In the last number of years siRNA has emerged as a key technique in understanding gene function. While siRNA has been used in lower organisms such as Caenorhabditis elegans, its use in mammalian cells, where gene manipulation is difficult, is where its greatest benefit has been realised. The advan ...

This chapter is intended as a handbook for anyone interested in using microarrays to study Toll-like receptor (TLR) function or any other biological question. Although microarray technology has developed into a standard tool at many laboratories disposal, most of the actual microarray ...

Forward genetics has led to many discoveries and particularly in the field of Toll-like receptors (TLRs), it has played an important role in identifying key components involved in the innate sensing of pathogens. With the mouse genome fully sequenced and the ability to generate many mutant phe ...

The mammalian protein–protein interaction trap (MAPPIT) is a two-hybrid technique founded on type I cytokine signal transduction. Thereby, bait and prey proteins are linked to signalling deficient cytokine receptor chimeras. Interaction of bait and prey and ligand stimulation re ...

Two-dimensional (2-D) gel electrophoresis concerted with protein identification by mass spectrometry (MS) is an extremely powerful method for comparative expression profiling of complex protein samples such as cell lysates. The highly resolutive 2-D electrophoresis allows ...

Purification of protein complexes and identification of the constituent components therein have been made relatively simple by the recent advances in proteomics. Uniting good biochemical and protein chemistry techniques with protein identification by mass spectrometry ( ...

Toll-like receptor (TLR) signalling involves five TIR adapter proteins, which couple to downstream protein kinases that ultimately lead to the activation of transcription factors such as nuclear factor-κB (NF-κB) and members of the interferon regulatory factor (IRF) family. TLRs pl ...

Although Toll-like receptor (TLR)-induced expression of several proinflammatory genes is required to provoke an efficient immune response, excessive or prolonged activation of TLR signaling can contribute to the development of septic shock and several inflammatory disease ...

Neutrophils are crucial components of our defence against microbial assault. They are short-lived cells, with regulation of their lifespan being a primary mechanism involved in the regulation of their function. Delay of apoptosis facilitates their clearance of pathogens, whilst a ...

Both ubiquitination and phosphorylation are crucial mediators involved in controlling the functions of numerous proteins belonging to the Toll-like receptor (TLR) signaling pathways. Altering the aforementioned post-translational events can be detrimental to the host su ...

Toll-like receptors (TLRs) are principal innate immune sensors critically involved in the recognition of evolutionary conserved microbial and viral structures called “pathogen-associated molecular patterns” (PAMPs). Although recognition patterns of many TLRs have been ...

Toll-like receptors (TLRs) and interleukin-1 receptor (IL-1R) play crucial roles in host innate immune response against microbial infections. These receptors share a conserved cytoplasmic domain, the Toll/interleukin-1 receptor (TIR) domain, which is required for signaling t ...

Continual advancements in computing power and sophistication, coupled with rapid increases in protein sequence and structural information, have made bioinformatic tools an invaluable resource for the molecular and structural biologists. With the degree of sequence inform ...

Toll-like receptor (TLR) ligand-binding domains comprise 18–25 tandem copies of a 24-residue motif known as the leucine-rich repeat (LRR). Unlike other LRR proteins, TLRs contain significant numbers of non-consensus LRR sequences, which makes their identification by computer dom ...

Protein–protein interactions regulate biological networks. The most proximal events that initiate signal transduction frequently are receptor dimerization or conformational changes in receptor complexes. Toll-like receptors (TLRs) are transmembrane receptors t ...

This chapter details the tools that are available to study Toll-like receptor (TLR) biology in vitro. This includes ligands, host cells, and readouts. The use of modified TLRs to circumvent some technical problems is also discussed.

Toll-like receptors (TLRs) are key regulators of the innate and adaptive immune response to bacterial, viral, and fungal pathogens. To date, 10 human TLRs and 13 mouse TLRs have been identified and they exhibit tissue-specific mRNA/protein expression patterns. Thus, it is essential that the T ...

Dendritic cells (DC) are the most potent antigen-presenting cells and form a promising new treatment modality. Fully activated DC loaded with antigen are very useful in stimulating immune responses, in particular those to combat cancer. Immature DC can either cause immunological tole ...

This chapter presents a detailed protocol for the generation of mature, monocyte-derived dendritic cells (DC) that are loaded or“pulsed” with tumor-associated peptide antigens for use as patient-specific immunotherapy. The protocol can easily be adapted for the treatment of patie ...

Recombinant adenoviral (rAd) vectors are highly suitable for efficient genetic modification of dendritic cells (DC). In certain cases, the high immunogenicity of rAd may be a disadvantage This chapter describes the essential aspects of optimal rAd-mediated gene transfer into DC, dis ...