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A novel anti-histone H1 monoclonal antibody, SSV monoclonal antibody, improves lung injury and survival in a mouse model of lipopolysaccharide-induced sepsis-like syndrome.

Kusano T, Chiang KC, Inomata M, Shimada Y, Ohmori N, Goto T, Sato S, Goto S, Nakano T, Kawamoto S, Takaoka Y, Shiraishi N, Noguchi T, Kitano S - Biomed Res Int (2015)

Bottom Line: Competitive ELISA revealed that SSV mAb binds to histone H1.Increased levels of histones H1, H3, and H4 and inflammatory cytokines (TNF-α, IL-1β, and IL-6) in plasma and lung tissue after LPS injection were ameliorated by SSV mAb.SSV mAb is shown to have anti-inflammatory activity and organ-protective effects, highlighting the importance of controlling histone H1 as well as H3 and H4 levels during inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterological and Pediatric Surgery, Oita University Faculty of Medicine, 1-1 Hasama-machi, Yufu, Oita 879-5593, Japan.

ABSTRACT

Background: Histones play important roles in both host defenses and inflammation related to microbial infection. A peptide mimotope (SSV) was identified from a novel histone H1 monoclonal antibody (16G9 mAb) that was shown to inhibit the mixed lymphocyte reaction. In the present study, an anti-SSV producing hybridoma was established. We investigated the effects of SSV mAb in a mouse acute inflammation model induced by intraperitoneal injection of lipopolysaccharide (LPS).

Methods: SSV mAb was generated and characterized. Mice were treated with SSV mAb or a control IgG antibody prior to LPS injection. Evaluation of survival rate and lung tissue on histological score was performed. The levels of inflammatory cytokines and histones H1, H3, and H4 in plasma and lung tissue were measured by ELISA.

Results: Competitive ELISA revealed that SSV mAb binds to histone H1. SSV mAb improved lung injury and prolonged the survival of LPS-injected mice. Increased levels of histones H1, H3, and H4 and inflammatory cytokines (TNF-α, IL-1β, and IL-6) in plasma and lung tissue after LPS injection were ameliorated by SSV mAb.

Conclusion: SSV mAb is shown to have anti-inflammatory activity and organ-protective effects, highlighting the importance of controlling histone H1 as well as H3 and H4 levels during inflammation.

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Related in: MedlinePlus

Effect of SSV mAb on plasma cytokine levels. (a) Plasma TNF-α levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (b) Plasma IL-1β levels in the control (dashed line) or SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (c) Plasma IL-6 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (d) Plasma IL-10 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05).
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fig5: Effect of SSV mAb on plasma cytokine levels. (a) Plasma TNF-α levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (b) Plasma IL-1β levels in the control (dashed line) or SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (c) Plasma IL-6 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (d) Plasma IL-10 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05).

Mentions: The plasma TNF-α level reached a peak 3 hr after LPS injection in both groups and then began to gradually decrease (Figure 5(a)). Plasma TNF-α levels were higher in the control group than in the SSV mAb group during the entire observation time, and the difference reached a statistical significance from 3 to 24 hr after LPS injection (P < 0.05). Plasma IL-1β levels also reached a peak 3 hr after LPS injection in both groups and then began to gradually decrease (Figure 5(b)). Plasma IL-1β levels were higher in the control group during the entire observation time, and the difference reached a statistical significance from 3 to 9 hr after LPS injection (P < 0.05). Plasma IL-6 levels also reached a peak 3 hr after LPS injection in both groups and then began to gradually decrease (Figure 5(c)). Plasma IL-6 levels were higher in the control group and the difference reached a statistical significance at 3 h after LPS injection (P < 0.05). In contrast to the pattern displayed by histone H1 and the other cytokines, we found that plasma IL-10 levels in the SSV mAb group gradually increased (Figure 5(d)). Plasma IL-10 levels in the control group, however, displayed the typical pattern, reaching a peak 3 hr after LPS injection and then decreasing gradually. Plasma IL-10 levels were higher in the SSV mAb group than in the control group during the entire observation time, and the difference reached a statistical significance 12 and 24 hr after LPS injection (P < 0.05).


A novel anti-histone H1 monoclonal antibody, SSV monoclonal antibody, improves lung injury and survival in a mouse model of lipopolysaccharide-induced sepsis-like syndrome.

Kusano T, Chiang KC, Inomata M, Shimada Y, Ohmori N, Goto T, Sato S, Goto S, Nakano T, Kawamoto S, Takaoka Y, Shiraishi N, Noguchi T, Kitano S - Biomed Res Int (2015)

Effect of SSV mAb on plasma cytokine levels. (a) Plasma TNF-α levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (b) Plasma IL-1β levels in the control (dashed line) or SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (c) Plasma IL-6 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (d) Plasma IL-10 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05).
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fig5: Effect of SSV mAb on plasma cytokine levels. (a) Plasma TNF-α levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (b) Plasma IL-1β levels in the control (dashed line) or SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point) were measured by ELISA. Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (c) Plasma IL-6 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05). (d) Plasma IL-10 levels in the control (dashed line) and SSV mAb (solid line) groups at the time of LPS injection and then 3, 6, 9, 12, and 24 hr later (n = 10 per time point). Values plotted are means ± standard deviation. *Statistically significant differences (P < 0.05).
Mentions: The plasma TNF-α level reached a peak 3 hr after LPS injection in both groups and then began to gradually decrease (Figure 5(a)). Plasma TNF-α levels were higher in the control group than in the SSV mAb group during the entire observation time, and the difference reached a statistical significance from 3 to 24 hr after LPS injection (P < 0.05). Plasma IL-1β levels also reached a peak 3 hr after LPS injection in both groups and then began to gradually decrease (Figure 5(b)). Plasma IL-1β levels were higher in the control group during the entire observation time, and the difference reached a statistical significance from 3 to 9 hr after LPS injection (P < 0.05). Plasma IL-6 levels also reached a peak 3 hr after LPS injection in both groups and then began to gradually decrease (Figure 5(c)). Plasma IL-6 levels were higher in the control group and the difference reached a statistical significance at 3 h after LPS injection (P < 0.05). In contrast to the pattern displayed by histone H1 and the other cytokines, we found that plasma IL-10 levels in the SSV mAb group gradually increased (Figure 5(d)). Plasma IL-10 levels in the control group, however, displayed the typical pattern, reaching a peak 3 hr after LPS injection and then decreasing gradually. Plasma IL-10 levels were higher in the SSV mAb group than in the control group during the entire observation time, and the difference reached a statistical significance 12 and 24 hr after LPS injection (P < 0.05).

Bottom Line: Competitive ELISA revealed that SSV mAb binds to histone H1.Increased levels of histones H1, H3, and H4 and inflammatory cytokines (TNF-α, IL-1β, and IL-6) in plasma and lung tissue after LPS injection were ameliorated by SSV mAb.SSV mAb is shown to have anti-inflammatory activity and organ-protective effects, highlighting the importance of controlling histone H1 as well as H3 and H4 levels during inflammation.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterological and Pediatric Surgery, Oita University Faculty of Medicine, 1-1 Hasama-machi, Yufu, Oita 879-5593, Japan.

ABSTRACT

Background: Histones play important roles in both host defenses and inflammation related to microbial infection. A peptide mimotope (SSV) was identified from a novel histone H1 monoclonal antibody (16G9 mAb) that was shown to inhibit the mixed lymphocyte reaction. In the present study, an anti-SSV producing hybridoma was established. We investigated the effects of SSV mAb in a mouse acute inflammation model induced by intraperitoneal injection of lipopolysaccharide (LPS).

Methods: SSV mAb was generated and characterized. Mice were treated with SSV mAb or a control IgG antibody prior to LPS injection. Evaluation of survival rate and lung tissue on histological score was performed. The levels of inflammatory cytokines and histones H1, H3, and H4 in plasma and lung tissue were measured by ELISA.

Results: Competitive ELISA revealed that SSV mAb binds to histone H1. SSV mAb improved lung injury and prolonged the survival of LPS-injected mice. Increased levels of histones H1, H3, and H4 and inflammatory cytokines (TNF-α, IL-1β, and IL-6) in plasma and lung tissue after LPS injection were ameliorated by SSV mAb.

Conclusion: SSV mAb is shown to have anti-inflammatory activity and organ-protective effects, highlighting the importance of controlling histone H1 as well as H3 and H4 levels during inflammation.

Show MeSH
Related in: MedlinePlus