Gamma-glutamyl transpeptidase is up-regulated on memory T lymphocytes.

The ectoenzyme gamma-glutamyl transpeptidase (GGT) hydrolyzes glutathione (GSH), is required for the maintenance of normal intracellular GSH levels and modifies the activity of GSH-containing adducts. Previous data suggested that this enzyme was present on mitogen-activated T lymphocytes. However, the level of GGT protein expression on human mononuclear cell subsets has not been determined. A novel mAb to human GGT, 3A8, was developed. 3A8 was used to show that the expression of GGT is, in fact, highest on resting T cells that express markers of the memory phenotype, specifically CD45RO and decreased expression of CD45RB. The peripheral blood of patients with rheumatoid arthritis was found to have expanded numbers of T cells expressing levels of GGT up to 10-fold higher than controls. In addition, the CD4(+) T cell subset with the capacity to migrate across a human endothelial cell monolayer expresses high GGT levels. GGT expression was up-regulated on peripheral blood T cells following activation in vitro by either superantigen, phorbol ester, or IL-15, a stimulatory cytokine synthesized in rheumatoid synovium. Resting peripheral blood T cells that express GGT have higher levels of intracellular thiols than those that do not. These observations suggest that GGT may play an important role in the regulation of lymphocytes that are at a particular developmental stage.     

1141174724Complement activation induces dysregulation of angiogenic factors and causes fetal loss

Complement is an important effector in pregnancy loss in the antiphospholipid syndrome. We now test the hypothesis that complement activation is a necessary intermediary event in the pathogenesis of recurrent spontaneous miscarriage in DBA/2-mated female CBA/J mice (CBAxDBA), a well-studied model of autoantibody-independent pregnancy failure. Blockade of C3 activation with Crry-Ig completely rescued pregnancies in CBAxDBA mice (Crry-Ig vs untreated 8.5 ± 6.3% fetal resorptions vs 28.0 ± 7.2%, P <  0.01). Inhibition of C5 cleavage with anti-C5 mAb and blockade of C5a receptors with a peptide antagonist also prevented pregnancy loss (8.6 ± 4.4% and 8.0 ± 5.9% resorptions, respectively, P <  0.01 versus control). Inhibition of the alternative pathway resulted in pregnancy outcomes similar to controls (8.4 ± 6.6% versus 9.2 ± 3.2% resorptions). In CBAxDBA matings, we observed elevated plasma levels of soluble VEGF receptor-1 (sVEGFR-1; a potent anti-angiogenic molecule), increased placental inflammatory infiltrates and defective development of placenta. Our complement inhibitory strategies blocked the increase in sVEGFR-1, prevented functional deficiency of VEGF and rescued pregnancies. We confirmed the importance of complement as a proximal effector in in vitro studies; monocytes stimulated with C5a released sVEGFR-1 which sequesters VEGF required for normal placental development. Our data indicate that complement activation leads to recruitment of inflammatory cells and production of inhibitors of angiogenesis (sVEGFR-1) which cause placental dysfunction and fetal injury. These studies identify key innate immune effectors that mediate poor pregnancy outcomes and provide novel and important targets for prevention of recurrent pregnancy loss in patients.     

Effect on platelet properties of exposure to temperatures below 20 degrees C for short periods during storage at 20 to 24 degrees C

Background : When platelet concentrates (PCs) are shipped over long distances, it is not always possible to ensure that their temperature is maintained at 20 to 24°C. In addition, PCs are not agitated as during routine storage. Study Design and Methods : Studies have been conducted to evaluate how exposure to temperatures below 20°C in the absence of agitation influences properties of platelets. In initial studies, exposure to 4°C for 3 or 5 hours or to 12°C for 5 or 17 hours on Day 2 of a 5- to 6-day storage period was associated with a loss of discoid shape. This was reflected by slightly lower but statistically different morphology scores after storage compared to those observed with control platelets that were stored only at 20 to 24°C. In addition, a qualitative difference in morphology was noted in controls and PCs held at 16°C for 17 hours. In more detailed studies, both the in vivo viability and in vitro properties of platelets exposed between Day 1 and Day 2 to either 12°C or 16°C for 17 hours were evaluated. The protocol involved a paired study design (n = 4 for each exposure temperature) with the simultaneous storage of two identical PCs, one exposed to 12 or 16°C and the other one maintained at 20 to 24°C throughout the 5-day storage. Results : Exposure to 12°C significantly reduced (p < 0.05 by paired t test) the in vivo recovery to 37.6 ± 13.8 percent (mean ± 1 SD) from 47.8 ± 11.5 percent and the survival time to 2.0 ± 0.3 days from 6.5 ± 1.4 days. On exposure to 16°C, the differences in viability from those of control units were much less but still significant. The in vivo recovery was 42.7 ± 3.8 percent compared to 49.2 ± 3.0 percent and the survival time was 3.5 ± 1.2 days compared to 6.6 ± 0.3 days. The loss of in vivo viability of the test platelets was associated with a loss of discoid shape, as reflected by morphology scores, extent of shape change, and mean platelet volume. In addition, platelet metabolism also appeared to be affected, as suggested by increased lactate production. All of the in vitro properties except for total ATP and residual glucose that were statistically different from those of controls on exposure to 12°C were also significantly different on exposure to 16°C. Conclusion : These findings demonstrate that platelets undergo substantial changes in in vivo viability and in vitro properties when they are exposed to temperatures below 20°C for short periods.     

A blinded clinical comparison of MR imaging and CT in neuroradiology

The sensitivity and specificity of magnetic resonance (MR) imaging and computed tomography (CT) were compared in blinded readings of images of a consecutive series of patients with subsequently proved diagnoses. Overall, MR imaging was less sensitive than CT because of its lower sensitivity in detecting benign tumors. With a similar experimental protocol, the effects of technical refinements or contrast media on the sensitivity of MR imaging can be determined.     

Percutaneous transluminal dilatation of the iliac artery: long-term results

One hundred fifty-four patients with stenosis of the iliac artery underwent percutaneous transluminal angioplasty (PTA). These patients were followed for 1-7 years. The long-term results of the PTAs were analyzed by computer, and life tables were generated for dilatations of the iliac arteries with unimpaired flow and for those with an obstruction in the outflow tract. The accumulative 7-year patency rate was 90%, which agrees with other reports. This study demonstrates that the long-term results of PTA of iliac arterial stenoses are competitive with reconstructive vascular surgery. PTA should be the treatment of choice in patients with iliac arterial stenoses.     

Complement receptors and the shaping of the natural antibody repertoire

Complement and complement receptors have been known for several decades to play important roles in immune effector mechanisms related to pathogen elimination and tissue inflammation. In addition, studies over the last 10 years have clearly demonstrated a key role for the complement C3d activation fragment receptor designated CR2 (complement receptor type 2) in the switched-isotype, high-affinity and memory humoral immune responses to T-dependent foreign antigens. More recent studies have extended those observations to include a key role for CR2 and C3d in the humoral immune response to T-independent foreign antigens. Conversely, as these studies have proceeded, a parallel series of analyses have linked defects in expression or function of complement C4 and other classical pathway activation pathway proteins, as well as CR2 and the closely related CR1, to the loss of self tolerance to nuclear antigens such as double-stranded DNA and chromatin in systemic lupus erythematosus. With regard to the topic of this issue, it is now becoming increasingly clear that CR2 also plays a major role in the development of the natural antibody repertoire. Specifically, in the absence of this receptor natural IgM and IgG develop in the naïve animal that demonstrate clearly altered recognition patterns for specific natural antibody targets. This repertoire change is important physiologically in at least one setting because these CR2-dependent natural antibodies are necessary for the recognition of ischemic self tissues. In addition, it is possible that certain of the phenotypes manifest by CR2-deficient mice may be strongly influenced not only by effects on later stages of B cell activation and maturation, as commonly thought, but also by alterations in the pre-existing pool of natural antibodies that are influenced by this receptor. This review will examine the evidence that has accumulated over the last few years supporting these hypotheses.     

Acute tubular necrosis is characterized by activation of the alternative pathway of complement

BACKGROUND: Studies in animal models have shown that the alternative pathway of complement is activated in the kidney after ischemia/reperfusion. In addition, mice deficient in complement factor B, a necessary component of the alternative pathway, are protected from ischemic acute renal failure. The purpose of this study was to determine whether alternative pathway activation also occurs during the development of ischemic acute tubular necrosis in the human kidney. METHODS: Biopsies were identified from nine patients with morphologically normal kidneys and seven patients with evidence of acute tubular necrosis by light microscopy. Immunofluorescence microscopy was used to quantify and localize the complement activation products C3d and C4d. The results were correlated with available clinical data. RESULTS: Similar to mice, small amounts of activated C3d were present along the tubular basement membrane in normal kidneys. However, kidneys from patients with acute tubular necrosis had C3d complement deposition along a significantly greater number of tubules, and many of the tubules were completely circumscribed. In contrast, C4d was not detectable, indicating that complement activation occurred primarily via alternative pathway activation. CONCLUSION: Complement activation occurs in human ischemic acute tubular necrosis. As in rodents, complement activation along the tubular basement membrane after ischemia appears to occur principally via the alternative complement pathway. Because of this, an inhibitor of the alternative pathway might limit complement activation and inflammation after ischemia/reperfusion, thereby protecting the kidney from ischemic acute renal failure.     

The alternative pathway of complement in disease: opportunities for therapeutic targeting

The alternative pathway of complement is receiving increasing attention as a therapeutic target because of recent findings in several animal models that support its essential role in tissue injury and disease pathogenesis. Although the contribution of alternative pathway activation to serum complement activation in vitro is relatively modest, its role in generating activated pro-inflammatory fragments at extra-vascular sites is substantial. Several potential mechanisms might underlie this exaggerated effect, including local synthesis of alternative pathway components, disease-induced alterations of regulatory proteins, and influx of inflammatory cells that contain alternative pathway components into sites of injury. This review examines several animal models in which the alternative pathway is centrally involved in disease pathogenesis and which suggest a potential role for alternative pathway inhibitors as therapies for human disease. It is also expected that several clinically relevant studies will be presented at the XXth International Complement Workshop that will identify additional areas of interest with regard to this pathway.