Purification and partial characterization of a cytotonic enterotoxin produced by Aeromonas hydrophila

1989 ◽  
Vol 35 (7) ◽  
pp. 719-727 ◽  
Author(s):  
A. K. Chopra ◽  
C. W. Houston
Keyword(s):  
Cholera Toxin ◽  
Cho Cells ◽  
Aeromonas Hydrophila ◽  
Chinese Hamster ◽  
Biologically Active ◽  
Partial Characterization ◽  
Cytotoxic Activities ◽  
Ileal Loop

This report describes the purification and partial characterization of a cytotonic enterotoxin produced by a human diarrheal isolate (SSU) of Aeromonas hydrophila. The extracellular enterotoxin was purified by (NH4)2SO4 precipitation, hydrophobic column chromatography, and chromatofocusing. The highly purified enterotoxin exhibited a molecular mass of 44 kDa and an isoelectric point in the range of 4.3–5.5 as determined by chromatofocusing. Western blot analysis using Aeromonas anti-enterotoxin revealed a single band at 44 kDa; however, cholera antitoxin failed to detect the enterotoxin antigen. This non-cholera toxin cross-reactive (non-CTC) enterotoxin was biologically active in vivo as determined by rabbit ligated ileal loop and rabbit skin vascular permeability assays. Biological activity also was expressed in vitro by this toxin as measured by the elongation of Chinese hamster ovary (CHO) cells. The enterotoxic activity associated with this molecule was neutralized completely by homologous antibodies but not by cholera antitoxin. The purified toxin preparation was free of hemolytic and cytotoxic activities as determined by its inability to lyse rabbit red blood cells or damage CHO cells, respectively. Furthermore, this toxin induced the elevation of cAMP in CHO cells suggesting thereby that the mechanism of action of Aeromonas non-CTC enterotoxin may be similar to heat-labile enterotoxins of Escherichia coli and Vibrio cholerae.Key words: Aeromonas hydrophila, cytotonic enterotoxin, cholera toxin, cAMP

Cytotoxicity and suckling mouse reactivity of Aeromonas hydrophila isolated from human sources

1981 ◽  
Vol 27 (10) ◽  
pp. 1019-1027 ◽  
Author(s):  
W. M. Johnson ◽  
H. Lior
Keyword(s):  
Tumor Cells ◽  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Aeromonas Hydrophila ◽  
Adrenal Tumor ◽  
Chinese Hamster ◽  
Suckling Mouse ◽  
Ileal Loop ◽  
Suckling Mice ◽  
Enterotoxin Production

Human isolates of Aeromonas hydrophila and Plesiomonas shigelloides were tested for their ability to produce cytotoxins and (or) enterotoxins. The incidence of cytotoxin production by A. hydrophila was 81% for isolates from stool and 44% for extraintestinal isolates. Chinese hamster ovary (CHO) cells were more sensitive to A. hydrophila cytotoxin than either Vero or Y-1 mouse adrenal tumor cells (Y-1). There was no evidence of cytotonic enterotoxin production by any of the isolates tested. Cytotoxin-containing filtrates from A. hydrophila were found to provoke a positive reaction in suckling mice. The response in mice was heat labile, and data supporting correlation of this activity with the cytotoxin produced by these organisms are presented. Following concentration, cytotoxin and material reactive in suckling mice were found to coelute from Sephadex G-25 and dose–response was demonstrated in mice. Antitoxin prepared to this material effectively neutralized cytotoxin, mouse reactivity, and rabbit ileal loop response. No evidence was obtained for either cytotoxin or enterotoxin production by P. shigelloides strains tested in this study.

scholarly journals Antiplasmodial and Cytotoxic Activities of Drimane Sesquiterpenes from Canella winterana

2010 ◽  
Vol 5 (12) ◽  
pp. 1934578X1000501 ◽  
Author(s):  
Mary H. Grace ◽  
Carmen Lategan ◽  
Flaubert Mbeunkui ◽  
Rocky Graziose ◽  
Peter J. Smith ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Cho Cells ◽  
Chinese Hamster ◽  
Structure Activity Relationship ◽  
Activity Relationship ◽  
Cytotoxic Activities ◽  
Strong Activity ◽  
Structure Activity ◽  
Bioassay Guided Fractionation

The hexane extract from the leaves of Canella winterana exhibited strong activity against the chloroquine sensitive (CQS) strain of Plasmodium falciparum (D10) in vitro (IC50 2.53 μg/mL). Bioassay guided fractionation of this extract has led to the isolation of 5 drimane-type sesquiterpenoids: 9-epideoxymuzigadial, 9-deoxymuzigadial, muzigadial, 3-β-acetoxypolygodial and the newly isolated hemiacetal, named muzigodiol, with IC50-values of 1.01, 2.19, 0.31, 2.77 and 7.43 μg/mL, respectively. The first four compounds were tested for their cytotoxicity using Chinese Hamster Ovarian (CHO) cells, where they showed IC50-values of 1.82, 33.69, 1.18, and 58.31 μg/mL, respectively. A structure-activity relationship is discussed.

scholarly journals The CHO Cell Clustering Response to Pertussis Toxin: History of Its Discovery and Recent Developments in Its Use

Toxins ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 815
Author(s):  
Mary C. Gray ◽  
Richard L. Guerrant ◽  
Erik L. Hewlett
Keyword(s):  
Pertussis Toxin ◽  
Cho Cells ◽  
Cellular Response ◽  
Molecular Mechanisms ◽  
Chinese Hamster ◽  
Biologically Active ◽  
In Vitro Assays ◽  
Cho Cell ◽  
Electron Microscopic

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.

scholarly journals In vitro conversion of proapoprotein A-I to apoprotein A-I. Partial characterization of an extracellular enzyme activity.

1983 ◽  
Vol 258 (19) ◽  
pp. 11430-11433 ◽  
Author(s):  
C Edelstein ◽  
J I Gordon ◽  
K Toscas ◽  
H F Sims ◽  
A W Strauss ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Extracellular Enzyme ◽  
Extracellular Enzyme Activity ◽  
Partial Characterization

scholarly journals Cloning and characterization of small circular DNA from Chinese hamster ovary cells.

1984 ◽  
Vol 4 (1) ◽  
pp. 173-180 ◽  
Author(s):  
S W Stanfield ◽  
D R Helinski
Keyword(s):  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Single Copy ◽  
Chromosomal Dna ◽  
Cho Cell ◽  
Ovary Cells ◽  
Chromosomal Sequences

Small polydisperse circular (spc) DNA was isolated and cloned, using BglII from Chinese hamster ovary (CHO) cells. The properties of 47 clones containing at least 43 different BglII fragments are reported. The majority of the clones probably contain entire sequences from individual spcDNA molecules. Most of the clones were homologous to sequences in CHO cell chromosomal DNA, and many were also homologous to mouse LMTK- cell chromosomal sequences. The majority of homologous CHO cell chromosomal sequences were repetitive, although a few may be single copy. Only a small fraction of cloned spcDNA molecules were present in every cell; most occurred less frequently than once in 15 cells. Localization studies indicated that at least a portion of spcDNA is associated with the nucleus in CHO cells.

Suppression of tumor formation in vivo by expression of the JE gene in malignant cells

1991 ◽  
Vol 11 (6) ◽  
pp. 3125-3131
Author(s):  
B J Rollins ◽  
M E Sunday
Keyword(s):  
Cho Cells ◽  
Chinese Hamster ◽  
Suppressive Effect ◽  
Tumor Formation ◽  
Host Animal ◽  
Early Growth Response ◽  
Discernible Effect ◽  
Early Growth Response Gene

The early growth response gene JE encodes a monocyte chemoattractant, MCP-1. The JE/MCP-1 protein attracts and stimulates human monocytes and induces monocyte-mediated inhibition of tumor cell growth in vitro. Expression of human or murine JE/MCP-1 in Chinese hamster ovary (CHO) cells completely suppressed their ability to form tumors in nude mice. Coinjection of JE/MCP-1-expressing cells with nonexpressing CHO cells or with HeLa cells also prevented tumor formation. Since JE/MCP-1 expression had no discernible effect on the tranformed phenotype of these cells in vitro, the suppressive effect depends on host animal factors. These factors are likely to be components of the inflammatory response, because JE/MCP-1-expressing cells elicited a predominantly monocytic infiltrate at the site of injection. Our results suggest that JE/MCP-1 protein may be useful in cancer therapy.

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