Abstract
Galectins (GAL) are β-galactoside binding proteins. The objective of this study was to assess the effect of PMA on the regulation of galectin gene expression in bovine blood and neutrophils. Blood was collected from 3 clinically healthy Holstein cows. Neutrophils were isolated by differential centrifugation. Blood and neutrophils were treated with 10ng/ml of PMA or maintained in phosphate-buffered saline for 30 minutes at 370 C. Pooled total RNA were transcribed to cDNA for real-time PCR. Specific primers for bovine LGALS -1, -2, -3, -4, -7, -8, -9, -11, -12, and their binding proteins (bp) LGALS3bp, T cell immunoglobulin and mucin domain 3 (TIM-3) were used. Housekeeping genes RPLP0 and UCHL5 served as internal controls. Fold changes were calculated using the Livak method. Total protein concentration was assessed using the Bicinchoninic acid assay. Secretion of GAL -1, -2, -3, -4, -8, -9 was assessed using bovine GAL specific ELISA. All tested LGALS were detected. Activation with PMA differentially modulated expression and secretion of GAL in blood and neutrophils. Expression levels of LGALS -1 and LGALS -3 was not affected. Expression of LGALS -7, -8, -9, -11, -12, and TIM-3 was differentially modulated. The secretion of GAL -3 was significantly increased in response to PMA. The most pronounced effect of PMA was observed on LGALS-9, TIM-3 expression, and the secretion of GAL -3. Thus, GAL may serve as biomarkers of cell activation with functional consequences and warrant further study.
Highlights
Lectin Galactoside-Binding Soluble (LGALS) gene codes for several galectins (GAL)
The objective of this study was to assess the effect of Phorbol 12-myristate 13-acetate (PMA) on the regulation of galectin gene expression in bovine blood and neutrophils
Galectins are known to act as pattern recognition receptors and Pathogen Associated Molecular Patterns (PAMPs) that regulate immune responses according to the level of pathogenicity of the invading microorganisms
Summary
Galectins are a family of animal lectins that typically bind β-galactoside carbohydrates with high specificity (Modenutti, Capurro, Di Lella, & Martí, 2019). They are ubiquitously distributed across the mammalian tissues, including the innate and adaptive cells (Stowell et al, 2008), and their differential regulation is essential for maintaining cellular functions (Laderach et al, 2012). Galectins are known to act as pattern recognition receptors and Pathogen Associated Molecular Patterns (PAMPs) that regulate immune responses according to the level of pathogenicity of the invading microorganisms (van Kooyk and Rabinovich, 2008, Sato et al, 2009) Their binding proteins, such as Galectin-3 binding protein (LGALS3bp), T cell Immunoglobulin, and Mucin-domain-containing- protein 3 (TIM-3) play an important role in health and homeostasis. Recent studies have shown that regulation of LGALS gene expression and secretion is part of an effective immune system needed to combat infection and control disease
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