The expression of interleukin-4 (IL-4) is viewed as the hallmark of a Th2 lymphocyte, whereas the subsequent action of IL-4 and IL-13, mediated through the STAT6 signaling pathway, is seen as a prerequisite for the full development of Th2 immune responses to parasites and allergens. G4 mice, whose IL-4 gene locus contains the fluorescent reporter eGFP, were used to quantify the number of Th2 cells that develop during Nippostrongylus brasiliensis-or allergen-induced immune responses under conditions where IL-4 or STAT6 was absent. Here, we show that deletion of IL-4 or STAT6 had little impact on the number or timing of appearance of IL-4-producing Th2 cells. These data indicate that in vivo differentiation of naïve CD4 T cells to Th2 status often occurs independently of IL-4 and STAT6 and that recently described pathways of Th2 cell differentiation may explain how allergens and parasites selectively induce Th2-mediated immunity.allergen ͉ cytokine ͉ asthma ͉ Nippostrongylus

ʈʈHookworms infect more than 700 million people worldwide and cause more morbidity than most other human parasitic infections. Nippostrongylus brasiliensis (the rat hookworm) has been used as an experimental model for human hookworm because of its similar life cycle and ease of maintenance in laboratory rodents. Adult N. brasiliensis, like the human hookworm, lives in the intestine of the host and releases excretory/secretory products (ESP), which represent the major host-parasite interface. We performed a comparative proteomic analysis of infective larval (

No abstract

Type II immune responses are associated with protection against helminth infections, as well as the pathology of allergic diseases that are initiated against innocuous antigens. Two integral cytokines associated with type II immunity are interleukin (IL-)4 and IL-13. The 4C13R transgenic dual reporter mouse has been created to allow identification of IL-4 and IL-13 producing cells by the production of two intracellular fluorescent molecules, AmCyan and DS-Red respectively. The expression of the distinct reporter proteins are under the normal transcriptional control of the Il4 or Il13 genes. This technology allows for the analysis of in situ IL-4 and/or IL-13 production by the relevant differentiated immune cell types without any effect on the endogenous cytokine genes or their effector activities in the mouse. Using this reporter system we have identified that in vitro generated CD4+ Th2 cells have divergent expression of IL-4 and IL-13, suggesting cytokine specific Th2 subsets. Additionally, IL-13 expression is delayed compared with IL-4 in this system. Reporter expression in vivo has also highlighted that IL-4/IL-13 double producers are only a subpopulation of CD4+ T cells, and cytokine expression profiles differ significantly between lymph node and effector tissues during type II responses. Studying the expression and regulation of these cytokines will allow us to understand their contribution in both disease and protection.

Dendritic cells are critical for the initiation of immune responses, however, their role in priming IL4-producing Th2 cells is controversial and remains an area of intense investigation. We found that after skin injection of the nematode parasite Nippostrongylus brasiliensis (Nb), a strong inducer of Th2 responses, Nb material is taken up by a population of dermal dendritic cells in lymph node, and that this population is necessary and sufficient for IL4 priming. Blocking the migration of dermal dendritic cells to the lymph node also prevented the priming of IL4-producing cells, and chicken ovalbumin (OVA)-loaded dDC from Nb-treated mice were sufficient to sensitize OVA-specific CD4+ T cells to IL4 production in vivo. Nb-loaded dermal dendritic cells were CD11b+, CD326neg, CD103neg, PDL2hi and IRF4+, and modestly upregulated expression of OX40L. However, OX40L and TSLP were not required for Th2 priming. Thus, our data suggest that the appropriate stimuli can induce DC to express the unique signals that direct CD4+ T cells to Th2 differentiation.

We have examined the role of dendritic cells using different models of Th2 allergic immune responses in the skin. In the first model we used the nematode parasite Nippostrongylus brasiliensis, which infects rodent hosts by penetrating through the skin. Material from labelled N. brasiliensis is taken up by migratory dendritic cells and transported to the draining lymph node where it initiates a strong Th2 immune response. We have characterized the surface phenotype of these dendritic cells in terms of lineage markers, and for expression of markers associated with Th2 immune responses. We find that these dendritic cells express markers that are compatible with a steady-state population of dermal dendritic cells. In addition, we have carried out dendritic cell transfer experiments to show that these migratory dendritic cells maintain the full capacity to prime Th2 immune responses in vivo. Additional models of Th2 immune responses, such as those initiated by the powerful allergen House Dust Mite and by application of the vitamin D analogue MC903, are currently being investigated. Our data indicate that dendritic cells can carry a sufficient set of signals to instruct the initiation of Th2 immune responses in secondary lymphoid organs.