Biken journal最新文献

An improved micromethod for infectivity assays and neutralization (N) tests of dengue (DEN) type 1-4 viruses was developed, using 96-well plates and the PAP (peroxidase-antiperoxidase) staining technique. The foci formed on BHK-21 cell monolayers in wells of the plate were readily countable under an ordinary stereomicroscope. This micromethod has the advantages over the micromethod of the Lab-Tek 8 chamber slide system of lower cost, requirement for smaller volumes of test sera and applicability to larger number of serum specimens for N tests of DEN viruses.

Lymphokine activated killer cells (LAK cells) or interleukin 2 (IL-2)-activated killer cells were induced by recombinant IL-2 (TGP-3) for clinical adoptive immunotherapy of malignant diseases. After incubation of peripheral blood lymphocytes (PBL) with IL-2 and normal human plasma for 1-2 weeks LAK cells were obtained that showed a maximum cytotoxicity against target cells, and did not need a toxic dose of IL-2 to enhance or maintain their cytotoxicity. Both autologous and allogeneic LAK cells were used in five clinical cases without any immune side effects, and were effective in three cases.

Serological follow-up studies for up to 4 years on the levels of IgG antibody to EBV-determined nuclear antigen (EBNA) were carried out on 36 patients with nasopharyngeal carcinoma (NPC). The serum levels of IgA antibody specific to EBV capsid antigen (VCA) were also measured in some of the patients. The titers of EBNA antibody were measured by enzyme-linked immunosorbent assay (ELISA) and those of IgA antibody to VCA were measured by the indirect immunofluorescence method. The EBNA antibody titers in most sera from the patients before radiation therapy were found to be at least 4 times the mean values in the sera of healthy control adults. Within 2 to 8 months after completion of therapy by 4-MV liniac X-ray irradiation with total doses of 60 to 80 Gy, the titers of EBNA antibody in the sera of 6 patients had returned to normal levels, and low levels of EBNA antibody were maintained for a long time after therapy. These serological data were associated with a good clinical prognosis without recurrence or metastases. But in 6 patients, the patterns of change in the EBNA antibody levels were different: the levels remained high after therapy or first decreased to the normal level and then rose to at least 4 times this level. These 6 patients showed recurrence or metastases. The patterns of change in the EBNA antibody levels were well correlated with those of change in the levels of IgA antibody specific to VCA.(ABSTRACT TRUNCATED AT 250 WORDS)

Beta-antigen is one of the major proteins of Mycobacterium tuberculosis. We purified this antigen from the unheated culture filtrate of Mycobacterium tuberculosis Aoyama B and obtained nine monoclonal antibodies against the beta-antigen. Nine monoclonal antibodies were divided into two groups according to their patterns on Western blotting. The result indicated the existence of two or more determinant groups against these monoclonal antibodies on the beta-antigen molecule. The interspecies reactivity of monoclonal antibodies among twenty-one species of Mycobacteria was also examined by dot blotting analysis. Two monoclonal antibodies, designated 4G5E10 and 5F3F2, showed a specificity restricted to the Mycobacterium tuberculosis complex, could be used for serodiagnosis of Mycobacterium tuberculosis infection.

Antigen solution could be injected into the cornea of sensitized mice using a fine needle and a stereoscopic dissecting microscope. The resulting corneal reaction was shown to be a reliable method in the detection and estimation of delayed-type hypersensitivity in mice that had been immunized with a water-in-oil emulsion containing an ovalbumin and a cell wall adjuvant. Unlike the delayed skin reaction in the ear lobe, this corneal reaction was not affected by a coexisting Arthus reaction.

The role of the tumor-unique determinant(s) on two syngeneic murine hepatoma cells in inducing in vivo protective immunity was investigated in comparison with that of the tumor-cross-reactive determinant(s). Induction of vaccinia-reactive helper T cells in C3H/He mice by intraperitoneal (i.p.) inoculation of viable vaccinia virus and then immunization with vaccinia-infected syngeneic MH134 or MH129 tumor cells resulted in the production of potent anti-MH134 or -MH129 antibody as well as the generation of in vivo protective immunity. Neither antibody reacted with other syngeneic plasmacytoma or fibrosarcoma cells, but both cross-reacted appreciably with the other hepatoma cells as well reacted strongly as with the tumor cells used for immunization. The absorptions of anti-MH134 and -MH129 antisera with the respective hepatoma cells abolished their reactivities with both the corresponding hepatoma cells and the other hepatoma cells. In contrast, the absorption of these antisera with the other tumor cells resulted in loss of their cross-reactivities with the other hepatoma cells, but not loss of their specific reactivity to the respective hepatoma cells. Although in these hematoma systems, the above-mentioned immunization protocol resulted in in vivo induction of protective immunity and generation of antibodies, in vivo immunity as observed by Winn assays was mediated by Lyt-1+2- T cells and was specific for each type of hepatoma cells. These results indicate that these two types of hepatoma cells bear two kinds of antigenic determinants, one kind unique to each hepatoma and the other kind cross-reactive with the other hepatoma cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunoglobulin M antibody to hepatitis B core antigen (IgM anti-HBc) was measured by radioimmunoassay in the sera of 96 HBV carriers. IgM anti-HBc was detected in 17 of 66 patients with chronic active hepatitis and in 4 of 11 with liver cirrhosis. This antibody was not present in asymptomatic carriers or in patients with chronic persistent hepatitis. Testing of sequential samples revealed that the presence of IgM anti-HBc indicated active replication of HBV and at the same time an immune response to the virus. The relationship between IgM anti-HBc and the response to interferon (IFN) therapy was also studied. Results showed that IgM anti-HBc is a useful marker of the efficacy of interferon therapy.

The effects of vinblastine alone and in combination with vinblastine, colchicine and concanavalin A on microtubules of Trypanosoma gambiense cultured in vitro were studied ultrastructurally. Trypanosomes treated with vinblastine at 20 micrograms/ml, showed fusion of the extracellular flagellum with the plasma membrane of the parasite. As a result, the axoneme with the paraxial rod in the extracellular flagellum was taken into the cytoplasm. Although the axonemal and pellicular microtubules in T. gambiense differ in function and origin, the axonemal microtubules of the extracellular flagellum that was taken into the cytoplasm could be converted to pellicular microtubules by treatment with a combination of vinblastine (20 micrograms/ml), concanavalin A (10 micrograms/ml) and colchicine (100 micrograms/ml).

A live attenuated mumps and trivalent measles-rubella-mumps (MRM) vaccines have been applied to 887 and 148 children with various underlying diseases at the vaccine clinic of Osaka University Hospital between 1975 and 1985, respectively. Clinical reactions after mumps vaccination occurred in only 7 children (0.8%) and those after MRM vaccination in 28 children (19%), but their underlying diseases were not deteriorated by either vaccination. Clinical follow up study revealed that 2 of the 430 children immunized with mumps vaccine had contracted the disease during 7 year period and 2 of the 123 children immunized with MRM vaccine had contracted clinical mumps or rubella during 3 year period. The seroconversion rates after mumps vaccination were 70% and 61% by the hemagglutination inhibition (HI) test and neutralization (NT) test, respectively, while 94% by the fluorescent antibody to membrane antigen (FAMA) test. Those after MRM vaccination were 87% for measles, 96% for rubella by the HI test and 89% for mumps by the FAMA test. Serological follow up study revealed that antibodies elicited by mumps vaccination were sustained without substantial decline for at least 7 years. These results suggest that a live attenuated mumps and MRM vaccines are safe and effective in children with various underlying diseases.

There have been many unsuccessful attempts to induce gametocytogenesis in vitro. In the present experiment, however, we found that RPMI-CS medium and RPMI-FS medium prepared by dissolving powdered RPMI 1640 medium in the culture supernatants of hybridoma cells, hybrid line D21 and 219.5, respectively, that produce anti-P. falciparum antibody induced gametocytogenesis. Gametocytogenesis was consistently observed from 3 days after addition of these media. The culture supernatant of anti-P. falciparum antibody producing hybridoma cells did not induce gametocytogenesis in the absence of RPMI 1640 medium. RPMI-MS medium, prepared by dissolving powdered RPMI 1640 medium in the culture supernatant of myeloma cells, SP2/O-Ag 14, which was used as a control, induced a few gametocytes.