Diagnostic agent for chemotaxis

Abstract

A method is described for determining the chemotactic activity of leukocytes in a sample, in which method the expression of a cell adhesion molecule is determined.

1 . A method for determining the chemotactic activity of leukocytes in a sample, 
 characterized in that 
 the expression of a cell adhesion molecule is determined.  
   
     
     
         2 . The method as claimed in  claim 1 , 
 characterized in that 
 the chemotactic activity of granulocytes, in particular neutrophilic granulocytes, is determined.  
   
     
     
         3 . The method as claimed in  claim 1  or  2 , 
 characterized in that 
 the cell adhesion molecule is selected from L-selectin, CD62L, LECAM-1, Mel-14, LAM-1, Leu-8, TQ1, LEC.CAM-1, DREG56, GMP-140, ELAM and CD11b.  
 
 
     
     
         4 . The method as claimed in one of the preceding claims, 
 characterized in that 
 the change in the density at which the cell adhesion molecule is expressed on the surface of the leukocyte cells, which change accompanies the chemotactic activity, is determined, and/or a binding molecule which is specific for the cell adhesion molecule is used for determining the expression.  
   
     
     
         5 . The method as claimed in one of the preceding claims, 
 characterized in that 
 the sample is preincubated with a chemotaxis stimulant, in particular with fMLP and/or interleukin-8.  
   
     
     
         6 . The method as claimed in one of the preceding claims, 
 characterized in that 
 the determination is carried out in a flow-through cytometer and/or a fluorescence label is used for the detection.  
   
     
     
         7 . The method as claimed in one of the preceding claims, 
 characterized in that 
 anticoagulated whole blood and/or leukocyte-rich plasma is used as a sample and/or the sample is a human sample.  
   
     
     
         8 . The method as claimed in one of the preceding claims, 
 characterized in that 
 the shape and/or size of the leukocytes, in particular the change in shape of the leukocytes, is/are also determined.  
   
     
     
         9 . The method as claimed any one of the preceding claims, 
 characterized in that 
 after having been incubated with a chemotaxis stimulant, in particular with fMLP and/or interleukin-8, the leukocytes are conducted through a porous membrane.  
   
     
     
         10 . The method as claimed in either  claim 8  or  9 , 
 characterized in that 
 the change in the shape and/or size of the leukocytes is determined in a flow-through cytometer.  
 
 
     
     
         11 . The method as claimed in one of the preceding claims, 
 characterized in that 
 the number of leukocytes which have migrated through a porous membrane, in particular through a porous membrane which is coated with collagen, fibrinogen and/or gelatin, is also determined.  
   
     
     
         12 . The method as claimed in  claim 11 , 
 characterized in that 
 the cell counting is carried out by adding latex beads in a flow-through cytometer.  
   
     
     
         13 . A method for determining the chemotactic activity of leukocytes in a sample, 
 characterized in that 
 the number of leukocytes which have migrated through a porous membrane, in particular a porous membrane coated with collagen, fibrinogen and/or gelatin, is determined.  
   
     
     
         14 . A reagent kit for determining the chemotactic activity of leukocytes, in particular of granulocytes, 
 characterized in that 
 it contains a specific binding partner for detecting the expression of a cell adhesion molecule, in particular an antibody directed against the cell adhesion molecule, such as an antibody directed against the CD62L cluster, and also a chemotaxis stimulant and, where appropriate, a direct or indirect fluorescence label and, where appropriate, also a collagen-coated porous membrane and, where appropriate, also customary buffers, auxiliary substances and/or additives.  
   
     
     
         15 . A method for diagnosing an immunological defect and/or a disturbance in leukocyte function, 
 characterized in that 
 the chemotactic activity of leukocytes in a sample from a patient is determined using a method as claimed in one of  claims 1  to  13 , and the chemotactic activity of a negative control sample is determined in another assay, and the presence or the absence of an immunological defect and/or a disturbance in leukocyte function is elucidated by comparing the values which are obtained, with use being made, in particular, of a reagent kit as claimed in  claim 14.