The Effect of Purified Lipopolysaccharide on the Bactericidal Reaction of Human Serum Complement

Allen, Rosalind J.; Scott, G. K.

doi:10.1099/00221287-117-1-65

Cited by 9 publications

(22 citation statements)

References 6 publications

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“…The chromatographic purification of LPS from P. leiognathi was successfully carried out by the same methods as reported for E. coli ML308 225 (Allen & Scott, 1980). A similar total lipid content (measured as fatty acid methyl esters) of 11 % was found.…”

Section: Results a N D Discussionmentioning

confidence: 62%

“…2 in the case of E. coli, but the profile for the P. leiognathi extract is much more complex than the corresponding profile derived from purified outer membranes. Thus the differential solubility technique which is so convenient for isolating outer membrane proteins from E. coli is not suitable for use with P. leiog na t h i.The chromatographic purification of LPS from P. leiognathi was successfully carried out by the same methods as reported for E. coli ML308 225 (Allen & Scott, 1980). A similar total lipid content (measured as fatty acid methyl esters) of 11 % was found.…”

mentioning

confidence: 49%

“…When added to a mixture of E. coli ML308 225 and normal human serum, the LPS from P. leiognathi caused a 50% inhibition of the complement-dependent bactericidal reaction. We have shown previously that this inhibition is due to LPS binding to the bacterial surface, and can also be observed with E. coli K 12 (Allen & Scott, 1980, 1981. The binding of LPS to Salmonella typhimurium outer membranes has also been reported (Jones & Osborn, 1977).…”

Section: Results a N D Discussionmentioning

confidence: 93%

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Comparative Biochemistry of the Cell Envelopes of Photobacterium leiognathi and Escherichia coli

1983

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Photobacterium leiognathi closely resembles Escherichia coli with respect to cell lysis by lysozyme, and the fractionation of outer and cytoplasmic membranes. The two organisms differ in their phospholipid contents and, more significantly, in outer membrane protein compositions. I N T R O D U C T I O NPhotobacterium leiognathi ( P . mandapamensis) is a luminescent marine bacterium found as the specific symbiont in the luminous organ of Leiognathid and Apogonid fish, as a marine intestinal commensal or free-living in sea water (Reichelt & Baumann, 1973;Nealson & Hastings, 1979). The genus shares many properties with the Enterobacteriaceae (Baumann & Baumann, 1977). In particular, electron microscopy of thin sections of this organism reveals a cell envelope which is morphologically very similar to terrestrial Gram-negative bacteria, with the characteristic membrane, cell wall, periplasmic space and outer membrane (Reichelt & Baumann, 1973). It would not be surprising if closer analysis of the cell envelope of P. leiognathi revealed considerable differences from typical Gram-negative bacteria, since the marine organism must cope with a constant high osmotic pressure. At the same time, there is some evidence for greater membrane permeability in this genus. We have observed that P. leiognathi has a relatively high sensitivity to a variety of antibiotics and chemical mutagens. In addition, the periplasmic b-lactamase of Photobacterium species does not generally display the 'crypticity' typical of this enzyme in most other Gram-negative bacteria (Richmond and Sykes, 1973), and usually taken as a measure of outer membrane impermeability to b-lactam antibiotics (K. Smith & S. Lamb, unpublished results). We have accordingly analysed various aspects of the biochemistry of the cell envelope of P. leiognathi with particular emphasis on the composition of the outer membrane, and using Escherichia coli for comparative purposes. METHODSGrowth qf bacteria. The maintenance and growth of Escherichiu coli ML308 225 have been described previously (Allen & Scott, 1979). Photobacterium Ieiognathi strain K-721 (Nealson & Hastings, 1977) was obtained from the Roche Research Institute of Marine Pharmacology in Sydney, and was grown in liquid cultures using minimal medium (Reichelt & Baumann, 1973) without CaClz and supplemented with 0.2% (w/v) yeast extract. This bacterial strain was maintained on plates made by adding 1.2% (w/v) agar to the liquid medium.Mfnihranpprepcrrcition. Membranes from E. coli ML308 225 in the late-exponential growth phase were prepared and fractionated by sucrose density gradient centrifugation as described previously (Allen &Scott, 1979). A 500 ml culture of P. leiognarhi in the late-exponential growth phase was harvested by centrifugation, resuspended in 20 ml deionized water and mixed with 2 mg egg white lysozyme (Sigma) in 20 ml 30 mM-Tris/HCI buffer pH 7.5. Following incubation at 37 "C for 15 min, the mixture was centrifuged for 3 x lo4 g-min to remove intact cells and then for 2 x lo6 g-min. The resulting...

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Section: Results a N D Discussionmentioning

confidence: 62%

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confidence: 49%

Section: Results a N D Discussionmentioning

confidence: 93%

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Comparative Biochemistry of the Cell Envelopes of Photobacterium leiognathi and Escherichia coli

1983

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“…Previous studies have shown that gram-negative organisms like Escherichia coli are more sensitive to killing by complement when they are undergoing logarithmic growth (2). In one study (5), simultaneous disruption of outer membrane integrity and inner membrane integrity occurred upon addition of complement to E. coli in the logarithmic phase.…”

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confidence: 99%

Growth Phase Influences Complement Resistance ofBordetella pertussis

Barnes

Weiss

2002

Infect Immun

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The BrkA protein of Bordetella pertussis inhibits killing by the antibody-dependent classical pathway of complement; however, susceptibility to complement can be highly variable. Log-phase bacteria grown in Stainer-Scholte (SS) broth plated on Bordet-Gengou (BG) agar were about 500 times more sensitive to killing by complement than stationary-phase SS-BG cultures. While always more susceptible to complement than the wild-type strain, a BrkA mutant displayed a similar growth phase variation in susceptibility to complement. Growth phase susceptibility to complement was also observed for a mutant constitutive for Bvg activation of BrkA, suggesting that modulation of virulence factor expression was not responsible for sensitivity to complement. Susceptibility was not due to differential antigenic expression, since serum adsorbed with complement-resistant, stationary-phase SS-BG cultures lacked bactericidal activity against B. pertussis harvested at all times during the growth cycle. These results suggest that log-phase susceptibility to complement is not due to variable expression of BrkA or antigenic differences and may be an inherent property of rapidly growing cultures. Implications for vaccine development are discussed.

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“…A portion of 0.1 to 0.5 ml of each antiserum was added per ml of serum. The solution was incubated for 2 h at 0°C and then centrifuged briefly to remove precipitated protein (1). An equivalent volume of heat-inactivated serum containing 0.02% sodium azide was added to controls.…”

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Effects of trospectomycin on serum sensitivity of Escherichia coli UC 9451

Cialdella

Ulrich

Liggett

et al. 1990

Antimicrob Agents Chemother

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Trospectomycin sulfate, a chemically synthesized analog of spectinomycin, exhibits a broad range of activity against both aerobes and anaerobes, including the etiological agents of sexually transmitted diseases. Its activity in vitro against Escherichia coli is considered only moderate. At subinhibitory levels, however, trospectomycin induced changes in a pathogenic strain of E. coli, UC 9451, which significantly increased its sensitivity to serum lysis. This strain of E. coli shows high-level resistance to serum in vitro, typically growing twofold within a 45-min incubation period. Following exposure to one-fifth the MIC of trospectomycin, >99% of the bacteria were killed in 25% serum within 15 min. Surviving bacteria were static in this level of serum for over 3 h. Killing was due to lysis mediated by both the classical and alternative complement pathways. The bacteria exposed to trospectomycin were enlarged in both diameter and length, but they still grew at rates comparable to those of untreated bacteria. No other visible morphological changes could be directly related to the increase in serum sensitivity. The profile of outer membrane proteins obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was identical for trospectomycin-treated or untreated bacteria. However, the relative proportion of four major outer membrane proteins varied considerably.

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The Effect of Purified Lipopolysaccharide on the Bactericidal Reaction of Human Serum Complement

Cited by 9 publications

References 6 publications

Comparative Biochemistry of the Cell Envelopes of Photobacterium leiognathi and Escherichia coli

Comparative Biochemistry of the Cell Envelopes of Photobacterium leiognathi and Escherichia coli

Growth Phase Influences Complement Resistance ofBordetella pertussis

Effects of trospectomycin on serum sensitivity of Escherichia coli UC 9451

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