Abstract
Galectins (Gals) are a family of animal lectins that bind β-galactosides through a carbohydrate recognition domain. Galectin-8 is a tandem-repeat galectin, secreted intracellularly and extracellularly. It is associated with neutrophil migration and has been studied as a possible therapeutic to combat inflammation. The objective of this study was to evaluate the translational and the transcriptional effects of recombinant Galectin-8 (rGal-8) on cow neutrophils. Blood was collected aseptically from Holstein-Friesian cows (n=10) from the North Carolina A&T State University Dairy Unit. Neutrophils isolated were treated with rGal-8 (2μg), or PBS (control) and were incubated at 37°C, 5% CO2 for 1 hour. Supernatant from treated neutrophils was evaluated for total protein concentration, and galectin-8 secretion using bovine Galectin-8 Enzyme Linked-Immuno-Sorbent Assay (ELISA) kit. Total RNA was extracted, reverse transcribed, and RT-qPCR was performed using the RT² Profiler Cow Innate & Adaptive Immune Responses Array with 84 genes. The Livak method was used to calculate transcript abundance and fold change (FC>2 considered significant). Total protein concentration increased (P=0.0361) after rGal-8 treatment compared to the untreated control. Galectin-8 secretion was not significantly different in control compared to treated group (P=0.5819). Out of the 84 genes, 81 genes were differentially expressed in response to rGal-8; 14 up-regulated, 5 down-regulated, 61 genes remained unchanged. Treatment with rGal8 induced the expression of IRF7. The top five up-regulated genes include FAS, CD40, CD86, IFNGR1, STAT1; down-regulated genes were TLR9, CD14, CCR6, TICAM1, and TLR1. Selected genes were probed to validate fold change; the levels of gene expression were comparable to data from RT2 array. Exposure of bovine neutrophils to rGal-8 modified expression of immune response genes. The functional significance of the change needs further studies.
Highlights
The innate immune system, primarily leukocytes, serve as the first line of host defense and plays a crucial role in early recognition and pro-inflammatory response (Medzhitov & Janeway, 2000)
Treatment of bovine neutrophils with exogenous Galectin-8 had no effect on the neutrophil count and viability (Table 2)
Our current study investigated the effects of recombinant Galectin8 on secretion and expression of innate and adaptive immune response genes in bovine neutrophils
Summary
The innate immune system, primarily leukocytes, serve as the first line of host defense and plays a crucial role in early recognition and pro-inflammatory response (Medzhitov & Janeway, 2000). Numerous receptors are present on the neutrophil surface, some of these receptors detect chemoattractants that allow neutrophils to migrate toward areas of inflammation (Paape et al, 2003). Bovine neutrophils are comparable to neutrophils of other species in their functionality, activation mechanisms, diapedesis, and modulation of immune response (Bassel & Caswell, 2018). The balance between microbial killing and host tissue damage is linked to neutrophil biochemical composition which is species specific (Bassel & Caswell, 2018). Galectin-8 acts as an extracellular matrix (ECM) protein that positively or negatively regulates cell adhesion, depending on the extracellular context and cell surface jmbr.ccsenet.org
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
