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北海道菌株的评估，该菌株是对犊牛腹泻有效的乳酸菌。

evaluation of HOKKAIDO strain, effective lactic acid bacteria for calf diarrhea.

作者信息

Ikehata Mari, Konnai Satoru, Okagawa Tomohiro, Abe Kentaro, Honma Mitsuru, Kitamura Toru, Maekawa Naoya, Suzuki Yasuhiko, Murata Shiro, Ohashi Kazuhiko

机构信息

Department of Disease Control, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

Department of Advanced Pharmaceutics, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

出版信息

Front Vet Sci. 2023 Apr 5;10:1145445. doi: 10.3389/fvets.2023.1145445. eCollection 2023.

DOI:10.3389/fvets.2023.1145445
PMID:37089407
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10113454/
Abstract

Calf diarrhea adversely affects growth and sometimes results in mortality, leading to severe economic losses to the cattle industry. Antibiotics are useful in the treatment against bacterial diarrhea, but not against viral, protozoan, and antibiotic-resistant bacterial diarrhea. Therefore, there are growing requirements for a novel control method for calf diarrhea. Probiotics have been considered promising candidates for preventive and supportive therapy for calf diarrhea for many years. A recent study has revealed that HOKKAIDO strain (Lp-HKD) reduces intestinal pathology and the severity of diarrhea in bovine rotavirus (BRV)-infected calves. Lp-HKD is known to enhance the function of human immune cells and expected to be used as probiotics for humans. Therefore, it is hypothesized that Lp-HKD modulates antiviral immune response in cattle and provide the clinical benefits in BRV-infected calves. However, the detailed mechanism of Lp-HKD-induced immunomodulation remains unknown. Thus, this study aimed to elucidate the immunomodulatory and antiviral effects of Lp-HKD in cattle. Cultivation assay of bovine peripheral blood mononuclear cells (PBMCs) showed that live and heat-killed Lp-HKD stimulates the production of interleukin-1β (IL-1β), IL-6, IL-10, and interferon-γ (IFN-γ) from PBMCs. Stimulation by heat-killed Lp-HKD yielded stronger cytokine production than stimulation by the live Lp-HKD. Additionally, CD14 monocytes were identified as major producers of IL-1β, IL-6, and IL-10 under Lp-HKD stimulation; however, IFN-γ was mainly produced from immune cells other than CD14 monocytes. Depletion of CD14 monocytes from the PBMCs cultivation strongly decreased cytokine production induced by heat-killed Lp-HKD. The inhibition of toll-like receptor (TLR) 2/4 signaling decreased IL-1β and IL-6 production induced by live Lp-HKD and IL-1β, IL-6, and IFN-γ production induced by heat-killed Lp-HKD. Furthermore, live or heat-killed Lp-HKD also activated T cells and their production of IFN-γ and tumor necrosis factor-α. Then, culture supernatants of bovine PBMCs treated with heat-killed Lp-HKD demonstrated antiviral effects against BRV . In conclusion, this study demonstrated that Lp-HKD activates the functions of bovine immune cells TLR2/4 signaling and exerts an antiviral effect against BRV through the induction of antiviral cytokines. Lp-HKD could be useful for the prevention and treatment of calf diarrhea through its immune activating effect.

摘要

犊牛腹泻会对生长产生不利影响，有时还会导致死亡，给养牛业造成严重经济损失。抗生素对细菌性腹泻的治疗有效，但对病毒性、原生动物性和抗生素耐药性细菌性腹泻无效。因此，对犊牛腹泻新的控制方法的需求日益增加。多年来，益生菌一直被认为是犊牛腹泻预防和支持性治疗的有希望的候选者。最近一项研究表明，北海道菌株（Lp-HKD）可减轻牛轮状病毒（BRV）感染犊牛的肠道病变和腹泻严重程度。已知Lp-HKD可增强人类免疫细胞的功能，并有望用作人类益生菌。因此，推测Lp-HKD可调节牛的抗病毒免疫反应，并为BRV感染的犊牛带来临床益处。然而，Lp-HKD诱导免疫调节的详细机制尚不清楚。因此，本研究旨在阐明Lp-HKD在牛体内的免疫调节和抗病毒作用。牛外周血单个核细胞（PBMCs）的培养试验表明，活的和热灭活的Lp-HKD均可刺激PBMCs产生白细胞介素-1β（IL-1β）、IL-6、IL-10和干扰素-γ（IFN-γ）。热灭活的Lp-HKD刺激产生的细胞因子比活的Lp-HKD刺激产生的更强。此外，CD14单核细胞被确定为Lp-HKD刺激下IL-1β、IL-6和IL-10的主要产生者；然而，IFN-γ主要由CD14单核细胞以外的免疫细胞产生。从PBMCs培养物中去除CD14单核细胞会强烈降低热灭活的Lp-HKD诱导的细胞因子产生。Toll样受体（TLR）2/4信号通路的抑制降低了活的Lp-HKD诱导的IL-1β和IL-6产生以及热灭活的Lp-HKD诱导的IL-1β、IL-6和IFN-γ产生。此外，活的或热灭活的Lp-HKD还激活了T细胞及其IFN-γ和肿瘤坏死因子-α的产生。然后，用热灭活的Lp-HKD处理的牛PBMCs的培养上清液对BRV表现出抗病毒作用。总之，本研究表明Lp-HKD激活牛免疫细胞的功能、TLR2/4信号通路，并通过诱导抗病毒细胞因子对BRV发挥抗病毒作用。Lp-HKD通过其免疫激活作用可能对犊牛腹泻的预防和治疗有用。

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