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To establish a model of experimental hypersensitivity pneumonitis (EHP) in mice and to examine the influence of genetic background on the pulmonary inflammatory response to Micropolyspora faeni, we determined the responses of C57BL/6, SJL/J, and C3H/HeJ mice to intratracheal (i.t.) injections of M. faeni. Recipient animals received lymph node cells (LNC), peritoneal exudate cells (PEC), and spleen cells (SC) from sensitized mice cultured in vitro with M. faeni. Controls included serum containing anti-M. faeni antibody; uncultured SC from M. faeni-sensitized donors, and M. faeni-cultured SC from ovalbumin (OA)-sensitized donors. Recipients were challenged i.t. with M. faeni or normal saline 48 hr after the cell or serum transfer. We developed a model of EHP in mice. Increasing amounts of i.t. M. faeni were associated with increasing extent of pulmonary inflammation with no difference between the mouse strains. There was substantial increase of the extent of pulmonary abnormalities in the animals receiving cultured SC. The number of transferred cells and the M. faeni concentration correlated with the extent of pulmonary histologic abnormalities. Cultured PEC and LNC could transfer EHP in C3H/HeJ mice only. Serum containing anti-M. faeni antibody, cultured SC from OA-sensitized donors, and noncultured SC from sensitized donors could not transfer EHP. We conclude that it is possible to adoptively transfer EHP.

Cross-protection studies were conducted with vaccines prepared from two isolates of Dermatophilus congolensis (designated strain 1 and strain 2). The vaccines were prepared as either heat-inactivated, washed, formalized filamentous phase bacterium, mixed with alum as an adjuvant, and inoculated intramuscularly (type A vaccine) or sedimented live filaments inoculated intradermally (type B vaccine). The vaccinated sheep were challenged with D. congolensis zoospores of one or other strain. Challenge sites were observed for the presence and severity of lesions. Serum antibody levels to D. congolensis were monitored after vaccination and challenge. Type A and B vaccines from both strains produced some reduction in the severity of lesions when sheep were challenged with strain 1 but not with strain 2. Unvaccinated control sheep developed more severe and persistent lesions when challenged with strain 2 than controls challenged with strain 1. Serum antibody levels to the type B vaccine prepared from strain 1 were significantly higher (P less than 0.05) than antibody levels to type B vaccine from strain 2. These findings showed there was significant variation in virulence and antigenicity between these two isolates of D. congolensis.

Vaccination against Dermatophilus congolensis was carried out in groups of lambs raised on optimal or energy deficient diets. The groups differed significantly in weight, body condition score and plasma total protein and albumin. All animals were then challenged with D. congolensis in a dose response infection model. The vaccine was effective in the well nourished animals, reducing the number of affected lambs in the vaccinated group and the severity of the lesions and increasing the minimum dose required to cause infection. In contrast, all of the vaccinated energy-deficient lambs developed lesions. There was some evidence of vaccine effect in these animals but this was not as marked as that seen in the well nourished lambs. The malnourished lambs, vaccinated and non-vaccinated, took longer to heal than the well nourished groups. Resistance to challenge was not associated with serum antibodies or skin test reactivity to D. congolensis antigens.

Sera from 54 two- to three-year-old Thoroughbred horses from an English racing stable were examined for precipitins to antigen extracts prepared from 18 species of moulds (fungi and thermophilic actinomycetes) isolated from the same stable. Twenty-seven horses exhibited serum precipitins to one or more antigens; sixteen of the mould antigens elicited positive reactions in sera from one or more horses. Significantly more precipitins occurred in sera of those horses stabled in a barn than among those stabled in individual boxes. This indicated a possible association between type of housing, level of exposure to airborne moulds and presence of serum precipitins. None of the horses had overt respiratory disease. This study agrees with reports of the presence of serum precipitating antibodies to mould antigens in clinically healthy horses and confirms that serological tests, therefore, are of little value in the diagnosis of chronic obstructive pulmonary disease (COPD) or 'heaves'.

Intradermal vaccination of live crude filaments (vaccine A) was compared with a vaccine (vaccine B) consisting of a 45 kD zoospore protein and mucoid material coating filaments in its ability to protect sheep from experimental Dermatophilus congolensis infection. Fourteen and 21 days after challenge, vaccine A sheep had fewer lesions (P less than 0.001) than the vaccine B sheep. The lesions on the vaccine A sheep were also less severe 14 and 21 days after challenge (P less than 0.05, P less than 0.01 respectively). In a second study, vaccine A was assessed for its ability to protect against natural challenge. Ten weeks after contact with sheep with active and generalised dermatophilosis no difference was found between the number of lesions present on the vaccine A and unvaccinated sheep and no differences were found in the number of sheep in each group with active lesions.

The host cell-mediated immune response was examined following experimentally-induced infection of rats with Dermatophilus congolensis, the causal agent of the skin disease dermatophilosis. Mononuclear cells (MC) isolated from Wistar rats 10 days following the induction of a third infection underwent a strong and specific proliferative response, as assessed by a [3H]thymidine incorporation assay, when cultured with various concentrations of inactivated D. congolensis cocci. Using specific monoclonal antibodies in an indirect fluorescent antibody test, this in vitro response was found to be characterised by a large expansion of the W3/25 (T-helper phenotype) population to form 56% of the total. Finally, the primed and stimulated MC were assessed for their ability to produce factors capable of inhibiting macrophage migration. The culture supernatants of D. congolensis-stimulated MC from infected rats caused significant migration inhibition of normal rat peritoneal exudate cells, whilst the supernatants of similarly-stimulated MC from naive rats failed to cause significant inhibition. The results show that a MC subpopulation becomes primed following experimentally-induced infection with D. congolensis and becomes activated after subsequent, in vitro, exposure.

Bacteria, isolated from the skins of clinically normal sheep, were tested for inhibitory activity against Dermatophilus congolensis grown in vitro. Out of 85 bacterial isolates, 19, mainly Bacillus spp., showed zones of inhibition when grown together with D. congolensis. The inhibitory activity was shown to be due to the metabolites released by the bacteria.

Over periods of 22 and 14 months, IgG antibody concentrations in serum samples obtained monthly from 14 mares and 19 foals, respectively, were measured by use of ELISA against antigens of the following environmental microbes: Aspergillus umbrosus, Penicillium brevicompactum, Rhodotorula glutinis, Absidia corymbifera, Aspergillus fumigatus, Humicola grisea, Micropolyspora faeni, and Thermoactinomyces vulgaris. The mares and foals were on pasture from early June until early October, then were stabled during the winter season until the following June. In the mares, increased antibody concentrations against most microbes were observed typically in midwinter and late spring when the horses were stabled; antibody concentrations against R glutinis, however, peaked in August. Concentrations differed between the summer and winter seasons and, in most instances, between 2 consecutive years and correlated with amounts of rainfall during the previous harvest season. In the foals, circulating passively acquired antibodies disappeared within 3 to 4 months after birth. During the first year of life, substantially increased autogenous antibody concentrations were observed only against R glutinis. Antibody concentrations against the other microbes increased gradually toward the end of the indoor season. In a group of foals transferred indoors in autumn, 6 weeks later than the other foals, antibody concentrations were lower when measured in December. Results supported the view that, to minimize exposure to microbial spores during the winter season, horses should be kept outdoors as much as possible and attention should be focused on improving the ventilation in stables and the quality of feeds and beddings.

Livingston-Wheeler's cancer treatment is based on the belief that cancer is caused by a bacterium she has named Progenitor cryptocides. Careful research using modern techniques, however, has shown that there is no such organism and that Livingston-Wheeler has apparently mistaken several different types of bacteria, both rare and common, for a unique microbe. In spite of diligent research to isolate a cancer-causing microorganism, none has been found. Similarly, Livingston-Wheeler's autologous vaccine cannot be considered an effective treatment for cancer. While many oncologists have expressed the hope that someday a vaccine will be developed against cancer, the cause(s) of cancer must be determined before research can be directed toward developing a vaccine. The rationale for other facets of the Livingston-Wheeler cancer therapy is similarly faulty. No evidence supports her contention that cancer results from a defective immune system, that a whole-foods diet restores immune system deficiencies, that abscisic acid slows tumor growth, or that cancer is transmitted to humans by chickens.

Dermatophilus congolensis infection of rabbits was used to investigate the effects of active immunity on epidermal challenge following vaccination. Rabbits (three groups of four) were vaccinated intradermally with live whole-cell preparations of D. congolensis strains SS18C and FD11 (groups SSVAC and FDVAC respectively); a third group (UNVAC) remained as unvaccinated controls. Two weeks after vaccination, separate 1.5-cm2 clipped and ether-swabbed skin sites were inoculated with a 10-fold dilution range (10(7) to 10(1) zoospores per cm2 of skin) of SS18C or FD11. Lesion scores at each site were calculated from the sum of individual scores (0 to 4+) for erythema, oedema and scab formation multiplied by the percentage of the inoculated area affected. A clear dose-response relationship between the size of inoculum and the severity of lesions was seen for both D. congolensis stains in the control group (UNVAC). In the SSVAC and FDVAC groups the lesions were less severe and developed more quickly. The number of zoospores required to cause infection in the vaccinated animals was up to 10,000-fold higher for homologous inoculated sites and 100-fold for heterologous sites. Serological analysis was carried out with an ELISA system. Vaccination and challenge resulted in increases in specific antibody against D. congolensis antigens. Cross-reacting antibody to the heterologous strain of D. congolensis used was demonstrated in both vaccinated groups but did not correlate with equal protection to homologous or heterologous challenge. The dose-response relationship demonstrated by this model enabled semi-quantitative analysis of the effects of vaccination on D. congolensis infective dose and severity of infection.(ABSTRACT TRUNCATED AT 250 WORDS)

We developed a hypersensitivity pneumonitis model by sensitizing rabbits intratracheally (IT) twice after two subcutaneous sensitizations (SS), and investigated the serial changes in the production of antibody against micropolyspora faeni (Mf), macrophage migration inhibition test (MIT), and histopathologic features. On the third day after the last IT, the proportions of neutrophils in bronchoalveolar lavage fluid (BALF) increased. The antibody levels of Mf specific-antigen in BALF decreased transiently, but those in serum began to increase. The number of AFC in spleen increased rapidly. Histopathologic examination revealed alveolitis and prominent acute pneumonia. On the seventh day after the last IT, the proportion of lymphocytes in BALF, the antibody levels of Mf specific-antigen in BALF, the number of AFC in BALF, and the percent inhibition of MIT were highest. Histopathologic features were characterized by the formation of prominent epithelioid granulomas. On the fourteenth day after the last IT, these features returned to the state of untreated group. These results indicate that the lesions in this HP model are formed by both type III and type IV immune reactions (Coombs-Gall). It was concluded that the seventh day after the last IT was most appropriate for the study of pathophysiology of human HP.

Serological crossreactivity was analyzed between M. leprae and strains of various species of Corynebacterium, Mycobacterium, Nocardia, Rhodococcus, Streptomyces, and related organisms. M. leprae shares antigens with most of these organisms, and sera from patients with lepromatous leprosy contain antibodies against them. The results demonstrate that M. leprae shares more antigens with the mycobacteria than with strains of the other tested genera, thus supporting the view that the leprosy organism belongs to the genus Mycobacterium. One precipitinogen (designated p beta) was found to be common to M. leprae and the streptomycetes, and sera from patients with lepromatous leprosy contain antibodies against this antigen.

Selection of a phage-stable strain of a new species of the rifamycin-producing organism was carried out. The phage-stable mutants were selected with respect to the virulent phage 2739 isolated from a lysogenic culture of the rifamycin-producing organism. Spontaneous phage-stable mutants formed at a rate of 0.8 per cent. Most of them belonged to the morphological colony type with a decreased activity level. No shifts in variation with respect to the property of the antibiotic production were noted under the action of phage 2739. 62 per cent of the phage-stable variants isolated from the secondary growth colonies after infection with the phage were lysogenic and liberated phage 2739 to the culture fluid. Induction of mutations with MNNG, UV and gamma(Co30) rays increased the frequency of the phage-stable mutanta by 1.5 times. Active phage-resistant mutants stable to the phage because of its adsorption and liberating no phage 2739 into liquid media during its cultivation were selected.

Three purified cell-wall glycoprotein antigens, a, b and c, sensitive to sodium periodate and resistant to pronase, were extracted from mycelium of Micropolyspora faeni with aqueous phenol or trichloracetic acid. Pronase-sensitive, sodium periodate-resistant protein antigens were extracted from mycelium with aqueous phenol. Immunoelectrophoresis was a critical method of assessing purity of preparation.Antibodies to glycoprotein antigens only occurred in clinically defined cases of farmer's lung disease, whereas antibodies to protein antigens also occurred in symptomless farmers. Precipitins to a occurred in all cases of farmer's lung disease, and this antigen was isolated and purified.Fractionation of pooled sera from cases of farmer's lung disease showed that precipitins were IgG globulins, whereas latex-agglutinating antibodies were IgG, IgA and especially IgM globulins. It is suggested that farmer's lung may involve a cytotoxic type II reaction, in which glycoproteins adsorbed to tissue cells react with IgG, IgA and IgM immunoglobulins in the presence of complement, causing cellular damage.

Extraction of mycelium or walls of Micropolyspora faeni with cold or hot aqueous phenol yielded a lipopolysaccharide consisting of lipid A, phosphate, galactose, arabinose, glucose, glucosamine, and a dideoxy sugar. Extraction with trichloroacetic acid (TCA) yielded an incomplete molecule lacking lipid A. Part of an O-chain was secreted into the culture medium. Phenol and TCA extracts gave three lines of precipitation with human serum from cases of farmer's lung disease, and one of these was given by the culture medium polysaccharide. Serologically-reactive sugars were arabinose, galactose and glucose. The lipopolysaccharide fixed on to red cells which agglutinated in the presence of specific antibody and lysed on the addition of complement. The lipopolysaccharide appeared to elicit mainly IgM antibodies in animals, but IgM and IgG antibodies in humans.

The reverse single radial immunodiffusion technique was used to detect Dermatophilus congolensis antibody in sera collected from animals previously infected to varying levels with D congolensis. Ammonium sulphate and trichloroacetic acid extracts of five different strains of D congolensis obtained from different geographical locations were used as antigens. All the extracts showed variations in their sensitivities in detecting D congolensis antibody in the various serum samples. Multiple antibodies were detected by some extracts while some showed negative antibody reaction to all extracts. Two extracts also showed cross-reactions with the serum from an animal that was infected by Nocardia species. Trichloroacetic acid extracts of all the strains were found to be more active serologically, detecting antibody in more sera and giving sharper and clearer reactions than ammonium sulphate extracts.

Two specific glycolipids identified as monomannosyl diglyceride (G1) and monoacylated glucose (G2), isolated from Nocardiopsis dassonvillei strains, were found to be biologically active. They elicited antibody response in rabbits when administered with some carrier components: lecithine, methylated bovine serum albumin (MBSA) and Freund incomplete adjuvant (FIA). Antibodies to the glycolipids were detected in antisera to G1 and in the sera against crude cell antigen of N. dassonvillei (N). The ability of the lipids to induceskin delayed hypersensitivity (SDH) as well as adjuvant and mitogenic effect were also studied. Generally glycolipid G1 seemed to be more active in antibody response whereas glycolipid G2 was a better stimulator of cellular response. Glycolipid G2 appeared to be distinct adjuvant and mitogenic factor.

Zoospore, filamentous and soluble antigens were prepared from Dermatophilus congolensis and examined for their ability to protect sheep from challenge with D. congolensis zoospores. In 1 experiment, sheep were vaccinated with Antigens A, B and C. The number of sheep protected in the group vaccinated with Antigen B was greater (P less than 0.05) than that in the unvaccinated group after challenge. The group vaccinated with Antigen B had a higher antibody response (P less than 0.05) to Antigen B than to Antigen A or C. In a second experiment, 2 groups of sheep were vaccinated with Antigen B. All sheep in this study developed lesions after challenge, but those on the vaccinated sheep were less severe (P less than 0.05) than those on the unvaccinated sheep. The antibody response to Antigen A, 28 days after vaccination, was higher (P less than 0.05) than the response to Antigen B.

In a pilot survey to compare the relative prevalence of three diseases in apparently healthy White Fulani Zebu (WFZ) cattle slaughtered in Nigeria, sera from 80 randomly selected animals with no significant gross lesions on ante mortem and post mortem inspection were examined for antibodies to Brucella abortus, Dermatophilus congolensis and bovine leukaemia virus. Of the samples screened, 5.0, 8.8 and 2.0% showed serological evidence for brucellosis, cutaneous streptothricosis and bovine leukosis respectively.

One hundred and forty-one strains of Actinomycetales and related organisms were investigated by immunodiffusion for the presence of antigens which cross-react with the antigens of M. tuberculosis H37Rv. The test strains comprised 86 different species names and 20 different genus names; they were mainly environmental organisms isolated from soil, plants, animals and such like. More than 90% of these strains were shown to have one or more antigens in common with the tubercle bacillus, and 77% had two or more antigens in common with this organism. Certain strains, of Nocardia and Rhodococcus, demonstrated abundant cross-reactivity with the tubercle bacilli, sharing up to 5 and 6 precipitinogens respectively.

Farmers were evaluated for the presence of farmer's lung disease by serologic methods and by clinical histories. From a large farming population screened serologically, 40 of 92 farmers with precipitating antibodies to the thermophilic actinomycetes consented to be evaluated for clinical evidence of farmer's lung disease. Each subject completed a standardized questionnaire which was reviewd by a trained observer. On the basis of the questionnaire and an indepth clinical history, the subjects were grouped into those farmers who had a history of farmer's lung disease (38%), those with no history (50%), or a doubtful group (12%). When these groups were compared for radiologic changes, pulmonary function abnormalities, and immune function, no differences were discernible. None of the parameters tested were useful in predicting which farmer would develop clinical farmer's lung disease. The parameters tested did not provide reliable criteria for differentiating those patients who present without acute symptoms but relate a history of recurrent pulmonary illnesses associated with moldy forage.

We have previously reported that the sonicate extrast from Actinomyces viscosus T 14 V (Av. sup) causes polyclonal B cell activation (PBA) in murine splenic B cells without any T cell assistance. However, there are some reports describing T cell dependency of PBA induced by A. viscosus in human peripheral blood B cells, and it is still controversial whether or not A. viscosus has the capability of exhibiting PBA without the help of T cells. In this report, we examined PBA in human peripheral blood B cells using the same Av. sup which induced T cell independent PBA in murine splenic B cells. The PBA was evaluated in terms of immunoglobulin (Ig) production by means of micro-ELISA method and cell proliferation by incorporation of 3H-TdR after incubation of cells for 5 or 7 days with Av. sup. Human peripheral blood B cells were highly purified by the following methods; The peripheral blood was collected by venipuncture and the peripheral blood mononuclear cells were harvested using gradient centrifugations. These cells were passed through a Sephadex G-10 column, and T cells were eliminated by rosette formation with sheep red blood cells. B cells were highly purified from these T cell-depleted cell populations by panning method with anti-human Ig antibody coated dishes followed by complement-dependent cytotoxisity using monoclonal antibody cocktails consisting of OKT 3, OKT 4 and OKT 8. These highly purified B cells could not respond to T cell mitogens such as PHA.(ABSTRACT TRUNCATED AT 250 WORDS)

The incidence of detection of antibodies against antigens derived from two thermophilic actinomycetes in patients clinically suspected of having Farmer's Lung was assessed. Approximately 25% of samples submitted over a three year period were found to be positive in either the ELISA test or the CIEP test for antigens derived from M. faeni and T. vulgaris. The ELISA test proved to be more sensitive than the CIEP test in the detection of antibodies directed against both organisms.

We studied the effect of single and multiple injections of Corynebacterium granulosum on weight and histology of lymph nodes and spleen, on peripheral white blood cell count, response of peripheral blood lymphocytes, lymph node, and spleen cells to phytohemagglutinin and pokeweed mitogen, survival of skin allografts, and lung metastases of a syngeneic fibrosarcoma in C3Hf/Bu mice. Corynebacterium parvum was used in some studies on antitumor activity. The weight of lymph nodes and spleen was markedly increased by single and multiple i.p. injections of C. granulosum, the peak enlargement occurring at Day 7 in lymph nodes and at Day 16 in spleen. Histologically, there was an extensive proliferation of nucleated cells in the enlarged organs. C. granulosum did not change the total white blood cell count but caused a temporary lymphopenia. In general, in vitro response to phytohemagglutinin and pokeweed mitogen of blood lymphocytes and spleen cells was decreased. Lymph node cell response to phytohemagglutinin was increased by small doses (0.025 mg) of C. granulosum, was not altered by a single large dose (0.5 mg), and was decreased by multiple doses. The response of lymph node cells to pokeweed mitogen was increased by all treatments. These changes in response to mitogens were demonstrable for about 2 months after treatment. Treatment i.v. with 0.1 or 0.25 mg of C. granulosum given before but not after grafting significantly prolonged the survival of grafted BALB/c skin. Smaller doses of this bacterium were not effective. Splenectomy of skin graft recipients did not prevent the effect of C. granulosum. Treatment i.p. or i.v. with this bacterium significantly decreased the number of lung metastases from i.v.-injected fibrosarcoma cells, even if the cells were injected 3 to 4 months later. The magnitude of this effect varied with the dose and frequency of injection of C. granulosum and C. parvum.

Counterimmunoelectrophoresis (CIE) has been compared in a diangostic laboratory with agar gel double diffusion (DD) as a routine procedure for detection of antibodies to pathogenic and allergenic fungi and actinomycetes. It was shown to be of particular value in detecting antibodies to Aspergillus fumigatus. Thus 72 of 106 sera in which precipitins were detected were positive by CIE alone. Some sera were positive only by CIE to antigens prepared from Histoplasma capsulatum, Allescheria boydii, Candida albicans and C. parapsilosis.

Models of hypersensitivity pneumonitis (HP) should exhibit progression of pulmonary histological abnormalities during continuing challenges. Strain II guinea-pigs were sensitized with Micropolyspora faeni and received 2, 4, or 8 weekly intratracheal (i.t.) particulate M. faeni challenges. Control animals received normal saline (NS). Four days after the last exposure, randomly selected microscopic fields of lung (200/animal) were judged to be normal or abnormal. If abnormal, the location and nature of the abnormalities were determined. Compared with NS treated guinea-pigs, those exposed to 2, 4 and 8 weekly M. faeni challenges exhibited more extensive (P less than 0.001) pulmonary histological abnormalities which involved both the intraalveolar and interstitial compartments. More extensive abnormalities in the 8 week group compared with the 4 week group were caused by increased extent of interstitial mononuclear cell infiltration. The extent of pulmonary interstitial histological abnormalities transiently (four challenges) decreases, but then increases, so that progressive pulmonary inflammation occurs during continuing challenges.

Immune responses to Rhodococcus equi were assayed in mares and foals on 7 studs in south-eastern Australia using skin test reactivity to the intradermal injection of culture filtrate and an indirect fluorescent antibody test. The prevalence of positive skin-test reactions did not differ between studs with a history of R. equi disease and those without but there were more mares with high antibody titres on studs with a disease history. A leucocyte extract prepared from mares that were skin-test positive was evaluated for its ability to protect foals exposed to experimental or natural challenge: 2 foals receiving leucocyte extract became skin-test positive and had resolving lesions present at post-mortem examination. All foals challenged experimentally developed serum antibody but only in those that became skin-test positive were the lesions resolving. In a field trial of leucocyte extract, using 450 foals over 2 foaling seasons, no significant difference was detected in morbidity or mortality rates between treatment and control groups. It is suggested that, since R. equi is so ubiquitous and most horses show immunological evidence of exposure, the development of clinical disease may be related to individual inability to cope with this organism. This may be due to inherited immunological unresponsiveness or to environmental factors which increase the challenge or decrease resistance.

A selective broth enrichment technique was used to study the distribution of Rhodococcus equi in soil and grazing animals. Rhodococcus equi was isolated from 54% of soils examined and from the gut contents, rectal faeces and dung of all grazing herbivorous species examined. Rhodococcus equi was not isolated from the faeces or dung of penned animals which did not have access to grazing. The isolation rate from dung was much higher than from other samples and this was found to be due to the ability of R. equi to multiply more readily in dung. Delayed hypersensitivity tests were carried out on horses, sheep and cattle, but only horses reacted significantly. The physiological characteristics of R. equi and the nature of its distribution in the environment suggested that R. equi is a soil organism.

Concentrated skin washings, even from vaccinated animals, failed to inhibit the motility of the infective zoospores of Dermatophilus congolensis, or to prevent them from germinating or infecting cattle; their constituent immunoglobulins did not attach to the flagella although IgA and IgG2 did bind to the cell bodies. It is concluded that the specific antibodies at the skin surface of ruminants are unlikely to have a role in zoospore immobilisation. Post vaccination sera rapidly immobilised and clumped the zoospores by means of a coat around the flagella, in which immunoglobulins, particularly IgM, were detected. IgM and IgG1 also attached to the cell bodies of the zoospores.

Antigens in coccidioidin were compared with purified subfractions via tandem immunoelectrophoresis (IEP) and by a combination of advancing line and crossed IEP. Rocket IEP was suitable for titrating the reactions and showing the relationship between column fractions. These techniques required multicomponent antisera produced by hyperimmunization over many months and by the use of known standard migration pairs. The IEP variations were used to chart the development of antisera against coccidioidin factors, to monitor antigen purifications, and to test the immunochemical homogeneity of an isolated antigen. Mannose-based heteroglycans of Cryptococcus neoformans were recovered from the culture filtrate. After precipitation of the major viscous glucuronoxylomannan (GXM) with ethanol or cetyltrimethylammonium bromide, the supernate is reserved because it contains a galactoxylomannan (GalXM). After removal of glucuronic acid from the GXM, the resulting xylomannan of serotype A was amenable to 13C-nuclear magnetic resonance (NMR) spectrometry; it revealed nonreducing xylose, alpha-1,3-mannose, and alpha-1,2/1,3 disubstituted mannose, thus confirming by an independent means what was previously known. The characterization sequence of GalXM included: (1) gas-liquid chromatography (GLC) of neutral sugars as peracetylated aldononitriles; (2) methylation-fragmentation GLC mass spectrometry to determine the glycosidic linkages; and (3) 13C-NMR showing similarities to mannan of Saccharomyces cerevisiae. Affinity chromatography of the GalXM on concanavalin A separated the galactoxylo component from an adsorbed mannoprotein. Selection of monoclonal antibodies (MAbs) relies on presumptive enzyme immunoassays (EIAs) or radioimmunoassays for rapid screening of clones and for determination of isotypes; however, higher resolution confirmatory tests are needed to obtain MAbs of desired specificity. MAbs against Candida tropicalis mannan were labeled with horseradish peroxidase to use for detecting mannan in serum. MAbs against the partially purified "m" factor of histoplasmin were characterized by the enzyme-linked immunoelectro-transfer blot technique (EITB), revealing unsuspected complexity in the antigen. Secreted proteins of Nocardia asteroides were isoelectrically focused; three proteins, identified by EITB as promising to be specific for that actinomycete, were cut out of gels and used to immunize mice for production of MAbs. The fimbriae of Actinomyces viscosus and A. naeslundii that mediate lactose-reversible coagglutination with Streptococcus sanguis have been used to evoke MAbs.(ABSTRACT TRUNCATED AT 400 WORDS)

Serum antibody titres to Dermatophilus congolensis demonstrated by an enzyme-linked immunosorbent assay (ELISA) in young steers and in adult cows from an Ayrshire herd showed a bimodal distribution and provided evidence of subclinical infection. Very high titres detected in sera from crossbred Galloway steers were indicative of recent or existing infection which may have been masked by concurrent ringworm. The ELISA is a sensitive and technically simple method which enables sera to be screened for evidence of infection by D congolensis which may otherwise pass unrecognised. Such infections may be of importance not only in the epidemiology of the disease in farm animals but also as a potential source of infection for man and his domestic pets.

The frequency of serologically detectable (SD) HLA-A, B, and C loci antigens in subjects with farmer's lung disease (N = 100) was compared with that of age- and sex-matched normal normal farmers with no precipitating antibodies to extracts of thermophilic actinomycetes. A subset of the farming population with antibodies to the thermophilic actinomycetes and no evidence of clinical disease (N = 55) was also agae- and sex-matched to the farmer's lung disease population. No significant associations between any of the SD HLA antigens tested and farmer's lung disease were found in the study. The data demonstrated that there was no association between SD HLA antigens and farmer's lung disease in random populations.

Fifty-eight sera of subjects with suspected farmer's lung disease (FLD) were tested by double immunodiffusion and ELISA for the detection of IgG and IgM antibodies against Micropolyspora faeni. By comparing the two methods, we obtained a high concordance of results for IgG, but not so high for IgM. ELISA is therefore a sensitive and specific test in determining antibody concentration.