Minimal residual disease detection with tumor-specific CD160 correlates with event-free survival in chronic lymphocytic leukemia.

In chronic lymphocytic leukemia (CLL), the detection of minimal residual disease (MRD) correlates with outcome in the trial setting. However, MRD assessment does not guide routine clinical management and its assessment remains complex. We incorporated detection of the B cell, tumor-specific antigen CD160 to develop a single-tube, flow cytometry assay (CD160FCA) for CLL MRD to a threshold of 10(-4) to 10(-5). One hundred and eighty-seven patients treated for CLL were enrolled. Utilizing the CD160FCA methodology, there was a high level of comparison between blood and bone marrow (R=0.87, P<0.001). In a validation cohort, CD160FCA and the international standardised approach of the European Research Initiative on CLL group demonstrated high concordance (R=0.91, P<0.01). Patients in complete remission (CR) and CD160FCA negative had longer event-free survival (EFS) (63 vs 16 months, P<0.01) and prolonged time to next treatment (60 vs 15 months, P<0.001) vs MRD positive patients; with a median time to MRD positivity of 36 months. In multivariate analysis, CD160FCA MRD detection was independently predictive of EFS in patients in CR and even predicted EFS in the good-risk cytogenetic subgroup. CD160FCA offers a simple assay for MRD detection in CLL and gives prognostic information across different CLL risk groups.

Phosphorus concentration and loading reductions following changes in fertilizer application and formulation on managed turf.

Excess phosphorus, particularly in surface waters can lead to severe eutrophication. Identifying source areas, quantifying contributions, and evaluating management practices are required to address current and future water quality concerns. A before-after study was conducted from 2003-2010 on a sub-watershed of Northland Country Club Golf Course in Duluth, MN to demonstrate the impacts of two different phosphorus management approaches (Period 1: traditional application and timing using commercially available synthetic blends; Period 2: reduced rate, low dose applications, and organic formulations). Outflow median dissolved reactive phosphorus (DRP) and total phosphorus (TP) stream concentrations were significantly less in Period 2 compared to Period 1. There was no statistical difference in the mean TP loading in Period 1 (0.25 kg ha(-1) year(-1)) compared to Period 2 (0.20 kg ha(-1) year(-1)) or between the DRP loading in Period 1 (0.15 kg ha(-1) year(-1)) compared to Period 2 (0.09 kg ha(-1) year(-1)). However, by switching to organic phosphorus formulations and reducing application rates by greater than 75%, substantial reduction in DRP and TP concentrations was achieved. Based on these findings it is recommended that turf managers (parks and recreation to golf courses) explore the feasibility of altering their fertility management related to phosphorus by including organic formulations, low dose applications, and overall rate reductions. Additionally, it is recommended that the fertilizer industry develop and make more readily available commercial blends with lesser to zero amounts of phosphorus.

The risk of antimalarials in patients with renal failure.

We present here a patient with end stage renal failure who received two weeks antimalarial prophylaxis at full dose leading to life threatening toxicity with severe acute megaloblastic anaemia, symptomatic pancytopenia and exfoliative dermatitis. Prompt recognition and treatment can rapidly reverse these fatal effects but more importantly, education of patients before travel is imperative in preventing such events.

A complication of steroid therapy in acute leukaemia--a case report.

Common complications associated with steroid therapy are well documented. We report a rare and fatal complication, in which oesophageal erosion secondary to the use of steroids was associated with pneumopericardium.

Invasive pulmonary aspergillosis.

In susceptible patients, invasive aspergillosis has a high incidence and a mortality of up to 80%. The diagnosis of this condition is difficult, especially in the early stages of the disease and, as a consequence, antifungal therapy, despite its expense and toxicity, is often initiated empirically. Until recently, there were very few effective antifungal agents for established invasive aspergillosis, but the introduction of two new drugs, voriconazole and caspofungin, has increased the treatment options. These newer antifungal therapies, combined with improved early diagnosis due to the introduction of newer microbiologic techniques, offer the hope that there will be a significant improvement in the substantial morbidity and mortality associated with invasive aspergillosis over the next 5 years.

Multiple co-stimulatory signals are required for triggering proliferation of T cells from human secondary lymphoid tissue.

Vaccine-based therapies are being developed for a variety of cancers and their efficacy will be determined by their ability to stimulate T cells in the secondary lymphoid tissue. We found that T cells isolated from human secondary lymphoid organs (LT-T), in contrast to peripheral blood T cells (PB-T) are hyporesponsive to cross-linked anti-CD3 mAb (CD3c) even in the presence of exogenous IL-2. Using mAb to trigger CD2 and CD28 co-stimulatory molecules, we found that such dual co-stimulation of LT-T induces profound and sustained responses including CD25 expression, IL-2 secretion and proliferation. Different levels of co-stimulation produced a hierarchical pattern of responses in LT-T, which correlated with the degree of CD3-TCR down-regulation. Mature antigen-presenting cells (APC) restored the capacity of LT-T to proliferate to stimulation of the CD3-TCR complex. Blocking studies demonstrated that optimal proliferation was critically dependent on co-stimulation via CD2 and CD28 engaged by their ligands on the APC. Therefore, LT-T have increased co-stimulatory requirements as compared to PB-T, i.e. multiple co-stimulatory signals coupled to CD3-TCR triggering. Furthermore, LT-T were found to be dependent on APC for survival, in contrast to PB-T. Clearly, LT-T do not behave in a comparable way to PB-T and in vitro experiments assessing novel cancer vaccines should therefore use LT-T as the most appropriate population of responder T cells.

Adenoviral transduction of human 'clinical grade' immature dendritic cells enhances costimulatory molecule expression and T-cell stimulatory capacity.

The therapeutic use of dendritic cells (DC) in antigen-specific anti-tumor vaccines, requires sufficient numbers of functional DC, the preparation of which should comply with the code of Good Manufacturing Practice. In addition, the expression of tumor specific antigen should be possible in these DC. As a preclinical step, the method reported here was developed in healthy volunteers. Monocytes (Mo) were isolated by leukapheresis from 12 donors, purified by elutriation and then cultured for 6 days in sealed bags in AIM-V serum free medium with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-13 (IL-13). Between 6x10(8) and 1x10(9) immature DC (iDC) could be differentiated from one leukapheresis. Cells displayed a characteristic iDC phenotype (CD1a(+), CD14(-), CD80(+), CD86(+), HLA DR(+), CD83(-)), and had potent allogeneic and antigen dependent autologous T cell-stimulatory capacity. Moreover, iDC could be further differentiated into mature DC by CD40 ligation as assessed by CD83 expression and the upregulation of HLA-DR and costimulatory molecules. After infection with a recombinant adenovirus encoding for beta-galactosidase (betaGal), 50% to 80% of iDC expressed betaGal without toxicity. Adenovirus infection increased the expression of both costimulatory molecules and CD83, and also increased allogeneic stimulatory capacity. Thus, the method developed here allows us to use large numbers of functional iDC as will be required for therapeutic uses in man. These DC can express a transgenic protein.

Functional role of PECAM-1/CD31 molecule expressed on human cord blood progenitors.

CD31/PECAM-1 (platelet endothelial cell adhesion molecule-1) is a 130 kDa integral membrane protein of the immunoglobulin gene superfamily with the distinctive feature of being expressed on several cell types associated with the vascular compartment. In the present study we report a novel, unique CD31 mAb termed IP28A which reacts with all CD34 molecule expressing hematopoietic progenitor cells and a subset of T, B and NK lymphocytes from human cord blood. Interestingly, we show that the number of CFU-GM and BFU-E was significantly augmented in cord blood progenitor cultures when purified IP28A mAb was added to rhSCF plus rhGM-CSF and rhEpo, respectively. Thus, these results are of relevance in the field of hematopoietic stem cell transplantation as they reveal an agonistic property of the IP28A/CD31 mAb on the differentiation of cord blood progenitor cells.

The primary afferent depolarizing action of kainate in the rat.

Dorsal roots (L3-L7) isolated from immature (1-9 day old) rats were depolarized selectively by kainate (1-100 microM). L-Glutamate (25-100 microM), but not L-aspartate, mimicked the action of kainate. N-methylaspartate had no activity on these preparations and quisqualate was thirty times less active than kainate. Depolarizations evoked by L-glutamate (100-1000 microM) faded rapidly in the presence of L-glutamate. Depolarizations evoked by kainate were depressed during the fade induced by L-glutamate. Certain electrically evoked C-fibre volleys in dorsal roots or leg nerves of rats at any age were selectively depressed or abolished in the presence of kainate. The effect of kainate was more selective than that of gamma-aminobutyric acid or capsaicin. Prolonged treatment of dorsal roots with kainate did not appear to be deleterious to C-fibres. It is suggested that certain primary afferent C-fibres possess kainate receptors which may be activated physiologically by L-glutamate released at their central terminations.