Immunotherapy for neuroblastoma using syngeneic fibroblasts transfected with IL-2 and IL-12

Cytokine-modified tumour cells have been used in clinical trials for immunotherapy of neuroblastoma, but primary tumour cells from surgical biopsies are difficult to culture. Autologous fibroblasts, however, are straightforward to manipulate in culture and easy to transfect using nonviral or viral vectors. Here we have compared the antitumour effect of fibroblasts and tumour cells transfected ex vivo to coexpress interleukin-2 (IL-2) and IL-12 in a syngeneic mouse model of neuroblastoma. Coinjection of cytokine-modified fibroblasts with Neuro-2A tumour cells abolished their in vivo tumorigenicity. Treatment of established tumours with three intratumoral doses of transfected fibroblasts showed a significant therapeutic effect with reduced growth or complete eradication of tumours in 90% of mice, associated with extensive leukocyte infiltration. Splenocytes recovered from vaccinated mice showed enhanced IL-2 production following Neuro-2A coculture, and increased cytotoxicity against Neuro-2A targets compared with controls. Furthermore, 100% of the tumour-free mice exhibited immune memory against tumour cells when rechallenged three months later. The potency of transfected fibroblasts was equivalent to that of tumour cells in all experiments. We conclude that syngeneic fibroblasts cotransfected with IL-2 and IL-12 mediate therapeutic effects against established disease, and are capable of generating immunological memory. Furthermore, as they are easier to recover and manipulate than autologous tumour cells, fibroblasts provide an attractive alternative immunotherapeutic strategy for the treatment of neuroblastoma

Landholder profiling and typologies for natural resource-management policy and program support: Potential and constraints

The use of landholder typologies to aid the development, implementation, and monitoring natural-resource management (NRM) policies and programs has increased considerably during the past decade. This article explores the potential for using such typologies for a variety of NRM and rural and regional development applications. Review of typology use further suggests that there is potential to reline the way that typologies are developed and applied to better aid NRM, farming systems analyses, and rural and regional development. Before typologies will be adopted more widely, a number of theoretical and methodologic issues must be addressed. These include the following questions: (1) Which criteria and methods should or can be used to classify landholders? (2) How should studies across spatial and temporal scales be integrated? (3) How should multiple and single industry studies be inte-rated to gain the most value from research? We argue that quantitative research techniques are well suited to provide an underlying structure for landholder typologies, and qualitative research techniques are useful for developing understanding of the nature of variation within and between landholder types. We argued further that because of the potential utility and breadth for the application of landholder typologies, a nested set of landholder typologies could be developed that are coordinated at the national. regional, and local geographic levels, with repeated measures used to track the evolution with time of landholder practices, management values, and socioeconomic characteristics

Intra-mitrocondrial degradation of Tim23 curtails the survival of cells rescued from apoptosis by caspase inhibitors

10 páginas, 7 figuras, 1 tabla -- PAGS nros. 545-554Caspase inhibition can extend the survival of cells undergoing apoptosis beyond the point of mitochondrial outer membrane permeabilisation (MOMP), but this does not confer long-term protection because caspase-independent death pathways emerge. Here, we describe a novel mechanism of mitochondrial self-destruction in caspase-inhibited cells, whose hallmark is the degradation of Tim23, the essential pore-forming component of the TIM23 inner membrane translocase. We show that Tim23 degradation occurs in cycling and post-mitotic cells, it is caspase-independent but Bax/Bak dependent, and it follows cytochrome c release. The proteolytic degradation of Tim23 is induced by MOMP and is mitochondrion-autonomous, as it also occurs in isolated mitochondria undergoing permeability transition. Degradation of Tim23 is selective, as expression of several other inner membrane proteins that regulate respiratory chain function is unaffected, and is not autophagic, as it occurs similarly in autophagy-proficient and -deficient (Atg-5 knockout) cells. Depleting Tim23 with siRNA is sufficient to inhibit cell proliferation and prevent long-term survival, while expression of degradation-resistant Tim23-GFP in mitochondria delays caspase-independent cell death. Thus, mitochondrial autodigestion of Tim23 joins the array of processes contributing to caspase-independent cell death. Because mitochondrial biogenesis requires a functional protein-import machinery, preventing Tim23 degradation might, therefore, be essential for repairing damaged mitochondria in chronic degenerative diseasesThis work was supported by Programme grant 064232 from the Wellcome TrustPeer reviewe

Epithelial and neuronal calbindin in avian intestine. An immunohistochemical study.

It is well known that calbindin immunoreactivity is highly concentrated in the duodenal absorptive cells of young birds. We have shown that in the adult intestine of three avian species, calbindin content is much more variable. In addition to absorptive cells, we have detected throughout the gut of both sexes of the domestic fowl and in the large intestine of the Japanese quail a second type of calbindin-positive epithelial cell which has the shape of a typical endocrine cell. These cells were particularly abundant in the large intestine, in contrast to the usual distribution of endocrine cells along the gut. Calbindin was also detected in the nervous system of the intestine. Calbindin-positive nerve fibres were rare in the duodenum and ileum, numerous in plexuses and nerve processes in both muscular layers and lamina propria of the large intestine in domestic fowl and Japanese quail. In the mallard, nerve fibres were rarely calbindin positive while definitively positive for VIP. Calbindin of the peripheral nervous system of the domestic fowl and Japanese quail comigrates with the duodenal calbindin (27,000 dalton) in SDS gel electrophoresis.Journal ArticleResearch Support, Non-U.S. Gov'tinfo:eu-repo/semantics/publishe