In order to assign the location of the antibody active sites, purified anti-DNP and anti-HAS rabbit γG antibodies were subjected to reduction and alkylation and their heavy and light polypeptide chains were separated by gel filtration. Subsequently, the reconstituents of heavy and light chains isolated from antibody molecules as well as the hybrid reconstituents of antibody polypeptide chains and normal γG counterparts were prepared. In comparison by radioimmunoelectrophoresis and passive hemagglutination of the antibody activity of these preparations, I.e. heavy chain, light chain and their specific or nonspecific (hybrid) reconstituents, following results were obtained. 1) Isolated light chains of anti-DNP or anti-HAS antibodies had no antibody activity. 2) On the contrary, isolated heavy chains obviously showed the antibody activity, though less active than original antibody γG. 3) Obvious enhancement of antibody activity of heavy chains was observed when they were combined with light chains derived from antibodies having same specificity. The enhancement is considered to be attributable to the reconstitution of heavy and light chains to form original γG molecules. 4) When light chains of normal γG globulin molecules were added to antibody heavy chains, the reconstitution of both chains into 7S γG globulin was observed to occur, but the activity of these reconstituents was the same as that of heavy chain alone. 5) The reconstituent consisting of normal heavy chain and antibody light chain did not show any antibody activities. 6) Reduction and alkylation of purified antibody without subsequent exposure to acid had little effect upon its antibody activities.

View full abstract

The prolongation of coagulation of antibody-containing blood under anaphylactic shock has long been observed by many investigators. The phenomenon might be the result following the antigen-antibody reaction which has occurred in vivo. The modification of the blood coagulation by antigen-antibody reaction was studied by means of thrombelastography which was designed for continuous observation through its all phases. The antibody-containing blood was obtained from rabbits immunized with egg albumin or bovine serum albumin. In vitro tests, shortages of r and r + k were observed in all samples mixed with corresponding antigen showing an accerelation of coagulation and also a marked diminution of ma was observed in the test of citrated blood. On the other hand, in vivo test, prolongations of r and r + k with diminished ma were observed at 10 min after the injection of antigen. The marked prolongation of r and r + k in vivo test would suggest an antagonistic mobilization of some anticoagulants against accerelation which was primarily caused by antigen-antibody reaction. An activation of fibrinolysis was not observed thrombelastographically in both vivo and vitro tests.

View full abstract

The aim of this work was to study the antigenic components present in Candida albicans and to clarify the relationships between the dermal and bronchial reactions of immediate and delayed type provoked with the fractions from the organisms in man. Organisms of Candida albicans were repeatedly extracted with 0.1 N KOH and then with 1 N KOH. These extracts were precipitated with HCl at pH 4.0 and the supernates were deproteinized with TCA and precipitated with ethanol. These crude polysaccharide fractions were used as crude extracts. The crude extracts were further purified by the treatment of proteolytic enzyme followed by filtration with Sephadex G-200 to obtain purified polysaccharide fraction (PS). Precipitates from original extracts with HCl at pH 4.0 were purified by zone electrophoresis to obtain purified protein fraction (Prot.). PS contained 0.77% N and 95.3% sugar and Prot. Contained 14.2% N and 2.7% sugar. Skin tests with these fractions revealed: PS elicited immediate reaction, Prot. Showed immediate, delayed, or both reactions, and crude extracts provoked immediate, intermediate, delayed, or some combinations of these reactions. The incidences of positive immediate skin reactions were 37.4% in asthma, 27.3% in allergic rhinitis, 7.1% in tubeculosis and 9.0% in other nonallergic diseases. There was a significantly high incidence of immediate neactions in the atopic group for PS or crude extract. However, the atopic individuals exhibited no greater incidence of delayed neactions than the nonatopic individnals for Prot or crude cxtrats' Sixteen inhalation tests were performed in 13 asthmatic patients. The bronchial and systemic reactions were studied by spirometry, bodyplethysmography, measurements of body temperature, respiration rates, white blood counts, etc. Two among 5 subjects which were challenged with PS showed immediate reaction only, 1 among 4 who inhaled Prot. Showed delayed reaction only. Some challenged with one of both fractions, crude extracts or commercially available antigen extracts, reacted both immediately and delayedly. One of 2 subjects with negative skin reactions showed also negative bronchial reaction when crude extracts were inhaled. It may be concluded from the above observations that the antigenic substances of immediate skin hypersensitivity are included in PS of the C. albicaus while Prot. Contains those of delayed. This seems to be true on the antigenicity of the components of Candida albicans for bronchial reaction. Immediate allergic reaction is kown to induce bronchoconstriction in asthmatic patients. It is likely that delayed type reaction with exudation, edema and cell infiltration in respiratory tract may also cause dyspnea in patients with hyperreactivity in the bronchus and bronchioles.

View full abstract

This report is given of the results of skin tests, inhalation tests and specific hyposensitization therapy with five species of Torii mold extracts (Aspergillus, Alternaria, Cladosporium, Candida, Penicillium) on infants and children with bronchial asthma who visited our clinic. The results were as follows: 1. Seventeen (16.2%) of 105 patients reacted positively to scratch tests with extracts of five species of molds. 2. Two hundred ninety (50.2%) of 578 patients reacted positively to intracutaneous tests with molds. 3. Inhalation tests were made on 168 patients who were skin sensitive to mold, and 73 (43.5%) of these displayed positive provocative reactions to one or more of the five mold extracts. Alternaria and Candida gave rise to positive provocative reactions rather more frequently than did the other extracts. 4. From the observation above, it can be assumed that about 20 per cent of asthmatic children are sensitive to molds. 5. The rate of positive reactors to inhalation tests with mold extracts showed a tendency to decrease with the advance in age. 6. Most mold-sensitive patients gave a history of aggravation of symptoms in autumn but slight difference was recognized as to seasonal aggravation among the different species of molds. 7. According to the results of inhalation tests with house dust and molds on the same subject, relatively many patients reacted positively to either house dust or mold extracts but only 26.5 per cent were sensitive to both. 8. Twenty-nine patients who were sensitive to molds were treated over one year period with various mold extracts. Consequently 65.5 per cent of these showed a remarkable improvement in symptoms. The incidence of side effects resulting from hyposensitization therapy was 17.1 per cent.

View full abstract

The development of direct fluorescent complement technique has been done for demonstration of antigen-antibody reaction, because of the specificity of complement fixation reaction. 1 . For the specificity of this technique, human C1 was purified by the repeating of precipitation in low ionic strength and solubilization of euglobulin in high ionic strength at neutral pH. Purified C1 thus obtained showed high specific hemolytic activity. 2. Several conditions for the conjugation of FITC to the purified C1 were examined. Using the fluorescence labeled C1, specific fluorescence was observed in the following systems; sheep erythrocyterabbit anti sheep erythrocyte serum and MM2 (ascites form of mouse mammary tumor)-xenogeneic and syngeneic anti MM2 sera system. 3. When fluorescent C1 was separated into the three subcomponent fractions by Lepow's method, the strongest fluorescence was recovered from Clq fraction. 4. Specific fluorescence on the sensitized cells was not influenced by the addition of EDTA. However, it is known that C1q is capable of binding to antibody molecules in spite of the presence of EDTA. This phenomenon was discussed from this point, in connection with the existence of the majority of FITC on C1q moiety of C1 molecule.

View full abstract