Human dendritic cells pulsed with specific lipopeptides stimulate autologous antigen-specific T cells without the addition of exogenous maturation factors.

SUMMARY: Serum-free culture conditions to generate immature human monocyte-derived DC (Mo-DC) were optimized, and the parameters that influence their maturation after exposure to lipopeptides containing CD4(+) and CD8(+) T-cell epitopes were examined. The lipopeptides contained a single CD4(+) helper T-cell epitopes, one of a number of human leucocyte antigen (HLA)-A2-restricted cytotoxic T-cell epitope and the lipid Pam2Cys. To ensure complete maturation of the Mo-DC, we examined (i) the optimal lipopeptide concentration, (ii) the optimal Mo-DC density and (iii) the appropriate period of exposure of the Mo-DC to the lipopeptides. The results showed that a high dose of lipopeptide (30 microm) was no more efficient at upregulating maturation markers on Mo-DC than a low dose (6 microm). There was an inverse relationship between Mo-DC concentration and the mean fluorescence intensity of maturation markers. In addition, at the higher cell concentrations, the chemotactic capacity of the Mo-DC towards a cognate ligand, CCL21, was reduced. Thus, high cell concentrations during lipopeptide exposure were detrimental to Mo-DC maturation and function. The duration of exposure of Mo-DC to the lipopeptides had little effect on phenotype, although Mo-DC exposed to lipopeptides for 48 rather than 4 h showed an increased ability to stimulate autologous peripheral blood mononuclear cells to release interferon-gamma in the absence of exogenous maturation factors. These findings reveal conditions for generating mature antigen-loaded DC suitable for targeted immunotherapy.

Body awareness therapy: a new strategy for relief of symptoms in irritable bowel syndrome patients.

AIM: To compare irritable bowel syndrome (IBS) patients with apparently healthy persons and to evaluate body awareness therapy, which is a physiotherapeutic remedy focusing on normalising tensions in the body, for the treatment of IBS with the hypothesis that altered body tension is associated with the syndrome. METHODS: Twenty-one IBS patients received body awareness therapy two hours weekly for 24 wk. At baseline as well as after 12 and 24 wk, they underwent examinations including resource oriented body examination in combination with body awareness scale evaluation and filled in gastrointestinal and psychological symptom questionnaires. Saliva cortisol was analysed. A group of 21 apparently healthy persons underwent the same examinations once. RESULTS: Compared to the apparently healthy group, IBS patients scored higher at baseline for gastrointestinal and psychological symptoms. They showed more often alterations in normal body tension patterns, as well as deviating cortisol slopes in saliva. After 24 wk of body awareness therapy, their gastrointestinal and psychological symptoms were reduced overall. Somatic symptoms decreased in parallel with depressive symptoms. Whole body pain score decreased, coping ability as well as biochemical stress markers improved. CONCLUSION: IBS patients scored higher for gastrointestinal and psychological symptoms, and presented with altered biochemical stress markers. Their body tension deviated compared to healthy controls. Furthermore, body awareness therapy gave relief of both somatic complaints, psychological symptoms and normalised body tension. These findings indicate that distorted tension constitutes an important part of the symptoms in IBS.