Successful treatment with corticosteroids for refractory deterioration of cryptococcal meningitis with an increasing cryptococcal antigen titer.

We report a 55-year-old man with Cryptococcus neoformans meningitis who showed refractory deterioration twice with an increased cerebrospinal fluid cryptococcal antigen titer during the course of treatment. Although the initial deterioration was temporarily improved by placement of a ventriculoperitoneal shunt, he experienced deterioration again. However, he improved after administration of systemic corticosteroids. The present case suggests that systemic corticosteroid can be a choice of treatment to rescue immunocompetent patients with Cryptococcus neoformans meningitis and severe deterioration, even if cerebrospinal fluid analysis shows an increased cryptococcal antigen titer.

Inadequacy of High-Dose Fluconazole Monotherapy Among Cerebrospinal Fluid Cryptococcal Antigen (CrAg)-Positive Human Immunodeficiency Virus-Infected Persons in an Ethiopian CrAg Screening Program.

A total of 817 human immunodeficiency virus-infected Ethiopians with CD4 <150 cells/mL underwent plasma cryptococcal antigen (CRAG) screening. CRAG prevalence was 6.2%. Of participants with plasma CRAG titers >1:640, 96% (27 of 28) had cryptococcal meningitis (cerebrospinal fluid CRAG-positive) whereas 50% (7 of 14) with 1:160-1:320 titers had meningitis. With fluconazole 1200 mg/d therapy, 68% of meningitis patients (23 of 34) died within 3 months. Plasma CRAG titers >1:160 predict meningitis, requiring more intensive antifungal therapy.

Occurrence of household mold and efficacy of sodium hypochlorite disinfectant.

The occurrence and distribution of mold on household surfaces and the efficacy of bleach-based (sodium hypochlorite, NaOCl) disinfectants on mold viability and allergenicity was documented. Household microenvironments prone to increased moisture were specifically targeted. Using the sticky tape method, 1330 samples were collected from non-porous indoor surfaces of 160 homes across the United States, and analyzed for mold. Homes were randomly selected and recruited via phone interviews. Culture and immunoassays were used to measure the viability and reduction of allergenic properties of Aspergillus fumigatus following 2.4% NaOCl treatment. All homes and 72.9% of surfaces tested positive for mold. Windowsills were the most frequently contaminated site (87.5%) and Cladosporium the most commonly identified mold (31.0%). Five-minute exposures to 2.4% NaOCl resulted in a >3 to >6-log10 reduction of culturable mold counts in controlled laboratory studies. Organisms were nonculturable after 5- and 10-min contact times on non-porous and porous ceramic carriers, respectively, and A. fumigatus spore-eluted allergen levels were reduced by an average 95.8% in 30 sec, as indicated by immunoassay. All homes are contaminated with some level of mold, and regrowth is likely in moisture-prone microenvironments. The use of low concentrations (2.4%) of NaOCl for the reduction of culturable indoor mold and related allergens is effective and recommended.

Human serum antibody reactivity towards Paracoccidioides brasiliensis antigens treated with sodium metaperiodate.

In this study, we evaluated the profile of anti-Paracoccidioides brasiliensis immunoglobulin isotypes in serum from patients with the acute and chronic forms of paracoccidioidomycosis, using the whole Paracoccidioides brasiliensis antigen and the antigen treated with sodium metaperiodate. All the immunoglobulin isotypes present in the serum from patients with the acute and chronic forms of paracoccidioidomycosis presented higher reactivity towards the whole antigen than to the antigen treated with metaperiodate (P < 0.05). The reactivity of IgG and IgM to the antigen treated with metaperiodate was greater in serum from patients with the acute form of the disease (P < 0.05), while IgA was more reactive in serum from patients with the chronic form (P < 0.05). There was greater reactivity of IgG1 and IgG2 to the whole antigen and the antigen treated with metaperiodate in the serum from patients with paracoccidioidomycosis than there was in serum from patients with other parasitic infections (P < 0.05). Furthermore, IgG1 from patients with the acute form recognized the 19kDa, 27kDa and 31kDa antigens in the western blot test. Thus, the results suggest that modifications to the epitopes of Paracoccidioides brasiliensis antigens may help to improve the immunodiagnosis of paracoccidioidomycosis.

Human serum antibody reactivity towards Paracoccidioides brasiliensis antigens treated with sodium metaperiodate / Reatividade de anticorpos de soros humanos a antígenos de Paracoccidioides brasiliensis tratados com metaperiodato sódico

In this study, we evaluated the profile of anti-Paracoccidioides brasiliensis immunoglobulin isotypes in serum from patients with the acute and chronic forms of paracoccidioidomycosis, using the whole Paracoccidioides brasiliensis antigen and the antigen treated with sodium metaperiodate. All the immunoglobulin isotypes present in the serum from patients with the acute and chronic forms of paracoccidioidomycosis presented higher reactivity towards the whole antigen than to the antigen treated with metaperiodate (P < 0.05). The reactivity of IgG and IgM to the antigen treated with metaperiodate was greater in serum from patients with the acute form of the disease (P < 0.05), while IgA was more reactive in serum from patients with the chronic form (P < 0.05). There was greater reactivity of IgG1 and IgG2 to the whole antigen and the antigen treated with metaperiodate in the serum from patients with paracoccidioidomycosis than there was in serum from patients with other parasitic infections (P < 0.05). Furthermore, IgG1 from patients with the acute form recognized the 19kDa, 27kDa and 31kDa antigens in the western blot test. Thus, the results suggest that modifications to the epitopes of Paracoccidioides brasiliensis antigens may help to improve the immunodiagnosis of paracoccidioidomycosis.

Neste trabalho, foi avaliado o perfil de isotipos de imunoglobulinas anti-Paracoccidioides brasiliensis em soros de pacientes com formas crônica e aguda de paracoccidiodomicoses usando antígeno total e tratado com meta-periodato. Todos os tipos de imunoglobulinas presentes nos soros de pacientes com formas aguda e crônica apresentaram alta reatividade ao antígeno total quando comparado ao tratado com meta-periodato (P < 0,05). Houve maior reatividade de IgG e IgM anti-antígeno tratado com meta-periodato em soros de pacientes com forma aguda da doença (P < 0,05), enquanto IgA foi mais reativa em soros da forma crônica (P < 0,05). Houve maior reatividade de IgG1 e IgG2 com antígeno total e tratado com meta-periodato em soros de pacientes comparados aos com outras parasitoses (P < 0,05). Além disso, IgG1 de pacientes com a forma aguda reconhecem antígenos de 19kDa, 27kDa e 31kDa por western blot. Assim, os resultados sugerem que alterações nos epitopos de antígenos de Paracoccidioides brasiliensis podem auxiliar no aprimoramento do imunodiagnóstico da paracoccidioidomicose.

Scanning electron microscopy of encapsulated and non-encapsulated Cryptococcus neoformans and the effect of glucose on capsular polysaccharide release.

Cryptococcus neoformans has a polysaccharide capsule composed primarily of glucuronoxylomannan (GXM). This study focuses on the morphology of both encapsulated and non-encapsulated organisms in the presence and absence of monoclonal antibodies (mAbs) and serum proteins, and the effect of glucose on capsular polysaccharide release. Examination of the encapsulated C. neoformans strains 24067 and 34873 by scanning electron microscopy (SEM) revealed globular cells covered with a loose fibrillar network which was most prominent during the early stationary phase. In the presence of GXM-binding mAbs or serum the capsule border became distinct and bud scars were evident in the fibrillar network. In contrast, SEM of strain 52817, a non-encapsulated mutant of 34873 revealed ovoid cells devoid of a fibrillar network with bud scars and small surface protrusions. mAb 2H1 bound to cells from strains 24067 and 34873 but not 52817. No GXM was detected in supernatants of 52817 culture. For several strains, there was significantly more GXM in culture supernatants using high glucose media. In summary, our results indicate: i) SEM methods for studying capsular structure in C. neoformans; ii) no reactivity by GXM-binding mAb with a non-encapsulated strain; iii) the presence of distinctive bud scars in both encapsulated and non-encapsulated cells; and iv) dependence of GXM concentration on glucose concentration in culture media. The implications of these results are discussed.

Structural changes in rat Pneumocystis carinii surface antigens after terbinafine administration in experimental P. carinii pneumonia.

Terbinafine is a synthetic antifungal agent which has recently been found to be highly effective against Pneumocystis carinii. This study evaluated the efficacy of terbinafine on rat P. carinii antigenic profile and the immune response by Western blot analysis, in comparison with atovaquone and co-trimoxazole in rats with pneumocystosis. Terbinafine was shown to target two specific major antigens, particularly those of 116 and 35-40 kDa. Antibodies reactive against these moieties were found in all rats treated with atovaquone and co-trimoxazole, but not in those treated with terbinafine. These surface antigen modifications could be related to disease severity and could provide additional information for monitoring the efficacy of this treatment.

Assay for detecting IgE and IgG antibodies against Candida albicans cell-wall mannan.

In the present study, we assayed mannan-specific IgE and IgG antibodies in samples of serum isolated from blood collected from adult patients with bronchial asthma, using a liquid-phase method with a polysaccharide, mannan (Mn), purified from Candida albicans (C. alb), and investigated the relationships of allergenicity among a crude extract of C. alb, purified Mn, and acid protease (AP), The correlations between the titers of anti-Mn A and anti-Mn B IgE and IgG were very strong, and the levels of inhibition of anti-Mn A IgE and IgG reactions by Mn A and Mn B were almost identical. Although no common allergenicity was observed between Mn A and AP because there was no correlation between the titers of anti-Mn A and anti-AP IgE, and no inhibition of the anti-Mn A IgE reaction by AP, both antigens were found to exist in crude C. alb. The level of inhibition of anti-crude C. alb IgG reaction by Mn A or Mn B was about 60%. Approximately 70% inhibition of the anti-Mn A IgE reaction was observed for eight different fungal allergen extracts, but no inhibition was observed for 11 of the other fungal allergen extracts tested. The above results indicate that common antigenicity was observed between Mn A and Mn B in the human IgE and IgG antibody production system, and the cross-allergenicity observed among some fungi was considered to be the result of the common antigenicity of Mn isoforms.

Influencia del tratamiento con zidovudina (AZT) sobre los niveles de antígeno polisacárido capsular de Cryptococcus neoformans en los fluídos biológicos / Influence of zidovudine (AZT) on levels of capsular antigen of Cryptococcus neoformans in body fluids

Fueron investigados los títulos de antígeno polisacárido capsular de Cryptococcus neoformans en el momento del diagnóstico en 25 pacientes con criptococosis asociada al SIDA. Siete pacientes recibirían en el momento del diagnóstico de la micosis 500-600 mg/día de zidovudina (AZT) y otros 18 no recobirían ninguna medicación antirretroviral. Todos ellos recibieron tratamiento antifúngico específico inmediatamente después de realizado el diagnóstico micológico. Los títulos de los pacientes tratados con AZT fueron más bajos que aquellos no tratados con ningún esquema antirretroviral, aunque la diferencia observada careció de significación estadística (p>0,05). El promedio del tiempo de sobrevida (tomando desde el momento del diagnóstico de la micosis hasta la muerte) fue significativamente más largo (504,43 ñ 160 días) (p = 0,002). Tampoco se observaron diferencias significativas entre los recuentos de linfocitos CD4+ y la prevalencia de diferentes infecciones intercurrentes en ambos grupos. Las diferencias observadas entre ambas poblaciones estudiadas puede demostrar indirectamente la eficacia de la terapéutica antirretroviral para retardar el daño inmunológico provocado por el VIH sobre el sistema inmune de los pacientes (AU)

Effects of tunicamycin on glycoprotein antigens of Aspergillus fumigatus.

Tunicamycin, which inhibits N-glycosylation of proteins, was used as a tool to determine the type of linkage which occurs in glycoprotein antigens of Aspergillus fumigatus. When A. fumigatus extracts were electrophoretically separated and blotted then probed with anti-Aspergillus patients' sera, differences in antigenic profiles were noted when tunicamycin-treated samples were compared with controls. Tunicamycin had no detectable effect on the cellular proteinases of A. fumigatus, most of which are glycosylated. Some enzymatic components were lacking when extracellular proteinases were compared with those of control samples. The major catalase component of A. fumigatus is a concanavalin A (ConA)-binding glycoprotein. In cultures grown in the presence of tunicamycin, partially-deglycosylated catalase components were obtained which could be distinguished from the native catalase by altered mobilities in polyacrylamide gels. The effect of deglycosylation on catalase antigens was monitored using an antiserum raised to a ConA-binding fraction of A. fumigatus mycelium. These antibodies bound both to the native glycoprotein and the partially deglycosylated material. These latter two were largely unaffected when incubated with an antiserum raised to a non-ConA-binding fraction of A. fumigatus which is essentially carbohydrate free. The ability to produce partially-glycosylated antigens of A. fumigatus offers a model to study the effect of basic structural modifications on both the enzymatic and antigenic activities of these molecules.

Experimental aspergillosis in guinea pigs: influence of itraconazole on fungaemia and invasive fungal growth.

The guinea pig model of experimental aspergillosis was used to evaluate the efficacy of itraconazole 2.5 and 5 mg kg-1 in preventing the invasive phase of the disease when animals were already loaded with Aspergillus conidia. Evaluations were made by recording the survival rates, culturing fragments of nine organs, examining seven organs by means of histochemistry and immunohistochemistry (mAb EB-A1 to Aspergillus galactomannan) and by serological titration of galactomannan. The data indicate that itraconazole is highly effective in preventing true invasive aspergillosis. Serological evaluations of antigenaemia suggest that low titres may only reflect fungaemia, while titres of 1:8 and above are suggestive of invasive disease.

Variations in the antigenic effects of tunicamycin on Candida albicans.

We report on the effect of subinhibitory doses of tunicamycin on Candida albicans cells (BP strain high responder NCYC 1466) in a defined medium favourable for expression of the mycelial phase. Tunicamycin inhibited the synthesis of some protein fractions ranging from 40 to 65 kDa, where the immunodominant antigens of C. albicans responsible for the antibody response to systemic mycosis were inhibited. By two-dimensional immunoelectrophoresis, antigen extracts from the cell cultures grown with tunicamycin showed a migration modification and a lower number of precipitation arcs with variation in their height and range.

The effect of sulconazole on the antigenic composition of two dermatophyte species.

Trichophyton rubrum and Trichophyton interdigitale have been grown in liquid culture in the presence of sulconazole. The antigenic activity of detergent extracts of intact organisms was analysed following SDS-PAGE and the probing of Western blots with homologous antisera raised in rabbits and with sera from patients with dermatophyte infections. Differences in protein-band patterns were noted; some bands present in control samples were absent in azole-treated samples and vice versa. These differences were reflected in antigenic band patterns, especially among components of approximate molecular weight of 30-40, 50-60 and 92-100 kDa.

Effect of successful treatment with amphotericin B on Histoplasma capsulatum variety capsulatum polysaccharide antigen levels in patients with AIDS and histoplasmosis.

PURPOSE: The purpose of this study was to establish the effect of induction and maintenance treatment with amphotericin B on levels of Histoplasma capsulatum var. capsulatum polysaccharide antigen (HPA) in the urine and blood of patients with acquired immunodeficiency syndrome (AIDS) and disseminated histoplasmosis. PATIENTS AND METHODS: This was a retrospective study of the effect of amphotericin B treatment on levels of HPA in the urine or serum from 70 patients with AIDS and disseminated histoplasmosis. All patients received initial intensive induction treatment with amphotericin B, and a subset continued to receive amphotericin B at less frequent intervals for maintenance therapy to prevent relapse. Treatment regimens varied in intensity and duration and specimens were obtained at irregular intervals. Urine and serum specimens were stored and retested for HPA in the same radioimmunoassay. RESULTS: HPA levels in serum decreased by at least 2 units during induction therapy in all 19 (100%) patients with initial levels of greater than or equal to 2.6 units and reverted to negative in 40.9% of those with initial levels of greater than or equal to 1.0 unit. HPA in urine decreased by at least 2 units in 84.8% and reverted to negative in 17.3% of patients. During induction treatment, HPA cleared more rapidly from serum than from urine. During maintenance treatment, HPA levels in serum decreased by at least 2 units in 100% and became negative in 66.7%. HPA in urine decreased by at least 2 units in 54.5% and reverted to negative in 20.0%. Rates of clearance of HPA from the serum and urine were similar, 0.01 unit/week compared with -0.04 unit/week, respectively, but less than rates during induction treatment. CONCLUSIONS: Successful therapy of histoplasmosis with amphotericin B is associated with reduction of HPA in body fluids. Periodic measurement of HPA levels offers a method for monitoring the response to therapy and for comparing new treatments for histoplasmosis.