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Local immune regulation of mucosal inflammation by tacrolimus.

van Dieren JM, Lambers ME, Kuipers EJ, Samsom JN, van der Woude CJ, Nieuwenhuis EE - Dig. Dis. Sci. (2009)

Bottom Line: Local tacrolimus treatment is effective in a subgroup of proctitis patients.Tacrolimus inhibits activation of several pivotal immune cells of the intestinal mucosa.Murine studies indicate that colonic application of tacrolimus induces local rather than systemic immune suppression.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Hepatology, Erasmus MC, University Medical Center, 's Gravendijkwal 230, 3015 CE, Rotterdam, The Netherlands. j.vandieren@erasmusmc.nl

ABSTRACT

Purpose: Tacrolimus is a potent immunomodulator that is effective in the treatment of inflammatory bowel disease (IBD). However, potential toxicity and systemic effects with oral intake limit its use. Local tacrolimus treatment is effective in a subgroup of proctitis patients. This study aimed to evaluate whether colonic mucosal immune cells are susceptible to locally applied tacrolimus in vitro. Our in vivo studies aimed at evaluating whether local tacrolimus treatment in mice would bring about local immune suppression and to compare colonic and systemic tacrolimus levels after locally and systemically applied tacrolimus.

Results: In vitro tacrolimus inhibited the activation of multiple cell types present in colonic tissue; lamina propria T cells, NKT cells, and both classical- and non- classical antigen presenting cells. However, the cytokine production of epithelial cells was not inhibited by tacrolimus at these concentrations. After rectal administration in mice, tacrolimus blood levels were comparable to those obtained by oral intake. However, rectally treated mice exhibited a 14-fold higher concentration of tacrolimus within their colonic tissue than orally treated mice. Moreover, rectally applied tacrolimus resulted in a local but not a systemic immune suppression in mice.

Conclusions: Tacrolimus inhibits activation of several pivotal immune cells of the intestinal mucosa. Murine studies indicate that colonic application of tacrolimus induces local rather than systemic immune suppression.

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Related in: MedlinePlus

Colonic application of tacrolimus in mice results in mainly local presence of the drug and local immune suppression. a Mice were treated with 150 μl of tacrolimus (0.1 mg/ml) either intrarectally or intragastrically. Eighteen hours later, mice were sacrificed and colons and blood were removed. Colons were divided into a proximal part and a distal part, and homogenized. Tacrolimus levels were measured in blood and the homogenates by means of ELISA. The first four bars reflect the concentration of tacrolimus present in the colon (ng per mg protein). The last two bars represent the concentration of tacrolimus present in the blood (ng per ml). Irrespective of the route of administration, at 18 h after tacrolimus treatment, comparable amounts of tacrolimus are detectable in the blood. Tacrolimus was present in the colon in higher amounts after rectal than after oral administration of the drug. b Mice were treated with 150 μl of tacrolimus (1 mg/ml) or saline intrarectally. At 18 h after treatment, lymphocytes were isolated from lamina propria, iliac lymph node, mesenteric lymph node, and spleen, and stimulated for 72 h with αCD3αCD28 antibodies. IL-2 production was measured in the supernatant by means of ELISA. Bars represent the average IL-2 production of at least four mice. Lamina propria-derived lymphocytes (LPL) and colon draining lymph node-derived lymphocytes (ILN) of tacrolimus-treated mice tend to produce less IL-2 upon polyclonal stimulation than lymphocytes derived from saline-treated mice, whereas no differences in IL-2 production were observed within systemically derived lymphocytes. Asterisks indicate statistical significance (P < 0.05). Error bars indicate the standard error of the mean
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Fig2: Colonic application of tacrolimus in mice results in mainly local presence of the drug and local immune suppression. a Mice were treated with 150 μl of tacrolimus (0.1 mg/ml) either intrarectally or intragastrically. Eighteen hours later, mice were sacrificed and colons and blood were removed. Colons were divided into a proximal part and a distal part, and homogenized. Tacrolimus levels were measured in blood and the homogenates by means of ELISA. The first four bars reflect the concentration of tacrolimus present in the colon (ng per mg protein). The last two bars represent the concentration of tacrolimus present in the blood (ng per ml). Irrespective of the route of administration, at 18 h after tacrolimus treatment, comparable amounts of tacrolimus are detectable in the blood. Tacrolimus was present in the colon in higher amounts after rectal than after oral administration of the drug. b Mice were treated with 150 μl of tacrolimus (1 mg/ml) or saline intrarectally. At 18 h after treatment, lymphocytes were isolated from lamina propria, iliac lymph node, mesenteric lymph node, and spleen, and stimulated for 72 h with αCD3αCD28 antibodies. IL-2 production was measured in the supernatant by means of ELISA. Bars represent the average IL-2 production of at least four mice. Lamina propria-derived lymphocytes (LPL) and colon draining lymph node-derived lymphocytes (ILN) of tacrolimus-treated mice tend to produce less IL-2 upon polyclonal stimulation than lymphocytes derived from saline-treated mice, whereas no differences in IL-2 production were observed within systemically derived lymphocytes. Asterisks indicate statistical significance (P < 0.05). Error bars indicate the standard error of the mean

Mentions: We compared colonic and systemic tacrolimus levels after oral administration, which is the common route for systemic therapy, and after intra-rectal administration. As demonstrated by Fig. 2a, blood levels of intra-rectal treated mice are similar to blood levels of orally treated mice after 18 h of treatment. Importantly, upon rectal administration, a 14-fold higher concentration of tacrolimus was reached within colonic tissue in comparison to oral administration (Fig. 2a).Fig. 2


Local immune regulation of mucosal inflammation by tacrolimus.

van Dieren JM, Lambers ME, Kuipers EJ, Samsom JN, van der Woude CJ, Nieuwenhuis EE - Dig. Dis. Sci. (2009)

Colonic application of tacrolimus in mice results in mainly local presence of the drug and local immune suppression. a Mice were treated with 150 μl of tacrolimus (0.1 mg/ml) either intrarectally or intragastrically. Eighteen hours later, mice were sacrificed and colons and blood were removed. Colons were divided into a proximal part and a distal part, and homogenized. Tacrolimus levels were measured in blood and the homogenates by means of ELISA. The first four bars reflect the concentration of tacrolimus present in the colon (ng per mg protein). The last two bars represent the concentration of tacrolimus present in the blood (ng per ml). Irrespective of the route of administration, at 18 h after tacrolimus treatment, comparable amounts of tacrolimus are detectable in the blood. Tacrolimus was present in the colon in higher amounts after rectal than after oral administration of the drug. b Mice were treated with 150 μl of tacrolimus (1 mg/ml) or saline intrarectally. At 18 h after treatment, lymphocytes were isolated from lamina propria, iliac lymph node, mesenteric lymph node, and spleen, and stimulated for 72 h with αCD3αCD28 antibodies. IL-2 production was measured in the supernatant by means of ELISA. Bars represent the average IL-2 production of at least four mice. Lamina propria-derived lymphocytes (LPL) and colon draining lymph node-derived lymphocytes (ILN) of tacrolimus-treated mice tend to produce less IL-2 upon polyclonal stimulation than lymphocytes derived from saline-treated mice, whereas no differences in IL-2 production were observed within systemically derived lymphocytes. Asterisks indicate statistical significance (P < 0.05). Error bars indicate the standard error of the mean
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Related In: Results  -  Collection

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Fig2: Colonic application of tacrolimus in mice results in mainly local presence of the drug and local immune suppression. a Mice were treated with 150 μl of tacrolimus (0.1 mg/ml) either intrarectally or intragastrically. Eighteen hours later, mice were sacrificed and colons and blood were removed. Colons were divided into a proximal part and a distal part, and homogenized. Tacrolimus levels were measured in blood and the homogenates by means of ELISA. The first four bars reflect the concentration of tacrolimus present in the colon (ng per mg protein). The last two bars represent the concentration of tacrolimus present in the blood (ng per ml). Irrespective of the route of administration, at 18 h after tacrolimus treatment, comparable amounts of tacrolimus are detectable in the blood. Tacrolimus was present in the colon in higher amounts after rectal than after oral administration of the drug. b Mice were treated with 150 μl of tacrolimus (1 mg/ml) or saline intrarectally. At 18 h after treatment, lymphocytes were isolated from lamina propria, iliac lymph node, mesenteric lymph node, and spleen, and stimulated for 72 h with αCD3αCD28 antibodies. IL-2 production was measured in the supernatant by means of ELISA. Bars represent the average IL-2 production of at least four mice. Lamina propria-derived lymphocytes (LPL) and colon draining lymph node-derived lymphocytes (ILN) of tacrolimus-treated mice tend to produce less IL-2 upon polyclonal stimulation than lymphocytes derived from saline-treated mice, whereas no differences in IL-2 production were observed within systemically derived lymphocytes. Asterisks indicate statistical significance (P < 0.05). Error bars indicate the standard error of the mean
Mentions: We compared colonic and systemic tacrolimus levels after oral administration, which is the common route for systemic therapy, and after intra-rectal administration. As demonstrated by Fig. 2a, blood levels of intra-rectal treated mice are similar to blood levels of orally treated mice after 18 h of treatment. Importantly, upon rectal administration, a 14-fold higher concentration of tacrolimus was reached within colonic tissue in comparison to oral administration (Fig. 2a).Fig. 2

Bottom Line: Local tacrolimus treatment is effective in a subgroup of proctitis patients.Tacrolimus inhibits activation of several pivotal immune cells of the intestinal mucosa.Murine studies indicate that colonic application of tacrolimus induces local rather than systemic immune suppression.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology and Hepatology, Erasmus MC, University Medical Center, 's Gravendijkwal 230, 3015 CE, Rotterdam, The Netherlands. j.vandieren@erasmusmc.nl

ABSTRACT

Purpose: Tacrolimus is a potent immunomodulator that is effective in the treatment of inflammatory bowel disease (IBD). However, potential toxicity and systemic effects with oral intake limit its use. Local tacrolimus treatment is effective in a subgroup of proctitis patients. This study aimed to evaluate whether colonic mucosal immune cells are susceptible to locally applied tacrolimus in vitro. Our in vivo studies aimed at evaluating whether local tacrolimus treatment in mice would bring about local immune suppression and to compare colonic and systemic tacrolimus levels after locally and systemically applied tacrolimus.

Results: In vitro tacrolimus inhibited the activation of multiple cell types present in colonic tissue; lamina propria T cells, NKT cells, and both classical- and non- classical antigen presenting cells. However, the cytokine production of epithelial cells was not inhibited by tacrolimus at these concentrations. After rectal administration in mice, tacrolimus blood levels were comparable to those obtained by oral intake. However, rectally treated mice exhibited a 14-fold higher concentration of tacrolimus within their colonic tissue than orally treated mice. Moreover, rectally applied tacrolimus resulted in a local but not a systemic immune suppression in mice.

Conclusions: Tacrolimus inhibits activation of several pivotal immune cells of the intestinal mucosa. Murine studies indicate that colonic application of tacrolimus induces local rather than systemic immune suppression.

Show MeSH
Related in: MedlinePlus