Bioluminescent Bb in vivo imaging in mice

General or non-medical topics with information and discussion related to Lyme disease and other tick-borne diseases.
Post Reply
RitaA
Posts: 2768
Joined: Thu 1 Jul 2010 8:33

Bioluminescent Bb in vivo imaging in mice

Post by RitaA » Fri 16 Jan 2015 8:00

This definitely qualifies as science, but I found it in a scientist's online profile as opposed to a published article, so I'm posting it here.

http://njms.rutgers.edu/departments/mol ... y/parveen/
Nikhat Parveen, Ph.D.

Associate Professor

Virulence factors of Pseudomonas aeruginosa and Lyme disease spirochete, Borrelia burgdorferi
Mouse is a natural host of B. burgdorferi and C3H mice show several manifestations of Lyme disease observed in humans. We have recently adapted firefly luciferase-based detection system for B. burgdorferi. Using a combination of bioluminescent B. burgdorferi and mouse model of infection, we will further analyze the contribution of each bacterial ligand-host receptor interaction in Lyme pathogenesis. Tissue colonization by the spirochetes will be monitored non-invasively by employing in vivo imaging system.
Have a look at this rather cool image:

http://njms.rutgers.edu/departments/mol ... 2_2010.jpg

Could this type of imaging eventually be used to test the effectiveness of various antibiotics -- at least in mice?

Now if only researchers could find a human volunteer or two who would be willing to be infected with bioluminescent Bb so scientists could monitor how the disease disseminates. Surely anyone who steadfastly believes that all Lyme disease infections are easy to cure with a short course of antibiotics would be willing to volunteer in the name of science. Then again, maybe not. ;)

On a more serious note, other scientists are also using in vivo bioluminescent imaging of Borrelia burgdorferi in their research:
Hyde, J.A., Weening, E.H., Chang, M.H., Trzeciakowski, J.P., Höök, M., Cirillo, J.D., and J. T. Skare. 2011. Bioluminescent imaging of Borrelia burgdorferi in vivo demonstrates that the fibronectin binding protein BBK32 is required for optimal infectivity. Mol Microbiol. 82(1): 99-113.
Here's some ongoing research that Jennifer Hyde and her team are conducting:

https://www.collectiveip.com/grants/NIH:8497620
In vivo dual Bioluminescence Reporter System of Infectious Borrelia burgdorferi
5R21AI101740-02

Amount
$205,273

Source
NIH

Investigators
Jennifer Hyde

Effective Date
2013-07-01

Expiration Date
2015-06-30

Categories
Human Genetics

Infectious Disease

--------------------------------------------------------------------------------

Abstract

DESCRIPTION (provided by applicant): It is well established that Borrelia burgdorferi, the etiologic agent of Lyme disease, modulates gene expression during infection as it moves between an arthropod vector and mammalian hosts. Several genes required for the establishment of mammalian infection are known with the prototypical gene being ospC. The lipoprotein OspC is absolutely required for mammalian infectivity, and although a putative ligand-binding domain is essential for infectivity, the exact function of OspC is not known. Subsequent studies indicated that ospC is coordinately regulated via a response regulator (Rrp2) that, together with RpoN, drives the expression of RpoS, which then promotes the transcriptional activation of ospC and other infectivity-associated borrelial genes. However, the activation of ospC is transient as it is repressed following infection. In this regard, if ospC expression is made constitutive, the spirochetes are rapidly cleared. Despite this observation, the kinetics of ospC expression, i.e., the amplitude and diminution within a living system over time, is not known. Recently we have used in vivo imaging to detect light emitting (i.e., luciferase [luc] expressing) infectious B. burgdorferi following needle inoculation in mice. The advantage of this approach is that B. burgdorferi can be visualized numerous times in live mice over time to track the infectious process. Given the sensitivity of this technique, an additional potential applicatio might be to assess the expression of targeted genes. To test this premise, we have fused the ospC promoter (PospC) to luc. Our Preliminary Data suggests that ospC is highly expressed early in the infectious process within skin, but is significantly reduced later in the infection, consistent with prior reports indicating that it is down regulated following colonization and dissemination. The utility of this approach will now be expanded to further study the spatial expression or ospC as well as other genes that are coordinately regulated with ospC via RpoS. To this end we propose the following Specific Aims: (1) Characterize the in vivo tissue tropism and temporal production of borrelial ospC utilizing a dual bioluminescence reporter system;and (2) Determine the in vivo expression patterns of genes involved in the Rrp2-RpoN-RpoS regulatory pathway. In the proposed studies, the fate of ospC transcription will be tracked following disseminated infection as well as genes involved in the infectious process to determine if RpoS regulation is highly coordinated or occurs at differential times as B. burgdorferi disseminates. The ability to visualize these regulatory patterns of specific borrelial promoter-luc constructs, focusing on the promoters of genes known to be involved in experimental mouse infection, should provide important insight into the hierarchy and/or temporal expression of these loci to establish and maintain an infectious focus. This exciting approach provides a powerful non-invasive, real time modality to evaluate the activity of a given promoter in a temporal and spatial manner.

Lorima
Posts: 914
Joined: Mon 29 Oct 2007 20:47

Re: Bioluminescent Bb in vivo imaging in mice

Post by Lorima » Fri 16 Jan 2015 13:58

Fascinating! Good find!
"I have to understand the world, you see."
Richard Feynman

User avatar
ChronicLyme19
Posts: 564
Joined: Mon 11 Aug 2014 17:42
Location: NY, USA

Re: Bioluminescent Bb in vivo imaging in mice

Post by ChronicLyme19 » Fri 16 Jan 2015 14:54

Oh that is very cool. But does it bioluminesce at all times, or only when it's active? Do round body forms still luminesce?
Half of what you are taught is incorrect, but which half? What if there's another half missing?

RitaA
Posts: 2768
Joined: Thu 1 Jul 2010 8:33

Re: Bioluminescent Bb in vivo imaging in mice

Post by RitaA » Fri 16 Jan 2015 17:51

Great questions!

Stay tuned for answers from researchers -- after they ask themselves the same questions, that is. In all fairness, they may already know, and so might a few members here. I'm hoping they will chime in.

Post Reply