj
2.0807 0 Td
(and)Tj
1.7291 0 Td
(the)Tj
1.4683 0 Td
(recombinant)Tj
5.3688 0 Td
(protein)Tj
3.1521 0 Td
(was)Tj
1.7008 0 Td
(expressed)Tj
4.0931 0 Td
(as)Tj
1.0148 0 Td
(described)Tj
4.0535 0 Td
(elsewhere)Tj
4.1046 0 Td
([)Tj
0.83 0.64 0.02 0 k
(39)Tj
0 g
(].)Tj
2.0749 0 Td
(This)Tj
1.9899 0 Td
(anti-)Tj
-35.325 -1.3039 Td
(gen)Tj
1.6554 0 Td
(was)Tj
1.7008 0 Td
(kindly)Tj
2.7609 0 Td
(provided)Tj
3.8154 0 Td
(by)Tj
1.1792 0 Td
(Fernando)Tj
4.1102 0 Td
(Goldbaum)Tj
4.507 0 Td
(from)Tj
2.2053 0 Td
(Fundacio)Tj
0 Tc
3.3165 0.051 Td
()Tj
-0.004 Tc
0.4479 -0.051 Td
(n)Tj
0.7654 0 Td
(Instituto)Tj
3.668 0 Td
(Leloir.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 316.5732 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Immunization)Tj
6.0992 0 Td
(protocol)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 299.5653 cm
BT
/F8 1 Tf
1 TL
-0.019 Tc
0 0 Td
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2.7099 0 Td
(were)Tj
2.0636 0 Td
(randomly)Tj
4.0081 0 Td
(separated)Tj
3.8778 0 Td
(into)Tj
1.7801 0 Td
(four)Tj
1.8539 0 Td
(groups)Tj
2.88 0 Td
(and)Tj
1.6837 0 Td
(vaccinated)Tj
4.3087 0 Td
(according)Tj
4.0478 0 Td
(to)Tj
0.9978 0 Td
(the)Tj
1.423 0 Td
(following)Tj
-31.6344 -1.2982 Td
(scheme:)Tj
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(non-vaccinated)Tj
6.2021 0 Td
(control)Tj
3.0161 0 Td
(\(n)Tj
1.0771 0 Td
(=)Tj
0.7881 0 Td
(4\):)Tj
1.2132 0 Td
(PBS;)Tj
2.0012 0 Td
(Group)Tj
2.7836 0 Td
(3Ag)Tj
1.7972 0 Td
(\(n)Tj
1.0828 0 Td
(=)Tj
0.7824 0 Td
(6\):)Tj
1.2189 0 Td
(IntiminC)Tj
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Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 489.0897 284.8818 cm
BT
/F8 1 Tf
1 TL
-0.0253 Tc
0 0 Td
(280)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 501.5622 286.5826 cm
BT
/F8 1 Tf
1 TL
-0.019 Tc
0 0 Td
(+)Tj
0.7823 0 Td
(EspB)Tj
2.1883 0 Td
(+)Tj
0.7824 0 Td
(BLS-)Tj
-33.9077 -1.3039 Td
(Stx2B;)Tj
2.6475 0 Td
(Group)Tj
2.7836 0 Td
(2Ag)Tj
1.8028 0 Td
(\(n)Tj
1.0772 0 Td
(=)Tj
0.7824 0 Td
(6\):)Tj
1.2189 0 Td
(IntiminC)Tj
ET
Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 339.4771 271.8992 cm
BT
/F8 1 Tf
1 TL
-0.0253 Tc
0 0 Td
(280)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 351.8929 273.5433 cm
BT
/F8 1 Tf
1 TL
-0.019 Tc
0 0 Td
(+)Tj
0.7823 0 Td
(EspB;)Tj
2.3981 0 Td
(Group)Tj
2.7836 0 Td
(Stx)Tj
1.4173 0 Td
(\(n)Tj
1.0772 0 Td
(=)Tj
0.7823 0 Td
(2\):)Tj
1.2189 0 Td
(BLS-Stx2B.)Tj
4.5354 0 Td
(The)Tj
1.7348 0 Td
(immunization)Tj
-31.9178 -1.2982 Td
(protocol)Tj
3.4753 0 Td
(consisted)Tj
3.804 0 Td
(on)Tj
1.2473 0 Td
(two)Tj
1.6611 0 Td
(doses)Tj
2.3244 0 Td
(15)Tj
1.1452 0 Td
(days)Tj
1.9331 0 Td
(apart,)Tj
2.4094 0 Td
(with)Tj
1.9389 0 Td
(100)Tj
/F14 1 Tf
1.6044 0 Td
[()]TJ
/F8 1 Tf
(g)Tj
1.1622 0 Td
(of)Tj
0.9808 0 Td
(each)Tj
1.9729 0 Td
(antigen)Tj
3.101 0 Td
(by)Tj
1.1509 0 Td
(intramuscular)Tj
5.6523 0 Td
(ro-)Tj
-35.5632 -1.2983 Td
(ute.)Tj
1.6328 0 Td
(The)Tj
1.7404 0 Td
(antigens)Tj
3.4469 0 Td
(were)Tj
2.058 0 Td
(diluted)Tj
2.9366 0 Td
(in)Tj
1.0035 0 Td
(1)Tj
0.6803 0 Td
(mL)Tj
1.5477 0 Td
(of)Tj
0.9864 0 Td
(PBS)Tj
1.7915 0 Td
(and)Tj
1.6837 0 Td
(emulsified)Tj
4.2236 0 Td
(in)Tj
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(1)Tj
0.6803 0 Td
(mL)Tj
1.5477 0 Td
(of)Tj
0.9864 0 Td
(mineral)Tj
3.2259 0 Td
(oil-based)Tj
3.7076 0 Td
(adju-)Tj
-34.8828 -1.3039 Td
(vant)Tj
1.8935 0 Td
(\(Montanide)Tj
4.8586 0 Td
(ISA206,)Tj
3.2655 0 Td
(Seppic,)Tj
2.965 0 Td
(France\).)Tj
3.3958 0 Td
(The)Tj
1.7348 0 Td
(control)Tj
3.0217 0 Td
(group)Tj
2.5285 0 Td
(was)Tj
1.6554 0 Td
(vaccinated)Tj
4.3086 0 Td
(only)Tj
1.9163 0 Td
(with)Tj
1.9332 0 Td
(PBS)Tj
1.7915 0 Td
(emul-)Tj
-35.2684 -1.2983 Td
(sified)Tj
2.262 0 Td
(in)Tj
1.0092 0 Td
(the)Tj
1.423 0 Td
(adjuvant.)Tj
ET
Q
q
1 j
1 J
0 w
11.9999 0 0 11.9999 200.0125 195.307 cm
BT
/F0 1 Tf
1 TL
-0.0033 Tc
0 0 Td
(Sample)Tj
3.5716 0 Td
(collection)Tj
ET
Q
q
1 j
1 J
0 w
10 0 0 10 200.0125 178.2992 cm
BT
/F8 1 Tf
1 TL
-0.004 Tc
0 0 Td
(Blood)Tj
2.5851 0 Td
(samples)Tj
3.3846 0 Td
(were)Tj
2.1203 0 Td
(collected)Tj
3.7417 0 Td
(from)Tj
2.2053 0 Td
(all)Tj
1.1452 0 Td
(animals)Tj
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