Neumann E, Ramos M G, Santos L M, Rodrigues A C P, Vieira E C, Afonso L C C, Nicoli J R, Vieira L Q
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil.
Braz J Med Biol Res. 2009 Apr;42(4):358-67. doi: 10.1590/s0100-879x2009000400008.
Lactobacillus delbrueckii UFV-H2b20 has been shown to increase clearance of bacteria injected into the blood of germ-free mice. Moreover, it induces the production of type 1 cytokines by human peripheral mononuclear cells. The objective of the present study was to investigate the production of inflammatory cytokines [interleukin-12 (IL-12 p40), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma)] triggered in vitro by live, heat-killed or lysozyme-treated L. delbrueckii UFV-H2b20 and in vivo by a live preparation. Germ-free, L. delbrueckii-monoassociated and lipopolysaccharide (LPS)-resistant C3H/HeJ mice were used as experimental models. UFV-H2b20 induced the production of IL-12 p40 and TNF-alpha by peritoneal cells and IFN-gamma by spleen cells from germ-free or monoassociated Swiss/NIH mice and LPS-hyporesponsive mice (around 40 ng/mL for IL-12 p40, 200 pg/mL for TNF-alpha and 10 ng/mL for IFN-gamma). Heat treatment of L. delbrueckii did not affect the production of these cytokines. Lysozyme treatment decreased IL-12 p40 production by peritoneal cells from C3H/HeJ mice, but did not affect TNF-alpha production by these cells or IFN-gamma production by spleen cells from the same mouse strain. TNF-alpha production by peritoneal cells from Swiss/NIH L. delbrueckii-monoassociated mice was inhibited by lysozyme treatment. When testing IL-12 p40 and IFN-gamma levels in sera from germ-free or monoassociated Swiss/NIH mice systemically challenged with Escherichia coli we observed that IL-12 p40 was produced at marginally higher levels by monoassociated mice than by germ-free mice (40 vs 60 ng/mL), but IFN-gamma was produced earlier and at higher levels by monoassociated mice (monoassociated 4 and 14 ng/mL 4 and 8 h after infection, germfree 0 and 7.5 ng/mL at the same times). These results show that L. delbrueckii UFV-H2b20 stimulates the production of type 1 cytokines in vitro and in vivo, therefore suggesting that L. delbrueckii might have adjuvant properties in infection in which these cytokines play a major role.
德氏乳杆菌UFV-H2b20已被证明可提高无菌小鼠血液中注入细菌的清除率。此外，它还能诱导人外周血单个核细胞产生1型细胞因子。本研究的目的是调查活的、热灭活的或经溶菌酶处理的德氏乳杆菌UFV-H2b20在体外以及活体制剂在体内引发的炎性细胞因子[白细胞介素-12（IL-12 p40）、肿瘤坏死因子-α（TNF-α）和干扰素-γ（IFN-γ）]的产生情况。无菌、德氏乳杆菌单联和耐脂多糖（LPS）的C3H/HeJ小鼠用作实验模型。UFV-H2b20可诱导无菌或单联瑞士/国立卫生研究院小鼠以及LPS低反应性小鼠的腹膜细胞产生IL-12 p40和TNF-α，以及脾细胞产生IFN-γ（IL-12 p40约为40 ng/mL，TNF-α为200 pg/mL，IFN-γ为10 ng/mL）。德氏乳杆菌经热处理不影响这些细胞因子的产生。溶菌酶处理可降低C3H/HeJ小鼠腹膜细胞的IL-12 p40产生，但不影响这些细胞的TNF-α产生或同一小鼠品系脾细胞的IFN-γ产生。溶菌酶处理可抑制瑞士/国立卫生研究院德氏乳杆菌单联小鼠腹膜细胞的TNF-α产生。当检测无菌或单联瑞士/国立卫生研究院小鼠经大肠杆菌全身攻击后血清中的IL-12 p40和IFN-γ水平时，我们观察到单联小鼠产生的IL-12 p40水平略高于无菌小鼠（40对60 ng/mL），但单联小鼠产生IFN-γ的时间更早且水平更高（感染后单联小鼠在4和8小时时分别为4和14 ng/mL，无菌小鼠在相同时间为0和7.5 ng/mL）。这些结果表明，德氏乳杆菌UFV-H2b20在体外和体内均可刺激1型细胞因子的产生，因此提示德氏乳杆菌在这些细胞因子起主要作用的感染中可能具有佐剂特性。
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