Heated histoplasmin as a control for non-specificity in the complement fixation test for histoplasmosis

We present data in support of the use of a heated histoplasmin control for the complement fixation (CF) test for histoplasmosis to assist in the detection of the presence of h or m antibody.     

Candida albicans antigens studied in complement fixation test and agar gel diffusion test

Summary The antigens prepared simultaneously from seven type ACandida albicans strains by cell disintegration by means of sonic vibrations, grinding with the sea-sand, autolysis and the antigens precipitated with acetone from fluid cultures were studied in complement fixation test and agar gel diffusion test against selected rabbit immune sera. The complement fixation test was performed in plexiglass plates as chessboard titration. The agar gel diffusion test was performed in petri dishes according toOuchterlony and according toBjörklund's specific inhibition of precipitation. The activity of studied antigens determined in complement fixation test was high. The antigen titers were ranging from 1/512 to 1/8192. The reaction observed in agar gel diffusion test were also strong and precipitation spectra consisted of 1–3 lines. The reaction pattern obtained in both tests depended on the character of the analysed reagents. TheBjörklund's modification was successfully applied to serological grouping of the strains under study. Sixteen human sera collected from patients with symptoms of Asthma bronchiale and Bronchitis chronica, from whose sputumCandida albicans was isolated, were tested in both tests. The researches made on human sera were intended as preliminary tests.Head of the department: Prof. Dr.Z. Przybylkiewicz.     

Procedures for the production and separation of H and M antigens in histoplasmin: Chemical and serological properties of the isolated products

Two strains of Histoplasma capsulatum were required to prepare maximum yields of H and of M antigen from histoplasmin. The antigens were separated and partially purified by a series of procedures yielding an overall recovery of 70 to 90% of the individual antigens. Stable products suitable for use as reference products were obtained when the final purification step employed DEAE-cellulose with phosphate buffer elution at increasing molarity and decreasing pH. A final step of purification of each antigen with slab acrylamide gel electrophoresis gave products which were highly reactive and specific in a variety of serological tests with sera from persons with proven cases of histoplasmosis and with natural infections of heterologous deep mycoses. These antigens were maximally active at concentrations of 2 to 16 g protein in the complement fixation, capillary precipitin, microimmunodiffusion, or immunoelectrophoresis tests; 0.5 g gave a maximum delayed cutaneous hypersensitivity reaction in homologously infected animals and caused no appreciable reaction in control animals. Although these antigens appeared to be specific when tested with sera from persons with natural infections, the M and H antigens demonstrated the presence of an additional antigen reacting with sera of rabbits immunized with cell membrane and cell particulate fractions of Blastomyces dermatitidis. After purification by electrophoresis, both the H and M antigens of some preparations showed some decomposition and loss of reactivity after storage at 5 C for more than six months. The overall results suggest that the purified H and M antigens of Heiner (12) have multiple serological reactivity and may function in precipitin reactions, complementfixing reactions, hemagglutination of formalin-fixed goose red blood cells, and as antigens for delayed cutaneous tests.     

The complement fixation test in vaccinia with antigens from the brain,the testis and the skin

Summary Eight antigens, prepared from the brain, the testis and the skin of rabbits after the tissues had been inoculated with vaccinia virus, have been compared in the complement fixation test with vaccinia-immune-sera. It was rare that a serviceable antigen could be prepared from the brain. Antigens from the testis and the skin were always active, but the activity of the former surpassed that of the latter. The best results were obtained with a saline extract 1 : 100 of freshly removed testis; when frozen this extract may be kept for some weeks. In the sera of vaccinated rabbits and of a monkey complement fixing antibodies could be detected regularly, even 3 months after vaccination. In sera of men, who had been revaccinated more than 3 months earlier, this was only rarely the case.     

Analytical and preparative isotachophoresis of histoplasmin purified derivative components.

Skin test-active components of histoplasmin have been difficult to purify in quantity in the past. Analytical isotachophoresis in polyacrylamide gels was used to develop a high resolution system to separate the components of histo-plasmin. The analytical system was then expanded to preparative-scale isolation of the skin test-active components of histoplasmin.