Adrian M. Shields

Resolution‐associated molecular patterns (RAMP): RAMParts defending immunological homeostasis?

The resolution of inflammation is central to the maintenance of good health and immune homeostasis. Recently, several intracellular stress proteins have been described as having extracellular properties that are anti‐inflammatory or favour the resolution of inflammation. We propose that these molecules should be defined as resolution‐associated molecular patterns (RAMPs). RAMPs are released at times of cellular stress and help to counterbalance the inflammatory effects of pathogen‐associated (PAMPs) and damage‐associated (DAMPs) molecular patterns. We propose that heat shock protein 10 (HSP10), αB‐crystallin (αBC), HSP27 and binding immunoglobulin protein (BiP) should be considered founding members of the RAMP family. A greater understanding of RAMP biology may herald the development of novel immunotherapies.

UBE2L3 Polymorphism Amplifies NF-κB Activation and Promotes Plasma Cell Development, Linking Linear Ubiquitination to Multiple Autoimmune Diseases

UBE2L3 is associated with increased susceptibility to numerous autoimmune diseases, but the underlying mechanism is unexplained. By using data from a genome-wide association study of systemic lupus erythematosus (SLE), we observed a single risk haplotype spanning UBE2L3, consistently aligned across multiple autoimmune diseases, associated with increased UBE2L3 expression in B cells and monocytes. rs140490 in the UBE2L3 promoter region showed the strongest association. UBE2L3 is an E2 ubiquitin-conjugating enzyme, specially adapted to function with HECT and RING-in-between-RING (RBR) E3 ligases, including HOIL-1 and HOIP, components of the linear ubiquitin chain assembly complex (LUBAC). Our data demonstrate that UBE2L3 is the preferred E2 conjugating enzyme for LUBAC in vivo, and UBE2L3 is essential for LUBAC-mediated activation of NF-κB. By accurately quantifying NF-κB translocation in primary human cells from healthy individuals stratified by rs140490 genotype, we observed that the autoimmune disease risk UBE2L3 genotype was correlated with basal NF-κB activation in unstimulated B cells and monocytes and regulated the sensitivity of NF-κB to CD40 stimulation in B cells and TNF stimulation in monocytes. The UBE2L3 risk allele correlated with increased circulating plasmablast and plasma cell numbers in SLE individuals, consistent with substantially elevated UBE2L3 protein levels in plasmablasts and plasma cells. These results identify key immunological consequences of the UBE2L3 autoimmune risk haplotype and highlight an important role for UBE2L3 in plasmablast and plasma cell development.

Pro-resolution immunological networks: binding immunoglobulin protein and other resolution-associated molecular patterns

Appropriate regulation and subsequent resolution of acute inflammatory events is critical to the prevention of autoinflammatory diseases. Indeed, the chronic inflammation observed in diseases such as RA is at least partially consequent on the failure of endogenous immunoregulation. Current RA therapies (e.g. anti-TNF-α inhibitors and MTX) inhibit components of the inflammatory disease process without directly promoting the resolution of inflammation. We propose that the next generation of RA therapeutics will complement and augment endogenous immunoregulatory and pro-resolution immunological networks, thus promoting the definitive resolution of inflammation rather than temporary immunological control. Of particular interest with respect to this therapeutic approach is binding immunoglobulin protein [BiP; also known as glucose-regulated protein-78 (GRP78)], a member of the recently defined resolution-associated molecular pattern (RAMP) family of molecules. In this review, we consider the preclinical evidence from experiments in mouse and man that suggests BiP and other members of the RAMP family have the potential to herald a new generation of immunotherapeutics.

A new-age for biologic therapies: long-term drug-free therapy with BiP?

Heat shock proteins (HSPs) and other members of the much broader stress protein family have been shown to play important roles in coordinating multiple phases of immunological reactions; from facilitating immunological recognition, to promoting and regulating immunological responses and finally augmenting the resolution of inflammation and return to immunological homeostasis. In this review, we consider the challenges facing the stress protein field as we enter 2012; in particular we consider the role that HSPs and stress proteins may play in the initiation and termination of immunological responses. Special attention is afforded to the resolution-associated molecular pattern, binding immunoglobulin protein (BiP, also known as glucose regulated protein-78). We review the evidence that resolution-promoting proteins such as BiP may herald a new generation of biologics for inflammatory disease and reflect on the challenges of achieving clinical remission in rheumatoid arthritis with novel therapeutics and correlating clinical remission with immunological parameters of resolution of inflammation.

Improved monitoring of clinical response in Systemic Lupus Erythematosus by longitudinal trend in soluble vascular cell adhesion molecule-1

BackgroundTo determine whether optimal use of serial measurements of serum levels of soluble cell adhesion molecules (CAM) can improve monitoring of disease activity in SLE.MethodsSerum levels of soluble CAM and conventional SLE biomarkers were measured in serial samples (n = 80) from 21 SLE patients during and after flare and correlated in longitudinal analysis with disease activity determined by ECLAM score. Blood samples from a second cohort of 34 SLE patients were subject to flow cytometry to correlate serum biomarkers with B cell subsets.ResultsBy adjusting for the baseline level (at the first visit), delta soluble vascular cell adhesion molecule-1 (sVCAM-1) showed stronger correlation with changes in ECLAM score and improved sensitivity and specificity for identifying SLE responders versus non-responders compared to conventional SLE biomarkers including anti-dsDNA antibody titre and complement C3. Multiple regression analysis identified delta sVCAM-1 as the best marker of SLE clinical response. sVCAM-1 levels were significantly correlated with CD95+CD27+ activated memory B cells, CD95+ plasmablasts and circulating plasma cell numbers in SLE patients.ConclusionSubtracting a baseline level of sVCAM-1 for each individual substantially improved its utility as a biomarker. Delta sVCAM-1 was superior to conventional SLE biomarkers for monitoring changes in disease activity. This suggests that serial monitoring of serum sVCAM-1 trends should be considered in SLE patients to document responses to treatment. We hypothesise that the correlation between activated B cell subsets and circulating plasma cell numbers with soluble VCAM-1 serum levels in SLE may relate to the important role of VCAM-1 in B lymphocyte survival and maturation in bone marrow and secondary lymphoid tissues.

Systemic gene transfer of binding immunoglobulin protein (BiP) prevents disease progression in murine collagen-induced arthritis

Summary

Right ventricular metastasis of transitional cell carcinoma of the renal pelvis: successful single stage surgical treatment.

A 58-year-old female presented with symptoms mimicking infective endocarditis and was diagnosed with a right ventricular metastasis from a transitional cell carcinoma (TCC) of the left renal pelvis. The patient was treated with concurrent removal of the cardiac tumour and radical left nephrectomy followed by adjuvant gemcitabine-cisplatin chemotherapy. To our knowledge, this is the 14th report of cardiac metastases from TCC and the only case where one-stage surgical management of primary and cardiac metastases from TCC has been successfully completed.

A2.10 SLE associated UBE2L3 haplotype modulates plasma cell differentiation via genotypic regulation of NF-κB

Background and objectives Genome-wide association studies have identified a strong association between a single risk haplotype of the UBE2L3 gene and systemic lupus erythematosus (SLE). UBE2L3 is an E2 ubiquitin-conjugating enzyme that critically regulates ubiquitination by the linear ubiquitin chain assembly complex (LUBAC) with consequent effects on NF-kB signalling and inflammatory responses. Herein, we dissect the role of UBE2L3 in regulating NF-kB signalling in human B-cells and monocytes and demonstrate the risk haplotype drives plasmablast and plasma cell expansion in patients with SLE. Materials and methods UBE2L3 genotype data from GWAS in SLE was imputed using 1000 Genomes reference data. UBE2L3 function and signalling pathways were studied in vitro in HEK293 cells, or ex vivo using B cells and monocytes from healthy individuals or SLE patients (NF-kB translocation by Imagestream, multicolour flow cytometry of B cell subsets) stratified by UBE2L3 genotype. Parameters of SLE disease activity were collected and correlated with flow cytometry results. Results Data from SLE GWAS, imputed to 1000 Genomes level identified rs140490 as the most strongly associated UBE2L3 SNP, located at -270bp of the promoter region (p = 8.6 × 10–14; OR 1.30, 95% CI: 1.21–1.39). Microarray/western blot studies found that the rs140490 risk allele correlated with increased UBE2L3 expression in human B cells and monocytes. Overexpression of UBE2L3 in combination with LUBAC in HEK293-NF-kB reporter cell lines led to marked upregulation of NF-kB activity, which was abolished by a dominant-negative mutant UBE2L3[C86S]. RNAi blockade of UBE2L3 antagonised TNF signalling by inhibiting IκBα processing. UBE2L3 expression was 3–4-fold elevated in peripheral blood plasmablasts and plasma cells (p < 0.0001), with increased UBE2L3 expression in plasma cells from SLE patients compared to controls (p = 0.01). The T/T genotype at rs140490 was associated with a significant expansion in plasmablast and plasma cell populations in SLE patients (both p < 0.001), and showed a trend to correlation with increased SLEDAI scores (p = 0.06). Imagestream analysis demonstrated that rs140490 genotype correlated with both basal NF-kB activation in healthy individuals, as well as the sensitivity of NF-kB to CD40 stimulation in B cells and TNF stimulation in monocytes. Conclusions The UBE2L3 risk haplotype exerts a critical rate-limiting effect on TNF and CD40 signalling through regulation of LUBAC and NF-kB activation. This is the first demonstration that a complex trait variant at UBE2L3 regulates both basal NF-kB activation and sensitivity of NF-kB to stimulation in ex vivo human cells, resulting in accelerated B cell differentiation in SLE.

Treatment of collagen-induced arthritis with lentiviral BiP improves clinical parameters of disease

Rheumatoid arthritis (RA) is a chronic autoimmune disease manifesting primarily as an inflammatory polyarthritis of the synovial joints. Gene therapy has the potential to permit the delivery of therapeutic genes specifically to sites of rheumatological pathology through the specific transfection or transduction of joint-trafficking cells or via transcriptional targeting, thus minimising systemic immunosuppression. Immunoglobulin binding protein (BiP) is a 78 kD endoplasmic reticulum (ER) stress protein with multiple intracellular roles regulating the ER stress response. At times …