Purification of Chlamydia trachomatis by a simple and rapid filtration method

Campbell, Steven C.; Yates, Peter; Waters, Fiona; Richmond, Shirley J.

doi:10.1099/00221287-137-7-1565

Cited by 8 publications

(8 citation statements)

References 6 publications

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Section: Propagation Of Chlamydia Pneumoniaementioning

confidence: 99%

“…pneumoniae, 96-41, was isolated from the brain following autopsy of an Alzheimer's disease patient and propagated in the HEp-2 cell line similar to the technique described for the C. pneumoniae respiratory isolate AR-39 [7]. For in vivo experiments, C. pneumoniae was concentrated using an established technique described previously [7]. In brief, a homogenate of C. pneumoniae-infected HEp-2 cells was passed through a filter membrane with a 0.8 m pore diameter to remove large particulate cellular debris and the organism was "captured" on a filter membrane with a 0.2 m pore diameter.…”

Section: Propagation Of Chlamydia Pneumoniaementioning

confidence: 99%

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Chlamydia pneumoniae induces Alzheimer-like amyloid plaques in brains of BALB/c mice

Little

Hammond

MacIntyre

et al. 2004

Neurobiology of Aging

206

197

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Amyloid deposits resembling plaques found in Alzheimer's disease (AD) brains were formed in the brains of non-transgenic BALB/c mice following intranasal infection with Chlamydia pneumoniae. The mice were infected at 3 months of age with C. pneumoniae isolated from an AD brain. Infection was confirmed by light and electron microscopy in olfactory tissues of the mice. C. pneumoniae was still evident in these tissues 3 months after the initial infection indicating that a persistent infection had been established. Amyloid beta (A␤) 1-42 immunoreactive deposits were identified in the brains of infected BALB/c mice up to 3 months post-infection with the density, size, and number of deposits increasing as the infection progressed. A subset of deposits exhibited thioflavin-s labeling. Intracellular A␤1-42 labeling was observed in neuronal cells. Experimental induction of amyloid deposition in brains of non-transgenic BALB/c mice following infection with C. pneumoniae may be a useful model for furthering our understanding of mechanisms, linked to infection, involved in the initiation of the pathogenesis of sporadic AD.

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Section: Propagation Of Chlamydia Pneumoniaementioning

confidence: 99%

Section: Propagation Of Chlamydia Pneumoniaementioning

confidence: 99%

Chlamydia pneumoniae induces Alzheimer-like amyloid plaques in brains of BALB/c mice

Little

Hammond

MacIntyre

et al. 2004

Neurobiology of Aging

206

197

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“…Experimental C. pneumoniae infection of mice. The C. pneumoniae AR-39 isolate (ATCC) was propagated in the HEp-2 cell line and concentrated by using a technique previously described by Campbell et al (12). C. pneumoniae was then divided into aliquots and frozen until dilution for experimental infection of mice under biosafety level 2 precautions.…”

Section: Methodsmentioning

confidence: 99%

Age Alterations in Extent and Severity of Experimental Intranasal Infection withChlamydophila pneumoniaein BALB/c Mice

et al. 2005

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The intracellular bacterium Chlamydophila ("Chlamydia") pneumoniae is a pathogen for several respiratory diseases and may be a factor in the pathogenesis of chronic diseases of aging including atherosclerosis and Alzheimer's disease. We assessed whether aging is coupled with increased burden of infection in BALB/c mice after intranasal infection by C. pneumoniae. Six-and twenty-month-old BALB/c mice were infected intranasally with 5 ؋ 10 4 inclusion forming units (IFU) or 5 ؋ 10 5 IFU of C. pneumoniae. Lung, brain, and heart tissue were analyzed for infectious C. pneumoniae and for Chlamydophila antigen by immunohistochemistry. At both doses, aging was associated with a decreased proportion of animals that cleared infection from the lung and greater burden of infectious organism within the lung. We observed dose-dependent spread to the heart/ascending aorta in animals infected with C. pneumoniae. In mice given 5 ؋ 10 4 IFU, spread to the heart by day 14 was only observed in old mice. By day 28, all animals inoculated with 5 ؋ 10 4 IFU showed evidence of spread to the heart, although higher C. pneumoniae titers were observed in the hearts from old mice. In mice inoculated with 5 ؋ 10 5 IFU, spread of C. pneumoniae to the heart was evident by day 14, with no discernible age effect. C. pneumoniae was also recovered from the central nervous system (brain and olfactory bulb) of all mice by day 28 postinfection, with higher C. pneumoniae titers in old animals than in young animals. Our results suggest that infection with C. pneumoniae may be more severe in old animals.With the life expectancy increasing over this century and predicted to continue to rise over the course of the next half century, it is expected that, by the middle of the 21st century, more than 20% of the population will be over the age of 65 (19).

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“…The organisms were purified on renographin gradients and filtered through 0-6 pm pore diam. polycarbonate membrane filters (Nucleopore) to exclude chlamydial reticulate bodies (Campbell et al, 1991). In one series of experiments a chlamydial suspension in 200 pl PBS (2 x lo5 c.p.m.)…”

Section: Methodsmentioning

confidence: 99%

“…Infected cell homogenates were prepared by shaking with glass beads. Purified chlamydiae were recovered from the infected cell homogenate by density gradient centrifugation (Howard et al, 1974) or by differential filtration with 0.6 pm and 0.2 pm pore diameter polycarbonate membrane filters (Campbell et al, 1991).…”

Section: Methodsmentioning

confidence: 99%

Chlamydia trachomatis infection of cultured motile cells after uptake of chlamydiae from the substratum

Campbell¹,

Yates

Richmond

1993

Journal of General Microbiology

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The ability of motile cells to remove small inanimate particles from solid substrata is well documented. We show here that motile cells will pick up and internalize infectious particles of the obligate intracellular parasite Chlamydia tvachomatis when they are adherent to the substratum over which the host cells move. Two cell types were used to assess chlamydial uptake ; a feeder independent human squamous cell carcinoma variant (AC3A cells) and the McCoy cell line, Purified chlamydial elementary bodies were attached to glass or collagen-coated glass by centrifugation. Suspensions of cells were then allowed to sediment on to the substrata to which chlamydiae had attached. Both types of cell picked up chlamydiae and transported them over their surface during the course of attachment and spreading. Stereoscopic images obtained by confocal microscopy demonstrated that chlamydiae were found mainly on the surface of non-spread cells. After the cells had spread on the substratum they began to move around forming tracks where the chlamydiae had been removed. Some cell-surface-attached chlamydiae were endocytosed and a proportion of these proliferated during the 48 h after plating. However, chlamydiae attached to the substratum lost infectivity by a simple exponential decay process within a few hours of incubation in the extracellular environment. Therefore, increasing numbers of non-viable organisms were probably endocytosed as the time of extracellular incubation increased. This mode of infection may be relevant to in uivo situations where cell migration occurs after damage to mucosal surfaces.

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Purification of Chlamydia trachomatis by a simple and rapid filtration method

Cited by 8 publications

References 6 publications

Chlamydia pneumoniae induces Alzheimer-like amyloid plaques in brains of BALB/c mice

Chlamydia pneumoniae induces Alzheimer-like amyloid plaques in brains of BALB/c mice

Age Alterations in Extent and Severity of Experimental Intranasal Infection withChlamydophila pneumoniaein BALB/c Mice

Chlamydia trachomatis infection of cultured motile cells after uptake of chlamydiae from the substratum

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