Short Oral Communications Vaccines VO10- DEVELOPMENT OF Brucella abortus STRAIN RB51 AS AN EXPRESSION VECTOR FOR HETEROLOGOUS EUKARYOTIC AND VIRAL PROTEINS AND AS A CARRIER FOR AIDS VACCINE. Yakir Ophir 1 , Gerhardt Schurig 2 , Ramesh Vemulapalli 3 , George N. Pavlakis 4 , Barbara Felber 4 , A.T.M. Shamsul Hoque 1 , Weila Wang 1 , Hana Golding 1 and Basil Golding 1 . (1) CBER, FDA, Bethesda, MD, USA. (2) VA-MD Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, USA. (3) Purdue University, West Lafayette, IN, USA. (4) NCI, NIH, Fredrick, MD, USA. HIV-1 infection is associated with a loss in T-helper cell responses prior to onset of AIDS. Therefore, therapeutic vaccines should be T-helper cell independent. Earlier, we demonstrated that heat-killed Brucella abortus conjugated to a V3-loop peptide from HIV-1 elicits neutralizing antibodies and CTL even in mice depleted of CD4+ T-cells. Currently we are attempting to express heterologous eukaryotic and viral genes in B. abortus RB51. It was previously reported that RB51 can express heterologous bacterial proteins and mice vaccinated with these recombinants developed Th1-like immunity against the expressed proteins. HIV-derived proteins (gag and pol) and ovalbumin (OVA) were selected for expression in strain RB51. These genes were cloned into three B. abortus expression vectors: pBBSODpro and pBBgroE under the sodC and groE promoter sequences for constitutive expression; and pNOF100 under the tightly regulated inducible tac promoter. The expression of the cloned proteins was analyzed in whole cell extracts of B. abortus RB51 by Western blotting. It was possible to obtain low level expression of pol in pBBSODpro and improved expression in pNOF100 when grown in LB medium supplemented with glycerol, but not in TSB medium. OVA was only expressed in pNOF100 when grown in LB medium supplemented with glycerol. Optimal expression of eukaryotic genes in Brucella will most likely require selective codon optimization of the foreign genes, and possibly further modifications of expression vectors. VO11- PROTECTION AGAINST Neospora caninum IN A GERBIL MODEL USING Brucella abortus STRAIN RB51 EXPRESSING N. caninum PROTEINS. S. Ramamoorthy, G. Schurig, D. Lindsay, R. Vemulapalli, S.M. Boyle, N. Srirangananthan. Virginia- Maryland Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, VA 24061, USA. N. caninum, the protozoan parasite, is of emerging importance as a cause of abortions in cattle. The attenuated B. abortus strain RB51 has been used as an effective vaccine for cattle brucellosis. Strain RB51 has also been used as a vector for heterologous protein expression. It was hypothesized that putative virulence factors of N. caninum could be expressed in strain RB51 to develop a combined vaccine for neosporosis and brucellosis. The GRA7, SRS2 and MIC3 genes of N. caninum were cloned separately into plasmid pBBR1MCS downstream of the Brucella groE promoter and used to transform B. abortus strain RB51. The recombinant RB51vaccine strains were inoculated individually and in combination into groups of three gerbils. Each gerbil received intraperitoneally a primary dose of 6x10 8 CFU/ml followed by a booster dose of 2x10 6 CFU/ml administered four weeks later. Five weeks post immunization, the gerbils were challenged with 2x10 6 N. caninum NC-1 tachyzoites. Four out of five gerbils survived in the groups administered GRA7 and SRS2 alone. Two out of three gerbils survived in the MIC3 82 <strong>Brucellosis</strong> <strong>2003</strong> International Research Conference
Short Oral Communications Vaccines group. Five out of five gerbils survived in the groups where all three antigens were combined and in the groups where GRA7 was combined with SRS2 and with MIC3. Brain, spleen, lung and liver tissues were fixed in formal saline and subjected to histopathological analysis. Tissues that had the largest number of lesions per field of examination were awarded the highest scores. The lowest score was found in the GRA7, MIC3 combination group followed by the SRS2, MIC3 group. These preliminary results indicate that this approach may be a practical method for prophylaxis against N. caninum. <strong>Brucellosis</strong> <strong>2003</strong> International Research Conference 83
- Page 1:
Brucellosis 2003 International Rese
- Page 5:
Welcome As Professor Paul Nicoletti
- Page 9:
Instructions to presenters KEYNOTE
- Page 13:
Scientific Program
- Page 16 and 17:
Scientific Program - Monday, Septem
- Page 18 and 19:
Scientific Program - Monday, Septem
- Page 20 and 21:
Scientific Program - Tuesday, Septe
- Page 22 and 23:
Scientific Program - Wednesday, Sep
- Page 24 and 25:
Scientific Program - Poster Session
- Page 26 and 27:
Scientific Program - Poster Session
- Page 28 and 29:
Scientific Program - Poster Session
- Page 30 and 31:
Scientific Program - Poster Session
- Page 32 and 33: Scientific Program - Poster Session
- Page 35: Abstracts
- Page 38 and 39: Keynote Lectures while B. ovis and
- Page 40 and 41: Keynote Lectures BRUCELLOSIS IN WIL
- Page 42 and 43: Keynote Lectures CLASSICAL AND NEW
- Page 44 and 45: Keynote Lectures COMPARISON OF THE
- Page 46 and 47: Keynote Lectures projects (localizo
- Page 48 and 49: Short Oral Communications Epidemiol
- Page 50 and 51: Short Oral Communications Epidemiol
- Page 52 and 53: Short Oral Communications Epidemiol
- Page 54 and 55: Short Oral Communications Human bru
- Page 56 and 57: Short Oral Communications Human bru
- Page 58 and 59: Short Oral Communications Human bru
- Page 60 and 61: Short Oral Communications Diagnosis
- Page 62 and 63: Short Oral Communications Diagnosis
- Page 64 and 65: Short Oral Communications Diagnosis
- Page 66 and 67: Short Oral Communications Immunolog
- Page 68 and 69: Short Oral Communications Immunolog
- Page 70 and 71: Short Oral Communications Immunolog
- Page 72 and 73: Short Oral Communications Immunolog
- Page 74 and 75: Short Oral Communications Immunolog
- Page 76 and 77: Short Oral Communications Vaccines
- Page 78 and 79: Short Oral Communications Vaccines
- Page 80 and 81: Short Oral Communications Vaccines
- Page 84 and 85: Short Oral Communications Taxonomy
- Page 86 and 87: Short Oral Communications Taxonomy
- Page 88 and 89: Poster Session and all male animals
- Page 90 and 91: Poster Session 6- SEROLOGICAL INCID
- Page 92 and 93: Poster Session situation of a long-
- Page 94 and 95: Poster Session 14- HIGH PREVALENCE
- Page 96 and 97: Poster Session samples tested posit
- Page 98 and 99: Poster Session time. To confirm the
- Page 100 and 101: Poster Session 25- PRESENTATION OF
- Page 102 and 103: Poster Session specific diagnosis i
- Page 104 and 105: Poster Session Urinalysis was notab
- Page 106 and 107: Poster Session sera were positive w
- Page 108 and 109: Poster Session controversial. While
- Page 110 and 111: Poster Session 41- RAPID DETECTION
- Page 112 and 113: Poster Session 44- DEVELOPMENT AND
- Page 114 and 115: Poster Session Brucella ovis, Bruce
- Page 116 and 117: Poster Session were isolated and ty
- Page 118 and 119: Poster Session the ELISA, whereas 7
- Page 120 and 121: Poster Session agglutination (SAT),
- Page 122 and 123: Poster Session investigations in ap
- Page 124 and 125: Poster Session 65- PHAGOCYTOSIS AND
- Page 126 and 127: Poster Session 68- DOES OXYGEN TENS
- Page 128 and 129: Poster Session 72- IMPLICATION OF F
- Page 130 and 131: Poster Session 76- THE AQUAPORIN GE
- Page 132 and 133:
Poster Session adaptation to starva
- Page 134 and 135:
Poster Session constituents that ar
- Page 136 and 137:
Poster Session and revaccinated ani
- Page 138 and 139:
Poster Session weeks after infectio
- Page 140 and 141:
Poster Session melitensis wild type
- Page 142 and 143:
Poster Session indicated that HS en
- Page 144 and 145:
Poster Session This study aimed to
- Page 146 and 147:
Poster Session definition. Reviewin
- Page 148 and 149:
Poster Session ApaI+0/MseI+G) were
- Page 150 and 151:
Poster Session 109- COMPARATIVE PRO
- Page 152 and 153:
Poster Session enterobactin. At the
- Page 154 and 155:
Poster Session apparent differences
- Page 157 and 158:
Author index A Abalos, P.----------
- Page 159 and 160:
Author index GonzálezCarreró, M I
- Page 161 and 162:
Author index Q Qasem, J A.---------
- Page 163 and 164:
List of participants ABERNETHY, DAR
- Page 165 and 166:
List of participants CARNERO OJEA,
- Page 167 and 168:
List of participants Fax: 301 319 9
- Page 169 and 170:
List of participants Phone: 1 97 98
- Page 171 and 172:
List of participants IOANNOU, IOANN
- Page 173 and 174:
List of participants LÓPEZ-GOÑI,
- Page 175 and 176:
List of participants NEUBAUER, HEIN
- Page 177 and 178:
List of participants E-mail: rajash
- Page 179 and 180:
List of participants SUAREZ-GUEMES,
- Page 181:
Sponsors Brucellosis 2003 Internati