Abstract
Chinese hamster ovary (CHO) cells respond to pertussis toxin (PT) with a novel clustering pattern, which is dependent on biologically active PT. Since its description in 1983, this cellular response has been refined and used extensively for detection and quantification of PT activity, as well as anti-PT antibodies. There are limitations, however, in the use of this phenomenon as originally described. They are: (1) a subjective, observer-dependent scoring system; (2) the requirement for 16–24 h incubation in order for the response to be clearly detectable; and (3) apparent interference from non-toxin materials. To overcome these limitations, a number of alternative in vitro assays for PT, using CHO cells or other cell types, have been developed and are described elsewhere in this publication. In addressing the challenges associated with the CHO cell assay, we discovered that changes in the electrical impedance-based “normalized cell index” of PT-treated CHO cells obtained with the ACEA xCELLigence instrument enable objective detection/quantification of the PT-induced effect in as little as 3–4 h. To the best of our knowledge, the molecular basis for this intriguing response remains unknown. We present here electron microscopic (EM) images of control and PT-treated cells, which suggest some potential molecular mechanisms.
Highlights
The morphologic response of Chinese hamster ovary (CHO) cells to pertussis toxin (PT) has been used extensively for quantification of PT and anti-PT antibodies
In 1980, Erik Hewlett joined the Division of Geographic Medicine in the Department of Medicine, University of Virginia School of Medicine. This newly formed unit was headed by Dr Richard Guerrant, an established investigator in the field of diarrheal diseases, especially those caused by cholera toxin (CT)
Analysis Facility, which provides specialized, expensive equipment in a central facility, for use by individual investigators, we were able to employ an ACEA xCELLigence instrument. This novel technology uses special multi-well, cell-culture plates (E-Plates) with embedded gold electrodes to monitor impedance produced by a population of plated cells
Summary
The morphologic response of Chinese hamster ovary (CHO) cells to pertussis toxin (PT) has been used extensively for quantification of PT and anti-PT antibodies. PT, at final concentrations was added to CHO cells, which had been plated and incubated overnight; measurements were begun at the time of toxin addition.
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