Novel T lymphocyte proliferation assessment using whole mouse cryo-imaging

Patiwet Wuttisarnwattana, Syed A. Raza, Saada Eid, Kenneth R. Cooke, David L. Wilson

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

New imaging technologies enable one to assess T-cell proliferation, an important feature of the immunological response. However, none of the traditional imaging modalities allow one to examine quantiatively T-cell function with microscopic resolution and single cell sensitivity over an entire mouse. To address this need, we established T-cells proliferation assays using 3D microscopic cryo-imaging. Assays include: (1) biodistribution of T-cells, (2) secondary lymphoid organ (SLO) volume measurement, (3) carboxyfluorescein succinimidyl ester (CFSE) dilution per cell as cells divide. To demonstrate the application, a graft-versus-host-disease (GVHD) model was used. 3D visualization show that T-cells specifically homed to the SLOs (spleen and lymph nodes) as well as GVHD target organs (such as GI-tract, liver, skin and thymus).The spleen was chosen as representative of the SLOs. For spleen size analysis, volumes of red and white pulp were measured. Spleen volumes of the allogeneic mice (with GVHD) were significantly larger than those of the syngeneic mice (without GVHD) at 72 to 120 hours post-transplant. For CFSE dilution approach, we employed color-coded volume rendering and probability density function (PDF) of single cell intensity to assess T-cell proliferation in the spleen. As compared to syngeneic T-cells, the allogeneic T-cells quickly aggregated in the spleen as indicated by increasing of CFSE signal over the first 48 hours. Then they rapidly proliferated as evidenced by reduced CFSE intensity (at 48-96 hours). Results suggest that assays can be used to study GVHD treatments using T-cell proliferation and biodistibution as assays. In summary, this is the first time that we are able to track and visualize T-cells in whole mouse with single cell sensitivity. We believe that our technique can be an alternative choice to traditional in vitro immunological proliferation assays by providing assessment of proliferation in an in vivo model.

Original languageEnglish (US)
Title of host publicationMedical Imaging 2014
Subtitle of host publicationBiomedical Applications in Molecular, Structural, and Functional Imaging
PublisherSPIE
ISBN (Print)9780819498311
DOIs
StatePublished - 2014
EventMedical Imaging 2014: Biomedical Applications in Molecular, Structural, and Functional Imaging - San Diego, CA, United States
Duration: Feb 16 2014Feb 18 2014

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume9038
ISSN (Print)1605-7422

Other

OtherMedical Imaging 2014: Biomedical Applications in Molecular, Structural, and Functional Imaging
CountryUnited States
CitySan Diego, CA
Period2/16/142/18/14

Keywords

  • Biodistribution
  • CFSE dilution
  • Cryo-imaging
  • Fluorescent imaging
  • Graft-versus-host disease
  • Lymphocyte proliferation assay
  • Secondary lymphoid organs
  • Spleen

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging

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  • Cite this

    Wuttisarnwattana, P., Raza, S. A., Eid, S., Cooke, K. R., & Wilson, D. L. (2014). Novel T lymphocyte proliferation assessment using whole mouse cryo-imaging. In Medical Imaging 2014: Biomedical Applications in Molecular, Structural, and Functional Imaging [90381U] (Progress in Biomedical Optics and Imaging - Proceedings of SPIE; Vol. 9038). SPIE. https://doi.org/10.1117/12.2042960