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A Novel Murine Anti-Lactoferrin Monoclonal Antibody Activates Human Polymorphonuclear Leukocytes through Membrane-Bound Lactoferrin and TLR4.
- Source :
-
BioMed Research International . 11/15/2015, Vol. 2015, p1-11. 11p. - Publication Year :
- 2015
-
Abstract
- Soluble lactoferrin (LTF) is a versatile molecule that not only regulates the iron homeostasis, but also harbors direct microbicidal and immunomodulating abilities in mammalian body fluids. In contrast, little is known about the function of membrane-bound LTF (mbLTF), although its expression on human polymorphonuclear leukocytes (huPMNs) has been reported for decades. Given that LTF/anti-LTF antibodies represent a potential diagnostic/prognostic biomarker and a therapeutic target in patients with immune disorders, we wished, in the present study, to generate a novel human LTF- (huLTF-) specific mAb suitable for detailed analyses on the expression and function of mbLTF as well as for deciphering the underlying mechanisms. By using the traditional hybridoma cell fusion technology, we obtained a murine IgG1 (kappa) mAb, M-860, against huLTF. M-860 recognizes a conformational epitope of huLTF as it binds to natural, but not denatured, huLTF in ELISA. Moreover, M-860 detects mbLTF by FACS and captures endogenous huLTF in total cell lysates of huPMNs. Functionally, M-860 induces the activation of huPMNs partially through TLR4 but independently of phagocytosis. M-860 is thus a powerful tool to analyze the expression and function of human mbLTF, which will further our understanding of the roles of LTF in health and disease. [ABSTRACT FROM AUTHOR]
- Subjects :
- *REACTIVE oxygen species
*ANIMAL experimentation
*CELL culture
*ELECTROPHORESIS
*ENZYME-linked immunosorbent assay
*FERRITIN
*FLOW cytometry
*GENETIC techniques
*IMMUNOLOGIC diseases
*MEMBRANE proteins
*MICE
*MICROSCOPY
*MONOCLONAL antibodies
*NEUTROPHILS
*POLYMERASE chain reaction
*PROBABILITY theory
*RESEARCH funding
*WESTERN immunoblotting
*DATA analysis software
*TOLL-like receptors
*DESCRIPTIVE statistics
*PRECIPITIN tests
*MANN Whitney U Test
Subjects
Details
- Language :
- English
- ISSN :
- 23146133
- Volume :
- 2015
- Database :
- Academic Search Index
- Journal :
- BioMed Research International
- Publication Type :
- Academic Journal
- Accession number :
- 113630053
- Full Text :
- https://doi.org/10.1155/2015/285237