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Crotoxin from Crotalus durissus terrificus is able to down-modulate the acute intestinal inflammation in mice.

Almeida Cde S, Andrade-Oliveira V, Câmara NO, Jacysyn JF, Faquim-Mauro EL - PLoS ONE (2015)

Bottom Line: Crotoxin (CTX) is the main component of Crotalus durissus terrificus snake venom and has an immunomodulatory effect.The CTX administration resulted in decreased weight loss, disease activity index (DAI), macroscopic tissue damage, histopathological score and myeloperoxidase (MPO) activity analyzed after 4 days of acute TNBS colitis.The analysis of distinct cell populations obtained from the intestinal lamina propria showed that CTX reduced the number of group 3 innate lymphoid cells (ILC3) and Th17 population; CTX decreased IL-17 secretion but did not alter the frequency of CD4+Tbet+ T cells induced by TNBS instillation in mice.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunopathology, Butantan Institute, São Paulo, São Paulo, Brazil.

ABSTRACT
Inflammatory bowel diseases (IBD) is the result of dysregulation of mucosal innate and adaptive immune responses. Factors such as genetic, microbial and environmental are involved in the development of these disorders. Accordingly, animal models that mimic human diseases are tools for the understanding the immunological processes of the IBD as well as to evaluate new therapeutic strategies. Crotoxin (CTX) is the main component of Crotalus durissus terrificus snake venom and has an immunomodulatory effect. Thus, we aimed to evaluate the modulatory effect of CTX in a murine model of colitis induced by 2,4,6- trinitrobenzene sulfonic acid (TNBS). The CTX was administered intraperitoneally 18 hours after the TNBS intrarectal instillation in BALB/c mice. The CTX administration resulted in decreased weight loss, disease activity index (DAI), macroscopic tissue damage, histopathological score and myeloperoxidase (MPO) activity analyzed after 4 days of acute TNBS colitis. Furthermore, the levels of TNF-α, IL-1β and IL-6 were lower in colon tissue homogenates of TNBS-mice that received the CTX when compared with untreated TNBS mice. The analysis of distinct cell populations obtained from the intestinal lamina propria showed that CTX reduced the number of group 3 innate lymphoid cells (ILC3) and Th17 population; CTX decreased IL-17 secretion but did not alter the frequency of CD4+Tbet+ T cells induced by TNBS instillation in mice. In contrast, increased CD4+FoxP3+ cell population as well as secretion of TGF-β, prostaglandin E2 (PGE2) and lipoxin A4 (LXA4) was observed in TNBS-colitis mice treated with CTX compared with untreated TNBS-colitis mice. In conclusion, the CTX is able to modulate the intestinal acute inflammatory response induced by TNBS, resulting in the improvement of clinical status of the mice. This effect of CTX is complex and involves the suppression of the pro-inflammatory environment elicited by intrarectal instillation of TNBS due to the induction of a local anti-inflammatory profile in mice.

No MeSH data available.


Related in: MedlinePlus

Effect of CTX on Th17 cells, ILC3 and IL-17 secretion of mice with acute colitis induced by TNBS.Cell suspensions were prepared from lamina propria of distinct experimental groups after 4 days of TNBS-induced colitis and that received or not the CTX. The TCRß+CD4+RORγt+, TCRß+CD4+IL-17+ and Lin-CD90+IL-17+ cells were analyzed by flow cytometry. (A, B and C) Strategy for the analysis of TCRß+CD4+RORγt+ and TCRß+CD4+IL-17+ cells in the lamina propria obtained from each group of mice. The results of TCRß+CD4+RORγt+(D) and TCRß+CD4+IL-17+ cells (E) expressed as the mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2–3 independent experiments. (F) Strategy for the analysis of the Lin-CD90+IL-17+ cells. (G) The results of Lin-CD90+IL-17+ cells were expressed as a mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2 independent experiments. (H) Secretion of IL-17 in colonic tissue homogenates determined by ELISA. The results represent the mean obtained in the individual samples/group ± SEM. * p<0.05, ** p<0.01 and *** p<0.001 (n = 4–5 animals/group).
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pone.0121427.g003: Effect of CTX on Th17 cells, ILC3 and IL-17 secretion of mice with acute colitis induced by TNBS.Cell suspensions were prepared from lamina propria of distinct experimental groups after 4 days of TNBS-induced colitis and that received or not the CTX. The TCRß+CD4+RORγt+, TCRß+CD4+IL-17+ and Lin-CD90+IL-17+ cells were analyzed by flow cytometry. (A, B and C) Strategy for the analysis of TCRß+CD4+RORγt+ and TCRß+CD4+IL-17+ cells in the lamina propria obtained from each group of mice. The results of TCRß+CD4+RORγt+(D) and TCRß+CD4+IL-17+ cells (E) expressed as the mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2–3 independent experiments. (F) Strategy for the analysis of the Lin-CD90+IL-17+ cells. (G) The results of Lin-CD90+IL-17+ cells were expressed as a mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2 independent experiments. (H) Secretion of IL-17 in colonic tissue homogenates determined by ELISA. The results represent the mean obtained in the individual samples/group ± SEM. * p<0.05, ** p<0.01 and *** p<0.001 (n = 4–5 animals/group).

Mentions: As previously described T cell populations and ILCs are involved in the colonic inflammation. Consistent with this, we observed an increase in TCRß+CD4+RORγt+, TCRß+CD4+IL-17+ and Lin-CD90+IL-17+ cells in the lamina propria from the TNBS group when compared with the control groups (ETOH and ETOH/CTX) (Fig 3D, 3E and 3G). The CD4+Tbet+ cell population was also increased in the TNBS group compared with control groups (Fig 4B). In addition to this, high levels of IL-17 and IFN-γ were verified in colonic tissue homogenates of the TNBS group of mice (Fig 3H and 4C).


Crotoxin from Crotalus durissus terrificus is able to down-modulate the acute intestinal inflammation in mice.

Almeida Cde S, Andrade-Oliveira V, Câmara NO, Jacysyn JF, Faquim-Mauro EL - PLoS ONE (2015)

Effect of CTX on Th17 cells, ILC3 and IL-17 secretion of mice with acute colitis induced by TNBS.Cell suspensions were prepared from lamina propria of distinct experimental groups after 4 days of TNBS-induced colitis and that received or not the CTX. The TCRß+CD4+RORγt+, TCRß+CD4+IL-17+ and Lin-CD90+IL-17+ cells were analyzed by flow cytometry. (A, B and C) Strategy for the analysis of TCRß+CD4+RORγt+ and TCRß+CD4+IL-17+ cells in the lamina propria obtained from each group of mice. The results of TCRß+CD4+RORγt+(D) and TCRß+CD4+IL-17+ cells (E) expressed as the mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2–3 independent experiments. (F) Strategy for the analysis of the Lin-CD90+IL-17+ cells. (G) The results of Lin-CD90+IL-17+ cells were expressed as a mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2 independent experiments. (H) Secretion of IL-17 in colonic tissue homogenates determined by ELISA. The results represent the mean obtained in the individual samples/group ± SEM. * p<0.05, ** p<0.01 and *** p<0.001 (n = 4–5 animals/group).
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Related In: Results  -  Collection

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pone.0121427.g003: Effect of CTX on Th17 cells, ILC3 and IL-17 secretion of mice with acute colitis induced by TNBS.Cell suspensions were prepared from lamina propria of distinct experimental groups after 4 days of TNBS-induced colitis and that received or not the CTX. The TCRß+CD4+RORγt+, TCRß+CD4+IL-17+ and Lin-CD90+IL-17+ cells were analyzed by flow cytometry. (A, B and C) Strategy for the analysis of TCRß+CD4+RORγt+ and TCRß+CD4+IL-17+ cells in the lamina propria obtained from each group of mice. The results of TCRß+CD4+RORγt+(D) and TCRß+CD4+IL-17+ cells (E) expressed as the mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2–3 independent experiments. (F) Strategy for the analysis of the Lin-CD90+IL-17+ cells. (G) The results of Lin-CD90+IL-17+ cells were expressed as a mean of the absolute number ± SEM. The samples were prepared from a pool of cells from 4–5 animals/group performed in duplicate. The results are from 2 independent experiments. (H) Secretion of IL-17 in colonic tissue homogenates determined by ELISA. The results represent the mean obtained in the individual samples/group ± SEM. * p<0.05, ** p<0.01 and *** p<0.001 (n = 4–5 animals/group).
Mentions: As previously described T cell populations and ILCs are involved in the colonic inflammation. Consistent with this, we observed an increase in TCRß+CD4+RORγt+, TCRß+CD4+IL-17+ and Lin-CD90+IL-17+ cells in the lamina propria from the TNBS group when compared with the control groups (ETOH and ETOH/CTX) (Fig 3D, 3E and 3G). The CD4+Tbet+ cell population was also increased in the TNBS group compared with control groups (Fig 4B). In addition to this, high levels of IL-17 and IFN-γ were verified in colonic tissue homogenates of the TNBS group of mice (Fig 3H and 4C).

Bottom Line: Crotoxin (CTX) is the main component of Crotalus durissus terrificus snake venom and has an immunomodulatory effect.The CTX administration resulted in decreased weight loss, disease activity index (DAI), macroscopic tissue damage, histopathological score and myeloperoxidase (MPO) activity analyzed after 4 days of acute TNBS colitis.The analysis of distinct cell populations obtained from the intestinal lamina propria showed that CTX reduced the number of group 3 innate lymphoid cells (ILC3) and Th17 population; CTX decreased IL-17 secretion but did not alter the frequency of CD4+Tbet+ T cells induced by TNBS instillation in mice.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunopathology, Butantan Institute, São Paulo, São Paulo, Brazil.

ABSTRACT
Inflammatory bowel diseases (IBD) is the result of dysregulation of mucosal innate and adaptive immune responses. Factors such as genetic, microbial and environmental are involved in the development of these disorders. Accordingly, animal models that mimic human diseases are tools for the understanding the immunological processes of the IBD as well as to evaluate new therapeutic strategies. Crotoxin (CTX) is the main component of Crotalus durissus terrificus snake venom and has an immunomodulatory effect. Thus, we aimed to evaluate the modulatory effect of CTX in a murine model of colitis induced by 2,4,6- trinitrobenzene sulfonic acid (TNBS). The CTX was administered intraperitoneally 18 hours after the TNBS intrarectal instillation in BALB/c mice. The CTX administration resulted in decreased weight loss, disease activity index (DAI), macroscopic tissue damage, histopathological score and myeloperoxidase (MPO) activity analyzed after 4 days of acute TNBS colitis. Furthermore, the levels of TNF-α, IL-1β and IL-6 were lower in colon tissue homogenates of TNBS-mice that received the CTX when compared with untreated TNBS mice. The analysis of distinct cell populations obtained from the intestinal lamina propria showed that CTX reduced the number of group 3 innate lymphoid cells (ILC3) and Th17 population; CTX decreased IL-17 secretion but did not alter the frequency of CD4+Tbet+ T cells induced by TNBS instillation in mice. In contrast, increased CD4+FoxP3+ cell population as well as secretion of TGF-β, prostaglandin E2 (PGE2) and lipoxin A4 (LXA4) was observed in TNBS-colitis mice treated with CTX compared with untreated TNBS-colitis mice. In conclusion, the CTX is able to modulate the intestinal acute inflammatory response induced by TNBS, resulting in the improvement of clinical status of the mice. This effect of CTX is complex and involves the suppression of the pro-inflammatory environment elicited by intrarectal instillation of TNBS due to the induction of a local anti-inflammatory profile in mice.

No MeSH data available.


Related in: MedlinePlus