Diagnostic specificity of a sandwich ELISA for Aspergillus-related diseases

A sandwich ELISA with specificity for a major antigen (Ag 7) of Aspergillus fumigatus has been compared with indirect ELISAs with use of crude antigenic (culture filtrate) extracts and found to have a sensitivity of 100% and specificity of 92.3% for antibody detection in sera of patients with allergic bronchopulmonary aspergillosis (ABPA) and aspergilloma. In the Ag 7 ELISA, all sera from groups with ABPA (21) and aspergilloma (15) had positive titers, and mean values for both these groups were significantly higher (p less than 0.001) than control levels. By comparison, in the indirect ELISAs, most sera of patients with ABPA were positive; the sensitivity was 81% to 90%, and the specificity was 87%. Sera from other disease groups including sera from 13 patients with farmer's lung, 16 with tuberculosis, and 10 individuals with positive prick test to either A. fumigatus or Alternaria alternata were also tested by both types of assay. Up to five of these sera elicited significant positive results in the indirect ELISA, and although two sera were also positive in the Ag 7-specific ELISA, these latter titers were both below the range of values for the sera from patients with ABPA. With the use of an IgG calibration curve, the sensitivity of the assay was determined as within the range of 1 to 10 microgram of specific IgG antibody per milliliter. The Ag 7-specific ELISA is therefore a highly specific, sensitive assay for antibody detection in Aspergillus-related diseases.     

Truncated immunoglobulin Dmu causes incomplete developmental progression of RAG-deficient pro-B cells.

Early stages of B cell development are dependent on the expression of a pre-B cell receptor (BCR), composed of a mu heavy chain (HC) in association with surrogate light chain (SLC) proteins and the signaling molecules, Igalpha and Igbeta. During the formation of the variable region of the mu chain by somatic gene rearrangement, a truncated form of the mu protein (called Dmu) is sometimes produced by the rearrangement of a D(H) segment to a J(H) segment using one of three reading frames (designated rf2). When a Dmu protein is formed, subsequent B cell development is blocked by down-regulation of further HC rearrangements, so that a full-length muHC cannot be formed. In this study, we demonstrate that in recombinase activating gene (RAG)-2-deficient B220(+) CD43(+) pro-B cells in which B lymphopoiesis has been arrested at fraction C, transgenic expression of Dmu promoted partial developmental progression to fraction C', but was unable to mediate the pro-B to pre-B cell transition to fraction D effected by full-length muHC protein. These data suggest that the intracellular signaling pathways engaged by the Dmu pre-BCR are insufficient to facilitate the expansion and/or survival of pre-B cells, and are distinct from those engaged by the pre-BCR-containing full-length muHC.     

Interleukin 1 alpha acts as an autocrine growth factor for RPMI 1788, an Epstein-Barr virus-transformed human B cell line

The Epstein-Barr virus-transformed B cell line RPMI 1788 constitutively produces autocrine growth factors with molecular masses of 17 kDa, 24 kDa and 35 kDa. All three molecular forms were completely neutralized with anti-interleukin (IL) 1 alpha antiserum. Although IL 1 alpha and IL 1 beta mRNA were both equally detectable by Northern blotting, no IL 1 beta activity was found in partially purified RPMI 1788 supernatant. The growth of low density-seeded RPMI 1788 cells is specifically dependent on the presence of either IL 1 alpha or IL 1 beta. Since no other cytokine was found to be capable of sustaining proliferation, this cell line is suitable for the identification and quantification of IL 1, even in the presence of other cytokines.     

Ex vivo expansion and prophylactic infusion of CMV-pp65 peptide-specific cytotoxic T-lymphocytes following allogeneic hematopoietic stem cell transplantation.

Cytomegalovirus reactivation and infection post-allogeneic hematopoietic stem cell transplant continue to cause morbidity and mortality. Current pharmacologic therapies are limited by side effects. Adoptive transfer of ex vivo generated cytomegalovirus-specific T cells has the potential to restore immunity, prevent cytomegalovirus, and circumvent the need for pharmacologic therapies. We have generated donor-derived cytomegalovirus-specific cytotoxic T cells using dendritic cells pulsed with the HLA-A2 restricted nonapeptide NLVPMVATV (NLV) derived from the cytomegalovirus-pp65 protein. These cytotoxic T cells have been given prophylactically to 9 recipients aged 4 to 65 years on or after day 28 post-allogeneic hematopoietic stem cell transplant. Only 2 of 9 recipients received T cell depletion in vivo or in vitro. There were no immediate adverse reactions to the infusions. During 97-798 days of follow-up, 2 recipients developed cytomegalovirus reactivation; neither developed cytomegalovirus disease or required pharmacotherapy. Three recipients developed acute graft versus host disease after infusion. Two recipients died, 1 from thrombotic thrombocytopenia purpura secondary to cyclosporine, 1 from complications of graft versus host disease. A transient increase in numbers of cytomegalovirus-specific T cells demonstrated by NLV-tetramer binding was seen in 6 recipients. Prophylactic adoptive transfer of NLV-specific T cells is safe and may be effective in preventing cytomegalovirus reactivation.     

Tetanus toxin-induced seizures cause microglial activation in rat hippocampus

Tetanus toxin (about 20 mouse LD50) injected into the ventral hippocampus of rats leads to brief seizures occurring intermittently over a period of weeks. Toxin injection leads to the appearance of activated microglia (detected with OX42 immunohistochemistry) in the hippocampus. After 7-14 days, many activated microglia are visible in CA1 area of dorsal hippocampus aligned with the pyramidal cell dendrites and having the morphology characteristic of 'rod cells'. Extensive cell loss is found in dorsal CA1, but not at the injection site, in about one third of injected rats.     

Involvement of 2p23 in pulmonary inflammatory pseudotumors

Pulmonary inflammatory pseudotumors (IP) are rare mesenchymal proliferations that have a polymorphic histology and an unpredictable biologic behavior. The histologic spectrum of IP has led to uncertainty as to whether this tumor has a reactive or neoplastic pathogenesis. Reports of extrapulmonary IP have identified clonal chromosomal aberrations involving 2p23 in the region of the ALK gene. Using fluorescence in situ hybridization with a probe flanking the ALK gene at 2p23 and immunostaining for the ALK gene product, we studied formalin-fixed, paraffin-embedded tissues of pulmonary IP and found a subset (33%) with 2p23 aberrations. We suggest that chromosomal rearrangements and ALK immunostaining may be helpful in the diagnosis of a group of pulmonary IP and should be investigated as a potential tool for predicting their future biologic behavior. An association with anaplastic large-cell lymphoma was also observed. HUM PATHOL 32:428-433.     

An upper limit to the number of sodium channels in nerve membrane?

1. A small volume of artificial sea water containing 300 nM tetrodotoxin (TTX) was applied successively to seven lobster nerve trunks and the cumulative uptake of toxin investigated by bio-assay.2. Light and electron microscopy indicated that the nerve trunks had a total axonal area of 0·7 × 10⁴ cm²/g.3. Sodium analysis gave a sodium space for the nerve trunks of 30%.4. The amount of toxin taken up by the cells in 1 g of nerve is less than 1·6 × 10^-11 moles.5. It is argued that there are probably fewer than 13 sodium channels/μ² axon in lobster nerve.     

Alloactivated long-term cultured human T lymphocytes express both HLA-DR and SB antigens but lack lymphocyte stimulation capacity

Cell populations obtained from mixed leukocyte cultures of 6- or 10-day duration were found specifically to restimulate primed lymphocytes detecting HLA-linked SB as well as HLA-D-associated antigens. After expansion in vitro (9-75 days) with medium containing interleukin 2, the cultured cells expressed the T lymphocyte markers detected in indirect immunofluorescence by monoclonal antibodies Lyt-3, OKT3, OKT4, OKT8, and had high levels of HLA-DR antigens. In addition, they were shown in cell-mediated lymphocytotoxicity specifically to express SB antigens of the donor B cell type. Despite their positivity for DR and SB antigens, such cultured T cells failed to restimulate either SB- or D-specific secondary lymphocyte proliferation. Homogeneous cloned populations of cultured T cells also lacked lymphocyte stimulation capacity. In contrast, B cell lines, which also expressed DR and SB antigens, were potent stimulators of both SB- or D-directed proliferation. These data show that the activated T lymphocytes which express both HLA-DR and SB antigens are by themselves unable to stimulate lymphocyte proliferation.     

Blood concentrations and clinical findings in nonfatal and fatal intoxications involving glutethimide and codeine

Blood concentrations and clinical findings were evaluated in twenty-six nonfatal and twelve fatal intoxications involving the combination of glutethimide and codeine ("loads"). The mean glutethimide concentration was 10 +/- 5 mg/L for nonfatal cases (range 2-18 mg/L) and 13.9 +/- 6.6 mg/L for fatal cases (range 4.6-26.4 mg/L). The mean codeine concentration for fatal intoxications was 1.21 +/- 1.17 mg/L (range 0.13-4.32 mg/L). Codeine concentrations were not measured in cases of nonfatal intoxication. Nine nonfatal cases required hospitalization on a medical ward (mean length of stay 3 +/- 3 days). Depressed level of consciousness was the most common abnormal physical finding (24 cases); 18 patients were lethargic but arousable with nonpainful stimulation and 6 patients with serum glutethimide concentrations of 10 mg/L or greater were comatose. The level of consciousness showed statistically significant correlation with the glutethimide concentration (P less than 0.01). Twenty-four nonfatal intoxications involved at least one other drug in addition to glutethimide and codeine (salicylates in 12 and acetaminophen in 4), while only 7 fatal cases involved at least one additional drug (acetaminophen and diazepam in 3 each). The finding of glutethimide should prompt a search for codeine and vice versa, especially when the presence of either does not in and of itself explain the clinical condition of the patient.