Sole_Source-SANKYO_2017_.pdf

Sole Source Justification

Germ-Free Work on Transgenic Mice at Sankyo Labo Service Corporation:

Re-derivation, Maintenance and Gnotobiotic Study

To investigate the role of commensal microbiota as a trigger for spontaneous autoimmune uveitis in an IRBP-specific T cell receptor transgenic mouse model that we had developed (refs.1, 2), we have been collaborating since 2012 with a Japanese vendor, Sankyo Labo Service, who provides services to operate the germ-free isolators in the germ-free facilities for long-term projects. Unfortunately, there is no such commercial source in the US: We had unsatisfactory experience in 2010-11 with Taconic Farm Inc, which was the only source for us then, but they failed germ-free re-derivation of our transgenic mice several times and told us that they were unable to provide the service. Sankyo Labo

Service is the only vendor for us that is capable of providing the service of re-deriving and maintaining the “germ-free” colony of our transgenic mice and is willing to continue to work as collaboration, which turns out to be long-term projects.

We recruited Japanese collaborators to fulfill the goal of our project: Dr. Katsuko

Sudo, an expert on experimental animals from Tokyo Medical University who serves as an advisor at Sankyo Labo Service, and started to work on germ-free transgenic mice and microorganisms that may serve as a trigger for uveitis. Dr. Sudo and her colleague at

Sankyo Labo Service have access to their germ-free animal facility and are the only expert collaborators who are capable of doing gnotobiotic reconstitution in the germ-free isolators as well as of growing any kinds of microorganisms. We covered part of the research expenses associated with breeding and maintenance of the germ-free colony and gnotobiotic study in Japan and transferred the fund of from NEI in the past several years.

To continue our collaboration to identify the causative microorganism(s) for spontaneous uveitis in our transgenic mice, we need to massively expand the germ-free colony by increasing the number of isolators to operate, as well as to re-derive more transgenic mice under the germ-free conditions. We will then need additional isolators for gnotobiotic reconstitution study with candidate microorganisms. We agree to cover part of the expenses associated with this germ-free-related work at Sankyo Labo Service, where our germ-free transgenic mice are already maintained.

References:

1) Horai R, et al. Breakdown of immune privilege and spontaneous autoimmunity in mice expressing a transgenic T cell receptor specific for a retinal autoantigen. Journal of Autoimmunity 44: 21-33 (2013).

2) Horai R, et al. Microbiota-dependent activation of an autoreactive T cell receptor provokes autoimmunity in an immune privileged site. Immunity 43: 343-353 (2015).

5/26/2017