Induction of hepatitis C virus-specific cytotoxic T lymphocytes in mice by immunization with dendritic cells treated with an anthrax toxin fusion protein.

As a novel and safe vaccine strategy, the anthrax toxin-mediated antigen delivery system composed of lethal factor (LF) fusion protein and protective antigen (PA) has been studied to prime hepatitis C virus (HCV) core-specific cytotoxic T lymphocytes (CTLs) in vivo. The core epitope fused to LF (LF-core) together with PA induces a negligible core-specific CTL response in mice, whereas core-specific CTL are effectively primed in mice by injecting dendritic cells (DCs) treated in vitro with LF-core and PA. These findings imply that LF fusion protein plus PA in combination with dendritic cells may be useful for a novel T cell vaccine against HCV infection.

[Evaluation of the usefulness of a serodiagnosis kit, the determiner TBGL antibody for tuberculosis: setting reference value].

The determiner TBGL antibody developed by Kyowa-Medex co., ltd. is a new serodiagnostic kit for tuberculosis. To set cut-off point suitable for the kit, TBGL antibody titer of serum from healthy subjects were analyzed in relation to age, sex, previous and family history on tuberculosis. Based on results of healthy subjects, cut-off point was set by using several analytical methods. We propose 2U/ml as a cut-off point for screening of patients with tuberculosis considering the diagnostic efficiency by receiver operating characteristics (ROC) curve analysis and 4U/ml for diagnosis of tuberculosis, which was determined by parametric method considering its specificity.

Roles of macrophages in programmed cell death and remodeling of tail and body muscle of Xenopus laevis during metamorphosis.

Examination was made of the involvement of macrophage phagocytosis in programmed cell death of tail and body muscle of the frog, Xenopus laevis, during metamorphosis by electron microscopy and immunohistochemical analysis. Electron microscopic observation revealed that macrophages were often found to be present in body and tail muscles at the most active stage of metamorphosis and to actively phagocytose apoptotic muscle fragments. Developmental changes in macrophages were examined using the macrophage-specific antibody, HAM56. Macrophages initially appeared in the early climax stage (stage 59), when the triiodothyronine (T3) level was high, increased rapidly during the process of muscle cell death, and assumed their greatest number at the late climax stage (stage 63/64). They decreased after stage 65/66, with a decrease in T3. Distribution and change in the number of macrophages were the same as those of muscle apoptotic bodies (sarcolytes) during metamorphosis, which suggests an interactive mechanism between macrophages and dying muscle cells. For clarification of this, study was made of the expression of HAM56 antigens that were X. laevis homologs of mouse attachmin, non-specific adhesion proteins in macrophages. The expression of HAM56 antigens in macrophages was found to increase with macrophage phagocytosis at the late climax stage, thus, macrophage differentiation would appear to take place during metamorphosis and HAM56 antigens may be essential for macrophage-dying muscle cell interactions.

Spectrin localizations in the chicken bursa of Fabricius at different stages of development.

Before and after hatching, light microscopic and ultrastructural observations of spectrin were performed immunohistochemically in the chicken bursa of Fabricius. Before hatching, the frequency of spectrin-positive cells was very low. Among the spectrin-positive cells spectrin was mostly detected in patchy or diffuse form in the cytoplasm and rarely seen at surface membranes. Although cortical lymphocytes were spectrin-negative, numerous medullary lymphocytes were spectrin-positive after hatching. In the medullary spectrin-positive cells, staining intensity was uniform. Spectrins showed ultrastructural heterogeneity after hatching. Although a new type of spectrin localization associated with surface membranes was observed, this type of spectrin localization was not prominent. The increased number of spectrin-positive cells, uniform staining intensity and the localization of spectrin associated with surface membranes seem to coincide with B cell differentiation.

Ultrastructural localizations of J chains in the chicken bursa of Fabricius at different stages of development.

Before and after hatching, J-chain positive cells (JPC) were observed by immunoelectron microscopy in the chicken bursa of Fabricius. JPC were mostly lymphocytes, but epithelial cells were also detected as JPC. During the embryonic stage, J chains were mostly associated as patches with surface membranes. Furthermore, there was a diffuse localization in the cytoplasm. After hatching, J chains showed a similar subcellular localization as was seen before hatching. However, J chains were frequently detected in the cytoplasm, and rarely on the surface membranes after hatching. Staining intensities by corresponding antisera were stronger in the hatched chickens than in embryos. From these findings one may conclude that J chains are synthesized even at an early stage of B cell differentiation during embryonic life and are continuously produced at the later differentiation stages of B-cell lineage. The increased amounts of J chains estimated by staining intensity seem to coincide with B cell maturation and may correlate with signalling of IgM synthesis.

[Two cases of invasive pulmonary aspergillosis in non-immunocompromised hosts].

Invasive pulmonary aspergillosis generally occurs in immunocompromised hosts such as patients with leukemia, and other malignancies, who are receiving anti-cancer chemotherapy. In this report, two non-immunocompromised patients who developed invasive pulmonary aspergillosis are presented. Case 1: A 63-year-old man complained of productive cough and fever. He received antibiotic therapy from his personal physician. This symptoms did not respond, however, and dyspnea developed. He was then transferred to our hospital, about one month after the onset. The chest X-ray showed a meniscus shadow suggesting an aspergilloma in the right upper lung field and an infiltrative shadow in the remaining right lung field. Case 2: A 78-year-old man was admitted because of dyspnea, productive cough and appetite loss over the previous three months. The chest X-ray showed a meniscus shadow in the left upper field, an infiltrative shadow in the left lower field and a right pleural effusion sign was also observed. Both cases were diagnosed as having aspergillosis, early in their illness, by the detection of aspergillus antigen in their sera and histopathological and cultural studies of specimens obtained by TBLB. Both improved with intravenous amphotericin B (30 mg/day) and intravenous ulinastatin (200000 IU/day) administration. On the examinations conducted during hospitalization, there was no evidence of any immunosuppressive diseases or immunoincompetent conditions such as leukemia, and other malignancies human immunodeficiency virus infection, diabetes or alcoholism.

[A case of Aspergillus niger pneumonia cured with an early diagnosis].

We have experienced a case with A. niger aspergilloma who developed Aspergillus pneumonia after bacterial pulmonary infection. Examinations of sputum cytology and detection of serum Aspergillus antigen were useful for an early diagnosis of his condition. Early and intensive antifungal chemotherapy mainly with intravenous amphotericin B brought about his complete remission.

J-chain-positive cells in immunodeficient chickens.

Before and after hatching, J-chain-positive cells (JPC) were observed by immunoelectron microscopy in lymphoid tissues from chickens that had been chemically bursectomized (Bx) in ovo. These JPC were detected in spleens both of normal and Bx birds. Subcellular localization of J chains showed more variations in normal than Bx chickens. From these findings, JPC could be divided into JPC subpopulations in chickens.

[A case of endobronchial fibroma associated with recurrent pneumonia].

We present a case of endobronchial fibroma in a 59-year-old man admitted for repeated pneumonia, successfully treated by endoscopic Nd-YAG laser. His chest X-ray showed an infiltrative shadow in the right lower lung field and a mass shadow within the truncus intermedius. Bronchoscopy revealed a polypoid mass with lobulated whitish surface, obstructing 90% of the lumen. A biopsy taken from the tumor was suggestive of fibroma histologically. Two previous case reports stated that endobronchial fibroma readily detaches from the bronchial wall during removal. The tumor was successfully removed without dropping any tumor fragment to obstruct the distal bronchus by means of biopsy forceps manually attached to an endoscope with endoscopic Nd-YAG laser. The resected tumor was mainly composed of collagen fibers with scanty spindle-shaped fibroblastic cells, which was considered consistent with endobronchial fibroma. Endobronchial fibroma is a rare benign lung tumor, and only seven cases have been reported in the Japanese literature. There was no recurrence at three years and nine months.

[Clinical studies on lung disease due to atypical mycobacteria in our hospital during the past 13 years].

Among the patients with pulmonary diseases admitted to our hospital during a 13-year period from 1977 to 1989, clinical examinations and laboratory data on admission and the following clinical courses of 102 cases where atypical mycobacteria had been identified three times or more in sputum cultures at the time of hospitalization were investigated. 1. The ratio of the number of cases positive for atypical mycobacteria to those positive for acidfast bacilli in sputum cultures tested on admission was fairly constant, 6.0 to 7.8% every year since 1981. In the cases associated with positive sputum cultures for atypical mycobacteria, M. avium complex was observed in 84% of the cases: M. kansasii, M. fortuitum and M. chelonae were found in 13, 2 and 1 cases, respectively, since 1984. 2. A total of 102 cases studied consisted of 66 male and 36 female patients; the mean age was 61.9 years. 3. Sputa became negative on culture in 19 (86.4%) out of 22 cases of primary infection. In all primary infection cases, roentgenographic findings did not worsen and prognosis was extremely good. In secondary infection cases, sputum cultures became negative in 25 (83.3%) out of 30 cases expectorating a small quantity of mycobacteria on admission, where x-ray findings worsened in only one case. In contrast, in 47 cases expectorating a large quantity of the bacilli at the time of admission, negative sputum cultures were attained in only 14 cases (29.8%) and x-ray findings worsened in 10 cases, and their prognosis were poor. 4. In general, sputum cultures turned negative within 3 months after admission. If sputum cultures remained positive thereafter, it was found very difficult to stop expectoration of the bacilli in these patients, and hence their prognosis are supposed to be greatly affected by the bacteriologic findings in early stages of the disease.

Age-related changes of J-chain-positive cells in chickens.

Localizations of J-chain-positive cells (JPC) were examined in chicken lymphatic tissues before and after hatching. The cells containing J chain were first detected in medullary areas of the bursa of FABRICIUS during the embryonic stage. These positive cells were partly detected in the developing small lymphatic follicles: perhaps on newly differentiating precursor B-cells. In addition to these lymphatic follicles, connective tissue of bursal fold were also detected as J-chain positive. Although similar localizations of JPC were again observed in hatched chickens, positive areas of follicular medulla were strongly stained for fluorescence with corresponding antisera than that of embryonic ones. These data may reflect differences in the physiology of lymphocytes in respect to functional development. JPC localizations were next compared between the B-cell subpopulations, mu-(microPC) and alpha-chain-positive cells (alpha PC). The J-chains detectable in the IgM molecules were also found in follicular medulla. However, these follicles were almost found to be negative for J-chains detectable in the alpha PC before hatching. Any strong stainings for J-chain in the alpha PC were, moreover, not be observed in bursa after hatching. The microPC localizations in hatched chickens were roughly equal with the pattern of JPC localization. These analyses revealed the presence of the cells having the chains of both mu and J. The results together with other recent studies further shown that bursal J-chain can be partly detected in newly differentiated lymphatic follicles lacking IgM-producing and suggest the possible presence of B-cell-differentiation sequence of Ig-J+----IgM+J+----IgA+J+.

Suppression of chicken J chain production by in ovo bursectomy.

The influence of in ovo bursectomy on the levels of J and mu chains in the sera of embryonic and hatched chicks was studied by the dot blotting immunoassay. The results indicated a decrease in the level of J chains during the embryonic stage after treatment with testosterone compared with normal chicks. Testosterone treatment caused a decrease in J-chain levels after hatching which was more marked in reduced-alkylated than in non-reduced sera. In contrast, testosterone caused no significant change in serum levels of mu chains, either in ovo or after hatching. Our findings further present the paradox that although removal of the bursa of Fabricius by testosterone treatment did not impair mu-chain synthesis, B-cell differentiation was suppressed. These observations indicate that in ovo bursectomy selectively inactivates B-cell differentiation as indicated by the induction of immunodeficiency, and results in the failure of J-chain production.

Ultrastructural immunochemistry of J chains in chickens.

Using ultrastructural immunochemistry, we have determined subcellular localization of J chain molecules in chicken splenic cells. The majority of J chain positive cells (JPC) were lymphoblast-like cells. The data show that J chains predominantly are localized in cytoplasm with a considerable amount distributed on the cell surface. In the cytoplasm, J chains were diffusely expressed. Furthermore, these J chain molecules were clearly seen as cluster type. In addition to the J chain localization of subcellular organelles as described above, J chains were partly found on perinuclear spaces. As J chains are key protein in B cell differentiation into immunoglobulin (Ig) producing cells, these findings might help for studying regulation of B cell differentiation in addition to revealing the molecular assembly of polymeric Ig.

[A case of tuberculous pleurisy associated with myoclonus and Quincke's edema due to isoniazid and isoniazid sodium methanesulfonate].

A case of tuberculous pleurisy associated with myoclonus and Quincke's edema due to isoniazid (INH) and isoniazid sodium methanesulfonate (IHMS) was reported. A 75-year-old man was admitted to our division because of chest discomfort and the left chest pain of one month's duration. A conventional chest roentgenogram revealed pleural effusion in the left thoracic cavity. The pleural specimen obtained from the left parietal pleura revealed caseating granuloma. Myoclonus suddenly appeared two months after the administration of antituberculous drugs for tuberculous pleurisy. Therefore, INH was discontinued. Three days later the patient's myoclonus disappeared and nine days later IHMS was newly administered. The patient abruptly developed myoclonus and Quincke's edema. IHMS was discontinued and 30 mg of prednisolone was simultaneously given. Two days later myoclonus disappeared and two days more later Quincke's edema was improved. The lymphocyte stimulation test using IHMS was positive. At that time, levels of serum vitamin B6 were within normal levels. These results suggest that myoclonus may result from epileptogenic action caused by INH or IHMS, and Quincke's edema may result from hypersensitive reaction associated with IHMS.

Acid phosphatase in lymphoid tissues of developing chick embryos.

Activity of acid phosphatase (ACPase) in bursa and thymus has been determined in developing chick embryos of either normal or treated with bovine serum albumin (BSA). From the study of light microscopical histochemistry, ACPase activity could be detected in cytoplasm. These ACPase activities were detected in both the follicular medulla and cortex in bursa. In thymus, moderate ACPase activities were also obtained in both the cortex and medulla. ACPase activity in the tissue homogenate was increased in the bursa but the thymic ones showed constant level during the incubation days examined in normal embryos. ACPase distribution observed at the distinct developing stage indicates that ACPase is a useful parameter for growth and development of chicken lymphoid tissues.

[A clinical evaluation of fluconazole in the treatment of deep mycosis].

Fluconazole is a novel triazole antifungal agent developed by Pfizer Inc. and available in both oral and intravenous forms. It is characterized by a long serum half-life of 25 to 30 hours and good absorbability into tissues. In the present study, fluconazole was given to 12 patients with deep mycosis orally, intravenously or by local infusion. The patients included 4 cases of candidemia, 1 case each of candidemia and candiduria, candiduria, esophageal candidiasis, Candida hepatic abscess, pulmonary cryptococcosis and septicemia due to unspecified yeasts and 2 cases of pulmonary aspergillosis. Clinical efficacies of fluconazole against these infections were excellent in 2 cases, good in 8 and fair in 2. None of the patients reported any side effects. From the results of the study, fluconazole appears to be a useful and safe drug for the treatment of deep seated mycosis.

Loss of J chain during primary immune responses in chicks.

J chain positive cells (JPC) were studied by direct immunofluorescence in embryonic and newly hatched chicks. The results indicated a decrease in the amounts of JPC in the embryonic spleen and bursa of Fabricius after in ovo antigenic stimulation with sheep erythrocytes (SRBC) compared with that of unstimulated lymphocytes. The level of thymic JPC in the control chicks and those subjected to antigenic stimulation was always about the same. Partial re-expression of the J chain in splenic lymphocytes was detected in newly hatched, antibody producing chicks, while the percentage of JPC in non-antibody producing chicks did not recover to the control level. Further evidence obtained indicated that the JPC decreases did not depend on the antigen dosage. After antigenic stimulation, J chain re-expression in cells of embryos and newly hatched non-antibody producing chicks was found to be essentially the same. These findings imply that the re-expression of J chain molecules is associated with immunoglobulin production. Furthermore, it seems plausible that the non-re-expression of the J chain occurred at the time of immunological unresponsiveness.

J chain-positive cells in bursectomized chicks.

Using embryonic chickens treated with testosterone propionate, the effects of congenital absence of the bursa of Fabricius determined by the frequency of J chain-positive cells was examined in the spleen, thymus and bone marrow at the embryonic and newly hatched stages. J chain-positive cells in the chicks without bursa were reduced in the spleen. No differences in the numbers of the cells were detected in the thymus and bone marrow. These results imply that removal of the bursa of Fabricius cannot entirely prevent the generation of J chain-positive B cells. Furthermore, these results partly suggest the important role of the bone marrow in the proliferation of some J chain-positive cells in chicks without bursa.

Ontogeny of lymphocytes expressing J chain in chickens.

The ontogeny of chicken lymphocytes expressing J chain (LEJ) was investigated in the embryonic bursa of Fabricius, the spleen, and the thymus. Simultaneous appearance of LEJ was detected in the bursa and spleen on Day 14 of incubation. These cells were detected later in the thymus. The LEJ were found to increase rapidly in the spleen from the 19th to 20th incubation day. In adult chickens, the highest percentage of LEJ was also found in the spleen. These cells were seen in the thymus at a lower frequency. Intermediate numbers were found in bursal and peripheral blood lymphocytes. The frequencies of the LEJ were similar to those of lymphocytes positive for cytoplasmic immunoglobulins (Ig) IgA and IgM, but were not related to the number of lymphocytes expressing surface Ig. It is possible to consider that the suitable site for LEJ is the spleen, on the basis of the rapid increase in the number of LEJ just before hatching and from the fact that the highest value is found in adult chickens. Furthermore, LEJ may participate in secretion of IgA or IgM but not be associated with the expression of surface Ig.