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. 2013 Oct 4;8(10):e75368.
doi: 10.1371/journal.pone.0075368. eCollection 2013.

Lactobacillus plantarum DK119 as a probiotic confers protection against influenza virus by modulating innate immunity

Min-Kyung Park  1 , Vu Ngo, Young-Man Kwon, Young-Tae Lee, Sieun Yoo, Young-Hee Cho, Sung-Moon Hong, Hye Suk Hwang, Eun-Ju Ko, Yu-Jin Jung, Dae-Won Moon, Eun-Ji Jeong, Min-Chul Kim, Yu-Na Lee, Ji-Hun Jang, Joon-Suk Oh, Cheol-Hyun Kim, Sang-Moo Kang
Affiliations

Lactobacillus plantarum DK119 as a probiotic confers protection against influenza virus by modulating innate immunity

Min-Kyung Park et al. PLoS One. .

Abstract

Lactobacillus plantarum DK119 (DK119) isolated from the fermented Korean cabbage food was used as a probiotic to determine its antiviral effects on influenza virus. DK119 intranasal or oral administration conferred 100% protection against subsequent lethal infection with influenza A viruses, prevented significant weight loss, and lowered lung viral loads in a mouse model. The antiviral protective efficacy was observed in a dose and route dependent manner of DK119 administration. Mice that were treated with DK119 showed high levels of cytokines IL-12 and IFN-γ in bronchoalveolar lavage fluids, and a low degree of inflammation upon infection with influenza virus. Depletion of alveolar macrophage cells in lungs and bronchoalveolar lavages completely abrogated the DK119-mediated protection. Modulating host innate immunity of dendritic and macrophage cells, and cytokine production pattern appeared to be possible mechanisms by which DK119 exhibited antiviral effects on influenza virus infection. These results indicate that DK119 can be developed as a beneficial antiviral probiotic microorganism.

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Conflict of interest statement

Competing Interests: The authors declare that Tobico Inc partially funded this research. They declare that DWM, EJJ, JHJ, and JSO are employees in Tobico Inc. CHK and JSO have patents. MKP, JSO, and CHK hold stock options in Tobico Inc. The authors declare that co-author Sang-Moo Kang is a PLOS ONE Editorial Board member. They confirm that this does not alter their adherence to all the PLOS ONE policies on sharing data and materials.

Figures

Figure 1
Figure 1. Intranasal co-administration of DK119 (109 CFU) and A/PR8 H1N1 virus confers protection.
BALB/c mice (n = 5 per group) were infected with A/PR8 virus (2.5 LD50) alone or co-administered with DK119 (109 CFU per mouse) and A/PR8 H1N1 virus, and body weight changes were monitored daily for 14 days. Virus (V): mice infected with A/PR8 virus, DK119(109)+V: mice infected with DK119 and A/PR8 H1N1 virus.DK119 (109): mice treated with DK119 (109 CFU per mouse) alone. Bars indicate standard errors.
Figure 2
Figure 2. Intranasal co-administration of a low dose of DK119 (108 CFU) and A/PR8 virus (H1N1) provides survival protection.
BALB/c mice (n = 6 per group) were infected once with A/PR8 virus (2.5 LD50) alone or co-administered with DK119 (108 CFU per mouse) and A/PR8 virus, and monitored daily for 14 days. Virus(V): mice infected with A/PR8 virus, DK119 (108)+V: mice infected with DK119 and A/PR8 virus.DK119 (108): mice treated with DK119 (108 CFU per mouse) alone did not show weight losses.
Figure 3
Figure 3. Pretreatment of a low dose of DK119(107 CFU) confers improved protection by preventing severe weight loss.
A group of mice (n = 6) was once treated intranasally with a low dose of DK119 (107 CFU per mouse). After 4 days of DK119 treatment, mice were infected with A/PR8 H1N1 virus (2.5 LD50) in the presence of DK119 (109 CFU per mouse). The control group was infected with A/PR8 virus without DK119 and no pretreatment. Virus(V): mice infected with A/PR8 H1N1 virus, DK119(107+109)+V: mice pretreated with DK119 and infected with DK119 and A/PR8 H1N1 virus.DK119(107+109): mice treated with DK119 (107 CFU and 109 CFU per mouse). Bars indicate standard errors.
Figure 4
Figure 4. Effects of DK119 (108 CFU) treatment on preventing weight loss against influenza H1N1 virus infection.
A group of mice (n = 6) was treated with DK119 (108 CFU/mouse) at 4 days and 1 day prior to infection with A/PR8 virus (2.5 LD50) in the presence of a low dose of DK119 (108 CFU per mouse). Virus(V): mice infected with A/PR8 virus, DK119 (108+108+108)+V: mice pretreated twice with DK119 and infected A/PR8 H1N1 virus in the presence of with DK119 (108 CFU per mouse). Bars indicate standard errors.DK119 (108+108+108): mice treated with DK119 alone (108 CFU/mouse at 3 different time points without virus infection).
Figure 5
Figure 5. Oral administration of DK119 (109 CFU) has protective effects against influenza virus infection.
Mice (n = 5) were administered DK119 (109 CFU/mouse daily) orally for 10 days before infection and for 14 days after infection with A/PR8 H1N1 virus (2.5 LD50), and their body weight changes were recorded. Virus(V): A/PR8 virus infection, DK119(109)+V: Oral administration with DK119 and then virus infection. Bars indicate standard errors. DK119(109): mice treated orally with DK119 alone (109 CFU/mouse daily for 14 days).
Figure 6
Figure 6. Oral administration of mice with a low dose of DK119 has protective effects against influenza virus (H1N1) infection.
Mice (n = 6) were orally administered DK119 (108 CFU/mouse daily) for 10 days before infection and for 14 days after infection with A/PR8 virus (1 LD50). Body weight changes were recorded. Virus(V): A/PR8 virus infection, DK119 (108)+V: Oral administration with DK119 and virus infection.
Figure 7
Figure 7. DK119 pretreatment promotes lung viral clearance after influenza virus infection.
Lung viral titers at day 4 post infection with H1N1 influenza virus A/PR8 are determined and presented in log10 units of limiting dilutions showing viral replication in chicken eggs. (10?8)+V: mice intranasally pretreated with DK119 (108 CFU) and infected with A/PR8 virus in the presence of DK119 (108). (10?9)+V: mice intranasally pretreated with DK119 (107 CFU) and infected with A/PR8 virus in the presence of DK119 (109). DK119 (oral)+V: mice orally pretreated with DK119 (109 CFU) for 10 days and then infected with A/PR8 virus. Virus: Naïve mice infected with A/PR8 virus (2.5 LD50). * (p<0.05), ** (p<0.001) statistical significances are indicated. Bars indicate standard errors.
Figure 8
Figure 8. Effects of DK119 pretreatment on cytokine production upon influenza virus infection.
Levels of cytokines in bronchoalveolar lavage fluids (BALF) collected from mice (n = 3) at day 4 post infection with A/PR8 virus are presented in pg/ml as determined using cytokine ELISA. (A) IL-4, (B) IL-6, (C) TNF-α, (D) IL-12, (E) IFN-γ. DK119 (10?8): Intranasal DK119 10?8 control in the absence of A/PR8 virus infection. DK119 (10?9): Intranasal DK119 10?9 control in the absence of A/PR8 virus infection. (10?7)+V: DK119 (10?7) intranasal pretreatment and then A/PR8 virus infection in the presence of DK119 (10?9). (10?8)+V: DK119 (10?8) intranasal pretreatment and then A/PR8 virus infection in the presence of DK119 (10?9). Virus: Naïve mice infected with A/PR8 virus (2.5 LD50). * indicates statistical significance compared to the control virus group (p<0.05). Bars indicate standard errors.
Figure 9
Figure 9. DK119 pretreatment reduces influenza virus-induced lung histopathology.
Inflammatory histopathology after H&E staining of lung sections was compared from mice that were infected with influenza A/PR8 virus (H1N1) with or without DK119 treatment. (A) Naïve: Naïve mice without A/PR8 virus and DK119, (B) Virus: mice that were infected with A/PR8 virus without DK119 treatment, (C) DK119+Virus: DK119 (107) intranasal pretreatment and then A/PR8 virus infection in the presence of DK119 (109). All images were obtained using the same camera setting and shown at a magnification of X400.
Figure 10
Figure 10. DK119 increases CD11c+ cell populations from BMDCs.
Bone marrow derived dendritic cells (BMDCs) were seeded in concentrations of 2.5×105 cells/ml and treated with 5×106 CFU/ml of DK119 or 2.5×108 CFU/ml for 2 days. The cells within a circle are CD11c positive cells and their percentages indicated. A representative figure in each is shown from three independent experiments. Control: No DK119, DK119 (20): 5×106 CFU/ml of DK119, DK119 (1000): 2.5×108 CFU/ml of DK119.
Figure 11
Figure 11. Intranasal administration of clodronate-liposomes selectively depletes CD11c+F4/80+alveolar macrophages in lungs and bronchoalveolar lavages.
The cells from the bronchoalveolar lavage (BAL) and lungs from mice (n = 3) were harvested4 days after clodronate injection. CD3+ and CD11b+ cells were excluded to determine the effect of clodronate on depleting alveolar macrophages with CD3−CD11b−CD11c+F4/80+markers. (A) Representative profiles of flow cytometry gating of CD3−CD11b−CD11c+F4/80+phenotypic cells. (B) Percentages of CD3−CD11b−CD11c+F4/80+cells. (C) Cellularity of CD3−CD11b−CD11c+F4/80+cells. (−) Clodronate or (−)Clod: No clodronate control mice, (+) Clodronate or (+)Clod: Clodronate treated mice. BAL: Bronchoalveolar cells, Lung: Cells collected from lung tissues. *P<0.05, **P<0.001, ***P<0.0001 (Student T-test). Bars indicate standard errors.
Figure 12
Figure 12. Effects of clodronate treatment on DK119-mediated protection against H3N2 influenza virus infection.
Groups of mice (n = 6) were two times treated with DK119 (108 CFU/mouse) at 4 and 1 day earlier, then either received clodronate or PBS 4 hours prior to infection with A/Philippines/82H3N2 virus (2.5 LD50) in the presence of a low dose of DK119 (108 CFU per mouse). DK119+H3N2: mice pretreated with DK119 were infected with A/Philippines/82 (H3N2) virus. DK119+clodronate+H3N2: mice pretreated with DK119 were intranasally administered clodronate prior to infection with A/Philippines/82 (H3N2) virus. H3N2+Clodronate: mice treated with clodronate 4 hours prior to infection.DK119+Clodronate: mice treated with DK119 at the same doses and then clodronate administration. Bars indicate standard errors.
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References

    1. Thompson WW, Shay DK, Weintraub E, Brammer L, Bridges CB, et al. (2004) Influenza-associated hospitalizations in the United States. JAMA 292: 1333–1340. - PubMed
    1. Hancock K, Veguilla V, Lu X, Zhong W, Butler EN, et al. (2009) Cross-reactive antibody responses to the 2009 pandemic H1N1 influenza virus. N Engl J Med 361: 1945–1952. - PubMed
    1. Jones M, Del Mar C (2006) Safety of neuraminidase inhibitors for influenza. Expert Opin Drug Saf 5: 603–608. - PubMed
    1. Oxford JS (2007) Antivirals for the treatment and prevention of epidemic and pandemic influenza. Influenza Other Respi Viruses 1: 27–34. - PMC - PubMed
    1. Beigel J, Bray M (2008) Current and future antiviral therapy of severe seasonal and avian influenza. Antiviral Res 78: 91–102. - PMC - PubMed

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