Search PubMed⌕ Search

Biomedical subjects

I Takashima

Publications and source records attributed to I Takashima.

At least 91 records · Page 5Linked to original sources

Immunoelectron microscopy of Chlamydia psittaci with monoclonal antibodies.

An immunoelectron microscopic study was performed to determine the distribution of antigenic components on particles of Chlamydia psittaci and infected cells using a number of monoclonal antibodies (MAbs). Of three anti-lipopolysaccharide (LPS) antibodies (4D5, A2 and 4G5), two antibodies (4D5 and A2) reacted with the surface of reticulate bodies (RBs) but not with that of elementary bodies (EBs). The other antibody (4G5) reacted with both EBs and RBs. Examination of infected cells in thin sections revealed that 4D5 and A2 combined with the membranes of both EBs and RBs. These results indicate that each LPS epitope localized at a different position in the chlamydial membrane. Most MAbs directed to protein antigens reacted on the surface of both EBs and RBs though 3E9 specific for the 90 kDa and 50 kDa protein components combined with RBs only.

Antibodies, Monoclonal↗

Effect of neutralizing monoclonal antibodies on Hantaan virus infection of the macrophage P388D1 cell line.

The effect of neutralizing monoclonal antibodies (N MAbs) on Hantaan virus infection of macrophages was investigated using P388D1 cells, a murine macrophage cell line. MAbs to the G1 protein (G1b) and the G2 protein (G2a and G2c) neutralized viral infectivity in P388D1 cells. N MAbs to G1b showed much higher neutralizing potency than those to G2a and G2c. With each N MAbs, two distinct effects were observed: neutralization of viral infectivity occurring at high concentrations and enhancement of that at low concentrations. Non-neutralizing MAbs, on the other hand, showed only enhancement of viral infectivity even at high concentrations without any inhibitory effects. The Fab fragments of N MAbs showed neither neutralizing nor enhancing activities. However, when the virus-Fab complexes were reacted with the anti-Fab antibodies, both neutralization and enhancement of viral infectivity were restored depending on the dose of Fab fragments. These results indicate that Hantaan virus infection of P388D1 cells is mediated by the Fc portion of the antibodies and neutralization is dependent on the concentration of N antibodies bound bivalently to the neutralization site on the virion.

Animals↗

Serological assessment of chlamydial infection in the koala by a slide EIA technique.

A rapid and simplified slide enzyme immunosorbent assay (EIA) was developed for the diagnosis of chlamydial infection in the koala. HeLa 229 cells infected with koala strain Chlamydia psittaci were fixed on the surface of multiwell slides and used as the antigen. The assay consisted of first reacting koala antiserum with the fixed C psittaci antigen, followed by reaction with biotinylated rabbit anti-koala IgG, ABC reagent and substrate. The chlamydial EIA antibody titres obtained were compared with those of a complement fixation (CF) test using koala strain C psittaci as antigen. Of 35 koala sera tested, 16 CF positive sera (greater than or equal to 1:8) also had a positive titre (greater than or equal to 1:200) in the slide EIA test (sensitivity 93.8%, 15/16). Nineteen CF negative sera were also negative in the slide EIA (specificity 100%, 19/19). Sixty-eight samples of koala blood were collected by ear-prick using a sampling paper method and were assayed by both tests. Sensitivity of the slide EIA was 100% (15/15) and specificity of the test was 96.2% (51/53). To simplify the slide EIA for use as a practical screening test, a 3-point serum dilution series (1:100, 1:200, 1:400) was used. This 3-point slide EIA was compared with the CF test using sheep strain chlamydial antigen. Thirty-nine sera were assayed by both tests. The sensitivity of the 3-point method was 85.7% (6/7) and the specificity was 71.9% (23/32) as compared with the sheep antigen CF test.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Kinetic studies on the appearance of antigens of Chlamydia psittaci during its developmental cycle.

The kinetics of the antigen production of Chlamydia psittaci strains Izawa-1 and Pigeon-1041 (P-1041) was examined every 6 hr after infection up to 48 hr, by the indirect immunofluorescent antibody technique using monoclonal antibodies (MAbs). All three genus-specific antigenic determinants on lipopolysaccharide (LPS) appeared during the whole growth cycle. Antigenic determinants on proteins were, on the other hand, detected at various time periods from the early to the late stages of infection. However, a cross-reactive antigenic determinant on protein recognized by a MAb 3E9 was also detected during the whole growth cycle, similar to that on LPS. The time of appearance of common antigenic determinants on proteins of Izawa-1 and P-1041 was examined using cross-reactive MAbs, and it varied depending on heterologous and homologous MAbs. From the relationship between the detection of antigenic determinants and the morphological changes of chlamydial particles revealed by electron microscopy during the growth cycle, the antigenic determinants on proteins of Chlamydia psittaci were divided into two groups; one was specific to the elementary body and the other was coexisting in both the elementary body and the reticulate body.

Animals↗

Susceptibility of laboratory and wild rodents to Rattus or Apodemus-type hantaviruses.

Adult Wistar rats (Rattus norvegicus), Apodemus agrarius, Meriones unguiculatus, Clethrionomys rufocanus, and Apodemus argenteus were inoculated with Rattus-type (SR-11, KI-262, and TB-314) or Apodemus-type (Hantaan 76-118) hantaviruses. Production of serum antibody to the inoculated virus (IAHA titres of 1:32 to 1:4 096) was obsersved in all rodent species 10 weeks after virus inoculation. Rattus-type virus was detected in some organs of all the rodent species employed except of Apodemus agrarius. Apodemus-type virus was found only in some organs of Apodemus agrarius. Newborn Wistar rats induced antibody in high titres to both Rattus- and Apodemus-type hantaviruses. Rattus-type virus was detected in all the organs examined for up to 6 weeks after inoculation, whereas Apodemus-type virus disappeared from all organs except of brain and lung tissues. The virulence of the three Rattus-type viruses to newborn rats was different. These findings indicate that the susceptibility of rodents may vary depending on the combination of rodent species and virus strains; they also suggest that the various species of rodents may be the reservoir animals of hantavirus infection in nature.

Animals↗

Nucleotide sequence of a gene encoding a new genus specific protein of Chlamydia psittaci.

DNA fragment No. 13 from the C. psittaci pigeon strain, has been cloned in the plasmid pUC19. Hybridization analysis revealed that the fragment maintained a chlamydial common sequence. Furthermore, nucleotide sequencing identified two partial open reading frames (ORF), 675b. p. and 530b. p. Expression of ORFs revealed that the second ORF encoded 25KD polypeptide, whereas the first ORF did not produce any antigenic product. The 25KD beta-galactosidase fusion protein reacted strongly with chlamydia-specific antibodies elicited against a number of different chlamydial strains. Gene Bank analysis showed that this cloned gene is not highly homologous with chlamydia or other organisms for which nucleotide sequences have already been published. This 25KD polypeptide may be an additional genus-specific antigen of Chlamydiae.

Amino Acid Sequence↗

Characterization of DNA fragment from Chlamydia psittaci avian strain which shows high homology with hypB gene of Chlamydia.

A study was performed to characterize DNA fragment No. 17 of C. psittaci strain P-1041 which encoded 42 KD beta-galactosidase fusion protein with type-specific antigenicity. Sequence determination identified a partial open reading frame that spanned about 1,200b. p. nucleotides. Screening the literatures for the nucleotide and deduced amino acid sequences revealed extensive similarity between the DNA fragment of P-1041 and two chlamydial hypB genes. This DNA showed 91.5% homology with C. psittaci GPIC hypB gene in nucleotide sequence and 96.4% homology in deduced amino acid sequence. The hypB gene of C. trachomatis serovar A and the P-1041 DNA fragment showed 81.2% and 91.3% homology in nucleotide and amino acid sequences, respectively. Dot enzyme-linked immunosorbent assay, for the products of deleted DNA fragments defined the coding region for type-specific antigenic polypeptide. In addition, the P-1041 DNA fragment carried a sequence highly homologous (greater than 49%) with other bacterial and plant genes called chaperonin which responds to various stress in cells. From these results, the P-1041 DNA fragment was found to be a part of hypB gene and to encode the region critical for type-specific antigenicity.

Amino Acid Sequence↗

Percutaneous treatments for biliary diseases.

Percutaneous treatment of biliary disease was administered in 173 cases, mainly among high-risk or elderly patients. Diagnosis was acute cholecystitis in 32 cases, acute cholangitis in 16, hepatic abscess in five, gallbladder stones in 28, common bile duct stones in 11, intrahepatic stones in five, malignant obstructive jaundice in 61, and benign biliary stenosis in 15. The treatment was successful in 158 of 173 cases (91.3%). Complications were encountered in 17 (9.8%). Most of these occurred during the initial period and were mild and controllable with conservative treatments. There were two deaths (1.2% of patients); one was not related to the procedure. These data suggest that percutaneous treatments are safe and effective even among elderly or high-risk patients with biliary diseases.

Adult↗

Characteristics of passive immunity against hantavirus infection in rats.

The protective effects of passively administered antibodies against hantavirus infection were studied in newborn rats. Death as well as infection were completely prevented from intraperitoneal challenge of strain SR-11 (SR) (2 x 10(3) FFU, 10(2.1) LD50), in newborn rats which received 0.1 ml of anti-SR rat serum (neutralizing antibody titer, 1:640) 4 hr before the virus challenge. In these rats, no virus was detected in the peritoneal macrophages, lung, kidney, and brain. The immune serum infusion before the virus challenge also conferred protection to rats against an intramuscular or subcutaneous challenge of strain SR, but did not protect the rats against intracerebral challenge of the virus. In the rats which received the immune serum after the challenge, infection was not prevented, although some of the animals were protected from the death. Virus titers in the lung, kidney, and brain of the rats were reduced by the transfer of the immune serum even as late as 72 hr after the challenge. Cross-protection in the rats which received the immune serum was strong between strains SR and KI-262 within the same serotype, but very weak between strains SR and Hantaan 76-118 of different serotypes.

Animals↗

Horizontal and vertical transmission of Japanese encephalitis virus by Aedes japonicus (Diptera: Culicidae).

Aedes japonicus (Theobald) was evaluated for competence as a vector of Japanese encephalitis virus and for its ability to transmit the virus vertically to its F1 larvae. Ae. japonicus supported the growth of the virus at 20 and 28 degrees C after feeding on a virus-blood mixture of 10(6.2) plaque forming unit (PFU)/ml. This species was able to transmit the virus to suckling mice after feeding on a virus-blood meal (10(6.2) PFU/ml) or a viremic chick (10(3.7) PFU/ml). Vertical transmission of the virus in Ae. japonicus was demonstrated with a minimum infection rate of 0.7%.

Aedes↗

Comparison of virulence between two strains of Rattus serotype hemorrhagic fever with renal syndrome (HFRS) virus in newborn rats.

Two strains of hemorrhagic fever with renal syndrome (HFRS) virus from Rattus, SR-11 and KI-262, showed virtually identical antigenicity but differed from prototype strain Hantaan 76-118 (Apodemus origin) in a neutralization test. Wistar newborn rats inoculated intraperitoneally (i.p.) with SR-11, which was isolated from a laboratory rat associated with an outbreak of HFRS, developed clinical signs such as ataxia and limb paralysis and died at about 18 days after inoculation. The LD50 of SR-11 in 1-day-old rats was 10(1.2) focus-forming units (FFU). In contrast, the animals inoculated i.p. or intracerebrally with 10(4) FFU of KI-262, which was from a wild rat in a dumping-ground area--an enzootic focus where no human cases have been recorded--did not show any significant clinical signs. The susceptibility of rats to SR-11 fatal infection was age-dependent. Virus titers in brains, lungs, kidneys, and livers of the rats inoculated with SR-11 were significantly higher than those in the same organs of the animals infected with KI-262. Necrosis of neurons in the brain tissue occurred in the rats infected with SR-11, while it was mild in the animals infected with KI-262.

Aging↗

Mosquito collection in endemic areas of Japanese encephalitis in Hokkaido, Japan.

A study was conducted during 1985 and 1986 to evaluate the roles of mosquito species as possible vectors of Japanese encephalitis (JE) virus in Hokkaido. The number of Culex tritaeniorhynchus was very low among the four pig farms where outbreaks of abortion caused by JE virus were observed in swine populations. At one farm near Sapporo, only one Cx. tritaeniorhynchus was found among a total of 510 mosquitoes collected during the survey period from July to October 1985, even when JE virus activity among sentinel pigs was revealed by seroconversion. At another farm in the south, no individuals of this mosquito species were found among 987 mosquitoes collected at the time of the outbreaks of abortion. Cx. pipiens pallens, Anopheles species, Aedes vexans nipponii, and Ae. japonicus were predominant over Cx. tritaeniorhynchus. Cx. tritaeniorhynchus, almost a solve vector species of JE virus in the southern part of Japan, is probably not a vector of the virus in Hokkaido. The collected mosquitoes (2,332 from 1985 and 1,403 from 1986) were processed for virus isolation but no JE virus was isolated. More extensive field studies are necessary to provide further information on the role of mosquito species other than Cx. tritaeniorhynchus in the transmission of JE virus in the northern limits of its range including Hokkaido.

Abortion, Veterinary↗

Role of maternal antibody in protection from hemorrhagic fever with renal syndrome virus infection in rats.

The effect of maternal antibody on the protection of newborn rats from infection of HFRS virus strain SR-11 was examined. Antibody to HFRS virus was transferred from immune dams to their offspring prenatally as well as postnatally. IgG antibody was detected in the sera of fetuses by IFA test (titers from 1:64 to 1:256) and in fetal fluids (1:32) obtained from the uteri of immune dams at 20 days after mating. In the sera of the newborn, IgG titers of maternal antibody ranged from 1:64 to 1:256 just after birth, reached a peak titer around 1:2,048 at 2 weeks after birth, then declined and disappeared at about 8 weeks of age. No IgA and IgM antibodies were detected in the sera of fetuses and newborns. After intraperitoneal challenge by strain SR-11 (10(2.2) LD50), death and infection of 2-day-old rats from immune dams were prevented by the presence of maternal antibody. The protective effect of maternal antibody remained in 8-week-old rats having an IFA titer of maternal antibody as low as 1:16, and even in some 10-week-old rats with negative tests for maternal antibody.

Aging↗

Immunotyping of Chlamydia psittaci by indirect immunofluorescence antibody test with monoclonal antibodies.

Monoclonal antibodies against pigeon and budgerigar strains of Chlamydia psittaci were used to classify the immunotypes of C. psittaci strains by an indirect immunofluorescence antibody (IFA) test. Thirty-three C. psittaci strains from pigeons and 24 from budgerigars were divided into three immunotypes (P-I, P-II, and P-III) and (B-I, B-II, and B-III), respectively. Two strains from human psittacosis patients were identified as P-III and B-I, coinciding with the epidemiological evidence of each human infection. Two strains from psittacine birds, a parrot and a parakeet, were identical to the B-II immunotype. Other mammalian strains were quite distinct from avian strains in their IFA reaction with the monoclonal antibodies.

Animals↗

Enhanced neutralization of flavivirus by monoclonal antibodies against Negishi virus.

Ten monoclonal antibodies against Negishi virus were classified into 3 groups and 7 types according to the reaction patterns to Negishi virus by the hemagglutination inhibition (HI) test and by several kinds of neutralization tests. When kinetic neutralization was applied to these monoclonal antibodies at the same HI titer of 1:64, the initial slope and the persistent fraction of the kinetic curve was varied in each antibody. In the double-kinetic neutralization test, neutralization did not proceed further when the second antibody was the same type as the first one. When the second antibody was a different type from that of the first one, neutralization proceeded further. The mixtures of 4 monoclonal antibodies classified as different groups and types remarkably enhanced neutralization in the kinetic assay. These results suggested that an assortment of antibodies is needed for effective neutralization of Negishi virus and that a multi-hit model is likely operating in the neutralization of Negishi virus.

Animals↗