Brucella: Molecular and Cellular Biology

Book Cover......Page 1
Title Page......Page 2
ISBN 1904933041......Page 3
Dedication......Page 4
Contents (with page links)......Page 6
Books of Related Interest......Page 8
Contributors......Page 9
Foreword......Page 17
References......Page 20
1. Introduction......Page 21
2. Variability in the Genome Organization......Page 22
3. Insertion Sequences......Page 24
4.1. Genes Involved in the Biosynthesis of Lipopolysaccharide Antigens......Page 28
4.2. Genes Coding for Outer Membrane Protein Antigens......Page 29
5. DNA Polymorphism in Genes Encoding Cytoplasmic and Periplasmic Protein Antigens......Page 35
6. Random DNA Polymorphism......Page 36
7. Conclusions and Future Directions......Page 37
References......Page 38
1. Introduction......Page 44
2.1. Simple Genus-specific PCR Assays Potentially Amenable for Preliminary Screening......Page 46
2.2. Simultaneous Testing for Multiple Disease Agents Including......Page 49
3. PCR-Based Identification and Differentiation of Brucella at the Species/Biovar Level......Page 50
3.1. One-step Single PCR Assays That Can Differentiate One Brucella Species From All the Others......Page 51
3.2. Multiplex PCR Assays for the Identification and Differentiation of Brucella Species and/or Biovars......Page 52
3.3. PCR-RFLP Assays for the Identification and Differentiation of Brucella Species and/or Biovars......Page 53
4. Differentiation of Brucella Strains and Isolates by PCR for Epidemiological Trace-back......Page 54
4.2. Repetitive Element PCR......Page 55
4.3. Infrequent Restriction Site-PCR......Page 57
4.4. Multi-locus VNTR Analysis......Page 59
5. Accelerated Identification with Real-time PCR......Page 63
References......Page 64
Abstract......Page 71
1.2. Diagnosis of Human Brucellosis......Page 72
2.1. Targets and Primers......Page 75
2.2. Conventional PCR Methods......Page 76
2.3. ELISA-PCR Assay......Page 81
2.4. Real-time PCR Methods......Page 82
References......Page 84
1. Introduction......Page 87
2. Genes Delimitation......Page 89
3.1. The pCDS Pages......Page 92
3.2. Searching the pCDS Database......Page 95
4.1. Comparison With Other α-Proteobacteria......Page 97
4.3. Hypothetical Proteins......Page 99
References......Page 101
1. Introduction......Page 104
2. Sequence and Characteristics of the Brucella Genomes......Page 105
3. Genomic Diversity: Polymorphisms and Insertion Sequences......Page 110
4. Genomics and Taxonomy......Page 111
5. Genomics and Diagnostics......Page 113
6. Differential Gene Expression......Page 115
7. Designing Arrays that Address Closely Related Sequences......Page 116
References......Page 118
Abstract......Page 122
1. Brucella melitensis Proteome......Page 123
3. Brucella abortus Proteome......Page 126
5. Brucella Secretomes......Page 129
6. Conclusions......Page 130
References......Page 131
Abstract......Page 133
2. The Regulatory Network......Page 134
2.1. Transcriptional Regulators: GntR, LysR, AraC and Other Families......Page 135
2.2. Signal Transduction......Page 142
2.3. Specialised Global Regulators......Page 156
2.4. Post-transcriptional Regulation......Page 161
3. Conclusion......Page 163
References......Page 164
Abstract......Page 172
1. Introduction......Page 173
2.3. Lipid A......Page 176
3. Genetics and Biosynthesis of Brucella LPS......Page 178
3.1. O-Polysaccharide......Page 180
3.2. Core Oligosaccharide......Page 182
3.3. Lipid A......Page 184
4. Assembly and Transfer to the Bacterial Surface......Page 193
5. Concluding Remarks......Page 194
References......Page 195
Abstract......Page 204
1. Introduction......Page 205
1.2. Signal Delivering System in Animal and Plant Pathogenic Bacteria......Page 207
1.4. The VirB System of Brucella abortus......Page 208
3. Biosynthesis of Cyclic β-1,2-Glucan......Page 209
5. B. abortus Cyclic β-1,2-Glucan Transporter......Page 212
6. B. abortus Genome Organization of the Genes Encoding the Proteins Involved in Cyclic β-1,2-Glucan Synthesis and Secretion......Page 213
8. Osmoregulation of Cyclic Glucan Accumulation......Page 214
9. Virulence and Cyclic Glucan......Page 215
10. Conclusions......Page 218
References......Page 219
Abstract......Page 224
1. Introduction......Page 225
2. Brucella BvrS/BvrR Two-Component Regulatory System......Page 226
3. The Pleiotropic Control of the BvrS/BvrR and its Role in Host- Interaction......Page 227
4. ChvG/ChvI in Plant-Associated Agrobacterium and Sinorhizobium......Page 231
5. Common Regulatory Strategies in α-2 Proteobacteria......Page 235
References......Page 239
1. Introduction......Page 243
2.1. Erythritol Metabolism in the S19 Vaccine Strain......Page 244
2.2. The ery Operon in Brucella......Page 245
2.3. Transport of Erythritol......Page 246
3. Erythritol Catabolism Genes in Other Bacteria......Page 249
4.1. The Iron Connection......Page 251
References......Page 253
Abstract......Page 256
1. Introduction......Page 257
2. Central Role of 2,3-Dihydroxybenzoic Acid in Iron Acquisition by the Brucella spp.......Page 258
3. Evidence For the Production of a More Complex 2,3-DHBA-Based Siderophore by the Brucella spp.......Page 260
4. Transport of Fe+++-2,3-DHBA and Fe+++-Brucebactin Into Brucella......Page 262
5.1. Utilisation of Heme as an Iron Source......Page 265
5.2. Utilisation of Ferric Dicitrate as an Iron Source......Page 266
5.3. Utilisation of Ferrous Iron......Page 267
6.1. Fur......Page 268
6.2. RirA and Irr......Page 269
6.3. DhbR......Page 270
References......Page 271
1. Introduction......Page 277
2.1. Intracellular Cholesterol Transport Pathway......Page 278
3.1. B. abortus Induces Macropinocytosis of Macrophages......Page 280
3.2. B. abortus Macropinosomes Avoid Fusion with Other Macropinosomes......Page 281
3.3. Accumulation of Lipid Rafts on B. abortus Macropinosomes......Page 282
3.4. Macropinosomes Containing B. abortus are Replicative Phagosomes......Page 283
3.5. Effect of Raft-binding or Disrupting Agents on Brucella Infection Into Macrophages......Page 284
4.1. Acceleration of Bacterial Internalisation by Pre-loading Cholesterol Into Macrophages......Page 285
4.2. Effects of Agents That Modulate Cholesterol Trafficking on Bacterial Internalisation and Intracellular Replication......Page 286
4.3. Role of Cholesterol Trafficking in the Establishment of B. abortus Infection......Page 287
5.1. Tail Formation of Cellular Prion Protein With Internalisation of B. abortus......Page 288
5.2. Surface Exposure of Hsp60 on B. abortus......Page 289
5.3. Interaction of PrPC With Hsp60 of B. abortus......Page 290
5.4. Effect of PrPC Deficiency on B. abortus Infection......Page 292
References......Page 294
Abstract......Page 300
1. Introduction......Page 301
2. Binding and Penetration to Cells......Page 303
3. Intracellular Trafficking......Page 307
4. The Replicating Niche......Page 313
5. Brucella Molecules Released During Intracellular Life: The LPS Case......Page 316
References......Page 320
Abstract......Page 327
1.2. Macrophages are Specific Targets of Brucella for its Multiplication......Page 328
2. Entry of Brucella into the Macrophage......Page 329
2.2. B. abortus Needs Functional VirB and Targets Prion Protein on Mouse Macrophages......Page 330
3. The Early Brucella-Containing Phagosome Avoids Lysosome Fusion: The Loop Line......Page 331
4. Acidification of the Early Brucella-Containing Phagosome is Essential for Intracellular Replication......Page 332
5.1. The Important Role of Hsp70 (DnaK)......Page 333
5.2. Other Stress Proteins Involved in Brucella Virulence......Page 334
6. The Type IV Secretion System VirB of B. suis is Induced in the Early Phase of Macrophage Infection......Page 335
7. Lysosomes Do Not Fuse With the Brucellosome: Escape Action......Page 336
8. Adaptation of Brucella spp. to the Environment of the Brucellosome......Page 338
8.2. Genes Essential for Intramacrophagic Multiplication of Brucella spp.......Page 339
8.3. Evidence for Low Oxygen Tension Within the Brucellosome......Page 341
8.4. The Virulence Genes of Brucella spp. Indirectly Characterise the Niche in the Macrophage......Page 344
9. Brucella spp. Inhibit Apoptosis of the Host Cell......Page 346
10. Intracellular Traffic of Brucella spp. in Macrophages. Where do the Membranes Come From?......Page 347
11. Conclusions and Future Prospects......Page 348
References......Page 349
Abstract......Page 355
1. Introduction to Cellular Immunity......Page 356
2. IFN-γ is Crucial for Survival of Brucellosis......Page 357
3. Role of IL-12 in Protective Immunity......Page 361
4. Which Cells Produce the Protective IFN-γ: NK Cells, γδ T Cells or αβ T Cells?......Page 362
5. How Do Nude Mice Control Infection?......Page 363
6. Control of Brucellae in Macrophages by ROI and Nitric Oxide......Page 364
7. Role of TNF-α in Cellular Immunity to Brucella......Page 365
8. Innate Resistance Mechanisms of Macrophages......Page 366
9. Which is it: CD4 or CD8 T Cells That Are Key to Protection?......Page 367
10. The Implications of Cross-Presentation for Brucella Antigens......Page 371
11. Why Do BALB/c Mice Turn off the Protective IFN-γ Response During The Plateau Phase of Infection?......Page 374
12. Acknowledgements......Page 375
References......Page 376
Abstract......Page 382
1. Contrasts in Vaccine Strategies for Animals and Humans......Page 383
2. Human Vaccination......Page 384
3. Epidemiology......Page 385
4. Host-pathogen Interactions in Primary Infection......Page 386
5.2. Animal Models of Adaptive Immunity......Page 387
5.3. Mechanisms of Vaccine-Mediated Immunity......Page 388
6. Nonhuman Primate Models and Studies with Live, Attenuated Strains of B. melitensis......Page 390
7.1. Murine Intranasal Challenge Model......Page 393
7.2. Construction of WR201 and Testing in Mice......Page 394
7.3. Testing of WR201 in an Aerosol Challenge Model in Rhesus Macaques......Page 395
7.4. Implications of Experience with WR201......Page 396
7.6. Development and Testing of wboA-Complemented Rough Purine Auxotrophs as a Vaccine Delivery Platform......Page 397
7.7. Development and Testing of hfq Mutants......Page 398
7.9. Other Brucella Mutants......Page 399
8. Molecular Approaches to Enhance Immunogenicity......Page 403
References......Page 404
Abstract......Page 413
1. Safety Versus Efficacy in Vaccination......Page 414
2. Control of Ovine Brucellosis......Page 415
3. Acellular Vaccines Against Brucellosis......Page 416
4. Adjuvants and Microparticles......Page 417
5. Microencapsulation of the HS Antigenic Extract from Brucella ovis......Page 419
6. Cell Mediated Immunity and Protection......Page 425
7. Additional Considerations and Concluding Remarks......Page 430
References......Page 431
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