Immunology: An Illustrated Outline _HOT_
Indirect immunofluorescence In this technique the section is incubated with the test antibody, which is then visualized by the addition of a second-layer fluorescent anti-antibody. The amplification produced by the second antibody increases the sensitivity of the assay, and by using class- or subclass-specific reagents, particular antibody isotypes can be identified in the test serum. This technique is of great value for identifying antibodies against tissue antigens, as illustrated below, where antibodies against a pancreatic islet of Langerhans in diabetic serum were identified using indirect immunofluorescence on a frozen section of pancreas.
Immunology: An Illustrated Outline
The phenomenon known as immunothrombosis has garnered increased attention over the last few years. Much work has been done to characterize the cross talk between hemostasis and the innate immune system. This thesis outlines the role of the vascular endothelial cells during immunothrombotic events as regulators of coagulation, platelet-, and leukocyte recruitment.
A newly developed method for investigating the interaction between endothelial cells and the blood compartment illustrated the procoagulant and proinflammatory effects elicited by tumor necrosis factor α activated endothelial cells upon exposure to whole blood. The method was utilized in evaluating treatment of endothelial dysfunction and disruption with a heparin conjugate. Damaged or hypoxic endothelial cells, in addition to basement membrane collagen, that were pretreated with the heparin conjugate prior to contact with blood were found to have reduced activation of coagulation, platelet-, and leukocyte recruitment; in contrast to unfractionated heparin, which had no effect on the aforementioned parameters. The treatment was then investigated in the setting of ischemia reperfusion injury during kidney transplantation and the heparin conjugate was found to bind cultured endothelial cells with high avidity under cold storage conditions. Furthermore, it was found to bind to the renal vasculature during static cold storage and was subsequently found to be beneficial with regard to early graft function in an experimental mouse model of syngeneic kidney transplantation. Recipients of kidneys treated with the heparin conjugate had reduced serum creatinine compared to controls 24 hours after transplantation. Lastly, the anticoagulant properties of the heparin conjugate were investigated in comparison to unfractionated heparin. While the conjugate exerted reduced capacity with regard to thrombin inhibition, it rapidly inhibited the binding of platelets to exposed collagen. The conjugate was furthermore found to preferentially locate to sites of endothelial cell activation at early stage during endotoxic shock in mice.
Some of you might be interested in a new paper which we have just published in Frontiers in Immunology (link). The paper outlines a new model built on cellular cooperation to explain the establishment of peripheral T cell tolerance. Unusually, the paper is wholly theoretical. The paper is also somewhat technical, since the model is formulated mathematically as a linear programing optimization problem that can be implemented as a multiplicative update algorithm, which shows a rapid convergence to a stable state. We (i.e. the authors) would welcome feedback on the paper, as it represents the first step in a more ambitious plan to build a broader theoretical framework underpinning the human immune system.
Publishing in Nature Communications, the team of scientists from Newcastle, Sheffield and Glasgow Universities together with colleagues from Imperial College and Diamond Light Source, outline the structure of the main protein, SlpA, that forms the links of the chain mail and how they are arranged to form a pattern and create this flexible armour. This opens the possibility of designing C. diff specific drugs to break the protective layer and create holes to allow molecules to enter and kill the cell. 041b061a72