Fig. 3.

Chemotactic activity of proteins after reversed-phase chromatography (C8 nucleosil RP-8) in recurrent tonsillitis. (a) Chromatogram (RP-8 column). Supernatant collected from tissue homogenization was concentrated and applied to a heparin–sepharose cartridge. The eluate was collected as one fraction and subjected to size exclusion chromatography. The figure shows the chromatogram and the collected protein fractions labelled with numbers 1–18. (b) Chemotactic index and concentrations of chemokines. The manually collected protein fractions 1–18 were then analysed for chemotactic activity. In each test series, both positive and negative controls were included at appropriate concentrations. FNLP (formyl-Nle-Leu-Phe) served as a positive control for chemotactic responsiveness of PMN, whereas PBS was used as a negative control. The data shown here are those corresponding to 30 µl fraction aliquots transferred to the Boyden chamber. All fraction volumes tested had been freeze-dried prior to testing. In addition, an ELISA was performed to identify the different chemokines. Corresponding fractions were pooled as fractions 1, 2, 3 and 4 and subjected to Mono-S column (see Fig. 4).