TY - JOUR
T1 - Genomic loss in environmental and isogenic morphotype isolates of burkholderia pseudomallei is associated with intracellular survival and plaque-forming efficiency
AU - Saiprom, Natnaree
AU - Sangsri, Tanes
AU - Tandhavanant, Sarunporn
AU - Sengyee, Sineenart
AU - Phunpang, Rungnapa
AU - Preechanukul, Anucha
AU - Surin, Uriwan
AU - Tuanyok, Apichai
AU - Lertmemongkolchai, Ganjana
AU - Chantratita, Wasun
AU - West, T. Eoin
AU - Chantratita, Narisara
N1 - Publisher Copyright:
© 2020 Saiprom et al.
PY - 2020/9
Y1 - 2020/9
N2 - Background Burkholderia pseudomallei is an environmental bacterium that causes melioidosis. A facul-tative intracellular pathogen, B. pseudomallei can induce multinucleated giant cells (MNGCs) leading to plaque formation in vitro. B. pseudomallei can switch colony morpho-types under stress conditions. In addition, different isolates have been reported to have varying virulence in vivo, but genomic evolution and the relationship with plaque formation is poorly understood. Methodology/Principle findings To gain insights into genetic underpinnings of virulence of B. pseudomallei, we screened plaque formation of 52 clinical isolates and 11 environmental isolates as well as 4 isogenic morphotype isolates of B. pseudomallei strains K96243 (types II and III) and 153 (types II and III) from Thailand in A549 and HeLa cells. All isolates except one environmental strain (A4) and K96243 morphotype II were able to induce plaque formation in both cell lines. Intracellular growth assay and confocal microscopy analyses demonstrated that the two plaque-forming-defective isolates were also impaired in intracellular replication, actin polymeriza-tion and MNGC formation in infected cells. Whole genome sequencing analysis and PCR revealed that both isolates had a large genomic loss on the same region in chromosome 2, which included Bim cluster, T3SS-3 and T6SS-5 genes. Conclusions/Significance Our plaque screening and genomic studies revealed evidence of impairment in plaque formation in environmental isolates of B. pseudomallei that is associated with large genomic loss of genes important for intracellular multiplication and MNGC formation. These findings suggest that the genomic and phenotypic differences of environmental isolates may be associated with clinical infection.
AB - Background Burkholderia pseudomallei is an environmental bacterium that causes melioidosis. A facul-tative intracellular pathogen, B. pseudomallei can induce multinucleated giant cells (MNGCs) leading to plaque formation in vitro. B. pseudomallei can switch colony morpho-types under stress conditions. In addition, different isolates have been reported to have varying virulence in vivo, but genomic evolution and the relationship with plaque formation is poorly understood. Methodology/Principle findings To gain insights into genetic underpinnings of virulence of B. pseudomallei, we screened plaque formation of 52 clinical isolates and 11 environmental isolates as well as 4 isogenic morphotype isolates of B. pseudomallei strains K96243 (types II and III) and 153 (types II and III) from Thailand in A549 and HeLa cells. All isolates except one environmental strain (A4) and K96243 morphotype II were able to induce plaque formation in both cell lines. Intracellular growth assay and confocal microscopy analyses demonstrated that the two plaque-forming-defective isolates were also impaired in intracellular replication, actin polymeriza-tion and MNGC formation in infected cells. Whole genome sequencing analysis and PCR revealed that both isolates had a large genomic loss on the same region in chromosome 2, which included Bim cluster, T3SS-3 and T6SS-5 genes. Conclusions/Significance Our plaque screening and genomic studies revealed evidence of impairment in plaque formation in environmental isolates of B. pseudomallei that is associated with large genomic loss of genes important for intracellular multiplication and MNGC formation. These findings suggest that the genomic and phenotypic differences of environmental isolates may be associated with clinical infection.
UR - http://www.scopus.com/inward/record.url?scp=85092243628&partnerID=8YFLogxK
U2 - 10.1371/journal.pntd.0008590
DO - 10.1371/journal.pntd.0008590
M3 - Article
C2 - 32991584
AN - SCOPUS:85092243628
SN - 1935-2727
VL - 14
SP - 1
EP - 18
JO - PLoS Neglected Tropical Diseases
JF - PLoS Neglected Tropical Diseases
IS - 9
M1 - e0008590
ER -