Isolation of Kurloff cells by Percoll density gradient centrifugation. Protein labeling with 35S‐methionine of these cells

Landemore, Gérard; Debout, Claire; Quillec, M.; Izard, Juan Francisco Martín

doi:10.1111/j.1768-322x.1984.tb00258.x

Cited by 22 publications

(7 citation statements)

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“…Therefore, the present results indicate that Kurloff cells follow the same dif ferentiation pathway as normal thymocytes. It was earlier shown that Kurloff cells with identical morphology can be recovered in both low-and highdensity fractions of spleen cells [Landemore et al, 1984], This supports our view that the different densities of Kurloff cells is not related to the pres ence of, or type of, Kurloff inclusion but is more a reflexion of the respective thymocyte population property. We think that Kurloff cells have a relation to the normal thymocytes, as indicated by trans mission electron microscopical studies [Bimes et al, 1963].…”

Section: Discussionsupporting

confidence: 79%

Kinetic Study of Kurloff Cells in Guinea Pig Thymus

Sandberg¹,

Hagelin²

1986

Int Arch Allergy Immunol

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One single injection of estradiol to male guinea pigs resulted in the appearance of Kurloff cells in the thymus and spleen in a maximal number after 2–3 weeks. In both organs, Kurloff cells of three categories were observed with different size and number of the inclusion(s). In the thymus, cells with small and medium-sized inclusions were present almost exclusively among low-density thymocytes. Cells with one large inclusion were initially present among low-density cells, but with time an increasing proportion were found among high-density thymocytes. This indicates that the different categories of Kurloff cells represent maturational stages and follow the normal differentiation step of thymocytes from low to high density. According to electronic cell volume distribution analysis, estradiol treatment was associated with a shift in cell volume towards larger cells, and this shift was correlated in time with the appearance of Kurloff cells with large inclusions. In thymus sections there was evidence for a massive and selective migration of Kurloff cells via interlobular lymph vessels. No difference in localization of Kurloff cells was noted at various times after estradiol treatment. Attempts to induce Kurloff cell formation from bone marrow, spleen or thymus cells during 2 weeks in vitro were unsuccessful, also when culturing the cells in serum from estradiol-treated animals. This negative result, the long lag phase of estradiol-induced Kurloff cell formation in vivo, and the reported lack of estradiol receptor in Kurloff cells indicate that the Kurloff cell induction by estradiol may be indirect.

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Section: Discussionsupporting

confidence: 79%

Kinetic Study of Kurloff Cells in Guinea Pig Thymus

Sandberg¹,

Hagelin²

1986

Int Arch Allergy Immunol

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“…Using highly purified KC [29], soluble extracts obtained after dissociative extraction in either 4 M GnHCI or 7 M urea were subjected to ion exchange chromatography and the macromolecules responsible for these histochemical affinities were identified: i) unbound major 30-35 kDa glycoproteins designated as major KC GP [30,35] among which a fraction containing Neu5Ac(oc2,6)-D-Gal/Gal-NAc(~I,4)GIcNAc oligosaccharide sequences were shown to correspond to acid phosphatase subunits [33,42]; and it) protease-resistant chondroitin-4-sulphate proteoglycans eluted at 0.55 M NaCI [31,36] which correspond to PASpositivity and TB-metachromasia of the KB, respectively.…”

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confidence: 99%

Characterization of the non‐sulphated highly anionic kurloff cell glycoconjugates by lectin‐ and immunoblotting: Evidence for the presence of α(2,8)‐linked polysialic acid‐bearing molecules

Letaïef

Landemore

Oulhaj

et al. 1993

Biology of the Cell

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Urea or guanidine hydrochloride-soluble extracts from highly purified Kurloff cells (KC) radiolabelled in vitro were subjected to DEAE-cellulose chromatography. Among the three anionic peaks obtained, a major and non-sulphated peak (designated as peak IV) strongly affected by glucosamine-labelling and eluted at about 0.3 M NaCl was analyzed. Gel filtration on Sepharose CL4B and 10% SDS-PAGE indicated its heterogeneous size. Peak IV consisted mainly of N-glycans as shown by its susceptibility to tunicamycin. Further insight into its chemical nature was obtained by examining its binding capacity to different lectins and by immunodot analysis. It strongly interacted with concanavalin A (Con A) after dot-blot or Western blotting. A large amount of these glycoproteins is not of the high-mannose type since Galanthus nivalis agglutinin reacted weakly with peak IV. Moreover, bindings to Phaseolus vulgaris and to wheat germ agglutinins suggest the presence of bisecting N-acetylglucosamine residues. Bindings to Sambucus nigra and to Ricinus communis agglutinins, dramatically lessened and increased respectively after desialylation, suggest the presence of Neu5Ac alpha 2,6Gal/GalNAc sequences. The absence of outer sialic acid residues linked alpha 2,3 to galactose was demonstrated following Maackia amurensis agglutinin negativity. The use of poly(alpha 2,8-sialyl) endo-N-acylneuraminidase combined with immunodot procedure with a monoclonal antibody that specifically recognizes alpha 2,8-linked polysialic chains revealed that peak IV contains oligosaccharidic epitopes common to polysialylated neural cell adhesion molecules.

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“…KC suspensions were obtained as previously described [16]. Their * CorresPondence and reprints purity was checked by phase contrast microscopy and their via- Initial experiments revealed that extraction of the cells by homogenization, sonication, incubation in 1070 Triton X-100 or by any combination of these procedures yielded the same gross recovery of Asase activity.…”

Section: Obtaining Of Kcmentioning

confidence: 99%

Kurloff cell lysosomal arylsulphatases: Presence of both cationic and highly anionic isoforms of the sole B class

Taouji¹,

Buat²,

Izard³

et al. 1994

Biology of the Cell

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Following the previous ultrastructural demonstration of the presence of arylsulphatase (Asase) activities in Kurloff cells (KC) and of their quasi-exclusive localization in the Kurloff body (KB), this work investigates their biochemical and zymographic properties after extraction from purified KC suspensions. Using the discriminative inhibitory conditions of both the Baum or Lee-Vaupel and Conzelmann methods, nitrocatechol sulphate hydrolyzing enzymes of the KC were assumed to belong to the B class of the type II Asase alone. After electrophoretic separation under non-denaturing conditions in a 4-23% polyacrylamide gel, they were characterized by 55 kDa and 62 kDa zymographic bands. After isoelectric focusing, 'classical' cationic isoforms (pI 8.5) and two anionic isoforms (pI 4.4 and 4.6) were observed on zymograms. As expected for class B Asase, the different zymographic forms of KC Asase were only recovered in the unadsorbed fraction after anion-exchange chromatography on DEAE-cellulose column equilibrated with high ionic strength buffer. Their Km (2.1 mM), their optimum pH (5.8) and their inhibitions by sulfite, phosphate, sulphate and ascorbic acid as well as their slight stimulation by AgNO3 were also characteristic of this class of Asase. Finally, chondroitin-4-sulphate was shown to potentially be a physiological substrate for these lysosomal enzymes.

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Isolation of Kurloff cells by Percoll density gradient centrifugation. Protein labeling with 35S‐methionine of these cells

Cited by 22 publications

References 0 publications

Kinetic Study of Kurloff Cells in Guinea Pig Thymus

Kinetic Study of Kurloff Cells in Guinea Pig Thymus

Characterization of the non‐sulphated highly anionic kurloff cell glycoconjugates by lectin‐ and immunoblotting: Evidence for the presence of α(2,8)‐linked polysialic acid‐bearing molecules

Kurloff cell lysosomal arylsulphatases: Presence of both cationic and highly anionic isoforms of the sole B class

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