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Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune disease primarily affecting the optic nerves and spinal cord, which is usually associated with anti-aquaporin-4 antibodies. Here, we present two individuals who were negative for anti-aquaporin-4 antibodies and were initially diagnosed with seronegative NMOSD. Each patient's clinical course and radiographic features raised suspicion for an alternative disease process. Both individuals were found to have pathogenic variants of MT-ND5 , encoding subunit 5 of mitochondrial complex I, ultimately leading to a revised diagnosis of a primary mitochondrial disorder. These cases illustrate the importance of biochemical and genetic testing in atypical cases of NMOSD.

The results obtained by Huntingdon Research Centre participating in a blind trial of the micromass assay for the prediction of teratogenic potential are presented. Twenty-five coded compounds were tested without S-9 mix using a pre-agreed protocol; three compounds were later tested with S-9. The data were assessed for sensitivity, specificity and accuracy using three separate sets of criteria based on either concentration (the <500 mug/ml rule (i) and the <50 mug/ml rule (ii)) or specific inhibition of cell differentiation at relatively non-cytotoxic concentrations (the 2-fold rule (iii)). The best in vivo/in vitro correlation was obtained using the 2-fold rule; the <500 mug/ml rule was the most sensitive but gave a high false positive rate and the <50 mug/ml rule was of low overall accuracy (60%). It is suggested that selective inhibition of differentiation of one cell type and cytotoxicity at low dose levels may also indicate risk of embryo-foeto toxicity, a factor to be considered with the pharmacokinetics of the compound. The teratogens procarbazine, methotrexate and caffeine were not detected; diphenhydramine and furazolidone initially classified as non-teratogens in vivo, were predicted as teratogens by the micromass assay.

Pyroglyphid mites in house dust are important allergens associated with asthma in Europe, but comparable studies of house dust mites in the homes of patients with asthma have not been done in the United States. We examined the distribution of mites and mite allergen in the houses of 20 mite-sensitive children with asthma in Atlanta and measured IgE antibodies to mite allergens in their sera. One or more dust samples from bedding, bedroom floor, television room floor, or television room furniture from 17/20 houses contained greater than 10,000 ng of antigen P1 equivalent per gram of fine dust; amounts ranged from 280 to 230,400 ng/gm. Allergen levels were higher in dust samples from furniture and bedding than from floors. Dust samples obtained from houses in June to September had more mites and mite allergen than those houses sampled in March to April; relative humidity in the room also was higher in June to September. Mite numbers and allergen in floor and furniture samples were correlated with relative humidity in the room and were high when relative humidity was greater than 50%; antigen P1 equivalent was greater than 10,000 ng/gm in 21/39 such samples. Dermatophagoides pteronyssinus was present in all houses and dominant in 11/20. D. farinae was found in 17 houses and was dominant in six. All children studied had high IgE antibody with either D. farinae or D. pteronyssinus RAST; 16 of the 20 children also had IgE to antigen P1. It is likely that the IgE antibody responses in these 20 children with asthma were a direct result of exposure to high levels of mite allergen.(ABSTRACT TRUNCATED AT 250 WORDS)

Monoclonal, two-site radioimmunoassays (RIAs) were developed to measure allergen Der p I of Dermatophagoides pteronyssinus or Der f I of D. farinae. Microtiter plates coated with monoclonal antibody (Mab) were incubated with mite extract, and bound allergen was detected with a second 125I-labeled Mab of different epitope specificity. The Mab RIAs were very sensitive (nanogram range) and highly specific. D. pteronyssinus extracts with different concentrations of Der p I demonstrated parallel binding curves, whereas a potent D. farinae extract demonstrated less than 5% of the Der p I binding in the same assay. Similar parallel curves were obtained with several D. farinae extracts in the Der f I assay, whereas D. pteronyssinus extract demonstrated little or no binding. The Mab RIAs were compared with an inhibition RIA that measured cross-reacting determinants on both Der p I and Der f I (antigen P1 equivalent [AgP1Eq]). The results demonstrated good quantitative agreement between these assays in commercial mite and house dust extracts (mean difference 1.57 +/- 0.5-fold). Thirty house dust samples with known mite counts, Der p I, and AgP1Eq content were also compared. The summed Mab RIA values for Der p I and Der f I demonstrated a very good correlation with AgP1Eq values (r = 0.86; p less than 0.001) and with assessments of total mite-allergen content by RAST inhibition (n = 21, r = 0.77; p less than 0.001). Furthermore, in samples with more than 10 mites per 100 mg of dust, the Der p I: Der f I ratio closely correlated with the ratio of the two mites counted by microscopy (n = 15, r = 0.89; p less than 0.001). The Mab RIAs can measure allergen levels in mite or dust extracts without the need for purified allergen or affinity-purified antibodies and can readily be standardized. These assays will be useful in epidemiologic studies of allergic asthma, to assess patients' exposure to mite allergens, and the effects of avoidance regimens. Because of the long-term stability and reproducibility of the reagents, Mab-based assays for specific allergens will also play an important role in the standardization of mite and other allergen extracts.

The membrane characteristics and in vitro function of the circulating B cells from patients with late-onset hypogammaglobulinaemia (CVID) and from patients with sex-linked hypogammaglobulinaemia (XLA) have been investigated using ox red blood cells coated with goat anti-human Ig. All seven patients with XLA had less than 2.0% SmIgM + ve cells and attempts to isolate these cells were unsuccessful. The majority of the 27 patients with CVID had greater than 3.0% SmIgM + ve cells. When cultured either with normal T cells and pokeweed mitogen (PWM) or with Epstein-Barr virus (EBV), isolated SIg + ve cells from eight patients with CVID produced significantly less IgM than normal cells and very little IgG or IgA. They also proliferated less well than normal cells and this correlated with IgM production. Stimulator capacity in the mixed lymphocyte reaction was normal in four out of seven cases. These properties of SIg + ve cells in the blood of patients with CVID are very similar to those of the normal 'immature' B cells found in adult bone marrow.

Most patients with late-onset bypogammaglobulinaemia have normal numbers of B cells. In addition, some of these patients have been shown to have circulating T cells which suppress immunolobulin production in vitro. From our present studies on a large series of patients it appears that these two findings distinguish two separate groups of patients. The majority of the patients have circulating B cells which are 'immature' in the sense that they produce IgM but very little IgG or IgA in vitro. These patients also fail to produce IgA in vivo but their T cells show normal function in vitro and normal T cell markers. Patients with the second form of the disease, including those with associated thymoma, have very few circulating B cells, and relatively preserved IgA production in vivo. In our series it was only this minority of patients whose T cells showed abnormal markers and increased suppressor activity in vitro.

Patients with asthma may develop acute symptoms after exposure to domestic or laboratory animal allergens; however, they are usually not aware of a direct relationship between their acute attacks and exposure to pollen or dust mite allergens. The present experiments were designed to study whether the differences in symptoms could be explained by differences in the number or size of particles carrying airborne allergens. Airborne particles were collected with a filter or on the stages of a cascade impactor, and allergens were measured by use of inhibition radioimmunoassays. In rat rooms and during disturbance of rat litter, a large proportion of rat urinary allergen (45.9%) was collected on the second stage of the impactor (mean size approximately 7 microns diameter). When sampled 15 to 35 minutes after disturbance, 16% of these medium-sized particles were still airborne. By contrast, during disturbance of house dust, a significantly larger proportion of dust mite, antigen P1 (80.6 +/- 11.8%; p less than 0.001) was collected on the first stage of the impactor, and in keeping with the apparent size of these particles (diameter greater than 10 microns), very little of this allergen (less than 4%) was still airborne when sampled 15 to 35 minutes after disturbance. With nebulized diluted rat urine, approximately 75% of the allergen was collected on the fourth and final stages of the cascade impactor in keeping with the expected size, 0.5 to 3 microns in diameter. These results demonstrate that natural exposure to both allergens is strikingly different from the conditions used for bronchial provocation.(ABSTRACT TRUNCATED AT 250 WORDS)

To study the long-term effects of avoiding domestic allergens, nine asthmatic patients who were allergic to dust mites lived in hospital rooms for two months or more. In all patients symptoms and early morning peak flows improved. In seven patients anti-asthma treatment could be reduced and it was possible to carry out repeated bronchial provocation with histamine. Five of these patients showed a progressive eightfold or greater increase in the concentration of histamine necessary to provoke a 30% fall in forced expiratory volume in one second (PD30). The increase in PD30 in the seven patients during their period of living in hospital was highly significant. Avoidance of important allergens seems not only to result in clinical remissions but in many cases also reduce bronchial hyperreactivity.

In order to study seasonal variation in dust-mite allergen, we obtained dust samples from bedding, carpet, and/or sofas in 12 houses in central Virginia, monthly, for 1 year. The houses included those of nine patients with asthma of whom six were allergic to dust mites. Dust samples were assayed with an inhibition radioimmunoassay for mite allergen that detects cross-reacting determinants on Der f and Der p I from Dermatophagoides farinae and D. pteronyssinus, respectively. The results are expressed as micrograms of antigen P1 equivalent (AgP1Eq). The results demonstrate that large seasonal variations in allergen, i.e., more than twentyfold, can occur in dust from all sites and are not restricted to the houses of allergic patients. However, dust from some sites, particularly sofas, remained "high" (greater than 10 micrograms AgP1Eq per gram), whereas dust from other sites remained "low" (less than 1 microgram AgP1Eq per gram) throughout the year. Levels of mite allergen generally started to rise in July about 1 month after the rise in humidity. In August to December, the mean levels of AgP1Eq in house dust were highly significantly increased relative to April to May. In keeping with this finding, in 31 of 37 sites, the highest level for the year was observed in August through December. In four sites, mite bodies were counted, and the numbers increased sharply in June to July; however, they decreased in September in parallel with falls in humidity but several months before the fall in mite allergen. Ryegrass-pollen allergen in 12 sites was also assayed in house dust, and pollen-allergen levels demonstrated a sharp increase in May or June that fell back to preseason values within 2 months. Dust was also obtained from the houses of 50 patients with acute or severe asthma. The results on these samples suggest that mite-allergic patients are more likely to have attacks in the fall at a time when their houses have greater than 10 micrograms AgP1Eq per gram of dust. The magnitude of changes observed seasonally within individual houses and of differences between houses within a close geographic area suggests that interpretation of the relationship between allergic symptoms and mite-allergen exposure will require measurement of mite-allergen levels in individual houses.

A series of patients was identified who demonstrated immediate positive skin tests to intradermal Trichophyton extract. These skin responses did not correlate with other fungal skin tests and were present both in atopic and nonatopic patients. The individuals demonstrating positive immediate skin tests included patients with urticaria, angioedema, asthma, and/or rhinitis, as well as five of 34 normal control subjects. Most skin test positive individuals had a history of local fungal infection and clinical signs suggestive of fungal infection. By use of Trichophyton tonsurans extract linked to Sepharose as the immunosorbent, it was possible to measure IgE antibodies in 26/30 sera from skin test positive individuals. With strongly positive sera, RAST bound up to 30% of the radiolabeled anti-IgE added. The results confirm that most skin test positive individuals have IgE-mediated hypersensitivity to Trichophyton. These observations support the older view that absorption of dermatophyte allergen through the skin should be considered as a possible cause of allergic disease.

Two species of mites of the genus Dermatophagoides are common in house dust and make a major contribution to the allergen content of house dust. As judged by skin tests and the radioallergosorbent test, these mites, D. pteronyssinus and D. farinae, demonstrate extensive cross-reactivity. When the major allergen from D. pteronyssinus, antigen P1, was compared with the equivalent allergen from D. farinae, these two proteins were found to have both species-specific and common antigenic determinants. With specifically purified antibodies directed against the common determinants, we developed a radioimmunoassay for these antigen P1 equivalent proteins in mite extracts as well as house dust extracts. The quantity of allergen in dust from 63 houses (255 samples) was measured, and values ranged from less than 100 to greater than 100,000 ng/gm of fine dust. The correlation between antigen P1 equivalent (ng/gm) and the number of mites per gram of dust (identified by microscopy) was very good (r = 0.74; p less than 0.001). Furthermore, this correlation was not affected by the species of mite in the dust sample. Of 9243 mites identified, 95% were of the genus Dermatophagoides, and of these, 16.4% and 18% could be positively identified as D. farinae and D. pteronyssinus, respectively. When the assay for antigen P1 equivalent was applied to isolated components of mite cultures of both species, significant allergen was found in the mite bodies, cuticles, and their excreta (fecal particles). For the house-dust samples, less than 10% of the allergen measured could be explained by the allergen content of mite bodies. Our results demonstrate that a radioimmunoassay for antigen P1 equivalent can provide an accurate and simple means of assessing the quantity of mite-derived allergen in dust-mite extracts, house-dust extracts, or house dust.

Proliferation of peripheral blood T lymphocytes from unsensitized donors in response to fractions of Trypanosoma brucei was observed to be monocyte-dependent. The activated T cells displayed enhanced 'helper' activity, but no 'suppressor' activity during pokeweed mitogen (PWM)-induced immunoglobulin (IgM and IgG) synthesis, and were resistant to the inhibitory effects of theophylline on E rosette formation. Whilst in vivo studies have failed to reveal excessive T cell proliferation in patients with sleeping sickness, these results suggest a possible role for T cells in the induction of hypergammaglobulinaemia characteristic of this disease. Trypanosome fractions were not inhibitory to PWM-induced proliferation, and actually enhanced immunoglobulin synthesis. Thus it is unlikely that direct inhibition by the parasite per se is a major factor in the generation of immunosuppression by the T. brucei subgroup.