25 research outputs found
Modeling Bacterial DNA: Simulation of Self-avoiding Supercoiled Worm-Like Chains Including Structural Transitions of the Helix
Under supercoiling constraints, naked DNA, such as a large part of bacterial
DNA, folds into braided structures called plectonemes. The double-helix can
also undergo local structural transitions, leading to the formation of
denaturation bubbles and other alternative structures. Various polymer models
have been developed to capture these properties, with Monte-Carlo (MC)
approaches dedicated to the inference of thermodynamic properties. In this
chapter, we explain how to perform such Monte-Carlo simulations, following two
objectives. On one hand, we present the self-avoiding supercoiled Worm-Like
Chain (ssWLC) model, which is known to capture the folding properties of
supercoiled DNA, and provide a detailed explanation of a standard MC simulation
method. On the other hand, we explain how to extend this ssWLC model to include
structural transitions of the helix.Comment: Book chapter to appear in The Bacterial Nucleoid, Methods and
Protocols, Springer serie
Proinsulin-mediated induction of type 1 diabetes in HLA-DR4-transgenic mice
Abstract Antigen-specific immunotherapy of autoimmune disease currently remains the only potentially curative approach. However, translation of promising pre-clinical results into successful clinical application has proven challenging. In part, this is because pre-clinical findings in mouse models have to be redesigned for human application due to differences in MHC II. To reduce the gap between pre-clinical and clinical studies, we have created a novel mouse model that expresses human HLA-DR4, but no endogenous MHC on antigen-presenting cells. Moreover, human B7.1 (CD80) is expressed in the pancreatic islets under the control of the rat insulin promoter. Although this model does not develop diabetes spontaneously, it is susceptible to the induction of type 1 diabetes by challenging mice with overlapping peptides derived from murine proinsulin-2 in adjuvant. Unlike the NOD model of spontaneous type 1 diabetes, but akin to the human condition, this model does not have a gender bias. Furthermore, similar to the human condition, the disease is characterised by a diverse leucocyte infiltration of the pancreatic islets and the formation of anti-proinsulin auto-antibodies. The model that we report here offers detailed insights into type-1 diabetes and is expected to prove instrumental when studying the mechanism of action in translational, antigen-specific immunotherapy
Class II major histocompatibility complex tetramer staining: progress, problems, and prospects
The use of major histocompatibility complex (MHC) tetramers in the detection and analysis of antigen-specific T cells has become more widespread since its introduction 11 years ago. Early challenges in the application of tetramer staining to CD4+ T cells centred around difficulties in the expression of various class II MHC allelic variants and the detection of low-frequency T cells in mixed populations. As many of the technical obstacles to class II MHC tetramer staining have been overcome, the focus has returned to uncertainties concerning how oligomer valency and T-cell receptor/MHC affinity affect tetramer binding. Such issues have become more important with an increase in the number of studies relying on direct ex vivo analysis of antigen-specific CD4+ T cells. In this review we discuss which problems in class II MHC tetramer staining have been solved to date, and which matters remain to be considered