Early events associated to the oral co-administration of type II collagen and chitosan: induction of anti-inflammatory cytokines

Abstract

Oral administration of an antigen can result in local and systemic priming or tolerance and the basis of this dichotomy is poorly understood. The intestinal microenvironment, and factors such as nature of the antigen, dose, genetic background, uptake and concentration of the antigen that gain access to the internal milieu via the mucosa influence these active immunologic processes. Chitosan is a biocompatible natural polysaccharide able to promote the transmucosal absorption of peptides and proteins. The aim of our work was to study the effect of the co-administration of type II collagen (CII) and chitosan during the initial contact of the antigen with the immune system. Sixteen hours after feeding we evaluated several molecular events in mucosal and in systemic lymphoid tissues. We determined in Peyer's patches (PP) and spleen cells the number and activation of T cells, the arrival of the antigens, and the cytokine profile. In PP we found a reduction in the cell number without changes in CD3(+) cells. In spleen, instead, we observed an increase in CD3(+) cells as well as the internalization of the CD3 complex. CII:chitosan-fed animals exhibited a reduced secretion of IL-2 with an increase of IL-10 in PP and spleen respectively. In addition, in PP, CII:chitosan-fed rats showed increased levels of mRNA for transforming growth factor-beta, IL-4 and IL-10. Together, our data suggest that the co-administration with chitosan modifies the uptake and/or the distribution of the relevant antigen, and promotes an anti-inflammatory environment early after feeding.

The biocompatible polysaccharide chitosan enhances the oral tolerance to type II collagen

“…Interestingly, while the sustained contact of CII or CII : chitosan with the mucosal immune system generated specific tolerance, chitosan itself produced some effect on B cell function. We cannot explain this finding fully, although the result is in agreement with our previous reports showing that chitosan conditions, after sustained administration, an anti‐inflammatory environment at the inductive mucosal sites [21,22]. Recently, novel and remarkable activities of polysaccharides on mucosal immunity have been described that include the settlement of the T cell repertoire and the release of IL‐10 [41,42].…”

“…We have demonstrated that oral administration of chitosan triggers the recruitment of immature dendritic cells [22], up‐regulates IL‐4 and TGF‐β mRNA expression [21,22], reduces IL‐2 levels and increases specific IL‐10 release [21] at the inductive mucosal sites. Considering that most of these factors have been associated with the induction of regulatory subpopulations [21,22,30,37,38], we evaluated the occurrence of T cells with regulatory characteristics. Upon restimulation in vitro with the relevant antigen, both the diluent and chitosan groups showed similar percentages of CD4 + IL‐10 + cells in lymph nodes or spleen (Fig.…”

“…Chitosan's ability to modulate oral immunity is particular. At the same dose evaluated in this work, it reduces the enzymatic degradation of several proteins including CII [21,45], recruits immature dendritic cells at the inductive sites [22], promotes the expression of TGF‐β and IL‐4 mRNAs [21] and stimulates the release of IL‐10 [21,22,46]. Compounds with modulatory properties might influence the tolerance induction in the mucosa [47].…”

Local and systemic activity of the polysaccharide chitosan at lymphoid tissues after oral administration

“…TNF-␣, IL-4, and IL-12 (4 g) and TGF-␤ (2 g) of total RNA were incubated with 0.5 g oligo(dT; Biodynamics, Buenos Aires, Argentina) for 5 min at 70°C and allowed to stand on ice for 5 min. [24]. Each sample was incubated in a thermal cycler (PTC-100 thermal cycler, M.J. Research, Watertown, MA) using one cycle at 94°C for 5 min; this was followed by 25 cycles for ␤-actin, 30 cycles for IL-4, or 35 cycles for TGF-␤, TNF-␣, and IL-12; each cycle consisted of 1 min at 94°C, 1 min at 55°C (␤-actin, TNF-␣, and IL-12) or 58°C (TGF-␤ and IL-4), and 1 min at 72°C, with a final extension at 72°C for 5 min.…”

Chitosan enhances nanoparticle delivery from the reproductive tract to target draining lymphoid organs

“…29, 30 These amine groups have been described to reversibly destabilize cell plasma membrane, 31 which enhances uptake of cargo co-administered with chitosan. 32 In addition, when orally administered to mice, chitosan is transported primarily associated with DCs or macrophages to LNs that drain the gastrointestinal mucosa. 18 In the present study, it is conceivable that chitosan opens tight junctions of the vaginal epithelium to facilitate NP transepithelial delivery while also promoting greater association between NPs and cells locally in the RT to cause accumulation distally in the dLNs.…”