Cry j I, a major allergen of Japanese cedar pollen, has pectate lyase enzyme activity.

In the course of analyzing the partial amino acid sequences of Cry j I, a major allergen of Japanese cedar (Cryptomeria japonica) pollen, we found a peptide fragment which has a significant homology to some pectate lyase isozymes secreted by plant pathogenic bacteria. Therefore, we investigated whether Cry j I has pectate lyase activity. Cry j I reacted with polygalacturonic acid, resulting in the release of unsaturated uronide products.

Epitopes on Cry j I and Cry j II for the human IgE antibodies cross-reactive between Cupressus sempervirens and Cryptomeria japonica pollen.

Forty sea from French patients allergic to Cupressus sempervirens pollen were tested for cross-reactivities against Cry j I, Cry j II (major allergens of Cryptomeria japonica pollen) and other pollen allergens from botanically related plants. Seventy-three per cent of the sera reacted with either Cry j I or Cry j II, or with both of them. These IgE cross-reactions were blocked effectively by mAb 046 (anti-Cry j I) or N26, T27 (anti-Cry j II), and weakly by mAbs 052, 027 and 026 (anti-Cry j I).

Suppression of murine IgE responses with ovalbumin-pullulan conjugates: comparison of the suppressive effect of different conjugation methods and different molecular weights of pullulan.

Ovalbumin (OVA)-pullulan conjugates were made using four different conjugation methods and eight different molecular weights of pullulan ranging from 4,200 to 600,000. Pretreatment of mice by the administration of conjugates made by using cyanulic chloride or cyanogen bromide and pullulan of molecular weight 40,000 or more, anti-OVA IgE antibody response was suppressed completely and anti-OVA IgM and IgG antibody response was enhanced. In contrast, by the administration of conjugates made by using an oxidation or thiol activation method, only partial suppression of anti-OVA IgE antibody response was achieved and no enhancement of anti-OVA IgM and IgG antibody production was observed.

The presence of immunoreactive parathyroid hormone-related protein in parathyroid adenoma cells.

Parathyroid hormone-related protein (PTHrP) was first identified in human malignant tumors associated with humoral hypercalcemia of malignancy. We immunohistochemically examined the distribution of PTHrP both in 7 normal parathyroid glands and in a 20 parathyroid adenomas. Sixty-five percent of parathyroid adenomas (13 cases) were positive for PTHrP, whereas only one normal parathyroid gland was positive for PTHrP in the area of transitional oxyphil cells.

Antigenic analyses of Sugi basic protein by monoclonal antibodies: II. Detection of immunoreactive fragments in enzyme-cleaved Cry j I.

The 4 anti-Cry j I mAbs showing an epitope specificity different from each other, 046, 029, 026 and 027, were selected to analyze the structure of the antigenic determinant for each mAb on a Cry j I molecule. Immunoreactive fragments in enzyme-cleaved Cry j I were detected by means of the adsorption on the mAb column and of the binding to the mAbs on Elisa. The mAb 026 was found to be reactive to the fragments containing a Cry j I N-terminal region obtained by V8 protease or pepsin digestion, but not to those by lysylendopeptidase digestion.

Antigenic analyses of Sugi basic protein by monoclonal antibodies: I. Distribution and characterization of B-cell-tropic epitopes of Cry j I molecules.

Using 23 monoclonal antibodies raised against Sugi basic protein (SBP, major allergen of Japanese cedar pollen), composed of Cry j I and Cry j II, analyses of B-cell-tropic epitopes of Cry j I were performed. The following results were obtained. (1) As far as the mAbs were used, no major cross-reactive determinants were detected between Cry j I and Cry j II molecules.

Growth of rat-mouse hybridoma cells in immunosuppressed hamsters. An easy and effective method to prepare monoclonal antibodies from heterohybridoma cell lines.

Rat-mouse hybridoma cells producing anti-mouse IgE antibodies were intraperitoneally or subcutaneously inoculated into newborn or suckling hamsters receiving rabbit anti-hamster thymocyte globulin from the day of birth twice a week for at least 3 weeks. The hybridoma cells were found to grow in the abdominal cavity of the hamsters as ascites tumor or in subcutaneous tissue as solid tumor without loss of antibody-secreting activities. For the production of ascites, 2-week-old hamsters were preferable to newborn hamsters.

Identification of the second major allergen of Japanese cedar pollen.

We isolated and characterized the second major allergen (Cry j II) from Japanese cedar pollen. We found that most patients with this pollinosis had IgE antibody to this protein in addition to IgE antibody to Cry j I; however, some sera reacted only with Cry j I or Cry j II. IgE-ELISA inhibition studies revealed that Cry j I and Cry j II had no cross-allergenicity.

Preparation of a diphtheria toxin-pullulan conjugate that elicits good IgG antibody production with poor IgE synthesis.

Diphtheria toxin is detoxified through conjugation with pullulan. The toxin-pullulan conjugate is easily purified by DEAE-Sephacel chromatography. The conjugate forms a transparent 'clear line' with anti-toxin antibodies on agarose plate, which offers a good indicator of conjugate formation.

Biological and immunological properties of Sugi basic protein-pullulan conjugate. II. Is the reduced ability to elicit the Arthus reaction based on the poor activation of complement by immune complex consisting of anti-Sugi basic protein and Sugi basic protein-pullulan?

Ability of Sugi basic protein (SBP)-pullulan conjugate to elicit the Arthus reaction was found to be markedly reduced, about 100 times lower than that of native SBP. To analyze this reduced ability, activation of complement by immune complex consisting of SBP-pullulan and anti-SBP antibodies was studied. Tests for complement consumption, C3 conversion and cleavage of factor B revealed that immune complex formed with SBP-pullulan is incapable of supporting efficient activation of the complement system.

Comparison of immune responses to diphtheria and tetanus toxoids of various mouse strains.

Immune responses of 11 mouse strains with known genetical characteristics and two outbred strains to diphtheria and to tetanus toxoids were compared. Both diphtheria and tetanus antitoxins were titrated by passive hemagglutination. From the pattern of the immune response, the mouse strains tested may be classified into four groups.

Biological and immunological properties of Sugi basic protein-pullulan conjugate. I. Suppressive effect on IgE antibody production and on IgE-mediated reactions.

Sugi basic protein (SBP), a major allergen of Japanese cedar pollen, conjugated to pullulan acquires suppressive activities for IgE, but not for IgG antibody production, and for IgE-mediated PK reactions in a SBP-specific manner. When either normal or IgE anti-SBP antibody-producing mice were treated with the conjugate, IgE anti-SBP antibody levels scarcely increased even after immunization with SBP + alum. This suppression was shown to be SBP specific and IgE selective.

N-terminal amino acid sequence of a major allergen of Japanese cedar pollen (Cry j I).

A purified preparation of a major allergen of Japanese cedar pollen, sugi basic protein (SBP, Cry j I), was separated into 5 subfractions of 50-45 kDa. All of the SBP subfractions were confirmed to be reactive to IgE antibodies from patients with Japanese cedar pollinosis, and also to mouse anti-SBP monoclonal antibodies. The sequences of 20 N-terminal amino acids of these 5 subfractions were found to be identical.

One-point method for serological diagnosis of leptospirosis: a microcapsule agglutination test.

This paper describes a simple and rapid microcapsule agglutination (MCA) test. The results obtained by this new test have been compared with those obtained by the microtitre MCA and the microscopic agglutination (MA) test. The procedures required for the new test are easier and can be performed more rapidly than those necessary for the microtitre MCA test.

Serodiagnosis of leptospirosis in China by the one-point MCA method.

The one-point MCA method is very simple to perform and useful as a screening test in diagnosing leptospirosis in routine clinical laboratories. The kit, sensitized with six serovars occurring in Japan, was also useful in detecting serum antibodies of patients with leptospirosis in China.

Epidemiological studies on leptospirosis in Chiang Mai (Thailand).

A total of 270 serum samples collected in Chiang Mai province were examined for antibodies against leptospira using the microscopic agglutination test (MAT). Four of 40 serum specimens from patients who visited the hospital with the common cold, were positive with a titre of 20. Twelve (10.

Immunopotentiative effect of polysaccharide from kefir grain, KGF-C, administered orally in mice.

Since a water-soluble polysaccharide (KGF-C) from the kefir grains was shown to have the property of retarding tumor growth in vivo when administered orally, the effect of KGF-C was examined on antibody responses to thymus-dependent antigen, sheep red blood cells (SRBC), and thymus-independent antigen, dinitrophenyl-Ficoll and trinitrophenyl-lipopolysaccharide. Antibody response in mice intubated with KGF-C was enhanced to low doses of SRBC, but not to optimal or high doses. The optimal dose of KGF-C required for the enhancement was 100 mg/kg body weight.

IgE antibody against hepatitis B surface antigen in mice.

Hepatitis B surface antigen (HBs antigen) was examined for elicitation of IgE production by injection into mice. The Prausnitz-Küstner (PK)-type skin test in the rat was employed for detection of IgE antibody to HBs antigen, because no sufficient purified HBs antigen was available as the challenging antigen for the passive cutaneous anaphylaxis (PCA) test in rats. The positive PK test was considered to be due to IgE antibody, since the active principle was inactivated by heating the sera at 56 C for 30 min, did not bind to protein A and was eliminated by anti-mouse IgE antisera.

Effects of pH and lysine during detoxification of a hemorrhagic principle of Habu snake (Trimeresurus flavoviridis) venom with formalin on the immunogenicity of the toxoid.

A hemorrhagic principle (HR1B) isolated from the venom of Habu snake (Trimeresurus flavoviridis) was detoxified with formalin at pH 5, 7 and 9. Higher pH shortened the period needed for detoxification and lowered the final formalin concentration required for complete detoxification. The extent of polymerization and the immunogenicity of the toxoid formed were dependent upon pH during detoxification; the lower the pH, the higher the extent of polymerization and the immunogenicity of the toxoid.

'Clear line' formation by immunodiffusion of tetanus toxoid and mixtures of distinct monoclonal antibodies.

Single monoclonal antibodies could not form any precipitin line with tetanus toxoid in Ouchterlony analysis. However, some mixtures of two distinct monoclonal antibodies formed a visible precipitin line (conventional line), while others formed a transparent, just barely visible line, which we call a 'clear line'.