In the case of MS we can determine whether a specific immune cell population is functioning normally or not before or after manipulation of a defined condition. We can then experiment on the sorted cell populations to evaluate functional differences between them. So we end up for example, with one test tube of A B C and D and another with A B F and G. We pass the cells through the flow channel but now we program the sorter to separate cell populations with defined characteristics of interest. We can then react a population of cells with a panel of antibodies, each with its own specific fluorochrome. The cell sorter works on a similar principle to the flow cytometer but only surface molecules can be interrogated so as to preserve the cells. The flow cytometer essentially destroys the cells which have been analyzed so that their function as live cells cannot be investigated further. This limits our ability to investigate the function of a cell population repeatedly. Unfortunately the cells are not preserved after using this technique so we can sample what is happening in a population of immune cells in MS but we cannot use the analyzed cells again. The results thus obtained can then be evaluated further with a specialized computer program. The analysis occurs at speeds of up to 1000 cells/sec so a large number of cells can be analyzed in a very short time. Because we can also look at molecules inside a cell as well we can determine whether these cells are similar or different in internal molecules which may indicate similar or different cell functions. So, for example we can distinguish between a cell with a surface that expresses A B C and D, etc., and differentiate it from a cell that expresses A B F and G. Different lasers excite different fluorochromes, so that more than 20 molecules can potentially be studied simultaneously in different cell populations. When the cell is exposed to a particular and narrow wavelength of light from a laser the relevant fluorochrome fluoresces at a defined wavelength and this fluorescence and its intensity are then read by the flow cytometer. A suspension of the cells is then passed in single file through a flow cell where it interacts with a laser beam. Each specific antibody is labelled with a small molecule called a fluorochrome – and different antibodies are labelled with different fluorochromes to distinguish between them. Antibodies against defined molecules on the surface of the interior of the cells are reacted with the cells. This technique is integral to and has revolutionized much of contemporary immunology.įirst, cells are prepared for the analysis. It has many innovative uses but we mainly use it at MSRI as a cell biology tool to study immune cells. Understanding these processes will also allow us to rationally design more specific treatments for MS.įlow cytometry is a technique used to study the surface and interior of immune or other cells. Flow cytometers and Cell sorters are invaluable pieces of laboratory equipment which allow us to dissect the processes in immune cells which lead to the development of MS.This is the goal of the research we do at MSRI. The key to understanding this process in MS is to define the stages of function and malfunction of T regulatory cells and the resultant dysfunction of unrestrained T effector cells.In the case of autoimmunity such as MS, the T regulatory cells may not only fail to function but may change to function like T effector cells which cause inflammation and tissue (myelin) damage. T regulatory cells normally act to suppress the function of other immune cells called T effector cells, which prevents the development of autoimmunity.The failure of T regulatory cells is the result of a long developmental process which precedes the onset of clinical MS and which involves genetic inheritance as well as environmental influences acting on these cells to ultimately disturb their normal function.This type of disease is primarily due to a defect in function of certain immune cells known as T regulatory cells. Restricted autoimmune diseases like MS are confined to one tissue or organ.They reflect a widespread derangement in immune function. Systemic autoimmune diseases like Lupus affect the whole body.MS is an autoimmune disease which damages myelin and starts a chronic degenerative process in the brain and spinal cord.Influenza viruses, 3D illustration showing surface glycoprotein spikes hemagglutinin and neuraminidase
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