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Á¦¸ñ ±â¸® ¹Ú»ç ºê¸¯¼Ò°³-doi:10.1016/j.fsi.2015.10.033
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ÀÛ¼ºÀÏÀÚ 2020-04-29
Effects of intracellular products of Bacillus subtilis VSG1 and Lactobacillus plantarum VSG3 on cytokine responses in the head kidney macrophages of Labeo rohita
Sib Sankar Giria, c, Shib Sankar Senb, Cheng Chia, Hyoun Joong Kima, Saekil Yuna, Se Chang Parka, V. Sukumaranc
a Laboratory of Aquatic Biomedicine, College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul-151742, South Korea
b School of life Sciences, Jawaharlal Nehru University, New Delhi-110067, India
c Dept. of Biotechnology, Periyar Maniammai University, Thanjavur-613403, Tamil Nadu, India
Corresponding author : Se Chang Park, V. Sukumaran
Abstract
The efficiency of intracellular products (ICPs) of the probiotics Bacillus subtilis VSG1 and Lactobacillus plantarum VSG3 in stimulating cytokine responses in the head kidney (HK) macrophages of Labeo rohita was investigated. The HK macrophages were incubated with ICPs and lipopolysaccharide (LPS), and the responses of cytokine genes, namely interleukin-10 (IL-10), IL-1¥â, IL-12p35, IL-12p40, IL-18, tumour necrosis factor-¥á (TNF-¥á), nuclear factor kappa B (NF-¥êB), cyclo-oxygenase-2 (COX-2), interferon-1 (IFN-1), and IFN-¥ã were assessed by quantitative real-time PCR (qRT-PCR) at 2, 6, 12, 24, and 48 hours post-stimulation (hps). Among the proinflammatory cytokines, a strong increase in the gene ________expression________ of IL-1¥â and TNF-¥á was displayed mainly at 2-6 hps with ICPs, as compared to that of the positive control (LPS) or the negative control (PBS) (P < 0.05). However, COX-2 and NF-¥êB showed higher ________expression________ at 2 and 24 hps, and 6 - 24 hps with ICPs, respectively. Antiviral cytokines IFN-1 and IFN-¥ã displayed strong ________expression________s (P < 0.05) at 6 - 12 hps, and 12-24 hps with ICPs, respectively. Upregulation of the anti-inflammatory cytokine, IL-10, was recorded at 6-24 hps with ICPs, as compared to that controls. ________Expression________s of cell-mediated immune factor genes (IL-12p35, IL-12p40, and IL-18) were also significantly upregulated at different time points, except 48 hps, in HK macrophages stimulated with ICPs. Furthermore, enhanced cellular (phagocytic activity and nitroblue tetrazolium assay) and humoral (lysozyme) immune parameters in stimulated cells confirmed the induction of the inflammatory response. Therefore, the results of this in vitro study indicate that the ICPs of B. subtilis VSG1 or L. plantarum VSG3 have great potential for stimulating the cytokine responses in fish, and are thereby potential immunostimulants to fish. Further studies could be conducted to explore its suitability as an adjuvant vaccine in aquaculture.
Keywords : Intracellular products; Bacillus subtilis VSG1; Lactobacillus plantarum VSG3; Head kidney macrophages; Cytokine responses; Labeo rohita
 
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