Abstract：
:We have developed and validated an inexpensive and equivalent method for measuring eosinophil adhesion by beta(2)-integrin to endothelial ICAM-1 using bovine serum albumin (BSA) as a surrogate for the immunoglobulin supergene. The number of adherent eosinophils on BSA or ICAM-1 coated microplates was quantified by residual eosinophil peroxidase activity. Non-stimulated eosinophils did not adhere to either BSA or ICAM-1. However, after IL-5 stimulation, either BSA or ICAM-1 caused comparable and concentration-dependent adhesion of eosinophils. Eosinophil adhesion was rapid and occurred within 15 to 30 min of incubation for either BSA or ICAM-1. Preincubation of cells with CD11b or CD18 antibody specifically decreased adhesion to either BSA or ICAM-1. IL-5, PAF and fMLP all induced adhesion of eosinophils to either BSA or ICAM-1 in a concentration-dependent manner, and the optimal IL-5, fMLP and PAF concentrations for adhesion to BSA were the same as for adhesion to ICAM-1. BSA-binding was specific for beta(2)-integrin; neither alpha-CD49d mAb directed against the alpha(4)-chain or alpha-CD29 directed against the common beta(1)-chain of VLA-4 blocked adhesion to BSA or ICAM-1 controls. The protein tyrosine kinase inhibitor, genistein, the phosphatidylinositol 3-kinase (PI-3 kinase) inhibitor, wortmanin, and mitogen-activated protein kinase kinase (MEK) inhibitor, U0126, all inhibited IL-5-induced eosinophil adhesion to either BSA or ICAM-1 comparably. These results indicate that BSA is a reliable and economical surrogate ligand for ICAM-1 adhesion to beta(2)-integrin-dependent adhesion to ICAM-1. Ligation characteristics of BSA are identical to those for soluble ICAM-1, and the assay is suitable for assessment of signal transduction pathways mediating adhesion.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Zhu X,Subbaraman R,Sano H,Jacobs B,Sano A,Boetticher E,Muñoz NM,Leff ARdoi
10.1016/s0022-1759(00)00192-7keywords:
subject
Has Abstractpub_date
2000-06-23 00:00:00pages
157-64issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(00)00192-7journal_volume
240pub_type
杂志文章abstract：:The potential value of fluorescein-conjugated human alpha 2-macroglobulin as a probe for studying macrophages in murine peritoneal exudate and spleen cell suspensions has been investigated using a fluorescence-activated cell sorter. These studies revealed that the number of alpha 2-macroglobulin-positive cells in the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90360-6
更新日期：1985-10-10 00:00:00
abstract：:Monoclonal antibodies (Mab) are the fastest growing group of biopharmaceuticals in development. For production in mammalian cells, the four polypeptide chains of the immunoglobulin diheterotetramer must be assembled prior to exit from the endoplasmic reticulum. Various recombinant Mab expression vectors have been deve...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.10.010
更新日期：2007-01-10 00:00:00
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journal_title：Journal of immunological methods
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abstract：:In this paper we provide both a theoretical and experimental analysis of the sensitivity of a DNA quantitation assay using a fluorescent chromophore which non-covalently binds dsDNA. It is well-known that the range of DNA concentrations available for fluorescence quantitation depends on the concentration of the chromo...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.09.011
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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abstract：:It is important to know the contribution of specific allergens to a complex allergenic extract and to have a dependable method to assess the effector activity of an extract specifically depleted of that allergen. We have previously shown that removal of the major peanut allergen, Ara h 2, from a crude peanut extract (...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.06.031
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90051-3
更新日期：1989-03-10 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90384-0
更新日期：1989-04-21 00:00:00
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journal_title：Journal of immunological methods
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doi：10.1016/0022-1759(80)90085-x
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90253-p
更新日期：1992-05-18 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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更新日期：1984-01-20 00:00:00
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journal_title：Journal of immunological methods
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90090-3
更新日期：1991-11-22 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90207-8
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journal_title：Journal of immunological methods
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更新日期：2001-12-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.09.022
更新日期：2008-12-31 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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更新日期：1995-06-14 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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journal_title：Journal of immunological methods
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pub_type： 信件
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90359-n
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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更新日期：1981-01-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90334-8
更新日期：1980-01-01 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90223-6
更新日期：1985-09-03 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90421-8
更新日期：1988-04-06 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
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更新日期：1985-05-10 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90468-b
更新日期：1990-03-27 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90153-9
更新日期：1978-01-01 00:00:00
abstract：:This report describes the use of biotin-labeled antibodies in sensitive enzyme immunoassay systems for the measurement of bacterial antigens. Biotinylated immunoglobulins could be reproducibly formulated using N-hydroxysuccinimide biotin ester to link biotin to the immunoglobulin. Binding of the biotinylated antibody ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(83)80021-0
更新日期：1983-02-11 00:00:00
