Antibody markers of basal cells in complex epithelia

Abstract

In the course of immunohistochemical studies it has become apparent that there is a distinct phenotype of keratin expression that is shared by basal epithelial cells in a variety of different tissues. A basal cell can be defined as a cell in contact with a basal lamina but with no free luminal surface; this distinguishes it from a simple epithelial cell, which has a free luminal surface as well as basal lamina contact, and from stratifying suprabasal keratinocytes, which have neither basal lamina contact nor free luminal surface. All basal cells, whether they are in glandular ductal or secretory epithelia, or in stratified squamous epithelia, express the keratin pair K5 and K14. In this paper we describe monoclonal and polyclonal antibodies that are monospecific for both keratins 14 and 5 or are specific for denaturation-sensitive epitopes unique to basal cells, including five new monoclonal antibodies: LL001 and LL002 (to keratin 14), 2.1.D7 (to keratins 5, 6 and 8), and LH6 and LH8 (conformation-specific basal cell markers). These antibodies have been used to monitor the distribution of the basal cell phenotype and to demonstrate the expression of keratins 5 and 14 in this cell type, in both stratified epithelia and mixed epithelial glands. The consistent association of this keratin pair with basal cells suggests a possible specific function for these keratin in reinforcing epithelia under physical stress, whilst expression of these keratins may conflict with the differentiated functions of most simple epithelial cells.

Detrimental Effects of IFN-γ on an Epidermolysis Bullosa Simplex Cell Model and Protection by a Humanized Anti–IFN-γ Monoclonal Antibody

“…Other antibodies used were as follows: mouse mAbs used were anti-K14 ( Purkis et al, 1990 ), anti-desmoplakin (11-5F) (gifted by David Garrod, The University of Manchester, Manchester, United Kingdom), anti–β-actin (clone AC-15, number A5441, Sigma-Aldrich, St. Louis, MO), anti–β-catenin (number ab16051, Abcam, Cambridge, United Kingdom; clone 14, number 610153, BD Biosciences, San Jose, CA), and anti–keratin 17 (number NCL-L-CK17, Leica Biosystems, Richmond, IL); rabbit mAbs or polyclonal antibodies used were anti–phospho-STAT1 (Tyr701) (clone D4A7, number 7649, Cell Signaling Technology, Danvers, MA), anti-STAT1 (clone D1K9Y, number 14994, Cell Signaling Technology), and anti-phospho-Histone H3 (Ser10) Antibody (clone MC463, number 04-817, Merck Millipore, Burlington, MA); secondary antibodies used were goat anti-rabbit IgG (H+L) Alexa Fluor 594 (number A-11037, Thermo Fisher Scientific); goat anti-mouse IgG (H+L) Alexa Fluor 568 (number A-11031, Thermo Fisher Scientific); Anti-Mouse IgG (H+L), HRP Conjugate (number W4021, Promega, Madison, WI); and Anti-Rabbit IgG (H+L), HRP Conjugate (number W4011, Promega); and antibody to human kappa light chain conjugated to horseradish peroxidase was used in ELISA (kindly provided by Prof PG Sveshnikov, Russian Research Center for Molecular Diagnostics and Treatment, Moscow, Russia).…”

A cell-based drug discovery assay identifies inhibition of cell stress responses as a new approach to treatment of epidermolysis bullosa simplex

“…The following mouse monoclonal antibodies were used as primary antibodies: anti-phospho-ERK1/2 (Thr202/Tyr204) (clone E10, #9106; Cell Signaling Technologies, MA, USA), anti-β-actin (clone AC-15, #A5441; Sigma-Aldrich Corporation, MO, USA), anti-desmoplakin (clone 11-5F; gift from David Garrod, University of Manchester, UK), anti-keratin 14 (clone LL001; Purkis et al, 1990 ), anti-keratin 10 (clone DE-K10, #M7002; Agilent Technologies, CA, USA), anti-E-cadherin (clone 36, #610182; BD Biosciences, CA, USA) and anti-Ki-67 (clone MM1, #NCL-L-Ki67-MM1; Leica Biosystems, IL, USA). The following rabbit monoclonal or polyclonal antibodies were also used as primary antibodies: anti-phospho-AKT (Ser473) (clone D9E, #4060), anti-AKT (clone C67E7, #4691), anti-phospho-ERK1/2 (Thr202/Tyr204) (clone D13.14.4E, #4370), anti-ERK1/2 (clone 137F5, #4695) and anti-YAP (clone D8H1X, #14074) were purchased from Cell Signaling Technologies (MA, USA).…”

Airway Basal Cells, Protectors of Epithelial Walls in Health and Respiratory Diseases

“…Basal cells are epithelial progenitor cells and are defined by the expression of transformation-related protein (TP63) and cytokeratins 5 and 14 (KRT5/14) ( 29 , 30 ). Beside these classical markers, Zhao et al demonstrated that YES-associated protein 1 (YAP) is essential for the maintenance of airway basal cell identity and that YAP is closely related to TP63 to regulate stem cardinal behavior and to help determine epithelial architecture ( 31 ).…”