Although several studies have focused on allergic sensitization by dendritic cells, to date it is still open under which conditions these antigen-presenting cells are able to induce an allergic immune response. Our study reveals that BMDCs pulsed with LPS-free ovalbumine did not induce allergic disease. However, when BMDCs were activated with low-dose LPS during pulsing with allergen, these cells expressed an inflammatory set of cytokines and co-stimulatory molecules like CD86 and OX40L. Moreover, activated cells were able to prime mice for massive eosinophilic inflammation of the lung, airway hyper-reactivity, IgE production and production of Th2 cytokines by lymphocytes. Blocking experiments showed that expression of OX40L is not involved in induction of Th2 response. Interestingly, BMDCs that were activated with high dose of LPS lose their Th2-sensitizing capacity. Instead these cells induce a Th17 type immune response. We conclude that presentation of allergen by dendritic cells generated with GMCSF is not sufficient to lead to induction of allergic immune response. Further activation of BMDCs is required to prime mice for allergic immune response. In this study, we show that LPS is a suitable stimulus. However, when cells were activated with high dose LPS they tended to induce a Th17 response. Several animal studies have shown that airway sensitization could be achieved by priming mice with allergen-pulsed dendritic cells. [1][2][3] Lambrecht et al. 2 have shown that these antigen-presenting cells are critically involved in both sensitization phase and in ongoing allergic immune response. 2,4 However, in all of these studies ovalbumine preparations were used that were contaminated with LPS. There is some evidence from the literature that a TLR-mediated signal is required for effective induction of Th2 immunity. 5,6 In particular, stimulation via TLR4 seems to be a suitable stimulus as shown for example by Eisenbarth et al. 7 These authors showed that treatment with ovalbumine via the airways is not sufficient to sensitize mice. Only when low doses lipopolysaccharide were mixed to OVA, mice were sensitized towards the allergen. 7 On the basis of this observation Nigo et al. 8 have shown that allergy-promoting activity of low-dose LPS led to activation of mast cells via TLR4. Moreover, a recent publication of Hammad et al. 9 showed that LPS activation of epithelia cells has a significant impact on allergic sensitization towards house dust mite allergens. From these experiments the question arises, whether direct activation of DCs with LPS is sufficient or an LPSinduced cytokine milieu due to activation of mast cells and/or airway epithelium is required to arm these cells for sensitization of mice. As there is some evidence that GMCSF is involved in induction of Th2 response in vivo the possibility must be considered that Th2 induction could be generally attributed to GMCSF-generated dendritic cells. 10,11 Despite several years of research, the question still remains, which phenotype BMDCs must acquire to indu...

Heterozygous mutations of the tissue-specific transcription factor hepatocyte nuclear factor (HNF)1beta, cause maturity onset diabetes of the young (MODY5) and kidney anomalies including agenesis, hypoplasia, dysplasia and cysts. Because of these renal anomalies, HNF1beta is classified as a CAKUT (congenital anomalies of the kidney and urinary tract) gene. We searched for human fetal kidney proteins interacting with the N-terminal region of HNF1beta using a bacterial two-hybrid system and identified five novel proteins along with the known partner DCoH. The interactions were confirmed for four of these proteins by GST pull-down assays. Overexpression of two proteins, E4F1 and ZFP36L1, in Xenopus embryos interfered with pronephros formation. Further, in situ hybridization showed overlapping expression of HNF1beta, E4F1 and ZFP36L1 in the developing pronephros. HNF1beta is present largely in the nucleus where it colocalized with E4F1. However, ZFP36L1 was located predominantly in the cytoplasm. A nuclear function for ZFP36L1 was shown as it was able to reduce HNF1beta transactivation in a luciferase reporter system. Our studies show novel proteins may cooperate with HNF1beta in human metanephric development and propose that E4F1 and ZFP36L1 are CAKUT genes. We searched for mutations in the open reading frame of the ZFP36L1 gene in 58 patients with renal anomalies but found none.

The handheld organizer or personal digital assistant (PDA) is rapidly becoming a popular organizational tool, and there is a need for evaluation of alphanumeric character entry on these devices. The Palm operating system, the most common PDA operating system on the market, uses two methods for character entry, an on-screen virtual keyboard and a single-character handwriting recognition system called Graffiti. An initial experiment was conducted to investigate the character entry rates of novice and expert users of the device for the two methods of input. Experts were found to reach an average rate of 21 words per minute (wpm) using Graffiti and 18 wpm using the virtual keyboard. Novices were able to use Graffiti at a rate of 7 wpm and the virtual keyboard at 16 wpm. These character entry rates are evaluated with respect to some theoretical limitations, a predicted rate of entry based on Fitts' and the Hick-Hyman laws for the virtual keyboard, and pen and paper printing for Graffiti. The potential gain for new character entry systems and opportunities for improvement are discussed.

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