Burkholderia pseudomallei 28450 Melioidosis The bacterium Burkholderia pseudomallei is a gram-negative saprophytic bacillus that is found predominantly in Southeast Asia and tropical Australia and is the causative agent of the emerging infection melioidosis. Infection of humans with B. pseudomallei may occur by various routes, including via wounds and existing skin lesions, aspiration of contaminated water during near-drowning, and inhalation of organisms. The clinical manifestations of melioidosis are not uniform, and patients can present with pneumonia, skin abscesses, soft tissue abscesses, or osteomyelitis/septic arthritis. Pneumonic melioidosis is presented in over 50% of acute cases of the disease, and disseminated pneumonia is associated with high mortality. Chronic melioidosis usually occurs following acute melioidosis, and relapsing melioidosis can result from the reactivation of a latent B. pseudomallei infection, often due to the withdrawal of antibiotics or the failure to complete prescribed courses of antibiotics (Harland et al., 2007). Cattle Bos taurus 9913 Chicken Gallus gallus 9031 chinchillas Chinchillidae 10150 Copper Pheasant Syrmaticus soemmerringii 9067 Deer mouse Peromyscus maniculatus 10042 Ducks Anas 8835 Ferret Mustela putorius furo 9669 Goat Capra hircus 9925 Guinea pig Cavia porcellus 10141 Hamster Mesocricetus auratus 10036 Horse Equus caballus 9796 Human Homo sapiens 9606 Monkey Platyrrhini 9479 Mouse Mus musculus 10090 Pig Sus scrofa 9823 Rabbit Oryctolagus cuniculus 9986 Rat Rattus 10114 Raven Corvus corax 56781 Sheep Ovis aries 9940 Vole Microtus ochrogaster 79684 B. pseudomallei DNA vaccine encoding Flagellin Protein VO_0011538 Research Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction Balb/c Six-week-old female Balb/c mice from groups 1–3 were injected intramuscularly (i.m.) with pcDNA3/fliC in PBS (50, 100 or 150 μg); whereas those from group 4 (the negative control) were injected (i.m) with vector DNA(150 μg; pcDNA3) in PBS, whilst those from group 5 (the positive control) were injected subcutaneously with recombinant flagellin antigens (50 μg) emulsified in complete Freund's adjuvant. Each animal from group 5 was subsequently boosted with the same dose of flagellin in incomplete Freund's adjuvant 14 days subsequent to the initial flagellin injection. The animals in groups 1–4 were boosted twice with an identical dose of plasmid DNA, once at 7 days and then again at 14 days after the initial injection (Chen et al., 2006). Subsequent intravenous challenge of the vaccinated Balb/c mice with 10(5)CFU of B. pseudomallei resulted in the number of bacterial cells detected in liver and/or spleen being significantly reduced in the flagellin plasmid DNA vaccinated mice. At 7 days subsequent to infection of B. pseudomallei, 5/6 (83%) of flagellin plasmid DNA vaccinated mice had survived (Chen et al., 2006). The immunized mice were infected via the tail vein with 105 CFU of mixed B. pseudomallei suspended in sterile PBS (50 μl). As control groups, the mice received either sterile PBS or vector DNA (Chen et al., 2006). B. pseudomallei Subunit LolC Protein Vaccine VO_0011436 Research Intraperitoneal injection (i.p.) Intraperitoneal injection (i.p.) Recombinant protein preparation BALB/c Female 5- to 6-week-old BALB/c mice were immunized with the purified recombinant LolC protein given with adjuvant (Harland et al., 2007). The recombinant LolC protein afforded significant protection against the B. pseudomallei challenge (Harland et al., 2007). Mice immunized with the LolC protein or appropriate controls were challenged with B. pseudomallei K96243 on day 70 (Harland et al., 2007). B. pseudomallei Subunit PotF Protein Vaccine VO_0011438 Research Intraperitoneal injection (i.p.) Intraperitoneal injection (i.p.) Recombinant protein preparation BALB/c Female 5- to 6-week-old BALB/c mice were immunized with the purified recombinant PotF protein given with adjuvant (Harland et al., 2007). Immunization with the PotF protein domain afforded significant protection against a subsequent challenge with B. pseudomallei (Harland et al., 2007). Mice immunized with the PotF protein or appropriate controls were challenged with B. pseudomallei K96243 on day 70 (Harland et al., 2007). FliC VO_0011303 1929918 CDD:169576 CDD:144315 CDD:144340 28450 ? flagellin 388 flagellin; Reviewed; PRK08869 >gi|1929918|gb|AAB51476.1| flagellin [Burkholderia pseudomallei] MLGINSNINSLVAQQNLNGSQGALSQAITRLSSGKRINSAADDAAGLAIATRMQTQINGLNQGVSNANDG VSILQTASSGLTSLTNSLQRIRQLAVQASNGPLSASDASALQQEVAQQISEVNRIASQTNYNGKNILDGS AGTLSFQVGANVGQTVSVDLTQSMSAAKIGGGMVQTGQTLGTIKVAIDSSGAAWSSGSTGQETTQINVVS DGKGGFTFTDQNNQALSSTAVTAVFGSSTAGTGTAASPSFQTLALSTSATSALSATDQANATAMVAQINA VNKPQTVSNLDISTQTGAYQAMVSIDNALATVNNLQATLGAAQNRFTAIATTQQAGSNNLAQAQSQIQSA DFAQETANLSRAQVLQQAGISVLAQANSLPQQVLKLLQ Protective antigen Flagellin encoding plasmid DNA was injected into Balb/c mice intramuscularly and this elicited both a humoral and a cellular immune response. Subsequent intravenous challenge of the vaccinated Balb/c mice with 10^5CFU of B. pseudomallei resulted in the number of bacterial cells detected in liver and/or spleen being significantly reduced in the flagellin plasmid DNA vaccinated mice. At 7 days subsequent to infection of B. pseudomallei, 5/6 (83%) of flagellin plasmid DNA vaccinated mice had survived [Ref1361:Chen et al., 2006]. LolC/E Burkholderia pseudomallei K96243 VO_0010921 3093214 53719887 BPSL2277 BX571965 YP_108873 272560 1 2742869 2744122 - putative lipoprotein releasing system transmembrane protein 9.43 41147.07 417 Similar to Escherichia coli lipoprotein releasing system transmembrane protein LolE SWALL:LOLE_ECOLI (SWALL:P75958) (413 aa) fasta scores: E(): 1.3e-42, 34.54% id in 414 aa, and to Ralstonia solanacearum probable lipoprotein releasing system transmembrane rsc1117 or rs05757 SWALL:Q8Y0C7 (EMBL:AL646062) (416 aa) fasta scores: E(): 1.4e-112, 71.46% id in 417 aa >gi|53717639:2742869-2744122 Burkholderia pseudomallei K96243 chromosome 1, complete sequence GTCATTCATAGCGCAGCGCCTCCGCCGGGCGCACCTTCGCGCCGCGCCAGCTCGGATAGAGCGTCGCGAG CGCGGACAGCGCGAAAGCGATCACGCCGATCTTGATCACGTCGCCCGCGACGAGCTCGGACGGCAGCTCG CTGATGAAGTACACCGACGGCGGCAGGAACTGCACGCCGAACGCATGCTCGATCATCGGAATGAGCCACG GAATGCTCCACGCGATCAGGCAGCCGAGCGCGACGCCCGTCGCGGTCCCGACGAAGCCGATCGTCACGCC CTGCACGACGAAGATCTTCATGATCGAGCCCGGCTGCGCGCCGAGCGTGCGCAGGATCGCGATGTCCGCC TGCTTGTTCGTCACGGTCATCACGAGCGACGACACGAGATTGAACGCCGCGACCGCGATGATCAGCGTGA GGATGATGAACATCATCCGCTTCTCGATCTGCACCGCGGAGAACCACGTCTTGTTCTGCTGCGTCCAGTC GCGGATGTACAGGTCGCCCGACAGCGTGTGCGCGAGCTCGCGCGCGACCTGCGGCGCCTTCTGCATGTCC GTGAGGCGCAGCCGCACGCCCGTCGGCGCGGGCAGCCTGAAGAGCGCCTGCGCATCCTGGATGTCGATCA TCGCAAGCGTGCTGTCGTATTCGTAGTGCCCGGACTCGAAGATGCCGACCACCGTGAACTGCTTCAGGCG CGGCATCATCCCGGCCGGCGTGATCGTGCCTTCCGGCGCGACGAGCGTGACCTTGTCGCCGACGCCCACG CCGAGATTGCCCGCGAGCGCGTTGCCGAGCACGATCCCGAACTGGCCGGGCGCGAGCGCGGTGAGCGCGC CCGCCTTCATGTCCTTGCCGATGTCGGACACCTGCGGCTCGAGCGACGGCTCGACGCCGCGCAGCATCAC GCCGCTCACCGCGTCCTGCCGCGTGAGCAGCGCCTGCGCGTCGACATACGGCGCCGCGCCGATCACCGAG CGGTTCAGGCGCGCTTCCTTCGCGGTCAGTTGCCAGTCGGGCATCGAGCCCGTCGGCGAGAAGATCTCGA CGTGCGCGAGCACCGACAGCATCCGGTCGCGCACCTCTTTCTGGAAGCCGTTCATCACCGACAGCACGAC GATGAGCGCCGCGACGCCGAGCGCGATCCCGAGCATCGACACGAGCGCGATGAAGGAAATGAAGCCGTTG CCGGTCGTGCGTTTGCCGGCGCGCGTGTAGCGCCAGCCGATCTGCCATTCGTACGGGAGTTTCA >gi|53719887|ref|YP_108873.1| putative lipoprotein releasing system transmembrane protein [Burkholderia pseudomallei K96243] MKLPYEWQIGWRYTRAGKRTTGNGFISFIALVSMLGIALGVAALIVVLSVMNGFQKEVRDRMLSVLAHVE IFSPTGSMPDWQLTAKEARLNRSVIGAAPYVDAQALLTRQDAVSGVMLRGVEPSLEPQVSDIGKDMKAGA LTALAPGQFGIVLGNALAGNLGVGVGDKVTLVAPEGTITPAGMMPRLKQFTVVGIFESGHYEYDSTLAMI DIQDAQALFRLPAPTGVRLRLTDMQKAPQVARELAHTLSGDLYIRDWTQQNKTWFSAVQIEKRMMFIILT LIIAVAAFNLVSSLVMTVTNKQADIAILRTLGAQPGSIMKIFVVQGVTIGFVGTATGVALGCLIAWSIPW LIPMIEHAFGVQFLPPSVYFISELPSELVAGDVIKIGVIAFALSALATLYPSWRGAKVRPAEALRYE Protective antigen LolC was evaluated as an antigen for a vaccine candidate against B. pseudomallei infection. Nonmembrane regions of the B. pseudomallei protein was expressed and purified from Escherichia coli and then evaluated as a vaccine candidate in an established mouse model of B. pseudomallei infection. When delivered with the monophosphoryl lipid A-trehalose dicorynomycolate adjuvant, the protein stimulated antigen-specific humoral and cellular immune responses. Immunization with the LolC protein domain afforded significant protection against a subsequent challenge with B. pseudomallei. LolC provided a greater level of protection when it was administered with immune-stimulating complexes complexed with CpG oligodeoxynucleotide 10103. Immunization with LolC also protected against a subsequent challenge with a heterologous strain of B. pseudomallei, demonstrating the potential utility of this protein as a vaccine antigen for melioidosis [Ref1360:Harland et al., 2007]. PotF Burkholderia pseudomallei K96243 VO_0010922 3095748 53721504 BPSS0467 BX571966 YP_110489 272560 2 636534 637655 - putrescine ABC transport system, binding exported protein 6.89 37606.43 373 Similar to Escherichia coli putrescine-binding periplasmic protein precursor PotF or b0854 SWALL:POTF_ECOLI (SWALL:P31133) (370 aa) fasta scores: E(): 3.3e-67, 49.3% id in 357 aa, and to Pseudomonas aeruginosa polyamine transport protein pa0301 SWALL:Q9I6J0 (EMBL:AE004468) (365 aa) fasta scores: E(): 6.3e-69, 49.59% id in 367 aa. Possible alternative translational start site >gi|53721039:636534-637655 Burkholderia pseudomallei K96243 chromosome 2, complete sequence ATCAGTGTCCCGTCTTCAATTGCGCCCACAGACGGTTCTCGAGCCGCAGGATGTCGGCGGGCATCGGCCG CATCAGCGTCATCTTCTTGAGCACGTCCTCGGGCGGGTAGACGGTCGGGTCCTGCGCGACGGCGGGCACG ACGAACTGTCGCGCCGCGCGGTTCGCGCTCGGGTAGAACACTTCGTTGGTGATCGCCGCGTTGACCTTCG GGTCCTCGATGTAGTTGATCCACTTCAGCGCGGCTTCCGCGTGCGGCGCATCCTTCGGGATCACCATCAC GTCGAACCACAGCAGGCCGCCTTCCTTCGGATTCGAGAACTTGATGTCGTACGGGCGCTTCGCCTCGGCC GAGCGGCGATGCGCGATGCCGACGTCGCCCGACCAGCCGAGCGCGACGCATACGTCGTTGTTCGCGAGAT CGTTGATGTAGCCGGACGAGTTGAATTGCGTGATGTACGGGCGGACTTTCTTCAGCACCTCGAACGCCGC CTGGTAATCGGCCGGGTTCGTGCTGTTCGGATTCCTGCCCATGTATTGTAGCGTCGCGGCGAACACGTCG ACCGCCTGATCGAGGAACGATACGCCGCAGCCCTTGAGCTTGGATACGTTCGCCGGATCGAGCACGAGCG CCCAGCTGTCGACGGGCGCGTTCTCGCCGAGCGCCTTCTTCACCGCCTGCACGTTGTAGCCGAGGCCGTC GGTGCCGAACGCCCACGGCACGCCGTACTGGTTGCCCGGATCGGCGTCGGCGATCATCTTCATCAGCGTC GGATCGAGGTTCGACAGGTTCGGCAGCTTCGATTTGTCGAGCTTCTGGTAGACGCCCGCCTGAATCTGCT TGGCCATGTAGTTCGACGTCGGCACGACGATGTCGTAGCCGGAGCTGCCCGCGAGCAGCTTCGCCTGCAG CGTGTCGTCGCTGTCGTAGTTGTCGTACTTGACGTGGATGCCGGATTGCTTCTCGAAGTTCGGGATCGTG TCCTTCGCGATGTAGTCCGACCAGTTGTACACGTTCAGCTCGGCGCCGGCCGCGTGCGCGGCCGGCAGGG CGAGCGCCGACGCGCCGGCGAGCGCGACGAGCGCTGCGCGCGCGACCGCATGACGAAGATGGCTCACACT CA >gi|53721504|ref|YP_110489.1| putrescine ABC transport system, binding exported protein [Burkholderia pseudomallei K96243] MSVSHLRHAVARAALVALAGASALALPAAHAAGAELNVYNWSDYIAKDTIPNFEKQSGIHVKYDNYDSDD TLQAKLLAGSSGYDIVVPTSNYMAKQIQAGVYQKLDKSKLPNLSNLDPTLMKMIADADPGNQYGVPWAFG TDGLGYNVQAVKKALGENAPVDSWALVLDPANVSKLKGCGVSFLDQAVDVFAATLQYMGRNPNSTNPADY QAAFEVLKKVRPYITQFNSSGYINDLANNDVCVALGWSGDVGIAHRRSAEAKRPYDIKFSNPKEGGLLWF DVMVIPKDAPHAEAALKWINYIEDPKVNAAITNEVFYPSANRAARQFVVPAVAQDPTVYPPEDVLKKMTL MRPMPADILRLENRLWAQLKTGH Protective antigen PotF was evaluated as an antigen for a vaccine candidate against B. pseudomallei infection. Nonmembrane regions of the B. pseudomallei protein was expressed and purified from Escherichia coli and then evaluated as a vaccine candidate in an established mouse model of B. pseudomallei infection. When delivered with the monophosphoryl lipid A-trehalose dicorynomycolate adjuvant, the protein stimulated antigen-specific humoral and cellular immune responses. Immunization with the PotF protein domain afforded significant protection against a subsequent challenge with B. pseudomallei [Ref1360:Harland et al., 2007]. Chen et al., 2006 journal Chen YS, Hsiao YS, Lin HH, Yen CM, Chen SC, Chen YL 2006 24 6 750-758 Vaccine Harland et al., 2007 journal Harland DN, Chu K, Haque A, Nelson M, Walker NJ, Sarkar-Tyson M, Atkins TP, Moore B, Brown KA, Bancroft G, Titball RW, Atkins HS 2007 75 8 4173-4180 Infection and immunity Nelson et al., 2004 journal Nelson M, Prior JL, Lever MS, Jones HE, Atkins TP, Titball RW 2004 53 Pt 12 1177-1182 Journal of medical microbiology