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Biomedical subjects

T Matuhasi

Publications and source records attributed to T Matuhasi.

At least 19 recordsLinked to original sources

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. Many parathyroid adenomas (12 cases) were composed of both PTHrP-positive cells and--negative cells, and these two different type of cells showed a tendency to form nodules respectively in parathyroid adenoma. Although both chief cells and oxyphil or transitional oxyphil cells were positive for PTHrP in parathyroid adenoma, oxyphil or transitional oxyphil cells were more responsible for PTHrP production than chief cells. Chief cells are thought to produce parathyroid hormone mainly in parathyroid adenoma. On the other hand, little is known concerning the function and role of oxyphil or transitional oxyphil cells. Our results suggest that oxyphil or transitional oxyphil cells in parathyroid adenoma may have some functional roles different from those of chief cells through the production of PTHrP.

Adenoma

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. (2) 21 of the 23 mAbs were specific for Cry j I, and the anti-Cry j II mAbs were classified into four groups by their fine specificities, suggesting that Cry j I bears at least four antigenic determinant regions. (3) Cry j I molecules were found to take a monomeric form in solution and to display no repeating antigenic epitopes on their surfaces. (4) Some of the determinants seemed to be located in the interior of a Cry j I molecule, and when the Ag is coated on a plastic plate, the determinants become exposed on its surface. (5) Binding of human IgE antibodies to Cry j I and Cry j II was blocked by some of the obtained mAbs, suggesting that these epitopes recognized by the mAbs might have an important role in human allergic response against the cedar pollen.

Allergens

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. The mAb 027 was found to be capable of binding to the fragments containing a linear structure of Asn-Ala-Gly-Val-Leu-Thr-Cys-Ser-Leu-Ser-Lys, which were generated by V8 protease, lysylendopeptidase or pepsin digestion. Furthermore, the synthetic peptide Asn-Ala-Gly-Val-Leu-Thr-Cys-Ser- Leu-Ser-Lys-Arg could bind to 027, but not to 026, and could inhibit the binding of 027 to Cry j I or to its immunoreactive fragments. No fragments capable of reacting to the mAbs 046 and 029 could be found in this study, suggesting that 046 and 029 recognize a conformationally constituted epitope of Cry j I molecule which is destroyed by enzymatic cleavage. The epitope recognized by the mAbs 027 or 026 was found to be located in conformationally hidden parts of the molecule which was exposed to react to the mAbs only after the physicochemical or enzymatic treatment.

Allergens

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. In 3-week-old hamsters, the hybridoma cells could scarcely survive. The antibody titers of the ascites were determined to be 10(5)-10(6) in the ELISA and in the ability to neutralize the skin-sensitizing capacity of mouse IgE antibodies. The rat monoclonal antibodies were easily separated from ascites, serum or cell culture supernatant with affinity chromatography using Affigel protein A-Sepharose and anti-hamster IgG-Sepharose columns. The described method could be efficiently applicable for the proliferation of other hybridomas, such as human-human, human-mouse or hamster-mouse, etc.

Animals

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. The toxin-pullulan conjugate induces both IgG1 and IgG2b antibody production with diminished IgE response, while the alum-precipitated conventional toxoid causes mainly increases in IgE as well as IgG1 antibody formation. The anti-toxin HA titre (IgG antibody) induced by the toxin-pullulan conjugate parallels the neutralizing activity of the immune-sera. These results suggest that the conjugation of toxin to pullulan is a very powerful method by which to develop a vaccine that induces neutralizing antibody with diminished IgE antibody synthesis.

Animals

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. Cry j II did not react with anti-Cry j I monoclonal antibodies. In SDS-PAGE under a non-reducing condition, Cry j II showed a band at the 37 kDa position, compared with the 45-50 kDa bands of Cry j I. N-terminal amino acid sequence of Cry j II was completely different from that of Cry j I.

Allergens

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. Previously, we have shown data suggesting that SBP-pullulan conjugate would be a good candidate for desensitization therapy against cedar pollinosis. The results presented in this paper provide additional support for the suggestion.

Allergens

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. The ability of SBP-pullulan conjugate to elicit PK reactions on passively sensitized rat skin was markedly reduced. Furthermore, the conjugate inhibited the reactions mediated by the IgE antibodies and native SBP when it was injected into the IgE antibody-sensitized sites 2 h prior to SBP challenge. The reduced ability of the conjugate to elicit PK reactions was not due to loss of antigenic determinants during the conjugation procedures because binding activity of the conjugate to anti-SBP antibodies was as strong as that of native SBP. These results indicate that SBP-pullulan conjugate may be useful for 'desensitization therapy' against the pollinosis.

Allergens

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. [1] Strains ddY (SPF) and ddY (conv) and those with haplotype H-2b, such as C57BL/6 and C57BL/10, were high responders to both toxoids. [2] Strains with H-2d, such as BALB/c, B10.D2 and DBA/2Cr, were intermediate responders to both toxoids. [3] Strains with H-2k, H-2a or, H-2m, such as C3H/He, B10.BR, B10.BR/SgSn, B10.A/SgSnJ and B10.AKM/O1a, were high responders to diphtheria toxoid but low responders to tetanus toxoid. [4] The strain with H-2h4, B10.A (4R), was a poor responder to both toxoids.

Animals

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. Peptide mapping analyses of the SBP subfractions showed similar patterns, with some differences which might in part be due to the existence of an N-linked carbohydrate chain. The N-terminal amino acid sequence of SBP was identical to the reported sequence of an allergen of mountain cedar which vegetated in North America.

Allergens

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 (10.4%) of the 115 samples in the Doi Saket district showed a positive reaction. Only 2 of 115 sera of school children in Chiang Mai city had antibodies. Specific serovars detected were Leptospira hebdomadis (5), L. australis (3), L. icterohaemorrhagiae (2), L. bataviae (2), and one each of L. canicola, L. javanica and L. pyrogenes. One case of mixed infection with L. hebdomadis and L. javanica, and L. autumnalis and L. australis were observed.

Adolescent

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.

Agglutination Tests

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. Furthermore, the new test is more sensitive than the MA test in the early stages of leptospirosis. This new test appears satisfactory as a screening test for the early diagnosis of leptospirosis.

Agglutination Tests

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. The time-course studies on KGF-C administration implied that KGF-C exerted its effect on the early events of anti-SRBC response. The enhancement was not due to the alteration of kinetics of anti-SRBC responses. Furthermore, the enhancing effect on antibody responses to thymus-independent antigens, such as dinitrophenyl-Ficoll and trinitrophenyl-lipopolysaccharide, was observed neither in nu/nu nor in nu/+ mice, and the effect on delayed-type hypersensitivity response to a low dose of SRBC in normal mice was also found. These findings suggest that the oral immune enhancement by KGF-C is elucidated probably through T-cell but not through B-cell participation.

Adjuvants, Immunologic

Detection by proton nuclear magnetic resonance of elevated lactate concentration in serums from patients with malignant tumors.

Proton nuclear magnetic resonance (1H-NMR) spectra of the serum specimens from patients with malignant tumors were compared with those from presumably healthy persons. We found that 87% of serum specimens from the patients yielded a common proton signal, which was ascribed to the methyl protons of lactic acid; whereas only 9% of serum specimens from the healthy persons tested gave this signal. On the basis of these results we concluded that the lactate level in the serum can be used as a criterion for the diagnosis of cancer in humans and that the determination of lactate concentration in the serum is easily performed by means of 1H-NMR.

Humans