Pneumocystis carinii: New separation method from lung tissue

Walzer, Peter D.; Rutledge, M E; Yoneda, K; Stahr, Benjamin J.

doi:10.1016/0014-4894(79)90088-2

Cited by 44 publications

(19 citation statements)

References 15 publications

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“…Oocysts were further purified by centrifugation on a Ficoll-sodium diatrizoate density gradient. This technique was modified from that used by Walzer et al (14) to separate Pneumocystis carinii cysts and trophozoites from homogenized rat lung.…”

Section: Discussionmentioning

confidence: 99%

Electrophoretic and immunoblot analysis of Cryptosporidium oocysts

1988

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Summary Cryptosporidium oocysts were recovered by density gradient centrifugation from diarrhoeal faeces of four human patients and one goat kid. Goat-derived oocysts were further treated with excystation medium and the excysted oocyst walls purified by isopycnic ultracentrifugation. Soluble extracts from intact oocysts and the oocyst wall preparation were analysed by SDS-PAGE. Fifty-one polypeptide bands were detected in intact oocyst preparations: 48 were in the range 14 000-200 000 molecular weight (MW), two bands were less than 14 000 MW and one band was above 200 000 MW. Twenty-one bands were detected in the oocyst wall preparation, all within the range 14 000-200 000 MW. Immunoblot analysis oi Cryptosporidium polypeptides using acute or convalescent human and goat sera revealed a large number of reactive bands. Varying degrees of heterogeneity were observed within and between the two semm groups. Nine ofthe 10 human sera and all ofthe goat kid sera reacted with a 23 000 MW and 32 000 MW antigen. A 15 500 MW antigen was also detected by all the goat and four ofthe 10 human sera. Both serum groups reacted with various antigens above 40 000 MW. Surface labelling of three human isolatesofC/^p/o^^^onW/um oocysts with '25i was performed using the Boltonand Hunter reagent. The solubilized preparations were separated by SDS-PAGE on 12% and 18% slab gels and autoradiographed. Common bands were seen at 15 500, 32 000. 47 500, 79 000 and 96 000 MW. Some variation in the molecular weight of polypeptides labelled with ^'^H was observed among the three isolates. These studies indicate that the 32 000 MW antigen appears to be a suitable candidate for the preparation of an antibody probe because it is common to many isolates and is located on the exterior of the oocyst wall.

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Section: Discussionmentioning

confidence: 99%

Electrophoretic and immunoblot analysis of Cryptosporidium oocysts

1988

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“…Cysts of P. carinii were prepared, essentially as described by WALZER et al [10]. Wistar male rats, weighing about 250 g, were immunosuppressed by adding dexamethasone 1 mg·l -1 , and tetracycline 1 mg·ml -1 to the drinking water.…”

Section: Antigenmentioning

confidence: 99%

Local antibodies against Pneumocystis carinii in bronchoalveolar lavage fluid

Laursen

Jensen²,

Andersen³

1994

Eur Respir J

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“…Rat P. carinii preparation. P. carinii organisms were isolated from lung tissue according to the method of Walzer et al (42) with minor modifications (28). Semifrozen lung tissue was finely diced with a scalpel and stirred in calcium-and magnesium-free Hanks balanced salt solution (HBSS) with a magnetic stirrer for 2 hr at 4 C. The smallest particles were filtered through a 20-mesh stainless steel screen and held at 4 C. The lung fragments retained on the screen were again stirred in HBSS.…”

Section: Methodsmentioning

confidence: 99%

Plasminogen Activator Production in a Rat Model of Pneumocystis carinii Pneumonia

Angelici

Contini

Spezzano

et al. 2001

Microbiology and Immunology

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Several studies have indicated that the serine protease urokinase-plasminogen-activator (uPA) is an important factor in host defense against pulmonary pathogens. To gain a better insight into the role of uPA in Pneumocystis carinii (P. carinii) pneumonia (PCP), we evaluated PA production in alveolar macrophages (AMs) obtained from rats with steroid-induced PCP. Treatment with cortisone acetate favored PCP in 91% of rats. In the bronchoalveolar lavage (HAL) samples of immunosuppressed rats both with and without PCP, we observed a decrease in uPA activity as well as a decrease in cell number. Urokinase-PA production by AMs was reduced in rats treated with cortisone alone. However, an increase in cell-associated uPA was observed in rats with PCP. This increase appears to be produced in response to P. carinii infection. In fact, when AMs obtained from untreated healthy or immunosuppressed uninfected rats were challenged with P. carinii, a significant increase in PA activity in cell Iysates was observed, though a lower response was obtained in cortisone-treated animals. Our results suggest that healthy AMs respond to the presence of P. carinii with an increase in uPA production and that this response in immunodepressed rat-AMs is partially impaired.

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Pneumocystis carinii: New separation method from lung tissue

Cited by 44 publications

References 15 publications

Electrophoretic and immunoblot analysis of Cryptosporidium oocysts

Electrophoretic and immunoblot analysis of Cryptosporidium oocysts

Local antibodies against Pneumocystis carinii in bronchoalveolar lavage fluid

Plasminogen Activator Production in a Rat Model of Pneumocystis carinii Pneumonia

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