Research program at the University of Strathclyde Centre for Biophotonics

Research program at the University of Strathclyde Centre for Biophotonics

The research program at the University of Strathclyde Centre for Biophotonics is focused on the understanding of physiological and disease processes at a cellular level. In this type of research many technical difficulties can arise, especially when trying to understand the cell behaviour as it might occur in the body. The Centre for Biophotonics develops new optical approaches for overcoming problems such as:-

  • Imaging living tissues with minimal damage for longer periods.
  • Imaging at high resolution at deeper levels in tissues in situ.

Research at the laboratory of Professor Paul Garside and Dr James Brewer focuses on using this technology to study the cellular interactions during the initiation of immune responses. These interactions are studied in a number of scenarios, including vaccination, the host responses to infectious disease and the initiation of autoimmunity.

The collaboration in this area between Paul and James was initially established when both were Wellcome Trust Career Development Fellows working on oral tolerance and Th1/2 polarisation respectively. In both of these situations, cellular interactions between immune system cells ultimately dictate the immunological outcome. At the time, these studies were limited to analysis of cells in vitro, or ‘snapshots’ of isolated tissue sections. These limitations lead to a common interest to understand the multiple molecular and cellular interactions underlying immunological processes in a 4-dimensional spatiotemporal manner in vivo.

The group and the Centre for Biophotonics have developed the use of multiphoton microscopy to study the real time behaviour of primed or tolerised antigen-specific T cells in vivo. They established systems for the analysis of T cell and Antigen Presenting Cells (APC) behaviour in intact lymph nodes and demonstrated that subtle differences in the clustering of T cells with APC during the early stages (8 and 20 h) of the induction of oral tolerance or adjuvant-induced priming are associated with distinct immunological outcomes (1), highlighted in Nature Immunology). Thus, slightly larger, longer-lived clusters are the norm in the induction of priming whereas tolerance is associated with smaller, shorter duration clusters. More recent collaborative work (2) has provided a mechanistic insight into the observed differences in T cell behaviour as they demonstrated that CTLA-4 increases T cell motility and prevents the formation of stable contacts between T cells and APC, providing a potential mechanism by which CTLA-4 may modulate the activation of T cells. Understanding the basic interactions of T cells and APCs and will be critically important in developing rationally designed and targeted therapeutic intervention for infectious and autoimmune diseases and we shall continue to apply our fundamental findings in these areas.

In autoimmunity, the group has performed studies transferring in vitro polarized, OVA-specific TcR transgenic T cells to question the importance of joint specific T cells in the initiation of arthritis. These studies suggest that induction of autoimmunity in this model is a highly dynamic process involving two distinct tissues, the joint and the draining lymph node (3). As such, a greater understanding of the dynamics of the real time interactions of T cell and APC in joint and draining lymph node will be an important development in this area.

These and other techniques have been used with Wellcome Trust support to determine the cellular basis for generalised suppression seen during malaria infection (4, 5). These studies have been highlighted in Science and Nature Immunology and also made mainstream press (Sunday Herald and The Times) and clearly demonstrated that Plasmodium prevents the induction of immunity to heterologous antigens by modulating DC function, explaining why vaccine strategies fail in malaria endemic regions.

James and Paul’s research team have made considerable progress in determining the cellular and molecular interactions underlying many of the immunological scenarios in this field of research.

James says “Volocity is a useful tool that allows us to produce highly complex 4-dimensional data in a format that is easy to communicate to an audience The measurement functions in Volocity Quantitation have given us the tools to produce meaningful quantitative data from these images that allow us to perform objective analysis of these complex data sets”

Please visit the Centre for Biophotonics if you would like to read more about their research.

References

  1. Zinselmeyer, B. H., J. Dempster, A. M. Gurney, D. Wokosin, M. Miller, H. Ho, O. R. Millington, K. M. Smith, C. M. Rush, I. Parker, M. Cahalan, J. M. Brewer, and P. Garside. 2005. In situ characterization of CD4+ T cell behavior in mucosal and systemic lymphoid tissues during the induction of oral priming and tolerance. J. Exp. Med. 201:1815-1823.
  2. Schneider, H., J. Downey, A. Smith, B. H. Zinselmeyer, C. Rush, J. M. Brewer, B. Wei, N. Hogg, P. Garside, and C. E. Rudd. 2006. Reversal of the TCR Stop Signal by CTLA-4. Science. 313:1972-1975.
  3. Maffia, P., J. M. Brewer, J. A. Gracie, A. Ianaro, B. P. Leung, P. J. Mitchell, K. M. Smith, I. B. McInnes, and P. Garside. 2004. Inducing Experimental Arthritis and Breaking Self-Tolerance to Joint-Specific Antigens with Trackable, Ovalbumin-Specific T Cells. J. Immunol. 173:151-156.
  4. Millington, O. R., C. Di Lorenzo, R. S. Phillips, P. Garside, and J. M. Brewer. 2006. Suppression of adaptive immunity to heterologous antigens during Plasmodium infection through hemozoin-induced failure of dendritic cell function. J Biol 5:1-22.
  5. Millington, O. R., V. B. Gibson, C. M. Rush, B. H. Zinselmeyer, R. S. Phillips, P. Garside, and J. M. Brewer. 2007. Malaria impairs T cell clustering and immune priming despite normal signal 1 from dendritic cells. PLoS Pathog 3:1380-1387.
3D rendered image of a lymph node showing T cells. Image was obtained by multiphoton microscopy of an intact lymph node