Alterations in nucleotide content of human lung fibroblasts infected with Mycoplasma pneumoniae

Upchurch, S; Gabridge, Michael G.

doi:10.1128/iai.38.2.631-636.1982

Cited by 5 publications

(5 citation statements)

References 35 publications

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“…In vitro observations of ciliostasis correlate well with clinical findings that mucociliary clearance in the respiratory tract can remain depressed for extended periods following M. pneu-moniae infection (4). Cytopathological changes are accompanied at the biochemical level by a transient rise and then decline in galactose uptake and utilization, reductions in the uptake of precursors for macromolecular synthesis, alterations in RNA and protein synthesis (25,26), decreases in the level of intracellular ATP (18), and changes in nucleotide content (35).…”

supporting

confidence: 68%

Interaction of Mycoplasma pneumoniae with HeLa cells

Krause

Chen

1988

Infect Immun

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The susceptibility of HeLa cells to Mycoplasma pneumoniae-induced injury was examined. Infections were initiated with relatively low mycoplasma doses, carried out in a culture medium incapable of supporting M. pneumoniae replication in the absence of host cells, and monitored for up to 10 days. Under these conditions, a time-and dose-dependent decline in the number of viable host cells compared with that of uninfected controls was observed. The effect of M. pneumoniae infection on host cell macromolecular synthesis was also evaluated. At high doses of infection, synthesis of both protein and RNA declined rapidly relative to that in control cells. At lower doses there was a biphasic response in protein synthesis, which was substantially lower than that in the uninfected control by day 1 postinfection, returned to control levels by day 4 postinfection, and was again less than that in control cells by day 7 postinfection. In contrast, no transient recovery was observed in RNA synthesis, which declined very gradually over 7 days in infected HeLa cells compared with that in uninfected control cells. The ability of HeLa cells to support the proliferation of M. pneumoniae under these experimental conditions was demonstrated by quantitation of mycoplasma CFU in the nonpermissive medium in the presence or absence of HeLa cells. A negligible increase in the number of M. pneumoniae was observed over 4 days when HeLa cells were absent, while CFU increased by almost 20-fold when M. pneumoniae was cultured in the presence of HeLa cells. The susceptibility and response in macromolecular synthesis in M. pneumoniae-infected HeLa cells differed from that recently described for a nontransformed culture of hamster trachea epithelial cells under the same experimental conditions (Y.-Y. Chen and D. C. Krause, Infect. Immun. 56:570-576, 1988), underscoring the importance of the choice of host cell for in vitro modeling of M. pneumoniae pathogenesis.

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confidence: 68%

Interaction of Mycoplasma pneumoniae with HeLa cells

Krause

Chen

1988

Infect Immun

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“…Initial studies were devoted to developing a protocol for the preparation of the target cells. Fibroblasts of respiratory tract origin (MRC-5 cells, originally derived from human fetal lung tissue) are well characterized and are known to be sensitive to respiratory pathogens (14). However, their attachment rate to most substrates is quite low (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The unique feature of this material is that it is permeable to most gases in the size range of carbon dioxide. Such membranes have been used with gas-sensing electrodes (14) and for delivering oxygen to cells in batch culture (15). Most recently they have been used to evaluate the in vitro cytotoxic potential of ozone, a common gaseous pollutant (16,17).…”

Section: Discussionmentioning

confidence: 99%

Gaseous oxide toxicity evaluated with cell monolayers on collagen-coated, gas-permeable teflon membranes.

Gabridge¹,

Gladd²

1984

Environ Health Perspect

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A system was developed to evaluate the cytotoxic potential of gaseous oxides in vitro. Target cells were MRC-5 human lung fibroblasts cultivated as monolayers on gas-permeable , FEP-Teflon membranes. Membranes were secured in Chamber/Dishes with a 25 mm diameter well. To promote attachment of fibroblasts to the membranes, the latter were incubated in collagen ( Vitrogen ) solutions for 10 min prior to plating the cells. The collagen pretreatment was significantly more effective than poly-L-lysine, fetal calf serum, polybrene and bovine serum albumin. Several types (mouse and calf) of acid-soluble and alcohol-soluble collagen fractions were evaluated, and all of them promoted cell attachment with equivalent efficiency. Cells on membranes were exposed to gases in a Plexiglass chamber with a gas flow of 2L/min. Sulfur dioxide caused a marked loss in cell viability (as indicated by ATP content of the monolayer) after 30 min exposure to 0.01% and 0.005%. A level of 0.001% did not affect viability, and none of the levels tested caused a sloughing of the monolayer after 90 min. Nitrogen dioxide induced a more modest drop in cell viability after 30 min exposure to 0.1%, while 0.005% and 0.05% were nontoxic. No cell sloughing occurred with NO2 exposures, and exposures to CO2 at levels of 20% for 90 min were nontoxic. This system, with cell culture monolayers on gas-permeable Teflon membranes, is simple and convenient. As such, it has potential application to cytotoxicity evaluations with numerous gases.

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“…The consequences of M. pneumoniae infection have been studied in vitro, primarily in tracheal organ culture, and include a gradual loss of ciliary activity, nuclear swelling, cytoplasmic vacuolization, and the eventual exfoliation of ciliated cells (7). These cytopathological changes are accompanied by metabolic abnormalities in host cells, including decreases in intracellular ATP levels (21), nucleotide content (44), carbohydrate utilization, amino acid transport, macromolecular syntheses (32,33), and oxidative metabolism (17,20).…”

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confidence: 99%

Parasitism of hamster trachea epithelial cells by Mycoplasma pneumoniae

Chen

Krause

1988

Infect Immun

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Hamster trachea epithelial (HTE) cells were employed as a model system for Mycoplasma pneumoniae pathogenesis. To more closely mimic natural infection, M. pneumoniae was forced to rely upon host cells (as opposed to the growth medium) for nutrients, and infections were initiated with relatively low mycoplasma doses and monitored for extended time periods. A time-and dose-dependent decline in the viability of infected cells was observed; however, viability never declined below 50% of that in uninfected controls. Protein and RNA synthesis actually increased above control levels in infected cells, despite a concomitant decrease in viability. This response was pronounced at higher multiplicities of infection but was only transient at lower doses. In parallel studies in which a culture medium capable of supporting M. pneumoniae growth was used, loss of viability was accelerated. With a low-dose infection a transient increase followed by a precipitous decline in macromolecular synthesis was observed, relative to that in uninfected controls. At higher doses, however, macromolecular synthesis decreased dramatically and in proportion to the loss of viability. The requirement for HTE cells for mycoplasma growth under the experimental culture conditions was demonstrated by quantitating viable mycoplasmas in the culture medium in the presence or absence of HTE cells over 4 days. The increase in mycoplasma number was negligible in the absence of HTE cells, while a 30-fold increase was observed in the presence of HTE cells. These findings demonstrate the feasibility of long-term, low-dose studies of M. pneumoniae pathogenesis with trachea epithelial cells and a nonpermissive culture medium. This * Corresponding author. suspension through a sterile 1.2-p.m-pore-size filter. CFU 570 on July 16, 2020 by guest http://iai.asm.org/ Downloaded from

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Alterations in nucleotide content of human lung fibroblasts infected with Mycoplasma pneumoniae

Cited by 5 publications

References 35 publications

Interaction of Mycoplasma pneumoniae with HeLa cells

Interaction of Mycoplasma pneumoniae with HeLa cells

Gaseous oxide toxicity evaluated with cell monolayers on collagen-coated, gas-permeable teflon membranes.

Parasitism of hamster trachea epithelial cells by Mycoplasma pneumoniae

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