The Cromer blood group system: a review.

The antigens of the Cromer blood group system reside on decay-accelerating factor (DAF), a protein belonging to the regulators of complement activation family. The blood group system consists of 12 high-prevalence and three low-prevalence antigens. The molecular basis for the antigens is known, and with the exception of IFC, each antigen is the product of a single nucleotide change in the DAF gene and has been localized to one of the four complement control protein (CCP) domains on the DAF protein. The RBCs of people with the Cromer null phenotype, Inab, lack DAF but do not appear to demonstrate increased susceptibility to hemolysis. Antibodies to Cromer antigens are rarely encountered, although there is evidence that the antibodies may cause accelerated destruction of transfused RBCs. There is no risk of HDN associated with Cromer system antibodies because the placenta is a rich source of fetally derived DAF, which is thought to adsorb the antibodies.

Sensitization to Empynase(pronase B) in exposed hospital personnel and identification of the Empynase allergen.

BACKGROUND: Empynase is a proteolytic enzyme that is widely used as an anti-inflammatory drug in Korea. We evaluated the prevalence of sensitization to Empynase in association with respiratory allergy symptoms in exposed hospital personnel, and identified the IgE-binding components in the Empynase extract, using sera with high levels of specific IgE antibodies. METHODS: A total of 154 hospital personnel (135 nurses and 19 pharmacists) who worked in a university hospital and 123 unexposed healthy control subjects were enrolled. A questionnaire was administered that addressed demographics, job category, history of atopic diseases, diverse symptoms including nasal and lower respiratory symptoms, and the association of symptoms with work. Skin prick tests (SPTs) to common aeroallergens and Empynase extract were performed. Empynase-specific IgE antibody was detected by ELISA, and ELISA inhibition tests were conducted. IgE-binding components were identified by SDS-PAGE and IgE immunoblotting. RESULTS: Forty-two subjects (27.3%) complained of work-related respiratory symptoms (WRRS). Five nurses (3.7%) and one pharmacist (5.3%) had work-related asthma symptoms, and 34 nurses (25.2%) and six pharmacists (31.6%) had work-related rhinitis symptoms. The prevalence of sensitization to Empynase on SPTs was 20.1%, and tended to be higher in pharmacists (31.6%) than in nurses (18.5%). It was estimated that 3.9-8.4% of hospital personnel had WRRS attributable to Empynase. The duration of exposure was longer in positive SPT responders than in negative responders (51.9+/-27.5 vs. 39.2+/-27.3 months, respectively; P<0.05), and the prevalence of Empynase-positive SPTs was significantly higher in subjects with asthma than in those without asthma (57.1% vs. 18.4%, respectively; P<0.05). The levels of Empynase-specific IgE antibodies were significantly higher in pharmacists (76.1+/-83.4 OD units) and nurses (56.3+/-103.0 OD units) than in normal controls (39.8+/-12.7 OD units; P<0.05). Seven subjects (two pharmacists and five nurses) had high serum levels of Empynase-specific IgE antibodies; six of these subjects had WRRS. ELISA inhibition tests were performed with the sera of these six subjects, revealing significant inhibition only with the addition of Empynase. Four strongly staining protein bands (sizes: 36, 33, 16, and 10 kDa) from Empynase extract were observed to bind to the IgE antibodies of sensitized subjects. Conclusion Exposure to Empynase powder may cause rhinitis and asthma in hospital personnel, and the pathogenic mechanism appears to be IgE mediated.

Protease activity in cockroach and basidiomycete allergen extracts.

Inherent proteolytic activity was estimated in cockroach and basidiomycete extracts by quantifying acid soluble peptides that were released by incubating extracts with 1% bovine serum albumin as measured by Lowry (Sigma). Reference proteases released 740 (Proteinase K, 0.1 U), 248 (Trypsin, 1.0 U), and 533 micrograms/ml (Pronase, 0.5 U) of soluble peptides. American whole body cockroach extract (0.1 mg dry weight) released 330 micrograms/ml of soluble peptides, representing 13 trypsin equivalent units (TEU)/mg. Extracts from spores of the mushroom Pleurotus ostreatus released 230 micrograms/ml (0.9 TEU/mg) and Pleurotus cap extract released 112 micrograms/ml (0.5 TEU/mg). Mycelium of Pleurotus and the mushroom Psilocybe cubensis and spores of Psilocybe and the puffball Calvatia cyathiformis showed negligible amounts of proteolytic activity. The protease inhibitor phenylmethylsulfonyl flouride reduced the proteolytic activity of American whole body cockroach extract by 80% (@1 mM) and the inhibitor ethylene diaminetetraacetic acid inhibited the proteolytic activity of Pleurotus spores by 95% (@1 mM). Loss of allergen activity as determined by RAST inhibition and immunoprinting correlated with protease activity. Thus, in the preparation and handling of allergen extracts, one should employ conditions that minimize proteolysis.

Isolation of monoclonal antibodies to chromatin and preliminary characterization of target antigens.

This paper describes the isolation of monoclonal antibodies to chromatin-associated protein antigens and their use in the characterization of such proteins by indirect immunofluorescence. Hybridomas were derived by fusion of the mouse myeloma Ag8653 with spleen cells from mice immunized with chromatin from human liver, rat liver or a human lymphoblastoid cell line. Hybrids were screened by solid-phase radioimmunoassay. The proportion of positive hybrids varied with the immunizing chromatin as follows: human liver 55/83, human lymphoblast 8/183 and rat liver 2/82. Fifteen antibodies derived from these fusions (7, 7 and 1 respectively) were subjected to further analysis. Most of these (11/13) were IgM and recognized both human and rat chromatin (12/15). Most of the target antigens were protease sensitive (8/13) and nuclease resistant. In fact the binding of five antibodies to lymphoblast chromatin was more than doubled by preincubation with DNAase I. The subcellular location of target antigens was examined by indirect immunofluorescence. Seven antibodies stained at least one of several cultured cell lines tested. Three gave staining patterns consistent with the in vivo association of the target antigen with chromatin recognizing, respectively, the interphase nucleus and metaphase chromosomes, the nuclear periphery and the mitotic spindle and other microtubule-containing structures. The remaining four all recognized antigens associated with the intermediate filament network.

Migration inhibition factor-like activity in inflammatory synovial fluids might be due to proteases.

Using an assay of macrophage migration, where the cells emigrate from an agarose droplet, it was found that the neutral proteases trypsin, chymotrypsin, Pronase and elastase have MIF-like activity. Appropriate enzyme inhibitors counteract this effect. To twelve synovial fluids from patients with inflammatory arthritis, which have MIF-like activity (migration index between 0-3 and 0-7) protease indhibitors (Trasylol, ovomucoid and soybean inhibitor) were added. Ten of the fluids lost some of their MIF-like activity with at least one inhibitor. Phenylmethylsulphonylfluoride counteracted totally the MIF-like activity of the two fluids tested. It is concluded that MIF-like activity of inflammatory synovial fluids is due, at least partially, to proteases.