Biomedical subjects

R Yanagawa

Publications and source records attributed to R Yanagawa.

At least 37 records · Page 2Linked to original sources

PubMed1990

Cloning and expression of a pili gene of Corynebacterium renale in Escherichia coli.

A plasmid gene library of Corynebacterium renale piliated strain No. 109P+ was prepared in Escherichia coli in order to study the chemical structure of the pili of C. renale. Of 3,000 recombinant clones tested, 5 reacted with anti-pili anti-serum. The gene products of these clones reacted with anti-pili monoclonal antibodies 8/4, 5/2 and B20/3 but lacked the reactivity with 13/4. SDS-PAGE analysis revealed that the expressed protein had a molecular mass of 48 kilodalton and deletion analysis showed that the encoding region for this protein was localized within a 1.4 kilobase gene including a promoter sequence. Immunoelectron microscopy showed that mouse antibodies raised to the expressed protein bound to the entire surface of the pili of C. renale. These results indicate that the cloned gene encodes a major structural protein of C. renale pili.

Animals↗

PubMed1988

Opsonic effect of monoclonal antibodies against Leptospira interrogans serovar copenhageni.

The opsonic effect of monoclonal antibodies (McAbs) against Leptospira interrogans serovar copenhageni strain Shibaura was examined in vitro using radiolabeled organisms and mouse peritoneal exudate macrophages. Four IgG McAbs (all IgG3) and 2 IgM McAbs were used, all showed different reactivities and were bound to homologous lipopolysaccharide (LPS). IgG3 McAbs at the sub-agglutinating concentration were opsonic, but the degree of the opsonic effect varied among IgG3 McAbs. Opsonization with IgG3 McAbs at the concentration showing the same level of ELISA binding showed that the highest opsonic effect was exerted by Sw-6 antibody. The other IgG3 McAbs showed a similar but lower opsonic effect. IgM McAbs, which were not opsonic at the sub-agglutinating concentration even in the presence of complement, showed opsonic effect at the agglutinating concentration.

Agglutination Tests↗

PubMed1988

Presence of a variety of antigenic variants belonging to at least 4 different serovars in a population of Leptospira interrogans serovar hebdomadis.

The presence of antigenic variants belonging to at least 4 different serovars, jules, kremastos and 2 new serovars, was demonstrated in a population of cloned Leptospira interrogans serovar hebdomadis. Thirteen antigenic variants were isolated from the clone of serovar hebdomadis by culturing in the presence of 13 monoclonal antibodies against 5 serovars, hebdomadis, worsfoldi, kambale, jules and kremastos Kyoto, respectively. These variants were divided into 4 groups, based on the reactivity with the above monoclonal antibodies. Four antigenic variants, one each from the 4 groups, were identified as jules, kremastos and 2 different new serovars. The variation frequency of the 13 antigenic variants ranged from 10(-5.48) to 10(-7.70) except for one, whose frequency was as high as 10(-1.31).

Agglutination Tests↗

PubMed1988

Broad reacting surface antigens in Leptospira biflexa serovar andamana.

Previous investigations had demonstrated that genus-specific and species-specific antigens of leptospires are deep-seated within the leptospiral cell. Conversely, the present study has shown that strain CH11, serovar andamana of the non-pathogenic species of Leptospira biflexa is endowed on its surface with two cross-reacting antigens; a newly recognized antigen common to L. interrogans and L. biflexa spp. and an antigenic determinant common to a previously described genus-specific protein antigen (GP-Ag). The serovar andamana, when thimerosal-treated, behaved like an interspecies-specific antigen, cross-reacting in the complement fixation test with sera from rabbits immunized with L. interrogans and L. biflexa spp. and with sera from subjects with leptospirosis from various serovars. In the immuno electron microscopic test, human leptospirotic sera bound to the surface of thimerosal-treated andamana and not to the surface of untreated andamana showing that an interspecies-specific antigen was located underneath the outermost layer of this serovar. In the same test, the monoclonal antibody GP-7 against the GP-Ag bound to the surface of untreated andamana and not to the surface of thimerosal-treated andamana, showing that an antigenic determinant, common to GP-Ag and different from the first one, was located on the outer membrane of andamana. In human leptospirotic sera, antibodies against the cross-reacting antigen of thimerosal-treated andamana demonstrated by the complement fixation test were formed earlier and in a higher percentage of sera than the serovar-specific antibodies against 16 L. interrogans serovars demonstrated by the microagglutination test.

Agglutination Tests↗

PubMed1988

Affinity constants of anti-leptospira monoclonal antibodies, numbers of antigenic determinants on leptospiras, and their influence on the microscopic agglutination test and ELISA.

The affinity constants of anti-canicola and anti-hebdomadis monoclonal antibodies and the numbers of antigenic determinants per organism of each serovar of leptospira were determined, and their influence on the results of microscopic agglutination test (MAT) and ELISA were examined. In the combinations of a monoclonal antibody and some serovar which showed higher affinity constants at levels of 10(7) to 10(8)/M order, both MAT and ELISA were positive except for one combination, while in the combinations which gave lower affinity constants at levels of 10(6)/M order, MAT was positive but ELISA was negative. The number of antigenic determinants seemed to have no significant influence on the results. The disagreement due to the difference in the threshold affinities of MAT and ELISA should be considered in the use of monoclonal antibodies in the taxonomy of leptospira.

Agglutination Tests↗

PubMed1988

Comparative studies of strains Ictero No. I and RGA as the type strain of Leptospira interrogans: agglutinin absorption test, protein and antigen profiles, and enzyme activities.

Strain Ictero No. I, the first isolate of Leptospira, isolated by Inada and Ido in 1914, was found to be sufficiently qualified to be the type strain of Leptospira interrogans rather than strain RGA. In an agglutinin absorption test, anti-Ictero No. I serum was not absorbed completely with strain RGA, and 25% of the homologous titer remained unabsorbed, while anti-RGA serum was completely absorbed with strain Ictero No. I. Thus, strain Ictero No. I was not serologically identical with strain RGA, and the two strains were considered to be different serovars. A protein band with a molecular weight of approximately 33,000 daltons was detected in strain Ictero No. I but not in strain RGA by SDS-PAGE. By Western blotting, this protein band was detectable with anti-Ictero No. I serum but not with anti-RGA serum. The presence of the 33K protein in strain Ictero No. I, but not in strain RGA, was confirmed by radioimmunoprecipitation using [125I]-labeled antigens, indicating that the protein antigen was surface-exposed. Only 8 of the 89 enzymes activities were different between strains Ictero No. I and RGA (line Sapporo). From the above results, we propose that strain Ictero No. I should be designated as the type strain of L. interrogans instead of strain RGA.

Agglutination Tests↗

PubMed1988

Restriction endonuclease DNA analysis of antigenic variants of leptospires selected by monoclonal antibodies.

The genome of antigenic variant CV (CT3)-1 derived from Leptospira interrogans serovar canicola was compared by cleavage with restriction endonucleases with the parent and serovar bafani, to which the variant was serologically most closely related. No differences were observed between the parent and variant in DNA restriction endonuclease patterns using eight restriction endonucleases. Serovar bafani was different in the patterns from the parent and antigenic variant CV (CT3)-1. The two antigenic variants derived from serovar hebdomadis, HV (H16)-1 and HV (H19)-1 which belonged serologically to serovars jules and hebdomadis, respectively, were compared by restriction endonuclease DNA analysis with the parent and serovar jules. No differences were observed between the parent and variants in DNA restriction endonuclease patterns using the same enzymes. But some differences were observed in DNA restriction endonuclease patterns between HV (H16)-1 and serovar jules. Thus, the antigenic variant selected from the parent by the anti-parent monoclonal antibody and serologically different from the parent, being identified either as a new serovar or as a known one, was found to be similar to the parent by the restriction endonuclease DNA analysis.

Antibodies, Monoclonal↗

PubMed1988

Expression of antigen genes of Leptospira interrogans serovar canicola in Escherichia coli.

Leptospira interrogans serovar canicola DNA was cloned into the plasmid pBR322 and introduced into E. coli. Eight out of approximately 10,000 transformants were found to express antigens of canicola by ELISA including colony ELISA blot test using anti-canicola antiserum. The canicola antigens expressed in the transformants reacted with the antisera against the serovars belonging to Canicola serogroup and other serogroups of L. interrogans. They did not react, however, with the antiserum against L. biflexa (with only one exception) nor with the antiserum against Leptonema illini. Thus, the recombinant DNA technique may provide alternative possibilities for preparing antigens of leptospires.

Antigens, Bacterial↗

PubMed1988