An adhesion assay using minimal shear force to remove nonadherent cells

John, J; Schroen, Daniel J.; Cheung, H. Tak

doi:10.1016/0022-1759(94)90391-3

Cited by 29 publications

(15 citation statements)

References 16 publications

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“…Nonadherent cells were removed by limited shear force. 40 Fluorescence of remaining cells was measured and the cell number determined using a standard curve of labeled cells. As controls, cyclosporin A (calcineurin inhibitor), ketokonazole (cytochrome P450 inhibitor), and lactacystin (proteasome inhibitor) were included at comparable concentrations.…”

Section: Ritonavir Inhibits Cell Adhesion and Expression Of Cell Adhementioning

confidence: 99%

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Antitumorigenic effects of HIV protease inhibitor ritonavir: inhibition of Kaposi sarcoma

Pati

Pelser²,

Dufraine³

et al. 2002

Blood

132

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Treatment of patients with human immunodeficiency virus (HIV) protease inhibitors such as ritonavir can result in increases in CD4 ؉ T-cell counts that are independent of a reduction in HIV-1 viral load. This lack of correlation between the 2 has led to the identification of additional effects of ritonavir that potentially alter HIV disease pathogenesis. Our previous studies indicated that ritonavir directly affects immune cell activation, proliferation, and susceptibility to apoptosis. We show here that ritonavir inhibited the activation and proliferation of primary endothelial cells and decreased the produc-

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Section: Ritonavir Inhibits Cell Adhesion and Expression Of Cell Adhementioning

confidence: 99%

“…A shear force assay, as previously described, 40 was adapted to measure adhesion of leukocytes to endothelial cells. HUVECs (passage 5) were grown to form a monolayer in 96-well plates.…”

Section: Adhesion Assaysmentioning

confidence: 99%

Antitumorigenic effects of HIV protease inhibitor ritonavir: inhibition of Kaposi sarcoma

Pati

Pelser²,

Dufraine³

et al. 2002

Blood

132

View full text Add to dashboard Cite

show abstract

“…6b) cells were treated for 4 h with 5 ng of TNF-␣ per ml and then mixed with KSIMM or HUT 78 cells, respectively, as a positive control for adhesion. Cells were then washed with minimal shear force (74). Plates were subsequently read with a Wallac Victor-2 fluorescence plate reader (Wallac, Gaithersburg, Md.).…”

mentioning

confidence: 99%

Human Herpesvirus 8-Encoded vGPCR Activates Nuclear Factor of Activated T Cells and Collaborates with Human Immunodeficiency Virus Type 1 Tat

Pati

Foulke

Barabitskaya

et al. 2003

J Virol

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“…In some experiments, eosinophils were suspended in buffer with saturating concentrations (10 g/ml) of anti-ICAM-1, anti-ICAM-2, and/or anti-ICAM-3 before introduction into the wells. At the end of the predetermined adhesion period, nonadherent eosinophils were removed by submersion and inversion of the 96-well plate for 10 min in a tank of HBSS with calcium (16). This permitted uniform, 1 ϫ g sedimentation of nonadherent cells.…”

Section: Adhesion Assaymentioning

confidence: 99%

“…The nonadherent eosinophils were allowed to "fall" out of the well for 10 min. The optimal time for inversion and sedimentation was determined to be 10 min after the method of St. John et al (16). We validated inversion-sedimentation time by placing CTO-labeled eosinophils in wells in the absence of prebound CD4 ϩ T cells.…”

Section: Activation Of Cd4 ϩ T Cells Induces Adhesion Of Eosinophilsmentioning

confidence: 99%

CD4+ T Cell and Eosinophil Adhesion Is Mediated by Specific ICAM-3 Ligation and Results in Eosinophil Activation

Douglas

Leff

Sperling

2000

The Journal of Immunology

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T cells and eosinophils, which are found in close proximity in asthmatic lungs, express many surface receptors that are counterligands. These data suggest that direct interactions between these cell types could play an important role in regulating airway inflammation in asthma. We examined the effect of selective adhesion between counterligands on human eosinophils and CD4+ T cells to determine 1) the existence of specific adhesive interactions and 2) if augmented specific adhesion to CD4+ T cells also caused augmented secretion of leukotriene C4 (LTC4) from eosinophils. A new method for binding of human CD4+ T cells to microwell plates was developed, which allowed for specific quantitative assessment of eosinophil adhesion to individual CD4+ T cells in culture. Adhesion of CD4+ T cells to eosinophils was minimal in unstimulated cells but increased after activation of T cells by PMA. Augmented adhesion was regulated substantially through binding of ICAM-3 and only minimally by ICAM-1. We further evaluated whether this specific adhesion up-regulated stimulated secretion of LTC4 from eosinophils. Adhesion with CD4+ T cells augmented eosinophil secretion of LTC4 caused by FMLP plus cytochalasin. Blockade of ICAM-3, as well as ICAM-1, inhibited completely the augmented secretion of eosinophil LTC4. We demonstrate that eosinophils and CD4+ T cells are capable of ligand-specific adhesion that is mediated predominantly by ICAM-3 ligation and that this binding causes augmented eosinophil secretion.

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An adhesion assay using minimal shear force to remove nonadherent cells

Cited by 29 publications

References 16 publications

Antitumorigenic effects of HIV protease inhibitor ritonavir: inhibition of Kaposi sarcoma

Antitumorigenic effects of HIV protease inhibitor ritonavir: inhibition of Kaposi sarcoma

Human Herpesvirus 8-Encoded vGPCR Activates Nuclear Factor of Activated T Cells and Collaborates with Human Immunodeficiency Virus Type 1 Tat

CD4+ T Cell and Eosinophil Adhesion Is Mediated by Specific ICAM-3 Ligation and Results in Eosinophil Activation

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