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HISTOPLASMIN SENSITIVITY OF YOUNG MEN IN ALASKA, HAWAII, THE PHILIPPINES AND PUERTO RICO.

The author describes results of a continuing study of skin sensitivity to histoplasmin among United States Navy recruits from Alaska, Hawaii, the Philippine Islands and Puerto Rico. For comparison, material is also included for several groups of recruits from different states of mainland United States.The evidence is interpreted as indicating that the frequent large reactions in Puerto Rican and Filipino recruits represent specific infection with Histoplasma, whereas the small reactions in recruits from Hawaii and the Philippines represent cross-reactions due to sensitization with a presently unknown agent or agents.

Alaska↗

Incidence of histoplasmin hypersensitivity in the Philippines.

Long-term residents of the Philippines were skin tested with histoplasmin skin test material. This study was conducted with 143 electric company (MERALCO) employees from Manila, Philippines. We found that 37 (26%) of the subjects were skin test positive. Characteristics of the positive group were: average age of 37 years; all except one were lifelong inhabitants of Metro Manila; 25 were male and 12 were female; one-half of the subjects reported extended contact with chickens. Despite these findings, histoplasmosis is considered to be a very rare disease in the Philippines. This survey indicates that Histoplasma capsulatum is sufficiently present in the Philippines to come in contact with one-fourth of the test population. This reinforces the hypothesis that histoplasmosis is present in the Philippines and is probably being misdiagnosed as granulomatous-inducing diseases such as tuberculosis, e.g., so-called "drug resistant" tuberculosis. We recommend larger surveys of this type and attempts to culture the etiologic agent from natural sources such as chicken and bat droppings.

Adult↗

Immunodiffusion studies on the antigens of Histoplasma capsulatum: comparison of mycelial histoplasmin with the yeast phase reagent Histolyn-CYL.

Immunodiffusion assays were performed to determine if H and M antigens associated with mycelial histoplasmin were present in the yeast phase reagent, Histolyn-CYL (H-CYL). Neither H nor M antigens could be detected in H-CYL. However, when H-CYL was reacted against human histoplasmal sera a positive reaction was evidenced in all instances in which a response was elicited with a reference mycelial immunodiffusion reagent.

Antibodies, Fungal↗

Effects of histoplasmin M antigen chemical and enzymatic deglycosylation on cross-reactivity in the enzyme-linked immunoelectrotransfer blot method.

The enzyme-linked immunoelectrotransfer blot (EITB) method was evaluated as a suitable method for detecting antibodies against M antigen of Histoplasma capsulatum by use of both glycosylated and deglycosylated M protein of histoplasmin (HMIN). Sera from patients with histoplasmosis, paracoccidioidomycosis, blastomycosis, coccidioidomycosis, and aspergillosis were tested by the EITB with glycosylated M protein of HMIN. This assay demonstrated 100% sensitivity with histoplasmosis serum samples, all of which reacted with the 94-kDa glycoprotein (M antigen). Although the EITB is highly sensitive, it is not specific for histoplasmosis when glycosylated M protein is used as an antigen. A total of 81% of paracoccidioidomycosis, 25% of blastomycosis, 33% of coccidioidomycosis, 73% of aspergillosis, and 16% of tuberculosis serum samples cross-reacted with M protein of HMIN and yielded patterns indistinguishable from those obtained with histoplasmosis serum samples. The EITB reactions with both untreated M antigen and M antigen altered by periodate oxidation or by deglycosylation with endoglycosidases were compared. Cross-reactions with heterologous sera in the EITB could be attributed to periodate-sensitive carbohydrate epitopes, as reflected by the increase in the test specificity from 46.1 to 91.2% after periodate treatment of M protein. The EITB for the detection of antibodies to M antigen is a potential diagnostic test for histoplasmosis, provided that periodate-treated M protein is used as an antigen.

Antibody Specificity↗

Purification, composition, and serological characterization of histoplasmin-H and M antigens.

To obtain purified H and M antigens suitable as primary standards in the serological diagnosis of histoplasmosis by agar gel double-diffusion tests, H and M reactive components of histoplasmin were fractionated by column chromatography by using Sephadex G-100, Sephadex G-200, and diethylaminoethyl cellulose. Six fractions from diethylaminoethyl cellulose were reactive in agar gel double-diffusion, complement fixation, and capillary precipitin tests. When examined by electrophoresis on acrylamide gel, one M antigen (fraction 2, molecular weight greater than 200,000) and two H antigens (fractions 5 and 6, molecular weight greater than 200,000) each gave essentially a single protein band. In agar gel double-diffusion and complement fixation tests with sera from patients with proven cases of histoplasmosis, blastomycosis, or coccidioidomycosis, these two fractions of H antigen and the one of M antigen reacted only with sera from proven or suspect cases of histoplasmosis and showed reactivity with those sera known to contain only the anti-H or anti-M antibody, respectively. Fraction 2 (M antigen) and fractions 5 and 6 (H antigens) had carbohydrate-to-protein ratios of 1.60, 0.77 and 0.78, respectively. Both antigens contained galactose, glucose, mannose, and hexosamine, with mannose being the predominant sugar. Fraction 2 was characterized by a high proline and glucose content, whereas fractions 5 and 6 contained higher concentrations of galactose, mannose, glycine, and alanine. Each of these products appeared to separate into two active fractions, one of a molecular weight greater than 200,000 and one of a molecular weight less than 35,000. The M antigen component of fraction 2 was still characterized by a high proline content, whereas the H antigen components of fractions 5 and 6 had a high content of glutamic acid, serine, glycine, and proline.

Antigens, Fungal↗

Evaluation of histoplasmin for the presence of H and M antigens: some difficulties encountered in the production and evaluation of a product suitable for the immunodiffusion test.

The development of a single histoplasmin suitable for use in immunodiffusion testing of a variety of human histoplasmosis sera depends not only upon the presence of both antigens, but on relative concentration of one to the other, and perhaps on the chemical characteristics of the H and M antigens of a given strain.

Antigens, Fungal↗

Rheumatoid factor: a cause of fals positive histoplasmin latex agglutination.

Ten of thirteen patients with positive histolatex agglutination titers of 1:32 or greater had no evidence of acute histoplasmosis. Three of these false positives had rheumatoid arthritis. A fourth had a rising mycoplasma complement fixation titer, and the fifth had a high titer of cold agglutinins. All of these are associated with abnormal immunoglobulin M production. To evaluate the role of rheumatoid factor in producing false positive histolatex agglutination, the histolatex test was performed on sera from 32 patients having rheumatoid factor at a titer of 1:40 or greater. Four of these sera agglutinated the histoplasmin-coated latex particles at titers of 1:32 or greater. Review of clinical records suggests the this reactivity is nonspecific. It is our purpose to call attention to rheumatoic factor as a cause of false positive histolatex agglutination.

False Positive Reactions↗

Histoplasmin and paracoccidioidin skin reactivity in infantile population of northern Argentina (1).

In order to estimate ages at which etiological agents of systemic mycoses initiate infection, histoplasmin and paracoccidioidin skin tests were performed in 344 children of both sexes, between 2 and 15 years old. They were selected from a statistically significant population sample Gral. San Martín city (Northeast Argentina). Tests were read 48 h after injection and considered positive if a 5 mm on larger induration was present. Circulating antibodies were also evaluated by agar gel immunodiffusion. The overall infection rate for H. capsulatum was 9.2%, belonging to children from 4 to 14 years old, without significant differences among sexes. Five children from 2 to 14 years old were positive to paracoccidioidin (1.6%). None of the children had specific antibodies neither signs of active mycosis. Results show H. capsulatum infection can be found from age 4, while for P. brasiliensis the lower limit was two years old. These findings may contribute to better knowledge on infantile fungal infection in a geographical region where no previous references can be found.

Adolescent↗

Epidemiologic surveys of histoplasmin and paracoccidioidin sensitivity in Brazil.

We report here the results obtained in epidemiologic surveys of histoplasmosis and paracoccidioidomycosis carried out in Brazil using the histoplasmin and paracoccidioidin delayed hypersensitivity skin tests. Most of these data have not been previously published in scientific journals and are now reported here in two tables respectively concerning histoplasmosis (88 surveys) and paracoccidioidomycosis (58 surveys). The guidelines to be followed in surveys of this nature are also commented upon.

Brazil↗

Tuberculin and histoplasmin sensitivity in six solomon islands populations.

Skin testing with antigens from Histoplasma capsulatum, Mycobacterium tuberculosis (PPD-S), and atypical mycobacteria (PPD-B, PPD-G, PPD-Y, and PPD-platy) was carried out among six population groups in the Solomon Islands between 1968 and 1972. There was no positive reaction to histoplasmin among any of the groups, suggesting that histoplasmosis is not endemic in the Solomon Islands. There were significant numbers of tuberculin reactors among each group. Largest mean reactions to PPD-S were present among the Lau and Ulawa, in whom reactions to PPD-S were larger than those to any other antigen tested. Thus significant infection with M. tuberculosis appears to occur in these populations. This was corroborated by radiologic survey. Among the Lau, large reactions to PPD-G and PPD-Y were also elicited, raising the possibility of multiple infection. Among the Aita, Baegu, Nagovisi, and Ontong Java, PPD-G elicited the largest reactions. PPD-G produced the second largest reactions among the Lau and Ulawa. PPD-S elicited the largest or second largest reaction among 4 of the 6 groups. A notable exception was the Aita, in whom PPD-S elicited the smallest mean reaction. The Aita also had the lowest prevalence of radiologic findings consistent with tuberculosis. These observations suggest that M. tuberculosis has been introduced into the Solomon Islands from outside sources, a hypothesis which may explain the variability in prevalence of tuberculosis-like disease demonstrated by chest film among the six groups. Genetic differences may also play a role in this variability. The study also demonstrated a high prevalence of "baseline" sensitivity to the atypical mycobacteria among the Solomon Islanders. This sensitivity may confer some immunity to infection with M; tuberculosis, but this protection is far from complete.

Adolescent↗

Prevalence of histoplasmin sensitivity in Pakistan.

A total of 575 subjects, comprising healthy men, women, and school children as well as pulmonary tuberculosis and leprosy patients in different areas of the Punjab region of Pakistan, were tested for histoplasmin skin sensitivity. All, with the exception of those who presumably had been exposed to the fungus in some other country, were found to be non-reactors to histoplasm. The preliminary findings suggest that histoplasmosis is not endemic in this area, as the general population seems to be free from exposure to the fungus Histoplasma capsulatum.

Adolescent↗

Skin sensitivity to capsulatum and duboisii histoplasmins in Nigeria.

A total of 1313 persons comprising 1087 healthy subjects and 226 hospital patients with bronchopulmonary disorders were tested for reactivity to histoplasmins prepared from Histoplasma capsulatum and H. capsulatum var. duboisii. The overall sensitivity rates to the two antigens were 3.5 and 3.0% respectively. The incidence of sensitivity in the hospital patients was significantly higher (8.85% for 'capsulatum' antigen and 6.64% for 'duboisii' antigen) than that in different categories of the healthy population (1.66-5.03% and 0.5-4.47%, respectively). The majority of persons who had a positive reaction to duboisii antigen also reacted positively to capsulatum antigen and vice versa.

Adolescent↗

Pulmonary calcification and skin sensitivities to tuberculin, histoplasmin and coccidioidin among Polynesians.

Pulmonary calcifications are fairly common in Western Samoa, but they cannot be attributed entirely to tuberculosis, since they are also produced by certain systemic mycoses and parasitic infestations. Furthermore, in many tropical areas, skin sensitivity to tuberculin is often due to certain unidentified weak sensitizing agents, rather than to the usual virulent tubercle bacilli.Skin sensitivity to histoplasmin and coccidioidin is absent in both Western Samoa and the Tokelau Islands; consequently, pulmonary calcifications found in these places cannot be attributed to either histoplasmosis or coccidioidomycosis.Both Western Samoans and Tokelauans exhibit non-specific reactions to tuberculin throughout their lives, but these reactions are clearer and more distinct in groups aged 15 years or more.The prevalence of non-specific sensitivity to tuberculin should be investigated further.

Adolescent↗

Evaluation of cation exchange chromatography for the isolation of M glycoprotein from histoplasmin.

Cation exchange chromatography was evaluated to purify the M antigen from histoplasmin (HMIN). Two H and M antigen-containing fractions, soluble (S) and precipitate (PP), resulted from the initial 0.025 M, pH 3.5 citrate buffer dialysis step. The PP fraction contained 62% of the M antigen activity and was resolubilized. Both fractions were chromatographed on CM Sepharose CL-6B. Polysaccharide C antigen was abundant in the S fraction and most of it did not bind to CM Sepharose. M antigen-enriched fractions were eluted with 0.5 M NaCl. Re-chromatography of the relevant S fraction (S-II) and PP fraction (PP-II) by linear gradient fast protein liquid chromatography (FPLC) removed protein and C impurities. M antigen purified by FPLC from the PP-II fraction was depleted of other antigens when Western blots were probed with anti-M, anti-H and anti-C monoclonal antibodies (Mabs). M antigen was identified as a 94 kDa glycoprotein containing a specific-protein epitope and an epitope that reacted with a Mab against the polysaccharide C antigen. M antigen can be purified from HMIN by tandem cation exchange chromatography of the precipitable fraction on an open CM Sepharose CL-6B column followed by linear gradient FPLC.

Blotting, Western↗

Histoplasmin sensitivity among a student population in Crema, Po Valley, Italy.

Histoplasmin skin tests were conducted on a student population in Crema, Po Valley, Italy. In this geographical area two autochtonal cases of disseminated histoplasmosis occurred in HIV-negative men. In the last year we tested 776 senior high school students, mean age 18 years, 335 males and 441 females. Histopalsmin skin sensitivity was 1.23%, with a mean area of induration of 5.78 mm. All subjects but one had never been abroad. This preliminary survey confirms the possibility of autochtonal histoplasmal infection in the Po Valley, Italy.

Adolescent↗