<p>Immunomodulation by <i>Lacticaseibacillus</i> sp. is a subject of increasing interest; however, the time-period of administration for its immunomodulatory effect to occur is not well established. In the present study, we examined the effect of <i>L. casei</i> CB054 on cytokine transcription ex vivo in mouse splenocytes and the bacterial microbiota profile at 24, 48, and 72&#xa0;h after oral administration of 1 × 10<sup>6</sup> UFC/g viable <i>L. casei</i>. Cytokine mRNA transcription was evaluated for <i>IL4</i>,<i> IL10</i>,<i> IL12</i>,<i> IFNɣ</i>, and <i>TNFα</i> at different time points by qPCR. Microbiota was analyzed using fecal samples collected at zero, 24, 48, and 72&#xa0;h after <i>L. casei</i> administration. A significant upregulation (<i>p</i> &lt; 0.05) for <i>IL10</i> at 72&#xa0;h and <i>IL12</i> at 48 and 72&#xa0;h in supplemented groups was observed. <i>IFNɣ</i> and <i>TNFα</i> showed similar significant upregulation patterns (<i>p</i> &lt; 0.05) at all time points evaluated, whereas <i>IL4</i> showed significant transcriptional downregulation during the supplementation period. After DNA extraction, the V3-V4 region from the 16&#xa0;S rRNA gene was sequenced, and reads were processed using the Divisive Amplicon Denoising Algorithm (DADA2). <i>Bacteroides</i> and <i>Lactobacillus</i> were the most abundant genera in the supplemented groups. At the phylum level, Actinobacteria and Bacteroidetes were reduced to 24&#xa0;h and 48&#xa0;h compared to 0&#xa0;h, whereas the phylum Firmicutes significantly increased at 72&#xa0;h compared to 0&#xa0;h. In conclusion, evidence suggests that <i>L. casei</i> CB054 has a specific, time-dependent immunomodulatory effect on splenocyte cytokine transcription, modulating the bacterial communities of the mouse gut microbiome.</p>

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Lacticaseibacillus casei CB054 short-term supplementation effect on murine splenocyte cytokine transcription and the gut bacterial microbiota

  • Vitória Sequeira Gonçalves Zorzi,
  • Jeferson Vidart Ramos,
  • Neida Lucia Conrad,
  • Renan Eugênio Araujo Piraine,
  • Francisco Denis Souza Santos,
  • Fábio Pereira Leivas Leite

摘要

Immunomodulation by Lacticaseibacillus sp. is a subject of increasing interest; however, the time-period of administration for its immunomodulatory effect to occur is not well established. In the present study, we examined the effect of L. casei CB054 on cytokine transcription ex vivo in mouse splenocytes and the bacterial microbiota profile at 24, 48, and 72 h after oral administration of 1 × 106 UFC/g viable L. casei. Cytokine mRNA transcription was evaluated for IL4, IL10, IL12, IFNɣ, and TNFα at different time points by qPCR. Microbiota was analyzed using fecal samples collected at zero, 24, 48, and 72 h after L. casei administration. A significant upregulation (p < 0.05) for IL10 at 72 h and IL12 at 48 and 72 h in supplemented groups was observed. IFNɣ and TNFα showed similar significant upregulation patterns (p < 0.05) at all time points evaluated, whereas IL4 showed significant transcriptional downregulation during the supplementation period. After DNA extraction, the V3-V4 region from the 16 S rRNA gene was sequenced, and reads were processed using the Divisive Amplicon Denoising Algorithm (DADA2). Bacteroides and Lactobacillus were the most abundant genera in the supplemented groups. At the phylum level, Actinobacteria and Bacteroidetes were reduced to 24 h and 48 h compared to 0 h, whereas the phylum Firmicutes significantly increased at 72 h compared to 0 h. In conclusion, evidence suggests that L. casei CB054 has a specific, time-dependent immunomodulatory effect on splenocyte cytokine transcription, modulating the bacterial communities of the mouse gut microbiome.