ADAM10 and Notch1 on murine dendritic cells control the development of type 2 immunity and IgE production.

BACKGROUND: Allergy and allergic asthma are significant health burdens in developed countries and are increasing in prevalence. Dendritic cells (DCs) initiate immune responses to common aeroallergens, and ADAM10 has been demonstrated to be important for the development of adaptive responses. This study's objective was to understand the role of ADAM10 on DCs in the development of allergic and anaphylactic responses. METHODS: In this study, we used mouse models of allergic airway inflammation (house dust mice and Alternaria alternata) and OVA-induced models of active anaphylaxis to determine the DC-specific function of ADAM10 and Notch signaling. To examine TH 1 and TH 17 immunity infection with Anaplasma phagocytophilum and Citrobacter rodentium respectively, were used. RESULTS: Mice, which have ADAM10 deleted from DCs, have dramatic reductions in IgE production and do not develop significant TH 2 immune responses. Further, ADAM10DC-/- mice are resistant to IgE-mediated anaphylaxis. This response is selective for TH 2 immunity as TH 1 and TH 17 immunity is largely unaffected. Notch1, a known ADAM10 substrate, when knocked out of DCs (Notch1DC-/- ) demonstrated a similar reduction in anaphylaxis and IgE. Without ADAM10 and Notch1 signaling, DCs were unable to make cytokines that stimulate TH 2 cells and cytokines. Anaphylaxis and allergic lung inflammation were restored in ADAM10DC-/- with the overexpression of the Notch1-intracellular domain, confirming the role of Notch signaling. CONCLUSIONS: Targeting ADAM10 and Notch1 on DCs represent a novel strategy for modulating TH 2 immune responses and IgE production.

Placental alkaline phosphatase in patients with seminoma measured by ELISA as compared to RIA.

Placental alkaline phosphatase (PLAP) has been shown to be a reliable tumor marker in the management of patients with seminoma. Radioimmunoassay (RIA) and Enzyme linked immunoassay (ELISA) procedures were compared for PLAP levels in serum of seminoma patients. The statistical analysis showed excellent correlationship between the two. It is evident from the results that performance of ELISA in terms of sensitivity specificity, precision, accuracy and reproducibility is equivalent to or better than RIA.

Total leukocyte acid phosphatase and its isoenzymes in patients with leukemia.

Leukocyte acid phosphatase and its isoenzyme composition was studied in leukemic patients to determine the specificity of different isoenzymes in leukemic leukocytes. It was found that leukocyte acid phosphatase content is significantly increased in ALL, AML, and CML patients, while CLL patients had decreased levels of acid phosphatase. The distribution and intensity of leukocyte ACP isoenzymes vary in respective leukemic condition. Thus isoenzyme 'O' was predominant in AML and CML, while isoenzymes 1, 2 and 3 predominated in ALL. The lack of predominance of isoenzyme 3 was a feature in CLL patients. It was concluded that the isoenzyme patterns, though promising, presented inconclusive picture for diagnosis purpose and further studies on immunochemical characteristics of these isoenzymes are warranted to ascertain their cell specificity.

Long term survival in a patient with multiple myeloma. A case report with review of literature.

A 37 year old man with symptomatic multiple myeloma diagnosed in April 1968 presented with generalised bony pains, extensive skeletal osteolytic lesions, monoclonal gammopathy and 90 percent atypical plasma cells in the marrow. He was given cyclophosphamide for six months with minimal response and then initiated on melphalan for one year. He was asymptomatic for 15 years thereafter and presented again in 1985 with a relapse of the disease. Over the next four years he was given various combinations of chemotherapy including cyclophosphamide, vincristine, melphalan and carmustine. He responded well on two more occasions only to relapse again. Recently, he has developed a symptomatic relapse with 36 percent plasma cells. This case report highlights the fact that there is a subset of patients with myeloma who survive beyond ten years, but remain symptomatic and respond slowly to chemotherapy.

Risk of hypoparathyroidism after surgery for carcinoma of the thyroid.

The reported incidence of postoperative hypoparathyroidism varies from 3% to 32%, and many surgeons are reluctant to perform a total thyroidectomy because of the risk of permanent damage to the parathyroid glands. We prospectively studied three groups of patients for evidence of acute parathyroid damage: group A consisted of 29 patients with total thyroidectomies; group B, 21 controls with partial thyroidectomies; and group C, 29 patients with total thyroidectomies that had been performed several months to years earlier. Functional status of the glands was measured by serum parathormone, calcium, and phosphorus levels. Although these levels fell postoperatively, they remained within normal limits. None of the long-term follow-up patients (group C) was found to have hypoparathyroidism, leading us to conclude that this risk may be overstated in the literature.

Nutrition and esophageal cancer.

Several case control studies in esophageal cancer employing a food-frequency dietary questionnaire suggested a fairly uniform dietary pattern which could be implicated in the causation of the disease. Although various micronutrients (vitamins and trace elements) have also been found to have either a positive or negative association, findings were more clear-cut for the different food items contributing the micronutrients than for the specific micronutrients themselves. The difficulties encountered in dietary interviews in case control studies are discussed. Nutritional supplementation has a theoretical rationale, but clinical experience with randomized trials did not support the same.

Clinical significance of serum spermine in breast cancer.

The diagnostic significance of three polyamines (putrescine, spermidine and spermine), of carcinoembryonic antigen and of phosphoglucose isomerase have been compared in sera of patients with breast cancer or benign breast disease and normal age matched controls. The results of the study indicate that the performance of spermine was more striking than that of any of the other markers. The estimation of spermine may prove to be a valuable parameter not only in detection but also in prediction of recurrence of the disease.

Terminal deoxynucleotidyl transferase in acute leukemias by direct immunofluorescence.

TdT (terminal deoxynucleotidyl transferase) can be detected by radio enzymatic assay, biochemical assay in cell extracts, serum or plasma, and intracellularly in the smear by indirect immunofluorescent methods. The IgG fraction of anti-TdT serum is conjugated with fluoresceinisothiocyanate and used directly on the cytospin smears of methanol fixed bone marrow/blood smears. The mice thymocytes and peripheral mononuclear cells of healthy donors were used as positive and negative controls, respectively, for TdT. 64% of our cases of ALL were found to be TdT+. The lymphoblasts of L1 morphology (FAB classification) were more frequently positive for TdT as compared to blasts with L2 morphology. 71% of our cALLa positive blasts in acute lymphoblastic leukemias were TdT+ve as compared to 58% of T-ALL blasts. 75% of PAS positive ALL cases were positive for TdT as well. Only 57% of the cases when acid phosphatase showed unipolar positivity (T type) were positive for TdT. 12% of cases with acute myeloid leukemia (6/47) were TdT+ve and 33% of CML in blastic crisis had TdT+ve blasts. Biochemical assay and IF assay for TdT were in good correlation in our study.

Clinical significance of heat labile Regan isoenzyme variant in Hodgkin's disease.

The activity of a heat labile variant of Regan type alkaline phosphatase (HLRI) was studied in the serum of 38 histologically proved patients with Hodgkin's disease. The activity was also studied in 40 normal healthy adults matched for age and sex as control. HLRI was significantly elevated in patients with Hodgkin's disease in comparison with normal controls (1.12 +/- 0.12 units/ml vs 0.09 +/- 0.01 units/ml; P less than 0.001). The mean values of the isoenzyme tended to increase progressively with increasing stages of disease, reflecting a significant linear trend (P less than 0.001). The serial measurement of this variant may serve as a useful parameter in assessing disease activity and response to chemotherapy.

Immunity in esophageal carcinoma.

In carcinoma of the esophagus, response to in vivo sensitization with recall antigens and DNCB was markedly depressed with 13% and 16% positivity respectively. Similarly, the number of T-cells was found to be significantly low (24 +/- 14) as compared to normal control (61 +/- 23). Blastogenesis index with PHA was only 1.75 +/- 1.04 in contrast to normal of 6.79 +/- 2.57. This depression was independent of serum albumin level and body weight. Cell-mediated immunity was further depressed following radiotherapy and did not improve following enteral alimentation for 3 weeks. In untreated patients, there was a significant rise in levels of IgA (298 +/- 184 mg/100 ml) as compared to normal (154 +/- 54 mg/100 ml). Levels of IgA did show a downward trend following enteral hyperalimentation. Circulating immune complexes and serum CEA level were elevated in almost 50% of patients. These data confirm the influence of tumor-related impairment of cell-mediated immunity while nutrition appears to affect IgA levels.

Detection of Regan variant type of alkaline phosphatase isoenzyme in liver tissue of Indian childhood cirrhosis.

Alkaline phosphatase (ALP) isoenzyme composition was studied in sera and liver from patients with Indian childhood cirrhosis (ICC). A typical pattern consisting of a fast-moving anodal preliver band with slower moving diffuse liver and placental bands followed by an intestinal band was consistently observed in sera of patients in all stages of ICC and in pregnant mothers of index ICC patients. ICC liver ALP was relatively heat-stable and inhibited by L-phenylalanine and L-leucine. The isoenzyme also had similar immunological determinants to placental ALP and adult intestinal ALP isoenzymes. Total serum ALP isoenzyme and its heat-stable component progressively increased in concentration from early to advanced stages of the disease suggesting that the diseased liver in ICC is the source of the abnormal isoenzyme.

Effect of activated charcoal on the pharmacokinetics of high-dose methotrexate.

The serum methotrexate (MTX) concentrations were estimated in two groups of patients after 6-hour iv infusion of 1 g/m2 of MTX and leucovorin rescue. One group (seven patients) received 25 g of activated charcoal orally at 12, 18, 24, 36, and 48 hours, while the other group served as a control (eight patients). Pharmacokinetic analysis revealed a significant reduction in the area under concentration curve from 18 hours until the serum MTX levels reached 10(-8) M. The MTX concentrations were significantly lower in the group that received charcoal than in the control group, at 18, 24, 36, 48, 60, and 72 hours from the end of infusion. First and second half-life values for the control and the charcoal-treated groups did not differ significantly. The data suggest that charcoal aids in the elimination of MTX from the body by interruption of the enterohepatic circulation of the drug. Activated charcoal did not cause any undesirable effects.

Occurrence of heat-labile Regan type of alkaline phosphatase in hematopoietic tumors.

The physiocochemical and immunological properties of alkaline phosphatase extracted from Hodgkin's nodes, non-Hodgkin's lymphoma nodes and leukemic leukocytes have been studied. The alkaline phosphatase from these three tumor types possesses the same biophysical and biochemical properties and immunological determinants as the placental alkaline phosphatase. However, it is more heat-labile than the placental isoenzyme. Immunological experiments indicate that, of these tumor types, Hodgkin's tumor contains the largest amounts of heat-labile Regan type of alkaline phosphatase.

Hepatitis B "e" antigen: an apparent association with lactate dehydrogenase isozyme-5.

Serums containing the "e" antigen of hepatitis B virus were subjected to electrophoresis in polyacrylamide gel. An extra band appeared in the lactate dehydrogenase isozyme pattern, but this band was undetectable in serums containing antibodies to the e antigenic determinant. Prior separation of the lactate dehydrogenase isozyme-5 fraction by chromatography of serum on minicolumns of diethylaminoethyl-Sephadex-A50 improved electrophoretic identification of the extra band. Neutralization with antibodies to the e antigen as well as by antibodies to the homologous d or y component of the hepatitis B surface antigen removed the extra band; antibodies to the lactate dehydrogenase isozyme-5 removed both the normal and the extra enzymatic band of isozyme-5. This feature of the e antigen provides an assay system for laboratory diagnosis of potential clinical usefulness and suggests its possible role in pathogenesis of hepatocellular injury.