Analysis and modelling of immune cell behaviour in lymph nodes based on multi-photon imaging

This thesis presents quantitative studies of T cell and dendritic cell (DC) behaviour in mouse lymph nodes (LNs) in the naive state and following immunisation. These processes are of importance and interest in basic immunology, and better understanding could improve both diagnostic capacity and therapeutic manipulations, potentially helping in producing more effective vaccines or developing treatments for autoimmune diseases. The problem is also interesting conceptually as it is relevant to other fields where 3D movement of objects is tracked with a discrete scanning interval. A general immunology introduction is presented in chapter 1. In chapter 2, I apply quantitative methods to multi-photon imaging data to measure how T cells and DCs are spatially arranged in LNs. This has been previously studied to describe differences between the naive and immunised state and as an indicator of the magnitude of the immune response in LNs, but previous analyses have been generally descriptive. The quantitative analysis shows that some of the previous conclusions may have been premature. In chapter 3, I use Bayesian state-space models to test some hypotheses about the mode of T cell search for DCs. A two-state mode of movement where T cells can be classified as either interacting to a DC or freely migrating is supported over a model where T cells would home in on DCs at distance through for example the action of chemokines. In chapter 4, I study whether T cell migration is linked to the geometric structure of the fibroblast reticular network (FRC). I find support for the hypothesis that the movement is constrained to the fibroblast reticular cell (FRC) network over an alternative 'random walk with persistence time' model where cells would move randomly, with a short-term persistence driven by a hypothetical T cell intrinsic 'clock'. I also present unexpected results on the FRC network geometry. Finally, a quantitative method is presented for addressing some measurement biases inherent to multi-photon imaging. In all three chapters, novel findings are made, and the methods developed have the potential for further use to address important problems in the field. In chapter 5, I present a summary and synthesis of results from chapters 3-4 and a more speculative discussion of these results and potential future directions.

A distributed, compact routing protocol for the Internet

The Internet has grown in size at rapid rates since BGP records began, and continues to do so. This has raised concerns about the scalability of the current BGP routing system, as the routing state at each router in a shortest-path routing protocol will grow at a supra-linearly rate as the network grows. The concerns are that the memory capacity of routers will not be able to keep up with demands, and that the growth of the Internet will become ever more cramped as more and more of the world seeks the benefits of being connected. Compact routing schemes, where the routing state grows only sub-linearly relative to the growth of the network, could solve this problem and ensure that router memory would not be a bottleneck to Internet growth. These schemes trade away shortest-path routing for scalable memory state, by allowing some paths to have a certain amount of bounded “stretch”. The most promising such scheme is Cowen Routing, which can provide scalable, compact routing state for Internet routing, while still providing shortest-path routing to nearly all other nodes, with only slightly stretched paths to a very small subset of the network. Currently, there is no fully distributed form of Cowen Routing that would be practical for the Internet. This dissertation describes a fully distributed and compact protocol for Cowen routing, using the k-core graph decomposition. Previous compact routing work showed the k-core graph decomposition is useful for Cowen Routing on the Internet, but no distributed form existed. This dissertation gives a distributed k-core algorithm optimised to be efficient on dynamic graphs, along with with proofs of its correctness. The performance and efficiency of this distributed k-core algorithm is evaluated on large, Internet AS graphs, with excellent results. This dissertation then goes on to describe a fully distributed and compact Cowen Routing protocol. This protocol being comprised of a landmark selection process for Cowen Routing using the k-core algorithm, with mechanisms to ensure compact state at all times, including at bootstrap; a local cluster routing process, with mechanisms for policy application and control of cluster sizes, ensuring again that state can remain compact at all times; and a landmark routing process is described with a prioritisation mechanism for announcements that ensures compact state at all times.