Two-step separation of human peripheral blood monocytes on discontinuous density gradients of colloidal silica-polyvinylpyrrolidinone

Nathanson, S. David; Zamfirescu, P.; Drew, SI; Wilbur, Stanley M.

doi:10.1016/0022-1759(77)90176-4

Cited by 73 publications

(16 citation statements)

References 27 publications

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“…Ig+ cells were removed bypassing the remaining cells through an anti-mouse Ig column as described before [32]. The nonadherent cells, 65-70 % CD4+, were separated according to density on a discontinous Percoll gradient (Pharmacia, Uppsala, Sweden) centrifuged at lo00 x g for 25 min as described [33,34].The high-densityfractionof cells (1.130 > e 1.078) was used for the experiments. As accessory cells, spleen cells were used which had been depleted of T cells using anti-CD8 (3.155), anti-CD4 (GK1.5) [35] and anti-Thy-1.2 (AT83) [36] antibodies and complement as above.…”

Section: Cellsmentioning

confidence: 99%

Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4⁺ T cells

1993

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The production of several lymphokines by freshly isolated CD4+ T cells has been analyzed at the single-cell level, after stimulation with staphylococcal enterotoxin B (SEB). High frequencies of cells producing interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) were induced, but very low frequencies of CD4+ T cells produced IL-4, IL-5 or IL-10 in response to SEB. Exogenously added IL-4 markedly altered the lymphokine profile induced during primary SEB stimulation. IFN-gamma production was reduced, while a high fraction of cells contained IL-10 and IL-4 after activation in the presence of IL-4. We further demonstrate that IL-4 and IL-10 or IFN-gamma production was selectively induced in resting, high-density CD4+ T cells during primary stimulation, by SEB + IL-4 or SEB. Under conditions where both IL-10 and IFN-gamma were produced, most cells contained only one of the two lymphokines.

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Section: Cellsmentioning

confidence: 99%

Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4⁺ T cells

1993

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“…There is an increasing number of investigations of distinct classes or subclasses of human blood cells [26], For blood of ani mals, however, there is a nearly total lack of application of this new gradient material. The main properties of Percoli: its nontoxicity, the negligible osmolality, the inability to penetrate cell membranes and the general gentleness against blood cells [19,25,28] ren der this substance extremely useful as a me dium which does not impair cellular enzyme activities. As to enzyme activities in human and dog blood cells, we previously reported on the advantageous separation technique in Percoli [22].…”

Section: In Contrast To Man a Comprehensive De Scription Of The Enzymentioning

confidence: 99%

Enzyme Activities in Rabbit, Guinea Pig, Rat and Mouse Blood Cells after Separation on a Discontinuous Percoli Gradient

Lindena¹,

Sommerfeld²,

Höpfel³

et al. 1983

Enzyme

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A technique is described which allows a fractionation of rabbit, guinea pig, rat and mouse blood cells using a discontinuous Percoli gradient. With only two steps of centrifugation a simultaneous isolation of thrombocytes, mononuclear cells, polymorphonuclear cells, and erythrocytes in nearly pure form is performed in a very short time, starting from only 1 ml of blood. In these morphologically almost homogeneous cell fractions the activities of 12 enzymes were determined. The enzyme pattern, which with respect to the number of enzymes, specific cell populations and species was not yet investigated to such an extent, revealed a manifold higher enzyme content in the few cases, in which comparative studies were made. This can be attributed mainly to the completeness of the cell disruption technique using a detergent and certainly also to the use of Percoli as a preferable gradient material.

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“…The main properties of Percoli are its nontoxicity, low viscosity, very low osmolality, and its inabil ity to penetrate cell membrane [30,31].…”

Section: Introductionmentioning

confidence: 99%

Enzyme Activities in Blood Cells of Man and Dogs after Separation on a Discontinuous Percoli Gradient

Lindena¹,

Sommerfeld²,

Höpfel³

et al. 1983

Enzyme

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From human and dog peripheral blood, thrombocytes (TRC), mononuclear cells (MNC), polymorphonuclear cells (PMNC), and red blood cells (RBC), were harvested after separation on a discontinuous Percoli gradient in a two-step centrifugation procedure, and 12 enzyme activities were determined in these highly purified cells. The enzyme activities measured are generally severalfold higher than previously reported, a fact which is attributed to the gentle and time-shortening isolation in Percoli, the cell disruption technique using detergent and the enzyme test conditions.

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Two-step separation of human peripheral blood monocytes on discontinuous density gradients of colloidal silica-polyvinylpyrrolidinone

Cited by 73 publications

References 27 publications

Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4⁺ T cells

Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4⁺ T cells

Enzyme Activities in Rabbit, Guinea Pig, Rat and Mouse Blood Cells after Separation on a Discontinuous Percoli Gradient

Enzyme Activities in Blood Cells of Man and Dogs after Separation on a Discontinuous Percoli Gradient

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Two-step separation of human peripheral blood monocytes on discontinuous density gradients of colloidal silica-polyvinylpyrrolidinone

Cited by 73 publications

References 27 publications

Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4+ T cells

Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4+ T cells

Enzyme Activities in Rabbit, Guinea Pig, Rat and Mouse Blood Cells after Separation on a Discontinuous Percoli Gradient

Enzyme Activities in Blood Cells of Man and Dogs after Separation on a Discontinuous Percoli Gradient

Contact Info

Product

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Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4⁺ T cells

Manipulation of the superantigen‐induced lymphokine response. Selective induction of interleukin‐10 or interferon‐γ synthesis in small resting CD4⁺ T cells