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. 2024 May 18;25(10):5514.
doi: 10.3390/ijms25105514.

Bifidobacterium longum and Chlorella sorokiniana Combination Modulates IFN-γ, IL-10, and SOCS3 in Rotavirus-Infected Cells

Felizardo Velderrain-Armenta  1 , Guadalupe González-Ochoa  1 , Patricia Tamez-Guerra  2 , Ricardo Romero-Arguelles  2 , César I Romo-Sáenz  2 , Ricardo Gomez-Flores  2 , Lilian Flores-Mendoza  1 , Ramona Icedo-García  1 , José G Soñanez-Organis  1
Affiliations

Bifidobacterium longum and Chlorella sorokiniana Combination Modulates IFN-γ, IL-10, and SOCS3 in Rotavirus-Infected Cells

Felizardo Velderrain-Armenta et al. Int J Mol Sci. .

Abstract

Rotavirus is the main cause of acute diarrhea in children up to five years of age. In this regard, probiotics are commonly used to treat or prevent gastroenteritis including viral infections. The anti-rotavirus effect of Bifidobacterium longum and Chlorella sorokiniana, by reducing viral infectivity and improving IFN-type I response, has been previously reported. The present study aimed to study the effect of B. longum and/or C. sorokiniana on modulating the antiviral cellular immune response mediated by IFN-γ, IL-10, SOCS3, STAT1, and STAT2 genes in rotavirus-infected cells. To determine the mRNA relative expression of these genes, HT-29 cells were treated with B. longum and C. sorokiniana alone or in combination, followed by rotavirus infection. In addition, infected cells were treated with B. longum and/or C. sorokiniana. Cellular RNA was purified, used for cDNA synthesis, and amplified by qPCR. Our results demonstrated that the combination of B. longum and C. sorokiniana stimulates the antiviral cellular immune response by upregulating IFN-γ and may block pro-inflammatory cytokines by upregulating IL-10 and SOCS3. The results of our study indicated that B. longum, C. sorokiniana, or their combination improve antiviral cellular immune response and might modulate pro-inflammatory responses.

Keywords: Bifidobacterium; Chlorella; gastroenteritis; rotavirus.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Rotavirus (RV)-infected cell assays. Relative expression level of SOCS3, IFN-γ, STAT1, STAT2, and IL-10 genes in HT-29 cells infected with rotavirus using uninfected HT-29 as the control. Data were analyzed by ANOVA with subsequent Tukey test using GraphPad Prism 10. Different letters indicate statistical significance between treatments. p values < 0.05 were considered statistically significant.
Figure 2
Figure 2
B. longum and rotavirus (RV) assays. The relative expression levels of SOCS3, IFN-γ, STAT1, STAT2, and IL-10 genes in HT-29 cells treated with B. longum before (pre-infection) and after (post-infection) rotavirus infection were analyzed by ANOVA with subsequent Tukey test using GraphPad Prism 10. Different letters indicate statistical significance between treatments. p values < 0.05 were considered statistically significant.
Figure 3
Figure 3
C. sorokiniana and rotavirus (RV) assays. The relative expression level of SOCS3, IFN-γ, STAT1, STAT2, and IL-10 genes in HT-29 cells treated with B. longum before (pre-infection) and after (post-infection) rotavirus infection. Data were analyzed by ANOVA with subsequent Tukey test using GraphPad Prism 10. Different letters indicate statistical significance between treatments. p values < 0.05 were considered statistically significant.
Figure 4
Figure 4
B. longum in combination with C. sorokiniana and rotavirus (RV) assays. The relative expression level of SOCS3, IFN-γ, STAT1, STAT2, and IL-10 genes in HT-29 cells treated with B. longum and C. sorokiniana before (pre-infection) and after (post-infection) rotavirus infection. Data were analyzed by ANOVA with subsequent Tukey test using GraphPad Prism 10. Different letters indicate statistical significance between treatments. p values < 0.05 were considered statistically significant.
Figure 5
Figure 5
Heat map showing the mRNA expression of SOCS3, IFN-γ, STAT-1, STAT-2, and IL-10 in B. longum pre-infected with rotavirus (pre-RV): HT-29 cells infected with RV and treated with B. longum (Bl); B. longum post-infected with RV (post-RV): HT-29 cells treated with Bl and infected with RV; C. sorokiniana pre-RV: HT-29 cells infected with RV and treated with C. sorokiniana (Cs); C. sorokiniana post-RV: HT-29 cells treated with Cs and infected with RV; Bl + Cs pre-RV: HT-29 cells treated with B. longum in combination with C. sorokiniana and then infected with RV; Bl + Cs post-RV: HT-29 cells infected with RV and treated with B. longum in combination with C. sorokiniana. Data were analyzed by three-way ANOVA with subsequent Tukey test using GraphPad Prism 10.
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