Effect of Lactobacillus rhamnosus on the development of B cells in gut-associated lymphoid tissue of BALB/c mice
Effect of Lactobacillus rhamnosus on the development of B cells in gut-associated lymphoid tissue of BALB/c mice
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鼠李糖乳杆菌对 BALB/c 小鼠肠道相关淋巴组织 B 细胞发育的影响
DOI：
10.1111/jcmm.15574
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发表时间：
2020
影响因子：
5.3
通讯作者：
Wang Chun-feng
中科院分区：
文献类型：
--
作者：
Shi Chun-wei;Zeng Yan;Yang Gui-lian;Jiang Yan-long;Yang Wen-tao;Chen Yi-qiu;Wang Jing-ying;Wang Jian-zhong;Kang Yuan-huan;Huang Hai-bin;Ye Li-ping;Cao Xin;Wang Chun-feng
亲爱的编辑，乳酸菌(LAB)附着在胃肠道内表面，通过抗原提呈细胞调节粘膜和全身免疫反应。1在免疫球蛋白和细胞因子的刺激下，LAB还可以影响相关免疫反应的调节。2乳酸菌可抑制巨噬细胞产生IL-12和转录IL-12p40mRNA。3研究表明LAB可诱导全身抗炎细胞因子的产生，如白介素10(IL-10)。实验室产生的可溶性因子抑制促炎细胞因子的产生。4-6口腔实验室作用于胃肠道粘膜，至少70%的免疫细胞定植于肠道相关淋巴组织(GALT)。7虽然骨髓(BM)是哺乳动物B细胞发生的主要淋巴器官，但GALT也被认为是不同物种B细胞发育的主要淋巴器官。8、9此外，微生物区系对哺乳动物GLAT中B细胞的发育起着至关重要的作用。9、10然而，LAB对GALT中B细胞发育和功能的影响还有待进一步剖析。将50只1周龄BALB/c小鼠随机分为两组，即PBS对照组和鼠李糖乳杆菌(LGG)组，每组25只。L以107cfu/10μ剂量隔日灌胃给药，连续2周，以硫代巴比妥钠染毒小鼠为对照组。分别于给药后7、14、21、28、35d处死小鼠进行分析(每个时间点n=5)。首先，用流式细胞仪检测了小鼠骨髓、肠道固有层和Peyer氏斑中B细胞的发育阶段，即B220+CD43+−IGD−(PRO-B)、B220+CD43−IgM−IGD−(Pre-B)、B220+CD43−IgM+IGD−(未成熟B)和B220+CD43−IgM+IGD+(成熟B)。对小鼠脾(SPL)和肠系膜淋巴结(MLN)的成熟B细胞进行了分析。其次，检测小鼠SPL、MLN和PPS中B细胞表面CD40、CD80和MHC-Ⅱ的表达。用酶联免疫吸附试验检测肠道灌洗液中分泌型免疫球蛋白A(SIgA)和血清免疫球蛋白IgM、IgA及免疫球蛋白G(Ig G)水平。图1A显示了B细胞不同发育阶段的门控策略。LGG干预后第7天，LGG组骨髓中前B细胞百分率明显低于对照组，分别为1.25±0.17%和2.28±0.18%(n=5；P<；0.01，P=0.0033)。LGG干预后第35天，LGG组骨髓中前B细胞和未成熟B细胞的比例明显低于对照组，前B细胞分别为42.56±6.34%和64.64±0.89%(P=0.0087)，未成熟B细胞分别为11.88±3.97%和28.46±0.83%(n=5；P<；0.01，P=0.0017)。而在干预后第35天，LGG组骨髓中成熟B细胞的百分比显著高于对照组，分别为21.71±4.19%和5.15±0.23%(n=5；P<；0.01，P=0.0043)。B细胞在骨髓中发育的代表性数据如图S1a-E所示。随着干预的停止，LGG组前B细胞和未成熟B细胞的比例明显下降，成熟B细胞的数量显著增加，提示LGG促进BM中前B细胞向成熟B细胞的发育。在新发现的小鼠B细胞发育的初级淋巴器官--肠道的LPL中也得到了类似的结果。在IGG干预后第21天和第28天，IGG组未成熟B细胞在…中的百分比显著下降
Dear Editor, Lactic acid bacteria (LAB) adhere to the inner surface of gastrointestinal tract and regulate mucosal and systemic immune response through antigen-presenting cells. 1 Under the stimulation of immunoglobulin and cytokines, LAB can also affect the regulation of related immune response. 2 Lactic acid bacteria can inhibit the production of IL-12 and transcription of IL-12p40 mRNA by macrophages. 3 Studies have shown that LAB can induce the production of systemic anti-inflammatory cytokines, such as interleukin-10 (IL-10). Soluble factors produced by LAB inhibit the production of pro-inflammatory cytokines. 4-6 Oral LAB act on mucosa of gastrointestinal tract, and at least 70% of immune cells are colonized in gut-associated lymphoid tissue (GALT). 7 Although bone marrow (BM) is the major primary lymphoid organ of B lymphogenesis for mammals, GALT is also identified as the primary lymphoid organ for B cell development in different species. 8, 9 Besides, microbiota play an essential role for B cell development in mammal GLAT. 9, 10 However, the effect of LAB on the development and function of B cells in GALT needs to be further dissected. In this study, fifty 1-week-old BALB/c mice were randomly divided into two groups, namely the PBS control group and Lactobacillus rhamnosus (LGG) group with 25 mice per group. Mice were orally administrated with LGG at the dose of 107 cfu/10 μL every other day for 2 weeks, and mice treated with PBS were used as control. At 7, 14, 21, 28 and 35 days after the treatment, mice were killed for analysing (n= 5 for each time-point). Firstly, developmental stages of B cells, that is B220+CD43+IgM− IgD−(pro-B), B220+CD43− IgM− IgD−(pre-B), B220+CD43− IgM+IgD−(immature B) and B220+CD43− IgM+IgD+(mature B), were detected in mouse BM, intestinal lamina propria (LPL) and Peyer's patches (PPs) by flow cytometry. Mature B cells were analysed in mouse spleen (SPL) and mesenteric lymph nodes (MLN). Secondly, the expression levels of CD40, CD80 and MHC-Ⅱ on B cells were detected in mouse SPL, MLN and PPs. Lastly, we examined the Secretory Immunoglobulin A (SIgA) level in intestinal lavage fluid and serum IgM, IgA and Immunoglobulin G (IgG) by ELISA. Figure 1A shows the gating strategy for different developmental stages of B cells. On the 7th day after LGG intervention, the percentage of pro–B cells in BM of LGG group was significantly lower than that of control group, with 1.25±0.17% vs 2.28±0.18%(n= 5; P< 0.01, P= 0.0033). On the 35th day after LGG intervention, the percentage of pre–B and immature B cells in BM of LGG group decreased significantly compared to control group, with 42.56±6.34% vs 64.64±0.89% for Pre–B cells (n= 5; P< 0.01, P= 0.0087) and 11.88±3.97% vs 28.46±0.83% for immature B cells (n= 5; P< 0.01, P= 0.0017), respectively. However, the percentage of mature B cells in BM of LGG group increased significantly compared to control group on the 35th day after LGG intervention, with 21.71±4.19% vs 5.15±0.23%(n= 5; P< 0.01, P= 0.0043)(Figure 1B). The representative data of B cell development in BM for flow cytometry were shown in Figure S1A-E. With the cessation of intervention, the percentage of pre–B and immature B cells decreased significantly, while the number of mature B cells increased dramatically in LGG group, indicating that LGG promotes the development of pro-B to mature B in BM. Similar results were obtained in LPL of intestine, a newly identified primary lymphoid organ for B cell development in mice. On the 21st and 28th days after LGG intervention, the percentage of immature B cells in LPL of LGG group decreased significantly …