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Gut health immunomodulatory and anti-inflammatory functions of gut enzyme digested high protein micro-nutrient dietary supplement-Enprocal.

Kanwar JR, Kanwar RK - BMC Immunol. (2009)

Bottom Line: Our results indicate that Enprocal creates neither oxidative injury nor cytotoxicity, stimulates normal gut cell proliferation, up regulates immune cell activation markers and may aid in the production of antibodies.Furthermore, through downregulation of proinflammatory cytokines, Enprocal appears to be beneficial in reducing the effects of chronic gut inflammatory diseases such as inflammatory bowel disease (IBD).Stimulation of normal human fetal intestinal cell proliferation without cell cytotoxicity indicates it may also be given as infant food particularly for premature babies.

View Article: PubMed Central - HTML - PubMed

Affiliation: BioDeakin, Institute for Technology & Research Innovation, Deakin University, Geelong Technology Precinct, Geelong, Victoria, Australia. jagat.kanwar@deakin.edu.au

ABSTRACT

Background: Enprocal is a high-protein micro-nutrient rich formulated supplementary food designed to meet the nutritional needs of the frail elderly and be delivered to them in every day foods. We studied the potential of Enprocal to improve gut and immune health using simple and robust bioassays for gut cell proliferation, intestinal integrity/permeability, immunomodulatory, anti-inflammatory and anti-oxidative activities. Effects of Enprocal were compared with whey protein concentrate 80 (WPC), heat treated skim milk powder, and other commercially available milk derived products.

Results: Enprocal (undigested) and digested (Enprocal D) selectively enhanced cell proliferation in normal human intestinal epithelial cells (FHs74-Int) and showed no cytotoxicity. In a dose dependent manner Enprocal induced cell death in Caco-2 cells (human colon adencarcinoma epithelial cells). Digested Enprocal (Enprocal D: gut enzyme cocktail treated) maintained the intestinal integrity in transepithelial resistance (TEER) assay, increased the permeability of horseradish peroxidase (HRP) and did not induce oxidative stress to the gut epithelial cells. Enprocal D upregulated the surface expression of co-stimulatory (CD40, CD86, CD80), MHC I and MHC II molecules on PMA differentiated THP-1 macrophages in coculture transwell model, and inhibited the monocyte/lymphocyte (THP-1/Jurkat E6-1 cells)-epithelial cell adhesion. In cytokine secretion analyses, Enprocal D down-regulated the secretion of proinflammatory cytokines (IL-1beta and TNF-alpha) and up-regulated IFN-gamma, IL-2 and IL-10.

Conclusion: Our results indicate that Enprocal creates neither oxidative injury nor cytotoxicity, stimulates normal gut cell proliferation, up regulates immune cell activation markers and may aid in the production of antibodies. Furthermore, through downregulation of proinflammatory cytokines, Enprocal appears to be beneficial in reducing the effects of chronic gut inflammatory diseases such as inflammatory bowel disease (IBD). Stimulation of normal human fetal intestinal cell proliferation without cell cytotoxicity indicates it may also be given as infant food particularly for premature babies.

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Inhibition of lymphocyte-epithelial (A) and monocyte-epithelial (B) cell adhesion by Enprocal D and other digested milk product controls. For adhesion assays, Caco-2 cell monolayers, grown in 96-well plates, were treated for 18 h with Enprocal D and other digested milk product controls. CMFDA-labeled Jurkat (A) or THP-1 (B) cells were then added. After incubation for 60 min at 37°C, nonadherent immune cells were removed by washing with HBSS and the monolayer-associated Jurkat cells or THP-1 was counted. Values are showed as the percent inhibition. All treatments were performed in triplicate and assay was repeated three times independently with similar results. The mean for representative experiment was calculated and presented as a mean ± SD values. ** Indicates a highly significant P < 0.001 value from the control with media only. *Indicates a significant P < 0.05 value from the control with media only.
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Figure 8: Inhibition of lymphocyte-epithelial (A) and monocyte-epithelial (B) cell adhesion by Enprocal D and other digested milk product controls. For adhesion assays, Caco-2 cell monolayers, grown in 96-well plates, were treated for 18 h with Enprocal D and other digested milk product controls. CMFDA-labeled Jurkat (A) or THP-1 (B) cells were then added. After incubation for 60 min at 37°C, nonadherent immune cells were removed by washing with HBSS and the monolayer-associated Jurkat cells or THP-1 was counted. Values are showed as the percent inhibition. All treatments were performed in triplicate and assay was repeated three times independently with similar results. The mean for representative experiment was calculated and presented as a mean ± SD values. ** Indicates a highly significant P < 0.001 value from the control with media only. *Indicates a significant P < 0.05 value from the control with media only.

Mentions: Enprocal D in a dose dependant manner significantly (P < 0.001) inhibited the adhesion of lymphocytes and monocytes to Caco-2 cells. Similar levels of inhibition of adhesion were observed with P2 (Fig 8A &8B).


Gut health immunomodulatory and anti-inflammatory functions of gut enzyme digested high protein micro-nutrient dietary supplement-Enprocal.

Kanwar JR, Kanwar RK - BMC Immunol. (2009)

Inhibition of lymphocyte-epithelial (A) and monocyte-epithelial (B) cell adhesion by Enprocal D and other digested milk product controls. For adhesion assays, Caco-2 cell monolayers, grown in 96-well plates, were treated for 18 h with Enprocal D and other digested milk product controls. CMFDA-labeled Jurkat (A) or THP-1 (B) cells were then added. After incubation for 60 min at 37°C, nonadherent immune cells were removed by washing with HBSS and the monolayer-associated Jurkat cells or THP-1 was counted. Values are showed as the percent inhibition. All treatments were performed in triplicate and assay was repeated three times independently with similar results. The mean for representative experiment was calculated and presented as a mean ± SD values. ** Indicates a highly significant P < 0.001 value from the control with media only. *Indicates a significant P < 0.05 value from the control with media only.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2667481&req=5

Figure 8: Inhibition of lymphocyte-epithelial (A) and monocyte-epithelial (B) cell adhesion by Enprocal D and other digested milk product controls. For adhesion assays, Caco-2 cell monolayers, grown in 96-well plates, were treated for 18 h with Enprocal D and other digested milk product controls. CMFDA-labeled Jurkat (A) or THP-1 (B) cells were then added. After incubation for 60 min at 37°C, nonadherent immune cells were removed by washing with HBSS and the monolayer-associated Jurkat cells or THP-1 was counted. Values are showed as the percent inhibition. All treatments were performed in triplicate and assay was repeated three times independently with similar results. The mean for representative experiment was calculated and presented as a mean ± SD values. ** Indicates a highly significant P < 0.001 value from the control with media only. *Indicates a significant P < 0.05 value from the control with media only.
Mentions: Enprocal D in a dose dependant manner significantly (P < 0.001) inhibited the adhesion of lymphocytes and monocytes to Caco-2 cells. Similar levels of inhibition of adhesion were observed with P2 (Fig 8A &8B).

Bottom Line: Our results indicate that Enprocal creates neither oxidative injury nor cytotoxicity, stimulates normal gut cell proliferation, up regulates immune cell activation markers and may aid in the production of antibodies.Furthermore, through downregulation of proinflammatory cytokines, Enprocal appears to be beneficial in reducing the effects of chronic gut inflammatory diseases such as inflammatory bowel disease (IBD).Stimulation of normal human fetal intestinal cell proliferation without cell cytotoxicity indicates it may also be given as infant food particularly for premature babies.

View Article: PubMed Central - HTML - PubMed

Affiliation: BioDeakin, Institute for Technology & Research Innovation, Deakin University, Geelong Technology Precinct, Geelong, Victoria, Australia. jagat.kanwar@deakin.edu.au

ABSTRACT

Background: Enprocal is a high-protein micro-nutrient rich formulated supplementary food designed to meet the nutritional needs of the frail elderly and be delivered to them in every day foods. We studied the potential of Enprocal to improve gut and immune health using simple and robust bioassays for gut cell proliferation, intestinal integrity/permeability, immunomodulatory, anti-inflammatory and anti-oxidative activities. Effects of Enprocal were compared with whey protein concentrate 80 (WPC), heat treated skim milk powder, and other commercially available milk derived products.

Results: Enprocal (undigested) and digested (Enprocal D) selectively enhanced cell proliferation in normal human intestinal epithelial cells (FHs74-Int) and showed no cytotoxicity. In a dose dependent manner Enprocal induced cell death in Caco-2 cells (human colon adencarcinoma epithelial cells). Digested Enprocal (Enprocal D: gut enzyme cocktail treated) maintained the intestinal integrity in transepithelial resistance (TEER) assay, increased the permeability of horseradish peroxidase (HRP) and did not induce oxidative stress to the gut epithelial cells. Enprocal D upregulated the surface expression of co-stimulatory (CD40, CD86, CD80), MHC I and MHC II molecules on PMA differentiated THP-1 macrophages in coculture transwell model, and inhibited the monocyte/lymphocyte (THP-1/Jurkat E6-1 cells)-epithelial cell adhesion. In cytokine secretion analyses, Enprocal D down-regulated the secretion of proinflammatory cytokines (IL-1beta and TNF-alpha) and up-regulated IFN-gamma, IL-2 and IL-10.

Conclusion: Our results indicate that Enprocal creates neither oxidative injury nor cytotoxicity, stimulates normal gut cell proliferation, up regulates immune cell activation markers and may aid in the production of antibodies. Furthermore, through downregulation of proinflammatory cytokines, Enprocal appears to be beneficial in reducing the effects of chronic gut inflammatory diseases such as inflammatory bowel disease (IBD). Stimulation of normal human fetal intestinal cell proliferation without cell cytotoxicity indicates it may also be given as infant food particularly for premature babies.

Show MeSH
Related in: MedlinePlus