Neutrophil chemotaxis assay

The phagocytosis of pathogens by neutrophils generally involves several steps of chemotaxis, conditioning, phagocytosis and sterilization, which is a very complicated process. Under the action of chemokines, neutrophils move toward the bacteria, and the bacteria acting on the opsonins tend to adhere to the neutrophils, causing the neutrophil membrane to invade and swallowing the bacteria through the pinocytosis. The phagocytosis packet is fused to the lysosome in the cell to form a lysosome, which kills the bacteria. However, if the body's chemokines are reduced or the phagocytic cells themselves do not respond to normal chemokines, phagocytic phagocytosis can be attenuated, making the body susceptible to infection. Basic Information Specialist classification: cardiovascular examination classification: blood examination Applicable gender: whether men and women apply fasting: not fasting Analysis results: Below normal: The chemotaxis of neutrophils is weakened, suggesting that there is a lack of complement C3 in the blood, which is common in repeated infections, gingivitis, otitis media, and neutropenia in peripheral blood. Normal value: Newborn mobile index: 2.0-2.5 Adult Mobile Index: 3.0-3.5 Above normal: no information yet. negative: Positive: Tips: The experimental conditions are different and the difference is very large, so the normal control is very important. Normal value The newborn's mobile index is about 2.0 to 2.5; for adults, it is about 3.0 to 3.5. The experimental conditions are different and the difference is very large, so the normal control is very important. Clinical significance There are many natural chemokines in the human body. When the chemokines in the human body are reduced or the phagocytic cells themselves are not responding to normal chemokines, the chemokines' chemotaxis is weakened and the body is easily infected. The neutrophil chemotaxis test can often be used as a test marker for neutrophil motility in patients with certain diseases. Neutrophil chemotaxis is reduced, mainly suggesting that there is a lack of complement C3 in the blood, which is common in repeated infections, gingivitis, otitis media, and neutropenia in peripheral blood. Deficiencies in neutrophil chemotaxis are also seen in che-diak-Higashi syndrome, retarded leukocyte syndrome, actin dysfunction, membrane glycoprotein deficiency, and high IgE syndrome. Low results may be diseases: neutropenia, Behcet's disease, otitis media, positive results may be diseases: neutropenia, Behcet's scleritis, otitis media matters needing attention (1) The preparation of agarose plate can also be carried out with Eagle culture solution and RP-MI-1640 culture solution, but serum (AB normal human serum or calf serum) must be added, and the final concentration is 10% to 20%. If there is no serum, the number of migrated cells is reduced and dispersed, and the chemotactic movement is weakened. (2) The neutrophils examined cannot be collected by dextran sedimentation because of their ability to inhibit neutrophil migration. (3) The number of neutrophils added to each well must be accurately quantified, and the amount is closely related to chemotaxis. If the number of cells is too much, the chemotaxis index is decreased, the number of cells is too small, and the chemotaxis index is not decreased. However, since the number of migrated cells is reduced and dispersed, it is often difficult to accurately measure the distance of cell migration. (4) The pore spacing is ideally 2.4 mm, and the pore spacing is too large, so that the chemokine can be excessively diluted and diluted in the agarose. (5) The culture time is 2 to 3 hours, and the cell migration reaches a peak. Culture in a 5% CO2 environment enhances the neutrophil chemotaxis. Inspection process (1) Plate punching and loading: The prepared agar plate is punched with a 2.4 mm inner diameter puncher, and the hole pitch is 2 to 4 mm. Six specimens can be tested simultaneously on each plate. Take three holes in each row and measure one specimen. 5 μl of mesoporous white blood cell suspension, 5 μl of outer well plus Escherichia coli 24 h liquid culture filtrate, and 5 μl of inner well plus Tc199 medium were used as controls. (2) Insulation: After adding the above-mentioned plate, add it to the wet box at 37 ° C, and incubate for 2 to 3 hours. (3) Fixed staining: 3 ml of methanol was added to each dish and treated for 30 min. After adding 3 ml of 47% neutral formalin solution, fix for 30 min. The agarose layer was peeled off, and the cells adhered to the bottom of the plate were stained with Wright, washed with water and dried. Not suitable for the crowd People with reduced hematopoietic function such as leukemia, various anemia, myelodysplastic syndrome, or people with thrombocytopenia should pay attention to blood draw, and should not take more or more blood. Adverse reactions and risks 1. After the blood is drawn, do not press the needle hole to avoid subcutaneous hematoma. If there is a small piece of bruise in the blood, it is slightly tender, please don't panic, you can do hot compress after 24 hours to promote the absorption of blood. The general small amount of congestion will gradually absorb in 3 to 5 days and the color will become lighter and return to normal. 2. After the blood draw, symptoms such as dizziness, vertigo, fatigue, etc. should be immediately supine, drink a small amount of syrup, and then undergo a physical examination after the symptoms are relieved.