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Relatedness analyses of Histoplasma capsulatum isolates from Mexican patients with AIDS-associated histoplasmosis by using histoplasmin electrophoretic profiles and randomly amplified polymorphic DNA patterns.

The present paper analyzes the histoplasmin electrophoretic profiles and the randomly amplified polymorphic DNA (RAPD) patterns of the fungus Histoplasma capsulatum isolated from Mexican patients with AIDS-associated histoplasmosis. Clinical isolates from Guatemala, Colombia, and Panama, as well as H. capsulatum isolates from different sources in nature, were also processed. All histoplasmin samples shared four antigenic fractions of 200, 49, 10.5, and 8.5 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). According to their percentage of relatedness, based on SDS-PAGE histoplasmin electrophoretic image analysis, H. capsulatum isolates were divided in two groups: group A contained all AIDS-associated isolates studied and two human reference strains from Mexican histoplasmosis patients without AIDS; group B included bat guano, infected bat, and cock excreta isolates from the State of Guerrero, Mexico, plus three human histoplasmosis strains from Guatemala, Panama, and Colombia. Polymorphic DNA patterns evaluated by RAPD-PCR showed three major bands of 4.4, 3.2, and 2.3 kb in most H. capsulatum isolates studied. Four groups were related by DNA polymorphisms: group I was formed by most of the AIDS-associated H. capsulatum isolates studied, one human histoplasmosis strain from Colombia, two human reference strains from Mexican patients without AIDS, and one human histoplasmosis strain from Guatemala. Group II consisted of only a single strain from Panama. Group III included three strains: one from a Mexican patient with AIDS and two isolated from nature in Guerrero (cock excreta and bat guano). The last, group IV, consisted of only one strain isolated from an infected bat, captured in Guerrero. A tight relationship between phenotypic and genotypic characterization was observed, and both analyses could be useful tools for typing H. capsulatum from different sources and geographic origins.

Acquired Immunodeficiency Syndrome↗

Histoplasmin reaction. Comparison of a polysaccharide antigen to the filtrate antigen.

This work was planned by taking into account all the knowledge accumulated from the immunological study of paracoccidioidomycosis. It aimed at comparing a polysaccharide antigen from Histoplasma capsulatum to a classic histoplasmin with the help of intradermal tests of delayed type of hypersensitivity. Tests were applied to 115 individuals in Santo Amaro, a town in the State of São Paulo. Positive results using classic histoplasmin were obtained in 46.0% cases whereas positive results using the polysaccharide antigen at its highest concentration were obtained in 51.30% cases. The major conclusion in this investigation is that it is possible to use the polysaccharide antigen as histoplasmin instead of the filtrate antigen.

Adolescent↗

[An intradermal test with histoplasmin and paracoccidioidin in 2 regions of Rio Grande do Sul].

From August to October 1992 a study of prevalence of cutaneous positivity to histoplasmin and paracoccidioidin was carried out in the cities of Cachoeira do Sul (Valley of Jacuí River) and Santo Angelo (western hillside of the PlateAn), Rio Grande do Sul, Brazil. Skin tests were made in 193 soldiers from Cachoeira do Sul and 161 soldiers from Santo Angelo, both groups of 17 to 19 years of age. In Cachoeira do Sul the prevalence of positive tests to histoplasmin was 89% and to paracoccidioidin 82%. In Santo Angelo 48% of the intradermical reactions were positive to histoplasmin and 39% to paracoccidioidin.

Adolescent↗

[Cutaneous sensitivity to histoplasmin in a Martinique hospitalized population].

OBJECTIVE: Histoplasma capsulatum infection is considered to be endemic in the West Indies. Nevertheless, few epidemiologic studies have been conducted in this area. The histoplasmin skin test reflects the frequency of asymptomatic forms of histoplasmosis. We studied the prevalence of positive skin tests in a population of the French West Indies (Martinique). MATERIALS AND METHODS: Forty one patients (24 females and 17 males), age range 29 to 90 years, were tested for histoplasmin skin sensitivity between August and October 1997, in the department of dermatology of Fort de France (French West Indies). Patients with immunosuppression or personal history of histoplasmosis were excluded. RESULTS: Five patients had a positive skin test (12 p. 100). No significative association was found between a positive skin test and diabetes, rural occupations or exposure to bats. DISCUSSION: Despite the small number of cases, related with difficulties in obtaining histoplasmin, our study showed a sensitivity level similar to medium endemic areas of the USA. The positive skin test rate is much higher than the rate reported before in West Indies, in a sample of the population under 25 years of age.

Adult↗

RELATIONSHIP OF HISTOPLASMIN AGAR-GEL BANDS AND COMPLEMENT-FIXATION TITERS IN HISTOPLASMOSIS.

Wiggins, Geraldine L. (Communicable Disease Center, Atlanta, Ga.), and Joseph H. Schubert. Relationship of histoplasmin agar-gel bands and complement-fixation titers in histoplasmosis. J. Bacteriol. 89:589-596. 1965.-Culture filtrates of various strains of Histoplasma capsulatum were studied with sera from patients with histoplasmosis, by use of the agar-gel and complement-fixation tests. It was found that the filtrates represented three types: those containing H and M components, those with H only, and those with neither H nor M. The same strain of H. capsulatum often produced culture filtrates of more than one type. Complement fixation showed that only the filtrates having both the H and M components were sufficiently sensitive as diagnostic antigens to detect the antibodies in histoplasmosis. H and M fractions prepared by ion-exchange chromatography were found to produce more than one band; therefore, it was concluded that the available fractions were not pure. The formation and identification of bands were affected by the concentration of the antigen and the arrangement of the wells containing the antigen and serum. The optimal concentration of the antigen varied, depending on the position of these reagents. The method found most practical for the identification of bands in unknown sera consisted of selecting the optimal concentration of the components with a box titration, by use of antigen having both the H and M components, placing known positive sera in wells adjacent to the unknown sera, and studying the sera for lines of identity. The M band was usually closest to the antigen well, and the H was nearest the serum well. These bands could be reversed by dilution, adjusting the H antibodies so that they equalled or exceeded the M antibodies. One human serum in which the bands were reversed showed the H bands at a higher serum dilution than that obtained with the M band. Patients with culturally proven histoplasmosis had the M band in their serum regardless of the stage of disease or length of duration; only 4 of 10 patients in the active stage of histoplasmosis also had the H band. The low prevalence of the H band would eliminate it as a diagnostic tool for detecting the active stage of the disease. Other bands obtained with histoplasmin antigen in the sera from proven cases were of no assistance in determining the clinical status of the patient.

Agar↗

A SURVEY OF HISTOPLASMIN SENSITIVITY IN IRAQ.

During the past two decades much has appeared in the medical literature concerning systemic fungus diseases, the reports originating from many areas of the world. None of the reports, however, deals specifically with the existence, prevalence, and distribution of histoplasmosis in Iraq, and a survey was therefore undertaken in 1963 to determine the level of histoplasmin sensitivity in that country. In this survey, 1715 persons, whose ages ranged from under one year to 76 years and who were representative of the whole country, were given a histoplasmin skin-test and a chest X-ray. The majority of the 1461 persons whose tests could be read were non-reactors and the nine (0.6%) positive reactors showed no clinical manifestation of the disease. The authors therefore conclude that histoplasmosis is probably not present in Iraq, or exists at such a low level that it has no public health significance.

Epidemiology↗

Epidemiologic skin test survey of sensitivity to paracoccidioidin, histoplasmin and sporotrichin among gold mine workers of Morro Velho Mining, Brazil.

Skin tests with paracoccidioidin, histoplasmin and sporotrichin were applied to 417 workers of Morro Velho Mining in the State of Minas Gerais, Brazil, with the main purpose of detecting the prevalence of paracoccidioidomycosis-infection, histoplasmosis capsulate-infection and sporotrichosis-infection. The rates of positivity to the skin tests were 13.43% for paracoccidioidin, 17.50% for histoplasmin and 13.67% for sporotrichin. Several epidemiological factors were investigated for a better interpretation of the results. Paracoccidioides brasiliensis, Histoplasma capsulatum var. capsulatum and Sporothrix schenkii were not isolated from the soil samples from the mines investigated.

Adult↗

Epidemiological survey of histoplasmine and paracoccidioidine skin reactivity in an agricultural area in Bolívar state, Venezuela.

Paracoccidiodomycosis and histoplasmosis are endemic diseases in the south of Venezuela, representing a public health problem. Prevalence of Paracoccidiodes brasiliensis and Histoplasma capsulatum infections were estimated in Monte Ralo, a rural community area of Bolivar state using paracoccidiodine and histoplasmine skin tests. Paracoccidiodine was intradermically injected to 173 persons and readings were made at 24 and 48 h afterwards to 167 persons (97.85%). Reaction was positive in 8.5% (n = 14) at 24 h post-injection and 13.2% (n = 20) at 48 h. Farmers showed the higher percentage of positivity at 24 and 48 h. One hundred-seventy five persons were intradermically injected with histoplasmine but 167 and 157 of them attended for reading of the dermal reaction at 24 and 48 h post-injection respectively. Tests were positive in 25.7% (n = 43) at 24 h and 42.7% (n = 67) at 48 h. Further studies are needed in surrounding places to delimit the endemic area of these mycosis in the Bolivar state. However, epidemiological vigilance of PCM and H should be considered by local health authorities.

Adolescent↗

Studies on the thermal degradation of the H and M antigens of lyophilized histoplasmin.

Lyophilized histoplasmin for the agar gel microimmunodiffusion test has been prepared as a candidate World Health Organization Biological Reference Reagent. It was subjected to elevated temperature for given periods of time and analyzed by the capillary precipitin test and the single radial immunodiffusion test to determine the stability of the H and M antigens. H antigen showed no fall in relative potency when incubated at 48 degrees C for 20 days. M antigen showed a fall in relative potency after storage at 37 degrees C and 48 degrees C, but the extent of the fall was greater in the radial immunodiffusion test than in the capillary precipitin test. Half-lives of the antigens could not be calculated from the Arrhenius equation because the response curves at each temperature followed different kinetics. However, as based on zero time data, M antigen of the lyophilized histoplasma showed a 20% drop in relative potency when stored at -20 degrees C for 2 years. Other analyses suggested that M antigen of liquid histoplasmin stored at 5 degrees C and of lyophilized histoplasma stored at -20 degrees C was degraded at equal rates.

Animals↗

Determination of molecular weight, isoelectric point, and glycoprotein moiety for the principal skin test-reactive component of histoplasmin.

A purified component designated HPD (histoplasmin-purified derivative) dII was isolated from two different cru-e histoplasmin lots by a combination of gel filtration and polyacrylamide disc electrophoresis (Sprouse, 1969). A 0.05-mug portion of HPD dII was reactive and specific in detection of delayed hypersensitivity in guinea pigs experimentally infected with Histoplasma capsulatum. The objective of this study was to characterize this skin test-reactive component for (i) homogeneity, (ii) molecular weight, (iii) isoelectric point, and (iv) composition. Sephadex chromatography, polyacrylamide disc electrophoresis, sucrose density gradient ultracentrifugation, acid and heat denaturation, and immunoelectrophoresis indicate that HPD dII is (i) homogeneous, (ii) of approximately 12,000 molecular weight, and (iii) a glycopeptide of approximately 60% carbohydrate and 40% proteinaceous composition. The marked acid and heat stability exhibited by the compound probably is attributable to the prominent carbohydrate moiety in the molecule. Isoelectric focusing indicated an isoelectric point of 5.68. This would suggest that dII is an acidic compound with either predominance of acidic amino acid residues in the molecule or, more probably, an abundance of electron donors in the carbohydrate moiety. In summary, HPD dII appears to be a glycopeptide of approximately 12,000 molecular weight, reactive in elicitation of delayed hypersensitivity of histoplasmosis.

Animals↗

Monoclonal antibodies against the M-protein and carbohydrate antigens of histoplasmin characterized by the enzyme-linked immunoelectrotransfer blot method.

Monoclonal antibodies (MAbs) of two different specificities were produced by immunizing mice with the semipurified M antigen of histoplasmin. One type, from clone CB4, was an immunoglobulin M that precipitated a polysaccharide present in histoplasmin and also formed immunoprecipitates with a cross-reactive polysaccharide present in extracts of Blastomyces dermatitidis and Coccidioides immitis. The second type of MAb, from clone EC2, was an immunoglobulin G that reacted in the enzyme-linked immunoelectrotransfer blot (EITB) assay with a doublet of proteins with an apparent molecular size of 70 to 75 kilodaltons. This molecule is proposed as the authentic M protein antigen that is recognized by M antibodies in sera from mice and rabbits immunized with Histoplasma capsulatum and from persons with histoplasmosis. The M factor also occurs in an abundant disulfide-bridged dimer which has a molecular size of 150 kilodaltons and is nonimmunoreactive under the conditions of sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Animals↗

Apparent false positive histoplasmin latex agglutination tests in patients with tuberculosis.

Serum specimens from patients admitted to a respiratory disease hospital were examined by the histoplasmin latex agglutination, the complement fixation, and the agar gel immunodiffusion tests. Of 300 sera examined, 21 (7.0%) gave an apparent false positive reaction at a dilution of 1:16 or greater. Fourteen (66%) of the 21 patients studied has culturally proven tuberculosis. One patient each had a diagnosis of hypertensive cardiovascular disease with congestive heart failure, infection with atypical mycobacteria (Runyon group III), chronic pneumonitis secondary to gunshot wound, and pulmonary abscess of unknown etiology; two had bronchogenic carcinoma; and one serum specimen came from an apparently healthy employee. The results of the histoplasmin latex agglutination test should be interpreted with caution, particularly if only one serological determination has been made and the titer is low.

Adult↗

Solid-phase competitive-binding radioimmunoassay for detecting antibody to the M antigen of histoplasmin.

A radioimmunoassay (RIA) was designed and compared with complement fixation and immunodiffusion tests for their relative ability to detect antibodies in sera of histoplasmosis patients. M antigen, purified from histoplasmin, was fixed to microtiter wells as the solid phase, and specific rabbit 125I-labeled anti-M globulin was the source of indicator antibodies. The optimal concentrations for the competitive-binding assay were 1.6 ng per well for M antigen and 650 ng per well for the 125I-labeled anti-M globulin. A panel of sera from 29 histoplasmosis patients and from patients with other mycoses was screened for RIA activity and in complement fixation and immunodiffusion tests that used histoplasmin and Histoplasma capsulatum yeast-form antigens. The sera of 22 histoplasmosis patients reacted in the RIA, 21 in the complement fixation, and 16 in the immunodiffusion tests. Sera of patients with other mycotic infections did not react in the RIA, with the exception of those of one blastomycosis patient and one candidiasis patient. The RIA could be modified to quantitate M antigen; as little as 125 pg could be detected. The evaluation of this panel of histoplasmosis patients' sera showed that the RIA was about equivalent in sensitivity to the complement fixation test. Some advantages of the RIA over the complement fixation test were that RIA was less prone to cross-reactions and gave better quantitation of low-titered sera. The RIA was a 1-day test, was not hindered by the anti-complementary activity of some sera, and could be modified to quantitate minute amounts of M antigen.

Antibodies, Fungal↗

[Histoplasmin and paracoccidioidin reactions in Serra de Pereiro. (Ceara State--Brazil)].

Intradermal tests using histoplasmin and paracoccidioidin antigens were performed in 138 persons from Pereiro, Ceará, Brazil. The results were positive in 61.5% and 32.5% with histoplasmin and paracoccidioidin antigens respectively. These results suggest infection by H. capsulatum and P. brasiliensis in the people living in the studied area. New studies are necessary to detect histoplasmosis and paracoccidioidomycosis as clinical diseases in that region.

Adolescent↗

[Histoplasmin skin test and focal hemorrhagic chorioiditis (author's transl)].

A histoplasmin skin test was performed on 56 patients with focal hemorrhagic choroiditis. The test was negative in 53 choroiditis patients and positive in three patients. The three patients with posi-ive skin test had been living for a long time in the eastern part of the U.S.A. where histoplasma capsulatum occurs endemically. The results of this study suggest that the infection with histoplasmin capsulatum is not the cause of focal hemorrhagic choroiditis in our area.

Choroiditis↗

[Histoplasmin reactivity in poultry farm workers in the province of Ciego de Avila, Cuba].

An epidemiological survey with histoplasmin skin test was performed in 392 poultry farmers and 265 workers considered without occupational risk of exposition to Histoplasma capsulatum, etiologic agent of histoplasmosis. The results were positive in 28.8% and 13.2% in both groups respectively. Statistically, there was a significant difference between the two groups, so it can be considered that poultry farmers are in occupational risk of infection with H. capsulatum. In the first group, the workers which are more closely related with chicken manure showed a higher reactivity to histoplasmin skin test. The working time in the farms seems to influence in the test reactivity too.

Adult↗

[Relationship of histoplasmin intradermal tests and antibodies levels detected by ELISA and immunodiffusion].

A prospective study was carried out in two groups of individuals: a group 1 (n = 40) included workers from a poultry farm, with potential occupational risk of exposure to Histoplasma capsulatum, etiologic agent of histoplasmosis, and a group 2 (n = 16), persons without occupational risk of exposure to the agent. Histoplasmin skin test was performed in both groups, and three sera were obtained from each individual: 1) before skin test was done, 2) 30 days after, and 3) 180 days after it. In both groups the histoplasmin skin test, even when the test was positive, was not a sufficient antigenic booster to provoke an increase in the H. capsulatum antibody levels capable to be detected by the serologic tests used (ELISA and Double Immunodiffusion). These results contribute to improve the interpretation of ELISA test values in the diagnosis of histoplasmosis.

Antibodies, Fungal↗

Histoplasmine and paracoccidiodine epidemiological study in Upata, Bolívar state, Venezuela.

OBJECTIVE: To determine the prevalence of histoplasmosis and paracoccidiomycosis in Upata, a city in Bolivar state, Venezuela. METHODS: Cutaneous tests on 397 volunteers, 204 with histoplasmine and 193 with paracoccidioidine. Epidemiological data were collected in a protocol sheet. Readings were performed at 24 and 48 h post-injection, by the same observer, and a positive reaction was defined as hardened papule of 5 mm or more. RESULTS: Histoplasmine tests were positive in 19.6% and 34.0% at 24 and 48 h after antigens injection with a significant increase of reactivity (P=0.0019). Paracoccidiodine tests were positive in 11.3% and 19.7% at 24 and 48 h respectively and this difference was also significant (P=0.004). Risk factors which could explain these levels of reactivity could not be clearly identified. Cultures and animal inoculation from soil samples for fungus isolation were negative. CONCLUSIONS: Paracoccidioidomycoses and histoplasmosis are endemic in Upata.

Adolescent↗