scholarly journals Pathogenesis of shigella diarrhea. VII. Evidence for a cell membrane toxin receptor involving β1 {arrow} 4-linked N-acetyl-D-glucosamine oligomers

1977 ◽  
Vol 146 (2) ◽  
pp. 535-546 ◽  
Author(s):  
GT Keusch ◽  
M Jacewicz
Keyword(s):  
Cell Membrane ◽  
Cholera Toxin ◽  
Liver Cell ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Cell Membranes ◽  
Proteolytic Enzymes ◽  
Cell Monolayer ◽  
Toxin Receptor ◽  
Toxin Uptake

The binding of ShigeUa dysenteriae 1 cytotoxin to HeLa cells in culture and to isolated rat liver cell membranes was studied by means of an indirect consumption assay of toxicity from the medium, or by determination of cytotoxicity to the HeLa cell monolayer. Both liver cell membranes and HeLa cells removed toxicity from the medium during incubation, in contrast to WI-38 and Y-1 mouse adrenal tumor cells, both of which neither bound nor were affected by the toxin. Uptake of toxin was directly related to concentration of membranes added, time,and temperature, and indirectly related to the ionic strength of the buffer used. The chemical nature of the membrane receptor was characterized by using three principal approaches: (a) enzymatic sensitivity; (b) competitive inhibition and (c) receptor blockade studies. The receptor was destroyed by proteolytic enzymes, phospholipases (which markedly altered the gross appearance of the membrane preparation) and by lysozyme, but not by a variety of other enzymes. Of 28 carbohydrate and glycoprotein haptens studied, including cholera toxin and ganglioside, only the chitin oligosaccharide lysozyme substrates, per N-acetylated chitotriose, chitotetraose, and chitopentaose were effective competitive inhibitors. Greatest inhibition was found with the trimer, N, N', N" triacetyl chitotriose. Of three lectins studied as possible receptor blockers, including phytohemagglutinin, concanavalin A, and wheat germ agglutinin, only the latter, which is known to possess specific binding affinity for N, N', N" triacetyl chitotriose, was able to block toxin uptake. Evidence from all three approaches indicate, therefore, existence of a glycoprotein toxin receptor on mammalian cells, with involvement of oligomeric β1{arrow}4-1inked N-acetyl glucosamine in the receptor. This receptor is clearly distinct from the G(M1) ganglioside thought to be involved in the binding of cholera toxin to the cell membrane of a variety of cell types susceptible to its action.

Destruction of Cholera Toxin Receptor on HeLa Cell Membrane Using Microbial Endoglycoceramidase

1996 ◽  
Vol 328 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Kenji Yamamoto ◽  
Tomokazu Nagano ◽  
Hidehiko Kumagai ◽  
Yoshiko Okamoto ◽  
Shuzo Otani
Keyword(s):  
Cell Membrane ◽  
Hela Cell ◽  
Cholera Toxin ◽  
Toxin Receptor

Comparison of the binding of 59Fe- and 239Pu-transferrin to rat liver cell membranes

1990 ◽  
Vol 9 (1) ◽  
pp. 17-24 ◽  
Author(s):  
F. Planas-Bohne ◽  
W. Rau
Keyword(s):  
Molecular Weight ◽  
Cell Membrane ◽  
Membrane Protein ◽  
Rat Liver ◽  
Liver Cell ◽  
Cell Membranes ◽  
Apparent Molecular Weight ◽  
Protein S ◽  
Membrane Protein Complex ◽  
Isolated Cell

The binding of the 59Fe and 239Pu complexes of transferrin and 125I labelled transferrin [Tf (125I) ] to isolated cell membranes of rat liver has been studied. Transferrin forms a complex with an integral protein of the membrane which has an apparent molecular weight of about 180 kDa and is stable only at pH 7.4. Iron-59 is eluted from Sephacryl S 300 columns together with Tf (125I) or the Tf-membrane protein complex while 239Pu seems to be bound to different membrane protein(s). After isolation of the Tf-binding protein from 35S-labelled membranes and incubation with one of the metal-Tf complexes 59Fe elutes from a Sephacryl S 300 column together with 35S at an apparent molecular weight of ca. 250 kDa while 239Pu is found in fractions of lower molecular weight. It is concluded from these results that there are Tf-receptors in the liver cell membrane to which iron transferrin may bind. Plutonium, however, seems to be dissociated from Tf and bound directly to other membrane proteins.

scholarly journals THE INTERACTION BETWEEN TOXOPLASMA GONDII AND MAMMALIAN CELLS

1972 ◽  
Vol 136 (5) ◽  
pp. 1157-1172 ◽  
Author(s):  
Thomas C. Jones ◽  
Shirley Yeh ◽  
James G. Hirsch
Keyword(s):  
Electron Microscopy ◽  
Toxoplasma Gondii ◽  
Cell Membrane ◽  
Hela Cell ◽  
Cell Cultures ◽  
Hela Cells ◽  
Generation Time ◽  
Mammalian Cells ◽  
Cell Types ◽  
Special Procedure

Macrophage, fibroblast, and HeLa cell cultures have been infected with Toxoplasma gondii, and observations have been made on parasite entry and fate. A special procedure was devised for studying the entry of toxoplasmas by electron microscopy. Toxoplasmas were centrifuged onto the cells in the cold; fixation 1–3 min after warming yielded specimens showing numerous examples of parasites in the process of entering cells. The mechanism of entry into macrophages, fibroblasts, and HeLa cells was in all cases by phagocytosis. Micropseudopods were extended by the cells to envelop the attached parasites in a typical phagocytic vacuole. Apparently the toxoplasmas stimulated this response of HeLa cells and fibroblasts, cell types not usually phagocytic. No instance was seen of penetration of toxoplasmas through the cell membrane, or of parasites located free in the cytoplasm. Essentially all of the toxoplasmas that entered HeLa cells divided with a generation time of 9 hr; the parasites formed large rosettes situated in vacuoles, eventually leading to host cell rupture. Macrophages took in larger numbers of toxoplasmas than did HeLa cells, but approximately half of the parasites inside of macrophages degenerated within a few hours. The surviving toxoplasmas in macrophages divided every 8 hr, forming rosettes and eventually rupturing the cells.

scholarly journals TRANSFORMATION OF A HISTOCOMPATIBILITY IMMUNOGEN INTO A TOLEROGEN

1973 ◽  
Vol 138 (3) ◽  
pp. 723-733 ◽  
Author(s):  
Stephen H. Nimelstein ◽  
Allan R. Hotti ◽  
Halsted R. Holman
Keyword(s):  
Structural Change ◽  
Cell Membrane ◽  
Spleen Cell ◽  
Liver Cell ◽  
Cellular Response ◽  
Cell Membranes ◽  
Humoral Response ◽  
Cellular Responses ◽  
Liver Cells ◽  
Liver Cell Membrane

H-2 antigens on spleen cell membranes absorb antibody to H-2 antigens and induce both humoral and cellular responses. Liver cell membrane H-2 antigens by contrast also absorb antibody but do not influence cellular response and are tolerogenic for the humoral response. This paper demonstrates that syngeneic liver cells contain a substance which can transform the properties of allogeneic spleen cell membranes into those of allogeneic liver cell membranes, i.e., transform a humoral immunogen into a humoral tolerogen. The process appears to be accompanied by cleavage of an antigen component from the spleen membrane and hence to result in a structural change in the H-2 antigen.

Use of Uranium-Labeled Antibodies for Electron Microscopic Localization of Various Antigens in Mammalian Cells

1967 ◽  
Vol 25 ◽  
pp. 136-137
Author(s):  
J. P. Petrali ◽  
E. J. Donati ◽  
L. A. Sternberger
Keyword(s):  
Endoplasmic Reticulum ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Specific Antiserum ◽  
Electron Microscopic ◽  
E Coli ◽  
Cytoplasmic Membranes ◽  
Viral Progeny ◽  
Ultrathin Sections ◽  
Electron Microscopic Localization

Specific contrast is conferred to subcellular antigen by applying purified antibodies, exhaustively labeled with uranium under immunospecific protection, to ultrathin sections. Use of Seligman’s principle of bridging osmium to metal via thiocarbohydrazide (TCH) intensifies specific contrast. Ultrathin sections of osmium-fixed materials were stained on the grid by application of 1) thiosemicarbazide (TSC), 2) unlabeled specific antiserum, 3) uranium-labeled anti-antibody and 4) TCH followed by reosmication. Antigens to be localized consisted of vaccinia antigen in infected HeLa cells, lysozyme in monocytes of patients with monocytic or monomyelocytic leukemia, and fibrinogen in the platelets of these leukemic patients. Control sections were stained with non-specific antiserum (E. coli).In the vaccinia-HeLa system, antigen was localized from 1 to 3 hours following infection, and was confined to degrading virus, the inner walls of numerous organelles, and other structures in cytoplasmic foci. Surrounding architecture and cellular mitochondria were unstained. 8 to 14 hours after infection, antigen was localized on the outer walls of the viral progeny, on cytoplasmic membranes, and free in the cytoplasm. Staining of endoplasmic reticulum was intense and focal early, and weak and diffuse late in infection.

scholarly journals Enterotoxigenicity of Mature 45-Kilodalton and Processed 35-Kilodalton Forms of Hemagglutinin Protease Purified from a Cholera Toxin Gene-Negative Vibrio cholerae Non-O1, Non-O139 Strain

2006 ◽  
Vol 74 (5) ◽  
pp. 2937-2946 ◽  
Author(s):  
A. Ghosh ◽  
D. R. Saha ◽  
K. M. Hoque ◽  
M. Asakuna ◽  
S. Yamasaki ◽  
...  
Keyword(s):  
Cholera Toxin ◽  
Vibrio Cholerae ◽  
Hela Cells ◽  
Histopathological Examination ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Toxin Gene ◽  
Dependent Manner ◽  
A Cell

ABSTRACT Cholera toxin gene-negative Vibrio cholerae non-O1, non-O139 strain PL-21 is the etiologic agent of cholera-like syndrome. Hemagglutinin protease (HAP) is one of the major secretory proteins of PL-21. The mature 45-kDa and processed 35-kDa forms of HAP were purified in the presence and absence of EDTA from culture supernatants of PL-21. Enterotoxigenicities of both forms of HAP were tested in rabbit ileal loop (RIL), Ussing chamber, and tissue culture assays. The 35-kDa HAP showed hemorrhagic fluid response in a dose-dependent manner in the RIL assay. Histopathological examination of 20 μg of purified protease-treated rabbit ileum showed the presence of erythrocytes and neutrophils in the upper part of the villous lamina propria. Treatment with 40 μg of protease resulted in gross damage of the villous epithelium with inflammation, hemorrhage, and necrosis. The 35-kDa form of HAP, when added to the lumenal surface of rat ileum loaded in an Ussing chamber, showed a decrease in the intestinal short-circuit current and a cell rounding effect on HeLa cells. The mature 45-kDa form of HAP showed an increase in intestinal short-circuit current in an Ussing chamber and a cell distending effect on HeLa cells. These results show that HAP may play a role in the pathogenesis of PL-21.

scholarly journals Binding and biologic activity of glucagon in liver cell membranes of chronically hyperglucagonemic rats.

1977 ◽  
Vol 252 (21) ◽  
pp. 7434-7436 ◽  
Author(s):  
C.B. Srikant ◽  
D. Freeman ◽  
K. McCorkle ◽  
R.H. Unger
Keyword(s):  
Liver Cell ◽  
Cell Membranes ◽  
Biologic Activity

scholarly journals A DISTINCTIVE CELL CONTACT IN THE RAT ADRENAL CORTEX

1972 ◽  
Vol 53 (1) ◽  
pp. 148-163 ◽  
Author(s):  
Daniel S. Friend ◽  
Norton B. Gilula
Keyword(s):  
Cell Membrane ◽  
Adrenal Cortex ◽  
Cell Membranes ◽  
Freeze Fracture ◽  
Typical Feature ◽  
Cell Contact ◽  
Cortical Cells ◽  
Cell Dispersion ◽  
Extensive Cell ◽  
Distinctive Cell

Extensive cell contacts which resemble septate junctions occur between cells in the three major zones of the rat adrenal cortex. Characteristically, they extend between small intercellular canaliculi and the periendothelial space, frequently interrupted by gap junctions and rarely by desmosomes. Zonulae occludentes have not been identified in the adrenal cortex. Along this distinctive cell contact, the cell membranes of apposing cells are separated by 210–300 a bisected by irregularly spaced 100–150-A extracellular particles which are often circular in profile. In lanthanum preparations, these particles appear to form a continuous chain throughout the intercellular space and are visualized as an alveolate structure in sections parallel to the plane of the cell membrane. The cell membrane in the area of septate-like contact does not differ from nonjunctional areas of the cell membrane in freeze-fracture replicas. The cell contact retains its integrity after cell dispersion and after the separation of cell membranes from disrupted cells. The intercellular particles also persist after brief extraction in lipid solvents. Besides adherence, possible functions of this adrenal contact include maintenance of the width of the extracellular space, the provision of channels between intercellular canaliculi and the bloodstream, and utilization as cation depots. Similar structures are also present between adrenal cortical cells of several other species and between interstitial cells of the testis. This type of cell contact may, in fact, be a typical feature of steroid-hormone-secreting tissues in vertebrates.

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