Cytochemical Observations on Proteins, Alkaline and Acid Phosphatases, Adenosine Triphosphatase, and 5′‐Nucleotidase in Chick Liver Cell Cultures Infected with Trichomonas vaginalis*, †
Authors: SHARMA N. N. , HONIGBERG B. M. 
Published: May 2007

SYNOPSIS.By means of the ninhydrin‐Schiff method for proteins a diffuse reaction as well as one localized in granular inclusions can be shown in the cytoplasm of fibroblasts, epithelial cells, and macrophages in trypsin‐dispersed chick liver cell cultures. Nuclei and nucleoli also take the specific stain. A progressive loss of cytoplasmic and nuclear staining occurs in the fibroblasts in cultures infected with a relatively pathogenic strain ofT. vaginalis.A loss occurs in epithelial cells in advanced stages of degeneration, but in less damaged cells, while the diffuse reaction disappears, the number and staining intensity of the cytoplasmic inclusions remain unchanged or possibly may increase somewhat. The intensity of the diffuse reaction and the number and size of the characteristic inclusions increase in the active, parasite‐free, experimental macrophages, but phagocytes with trichomonads closely applied to their external surfaces and those containing the flagellates within their cytoplasm typically retain only a few weak‐staining inclusions.

Similar distribution of alkaline and acid phosphatases occurs in preparations treated according to Gomori's and Burstone's methods, except that no nuclear staining is obtained with the latter. Activity of both enzymes is localized primarily in inclusions which are dispersed thruout the cytoplasm of fibroblasts and epithelial cells and tend to accumulate along the cell membranes and around the nuclei. In the course of infection withT. vaginalisthere is a progressive loss of alkaline phosphatase from both cell types; however, the acid phosphatase activity increases. In the control macrophages both enzymes are localized in mostly rather large, rounded cytoplasmic inclusions. The number of such inclusions increases in the parasite‐free experimental macrophages, but only a few weak‐staining granules remainin phagocytes with engulfed trichomonads and in those whose external surfaces are in direct contact with the parasites. The loss of the inclusions is less apparent in macrophages containing degenerated flagellates than in the ones with healthy trichomonads, but regardless of the condition of the parasites, the highest enzymatic activity is found around them.

ATPase and 5′‐nucleotidase are localized in small granules dispersed thruout the cytoplasm of fibroblasts and epithelial cells. The granules tend to accumulate along the periphery of the cells and around the nuclei. A diffuse cytoplasmic reaction is present in preparations processed for 5′‐nucleotidase. Nuclei and nucleoli give positive reactions for both enzymes. In the course of infection with trichomonads, activity of the 2 enzymes declines in both culture cell types. Control macrophages have diffuse cytoplasmic reaction for ATPase and 5′‐nucleotidase and these enzymes are localized also in rounded cytoplasmic inclusions. Activity of both enzymes increases in the parasite‐free experimental phagocytes, but little if any diffuse staining and only a few characteristic inclusions are left in macrophages with engulfed healthy trichomonads and in those whose external surfaces are invested with the flagellates.

The ninhydrin‐Schiff‐positive inclusions found in the macrophages appear to be the same as some of those which have acid phosphatase activity and may well be identical with the glycolipoprotein bodies noted by us previously. On the grounds of their chemical constitution and behavior it seems likely that the inclusions are lysosomes.