Abstract
The aim of the study was to evaluate the effect of methyl donor supplementation on mRNA expression of galectin genes in cow and calf neutrophils in vitro. Galectins play a crucial role in homeostasis and disease regulation. Ten adult Holstein cows and five neonatal Holstein calves (3 wk. old) were used for isolation and in vitro culture of neutrophils. Neutrophils isolated from the adult cows were divided into two groups. Neutrophils from Group 1 were incubated with three concentrations of Lysine (Lys) to Met ratios of (3.6:1, 2.9:1, or 2.4:1). Neutrophils from Group 2 were also incubated with three concentrations of Lys to Met ratios and 50 μg /mL of LPS. Isolated neutrophils from calves were incubated with Met, Choline, and Taurine (37 °C, with 5% CO2 or 4 hours). Met did not affect LGALS expression in Group 1 cows. In Group 2, however, relative mRNA expression of LGALS9 was higher in neutrophils treated with 3.6 Lys: Met and 2.9 Lys: Met concentrations compared to 2.4 Lys: Met (0.33, 0.36 and 0.16 respectively) (P<0.05). A Met X LPS interaction was also observed in regard to the expression of LGALS8. LGALS8 transcription decreased when neutrophils were treated with LPS at 3.6 Lys: Met (No LPS=0.89 LPS= 0.56) (P<0.05). On the contrary, LGALS8 expression increased at 2.9 Lys: Met in the presence of LPS (No LPS=0.52 LPS= 1.95)(P<0.05). Treatment with LPS alone increased the expression of LGALS4 (P<0.05) and tended to increase LGALS12 (P<0.10) compared to control. In calves, LGALS-2, -7, and -9 were undetected. LGALS-1, -3, - 4, -8, and -12 were all reduced in neutrophils treated with 2.4 and 2.6 Lys: Met compared to 3.6 Lys: Met. Increasing Choline concentration tended to reduce LGALS1 (P<0.05) but reduced LGALS8 (P<0.05). Increasing taurine reduced LGALS1 and tended to reduce LGALS12. These results shed light on the modulation of galectin expression in cow and calf neutrophils in response to methyl donors.
Highlights
An animal with a properly functioning immune system is protected from a variety of pathogenic organisms, including viruses, bacteria, and parasites (Kawai and Akira 2010)
Because galectin expression is closely linked to immune-modulation and inflammation (Ekwemalor, Asiamah et al 2017, Asiamah, Adjei-Fremah et al 2018), it is imperative to explore whether galectin gene expression in neutrophils is affected with Met supplementation in cows and calves
We hypothesized that supplementation with methionine, taurine, or choline would benefit the innate immune response of cows and calves at least in part by altering the expression of galectin genes associated with neutrophils activity and inflammation
Summary
An animal with a properly functioning immune system is protected from a variety of pathogenic organisms, including viruses, bacteria, and parasites (Kawai and Akira 2010). Numerous studies focused on elucidating the molecular mechanisms leading to metabolic and infectious diseases are emerging(Adjei-Fremah, Ekwemalor et al 2016, Adjei-Fremah, Jackai et al 2016, Asiamah, Adjei-Fremah et al 2016, Asiamah, Adjei-Fremah et al 2016). Our understanding of this phenomenon will lead to the design of effective management practices in order to reduce cow and calf health disorders. We hypothesized that supplementation with methionine, taurine, or choline would benefit the innate immune response of cows and calves at least in part by altering the expression of galectin genes associated with neutrophils activity and inflammation. The effect of Met, CHO, and TAU supplementation in vitro on the mRNA expression of galectin genes in neutrophils isolated from neonatal calves was determined
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