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Lysosomal Enzyme Activity in Poliovirus-infected HeLa Cells and Vesicular Stomatitis Virus-infected L Cells: biochemical and Histochemical Comparative Analysis with Computer-aided Techniques

Institut für Virologie der Universität Würzburg, Würzburg, Germany, and Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Germany

The behaviour of lysosomal enzymes in poliovirus-infected HeLa S3 cells and vesicular stomatitis virus (VSV)-infected L cells was investigated both biochemically using enzyme assays, and histochemically using acid phosphatase dependent staining. The presence of the enzyme was shown histochemically under the light microscope by its reaction with naphthole-AS-BI-phosphate and a coupling reaction with diazotized pararosaniline. The light absorption of this stain in infected and uninfected cells was measured on a Universal Micro Spectrophotometer (UMSP-1) and recorded on-line as gray value cell images in a PDP-12 computer. These scanned images were analyzed by FORTRAN programs on a UNIVAC1108. The histochemically obtained distributions of the lysosomal enzyme are comparable to the results of the biochemical analysis. Lysosomes of poliovirus-infected cells displayed a release of lysosomal enzymes into the cytoplasm starting at 3 h after infection; VSV infection did not produce this type of effect. This investigation shows that it is possible to extract and demonstrate specific virus dependent changes using computer-aided cytophotometric techniques.

Received 7 May 1974; accepted 14 August 1974.