Other fungal antigens.
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The transfer of delayed hypersensitivity to Coccidioides immitis and Candida albicans antigens with immunogenic RNA extracts was studied in a mouse model. Sensitivity was measured by skin tests and footpad swelling responses. Immunogenic RNA converted normal spleen cells in vitro so that they produced antigen-specific delayed hypersensitivity in mice that were given injections of the cells. RNase reduced the rate of, but did not abolish, in vitro interaction of immunogenic RNA extracts with lymphocytes. Immunogenic RNA transferred sensitivity on direct intraperitoneal inoculation into mice. The transfer ability was resistant to RNase preparations active against both single- and double-stranded RNA. Sedimentation gradient fractions of the immunogenic RNA were assayed by intraperitoneal injection, and converting activity was found in two fractions, greater than 33S and 6S-13S. After treatment with RNase, all activity was shifted to the less than 6S fraction. Two fractions of the immunogenic RNA in its native state (greater than 33S and 6S-13S) were also able to convert spleen cells. The data indicate that the transfer of delayed hypersensitivity by immunogenic RNA preparations is associated with RNA but may not require the intact RNA molecule.
In a mouse model, cell-mediated immunity to Coccidioides immitis, as assayed by the delayed hypersensitivity skin test, was transferred with whole immunogenic RNA extract and its greater than 33S and 6S-13S sedimentation fractions. Both fractions were cleaved by RNase, but the products retained their transfer activity. The greater than 33S fraction of immunogenic RNA extract was inactivated by pronase, whereas the 6S-13S fraction was resistant to the proteolytic enzyme; however, after RNase treatment the latter fraction was sensitive to pronase. This finding suggests a protective role for RNA. Dialysis of immunogenic RNA extract yielded a dialysate with a ratio of absorbance at 260 nm to that at 280 nm (A260:A280) of 1.02. Similarly, the dialysis product of RNase-treated RNA is active and has an A260:A280 ratio of 1.34. The data indicate that at least part of the active moiety of immunogenic RNA extracts is an RNA-associated, pronase-labile peptide or nucleopeptide. Furthermore, it is possible that the dialyzable transfer factor may be the same peptide or nucleopeptide cleaved from immunogenic RNA during preparation of the transfer factor.
Mycological and serological studies were carried out as part of a survey of respiratory disease in Scottish maltworkers. 70% of stained sputum smears from 574 workers showed the presence of higher plant cells and/or myclelia, and the spores of common environmental fungi. Penicillium spp. (90%), Rhizopus stolonifer (48%) and yeasts (53%) were the dominant fungi in 699 sputum cultures, and showed a similar proportional distribution in 327 samples of grain, malt, culms and dusts from fifty-six maltings. 57% of 711 men were serologically positive for fungi, 22% for Aspergillus fumigatus, 20% for A. clavatus, 10% for A. niger, 16% for Cladosporium herbarum and over 3% for Rhizopus stolonifer, 6% of 132 men were positive for Penicillium cyclopium. No precipitating antibodies to antigens from Alternaria tenuis, Aureobasidium pullulans, Candida albicans, Geotrichum candidum, Rhodotorula glutinis or Trichoderma viride were detected in tests of forty sera. Sera from the 5.2% of men with symptoms of extrinsic allergic alveolitis showed increased reactivity to mycelial antigens from Aspergillus clavatus. The fungus was cultured from 21% of maltings, 7% of all environmental samples and from the sputa of 8% of maltworkers.
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Non-specific positive reactions have been revealed in enzyme-linked immunosorbent assays (ELISA) of sera for the presence of fungal antigen. These false positives were recognized by their occurrence in tests for both Candida albicans and Aspergillus fumigatus antigens and by their response to dithiothreitol, combined with their reaction with non-immune rabbit globulin. A scheme is proposed which differentiates between true and false positive reactions. Use of fractionated anti-fungal globulin in conjugates reduced the incidence of false positive results in sera from hospitalized patients and eliminated them from sera of healthy subjects. The test scheme was applied to two panels of sera containing samples from patients with (a) invasive candidosis and (b) invasive aspergillosis. The relevance of ELISA tests for the detection of fungal antigen in human serum is discussed.
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Peripheral blood lymphocytes from 59 patients with dermatophytosis and from nine young healthy women were studied by the lymphocyte transformation test (LT) using mitogens and bacterial as well as fungal antigens. The latter included Candida albicans (CA) and four dermatophyte species, viz. Trichophyton rubrum (TR), Trichophyton mentagrophytes (TM), Epidermophyton floccosum (EF) and Microsporum canis (MF). Most of the patients showed normal transformation in response to mitogens and non-dermatophyte antigens, indicating that they have no functional T-cell deficiency. Dermatophyte antigens act as stimulators in LT. In general, patient lymphocytes responded more strongly to these antigens than lymphocytes from controls. In most patients suffering from TM infections, response to the TM antigen was significantly stronger (p less than 0.05) than that in the other patients, indicating that this antigen preparation shows species specificity. In patients with Trichophyton (TR + TM) infections, response to the corresponding antigens was significantly stronger than that in the other patients, which suggests the existence of genus specificity. Any differences between patients suffering from chronic TR infections and those with acute TR infections were not observed, a finding which is in contrast to those obtained in other studies. However, a few patients with chronic TM infections responded weakly to mitogens and non-dermatophyte antigens. LT in four patients with id-reaction to TM infection was not found to differ from that in the remaining TM patients.
Lymphocyte, granulocyte, and macrophage function were studied simultaneously in 26 untreated patients with systemic lupus erythematosus (SLE) and 26 matched controls. The overall cellular response, as assessed in vivo by skin tests and sensitization to dinitrochlorobenzene, was diminished in the SLE group. Although T cells were reduced in number in patients with SLE, their function appeared unimpaired, as shown by normal lymphocyte transformation to phytohemagglutinin and bacterial and fungal antigens. The depressed cutaneous reactivity observed in vivo was explained by the finding of major deficits in the efferent limb of cellular immunity. Initial rate of phagocytosis of both polymorphonuclear neutrophils and macrophages was significantly reduced in patients with SLE as compared with normal persons. Because of cell interaction, the presence of an intrinsic macrophage defect in SLE may contribute to the alleged T- and B-cell dysfunction in that disease.
We investigated the possibility of decreased cellular immunity (C.I.) in a substantial group of atopic asthmatic patients with respiratory tract infections during autumn and winter. Two-hundred and ten atopic asthmatic patients were selected, and the following tests for cellular immunity were performed. Skin tests with 11 bacterial and fungal antigens. Normal average value of the skin reactions is 2.5 mm. E rosettes with sheep erythrocytes. Normal value for patients of this age is 60 +/- 8%. Immunoglobulins A, G and M and complement fractions C3 and C4 were determined by radial immunodiffusion. IgE was determined using PRIST (Phadebas, Pharmacia). IgE was elevated in all except two of the patients. IgG, IgA and IgM, C3 and C4 were normal. Twenty five of the 85 patients had some depression in cellular immunity (decreased E rosettes and/or decreased skin reactions). Patients with depressed cellular immunity were given 2.5 mg Levamisol three times a week for three months. Their progress was evaluated clinically and immunologically and a positive correlation was found between clinical improvement and increased C.I. in 18 of the 25 patients. We consider Levamisol to be useful in approximately 8.5% of all asthmatic patients.
During primary Trichophyton mentagrophytes infection of strain 2 guinea pigs, the colony-forming units (CFU) of fungi present within the lesion peaked between days 7 and 14, whereas the severity of the lesion itself peaked between days 11 and 16. Concomitant with the latter peak, a pronounced depression in the in vitro mitogenic activity of spleen cells (SPC) and lymph node cells (LNC) was observed. Only after resolution of the primary infection (day 21) did LNC show increased deoxyribonucleic acid (DNA) synthesis in the presence of fungal antigens. During cutaneous reinfection, there was no distinct peak fungal load and CFU appeared to decrease steadily during the accelerated course of a reinfection disease. LNC from guinea pigs with severe, ulcerated reinfection lesions generally exhibited a heightened response to fungal antigen in vitro. LNC from guinea pigs with mild reinfection dermatophytosis had depressed in vitro reactivity to mitogens and dermatophyte antigen. The suppression of blastogenic activity during dermatophyte infection appeared to be associated with autologous serum components, since increased DNA synthesis resulted when SPC or LNC were cultured with fetal calf serum. The depressed in vitro DNA synthesis of lymphocytes (cultured with dermatophyte antigens) that were harvested during reinfection was not accompanied by an impaired ability of infected guinea pigs to respond with a delayed-type hypersensitivity skin test in vivo. These results support the hypothesis that experimental T. mentagrophytes dermatophytosis is a cell-mediated hypersensitivity disease that can be modified by immunosuppressive control mechanisms elaborated or induced by the fungus.
Circulating antigen of Aspergillus fumigatus was demonstrated in the sera of experimentally infected, cortisone-treated mice and rabbits by enzyme-linked immunosorbent assay (micro-ELISA), confirming earlier results where fungal antigen had been detected by counter-immunoelectrophoresis (CIE). Peaks of detection of circulating antigen by CIE and micro-ELISA in mice were not simultaneous suggesting that the nature of the predominant antigens may have altered during the course of infection. CIE failed to detect fungal antigen in infected rabbits whereas micro-ELISA monitored antigenemia until death. Both CIE and micro-ELISA demonstrated the rapid clearance of intravenously inoculated Aspergillus-antigen from the rabbit circulation.
Factors which set up an allergy of the digestive tract are alimentary antigens, fungal or bacterial antigens, drugs, among other things. The allergy is favored by achlorhydria, lack of pancreatic or duodenal enzymes or inflammations of the gastro-intestinal mucosa. The origin of a digestive allergy quite decisively depends on the A immunoglobulins. They can combine with high molecular weight antigens and so inhibit the transit of the active antigen. In the diagnosis, the authors have found a test particularly useful in which a few drops of antigen solution are administered sublingually. Therapy includes restoration of disturbed digestive functions, an individually planned diet and the avoidance of alcohol and strong spices.
Sera from 35 apparently normal humans, 37 compromised human patients, 30 hedgehogs and 30 sheep, were examined for precipitating antibodies to four opportunistic fungi - Absidia corymbifera, Aspergillus fumigatus, Candida albicans and Rhizopus arrhizus-using counterimmunoelectrophoresis (CIE). Precipitins to A. fumigatus were almost exclusively confined to specimens obtained from the compromised human group (51% of those examined) while Candida precipitating antibodies were detected in the sera of both normal (26%) and compromised (49%) humans and in 10% of the hedgehog specimens. Serum precipitins against the two phycomycetes included in the investigations were rare. Because of the complexity of most fungal antigen extracts, it appears the two phycomycetes included in the investigations were rare. Beause of the complexity of most fungal antigen extracts, it appears essential that sera be tested against a number of different antigen concentrations if CIE is to be used with confidence in fungal serology.
Although hypersensitivity has been postulated, the etiology of Sweet's syndrome has never been elucidated. In the present case intradermal tesing with viridans streptococcus led to an immediate reaction and reproduced a lesion clinically and histologically identical to that of the primary disease. Other bacterial and fungal antigens failed to elicit any response. These findings provide further support for the role of hypersensitivity in the pathogenesis of this disorder. It would appear that immediate or humoral immunit
Mycotoxicosis is a term used to define a toxic reaction due to the ingestion of toxins produced by fungi. Oral ingestion, however, may not be the sole means of exposure. We have recently observed ten patients who had inhaled massive amounts of fungi, which resulted in an apparent toxic pulmonary reaction. Immunologic studies showed no sensitivity to various fungal antigen preparations and histologic study of the lung showed a multi-focal acute process, with primary involvement of the terminal bronchioles containing large numbers of various spores. Cultures from lung biopsy material revealed at least five fungal organisms. A one to ten year followup indicates that avoidance of massive reexposure to fungal dust is the key to the prevention of recurrent pulmonary mycotoxicosis.