, Volume 7, Issue 4, pp 363-375

Demonstration of specific receptors for fluoresceinated casein on human neutrophils and monocytes using flow cytometry

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Abstract

-Casein is chemotactic for human neutrophils (PMNs) and monocytes. The binding of fluorescein (FITC) -conjugated casein (mixture ofα,β,and κ- casein) and purifiedα-casein to PMNs, monocytes, and lymphocytes was analyzed using flow cytometry. These studies demonstrate that 75–95% of PMNs and 46–85% of monocytes have membrane receptors for casein while lymphocytes lack these receptors. The binding of FITC-casein and FITC-α-casein was specific and was blocked only by unlabeled casein andα-casein, but not by ovalbumin, bovine or human serum albumin,β-casomorphin, C5a, or formyl-methionyl-leucylphenylalanine (fMLP). The binding of FITC-casein was reversible when PMNs were stained with this fluorescent agent and subsequently incubated with unlabeled casein. Double-labeling studies of mononuclear cells using FITC-casein and the OKM1 monoclonal antibody in conjunction with a rhodamine conjugated anti-Ig second antibody demonstrate that mononuclear cells binding FITC-casein also stain with the OKM1 monoclonal antibody, indicating a specificity for monocytes.

Supported in part by USDHHS grant CA20819 from the National Cancer Institute, and NU-05459 from the Division of Nursing.
Recipient of Senior Investigator Award from the National Arthritis Foundation.