In Vivo Imaging Demonstrates That Borrelia burgdorferi ospC Is Uniquely Expressed Temporally and Spatially throughout Experimental Infection.
Borrelia burgdorferi is a spirochetal bacterium transmitted by the Ixodes tick that causes Lyme disease in humans due to its ability to evade the host immune response and disseminate to multiple immunoprotective tissues. The pathogen undergoes dynamic genetic alterations important for adaptation from the tick vector to the mammalian host, but little is known regarding the changes at the transcriptional level within the distal tissues they colonize. In this study, B. burgdorferi infection and gene expression of the essential virulence determinant ospC was quantitatively monitored in a spatial and temporal manner utilizing reporter bioluminescent borrelial strains with in vivo and ex vivo imaging. Although expressed from a shuttle vector, the PospC-luc construct exhibited a similar expression pattern relative to native ospC. Bacterial burden in skin, inguinal lymph node, heart, bladder and tibiotarsal joint varied between tissues and fluctuated over the course of infection possibly in response to unique cues of each microenvironment. Expression of ospC, when normalized for changes in bacterial load, presented unique profiles in murine tissues at different time points. The inguinal lymph node was infected with a significant B. burgdorferi burden, but showed minimal ospC expression. B. burgdorferi infected skin and heart induced expression of ospC early during infection while the bladder and tibiotarsal joint continued to display PospC driven luminescence throughout the 21 day time course. Localized skin borrelial burden increased dramatically in the first 96 hours following inoculation, which was not paralleled with an increase in ospC expression, despite the requirement of ospC for dermal colonization. Quantitation of bioluminescence representing ospC expression in individual tissues was validated by qRT-PCR of the native ospC transcript. Taken together, the temporal regulation of ospC expression in distal tissues suggests a role for this virulence determinant beyond early infection.
Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M Health Science Center, Bryan/College Station, Texas, United States of America.
Jonathan T
JT
Skare
Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M Health Science Center, Bryan/College Station, Texas, United States of America.
Dana K
DK
Shaw
Department of Medical Physiology, College of Medicine, Texas A&M Health Science Center, Bryan/College Station, Texas, United States of America.
Jerome P
JP
Trzeciakowski
Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M Health Science Center, Bryan/College Station, Texas, United States of America.
Jenny A
http://orcid.org/0000-0001-9442-9980
JA
Hyde
D000704:Analysis of Variance; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D025065:Borrelia burgdorferi / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D005260:Female; D008163:Luminescent Measurements / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D020133:Reverse Transcriptase Polymerase Chain Reaction
D001426:Bacterial Proteins