A suppressor factor regulating the generation of anti-thyroglobulin(Tg)antibody forming cells in the peripheral blood lymphocytes of chronic thyroiditis patients was isolated from culture supernatants of high dose Tg-stimulated T-cells from normal individuals. The same culture supernatant obtained from the patient T-cells to suppress the Tg-specific antibody response. Thus the regulatory mechanism against anti-Tg antibody production seemed to be impaired in patients with chronic thyroiditis. The culture supernatant was fractionated into 4 fractions by column chromatography. The fraction II(MW43000-67000)showed distinct suppression of the autoantibody formation, but not of anti-sheep erythrocyte nor of anti-tetanus toxoid antibody formation from the antologus lymphocytes, indicating that fraction contained the suppressor factor specific Tg antigen. Suppressor activity of Fr II was abrogated by absorbing it in Tg antigen, but not in liver extract antigen, suggesting the presence of an antigen-binding site in the molecular structure of the factor. Fr II obtained from 2 donors was tested on the lymphocytes from several patients and showed suppression in 2 out of six and in 4 out of ten combinations respectively. Although this suggested the exsistence of some allogeneic barrier between Fr II suppressor factor and the patients' lymphocytes, the identity of any HLA antigen was not related to the presence or absence of suppression. When statistically analysed. However the fact that no suppressive effect was observed in HLA-DR antigen mismatched combinations and that the identity of DR antigen was weakly related to the suppression(p=0.16)suggested that the suppressive effect of Fr II was associated with the HLA antigen encoded by HLA-locus close to HLA-DR, but not with the DR antigen itself.

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Antilymphocyte antibodies in sera of patients with systemic lupus erythematosus were studied for their binding characteristics and antigenic determinants using peripheral blood flesh T cells, PHA blasts and various human lymphoid cell lines. The following observations were made: 1. SLE sera contained antibodies which reacted with PHA blasts, and their titers were higher in the more active stages of the disease. 2. SLE sera, which reacted with PHA blasts, were further studied for their reactivities with various human lymphoid cell lines. Almost all sera reacted with Tac Ag^+, la Ag^+ T cell lines. About one half reacted with Tac Ag^-, laAg^+B cell lines, But few sera reacted with Tac Ag^-, IaAg^-T cell lines. 3. Antibodies to activated T cell lines were of IgG and/or IgM classes. Stronger reactivity of the antibodies to the target was obtained at 15℃ than at 4℃. 4. Blocking experiments with monoclonal antibodies and absorption tests suggested that SLE sera contained anti-Tac antibody and/or anti-Ia antibodies.

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The present study concerns the idiotype of autoanti-thyroglobulin(Tg)antibodies induced experimentally in F1 hybrid rabbits and the idiotypic manipulation of the production of these antibodies. The following results were obtained: 1. At least some of the anti-Tg antibodies produced from different F1 hybrid rabbits carried the same or similar idiotypes. 2. F1 hybrid rabbits produced anti-anti-idiotypic antibodies when immunized with anti-idiotypic antibodies against anti-Tg antibody. 3. Anti-Tg antibodies were detected in serums of F2 hybrid rabbits born from a doe which had bee immunized with anti-idiotypic antibodies. These results suggest that the production of autoanti-Tg antibody can be idiotypically manipulated.

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Various respiratory diseases have been reported in relation to Aspergillus fumigatus(Af). Factors which designate the type of respiratory desease were suspected to depend not only on the inhalation dose of Af but also on patients' patterns of allergic responses to Af antigen. Therefore, humoral and cellulat immunity of patients with allergic bronchopulmonary aspergillosis(ABPA)and aspergilloma were studied in relation to Af. Most of these patients showed positive immediate type reaction in skin tests to Af antigen, although specific IgE for Af antigen was not detected in patients with aspergilloma. Late type and delayed type skin were positive in 4 of 6 patients with ABPA and aspergilloma. Skin tests to PPD were negative in 2 patients with ABPA and 2 patients with aspergilloma having old tuberculous cavities in lungs, Precipitating antibodies to Af antigen due to double diffusion method were positive in all patients. Specific IgG Af antigen was also measured quantitatively With enzyme linked immunosorbent assay. Increased levels of specific IgG were found in ABPA or aspergilloma with strongly positive precipitins. Lymphocyte blastogenesis induced by Af antigen was studied to find evidence of cellular immunity. High responsiveness of peripheral blood lymphocytes were found in patients with ABPA and aspergilloma. Especially, patients with negative PPD skin reactions showed higher lymphocyte responsiveness than patients with positive PPD skin reactions. Negative conversion of PPD skin test, might be induced by lymphokine, also suggested the existence of active granulomatous lesion in the lungs of patients with ABPA and aspergilloma. Elevated levels of specific IgG correlated with the high responsiveness of lymphocytes to Af antigen indicating the intimate relationshop between type III and type IV allergy. As a direct proof of commitment of cellular immunity in the lungs of ABPA sufferers. increased populations of lymphocytes and eosinophils were found and these lymphocytes responded to Af antigen at a high level than the peripheral blood lymphocytes of each patient with ABPA. These data indicate that cellular immunity as well as humoral immunity play an important role in the pathogenesis of ABPA and aspergilloma.

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One point mutation in the I-A subregion of C57BL/6 major histocompatibility complex causes the mutant, B6.C-H-2^<bm12>, to lose its ability to respond to beef insulin. F_1 hybrids of two nonresponder mice, B10.A and B6.C-H-2^<bm12>, were found to be good responders to beef insulin suggesting gene complementation. T cell clones were isolated from(B10.A× B6.C-H-2^<bm12>F_1 mice that had been immunized with beef insulin. These F_1 T cell clones responded to beef insulin but not the other immunized antigen, PPD, or the unrelated antigen, OVA. When these clones were tested with the antigen pork insulin, which differs from beef insulin only in the animo acids in positions A8-A10(A chain loop), a few clones failed to respond. From this we can say that these clones recognize an insulin determinant associated with the A chain loop. The remaining T cell clones responded to beef or pork insulin and therefore can be said to recognize a non-A chain loop determinant(A4 or B chain). Recognition of A chain loop on the beef insulin molecule is unrelated to the expression of Ia. W39 on antigen presenting cells because these F_1 mice do not express Ia. W39 on their cell surfaces.

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Suppressor cells in delayed hypersensitivity reactions to soluble protein antigens, which were induced in vitro from BALB/c splenoytes. were investigated. Transfer of in vitro cultured splenocytes into syngeneic recipients resulted in suppression of the hosts' delayed hypersensitivity reactions in an antigen-specific manner. These suppressor cells were characterized as B-cells by their adherence to nylon-wool column, resistance to treatment of anti-Thy1. Ly1 and Ly2 antibody in the presence of complement, adherence to anti-mouse immunoglobulin antibody-coated dish, and nonadherence to noncoated plastic dish. In addition to the radiation sensitivity, these suppressor B cells showed the capability of binding to a Sepharose column which was coated with primed antigen, but to one coated with irrelevant antigen. Thus, it was demon-stated that out in vitro induced suppressor cells of delayed hypersensitivity reactions were antigen specific B-cells. The possible mechanisms by which these suppressor B-cells exert such suppressor activity in vivo were discussed.

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Counterflow centrifugal elutriation (CCE)technology was developed and applied for the separation and isolation of target cells according to sedimentation velosity based on the size, density and shape of the cells. In this study a precise procedure for the isolation of target cells from other cells was established using the elutriator manufactured in co-operation with Hitachi Koki Co., Ltd. The study was further extended to make a fluorescence polarization analysis of the cell dynamics of the fractionated homogeneous cell population obtained through CCE. Mononuclear cells(approximately 2.0×10^8 cells)were obtained from 200ml of heparinized blood through a method of Ficoll-Triosil density gradient centrifugation and subsequently CCE was applied for the separation of the mononuclear cells into a homogeneous cell population. The total cell count and purity of monocytes collected into the final fraction were 1.45×10^7 cells and 81%, respectively. The function of the isolated monocytes in the tests for generation of O^-_2 and chemiluminescence, and cell aggregation was found to be normal. The analysis of lymphocyte fractions with monoclonal antibodies revealed that T^+_4 lymphocytes were rich in the earlier fractions while T^+_8 lymphocytes were predominantly found in the later part of the fractions. The intracellular fluidity of lymphocytes stimulated with PHA was measured by the fluorescence polarization method. The lymphocyte blastogenesis induced by PHA was simultaneously analysed in flow cytometry using an ethidium bromide staining method. It was found that the cell fractions showing higher intracellular fluidity corresponded with a higher value of blastogenesis.

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Sulfuric acid mist, a potent air pollutant, is known to accelerate sensitization and to induce asthma. However the mechanism of the acceleration is still debated. The purpose of the present study is to elucidate the mechanism using an experimental model of allergic respiratory disorder in guinea pigs. Guinea pigs were exposed to sulfuric acid mist, and were sensitized with heterogeneous albumin by inhalation. Their breathing curve patterns during the antigen inhalation were recorded. The measurement of in vitro histamine release from their lungs were then performed and the contents of histamine in the lungs were quantitated. The results were as follows: 1. Histamine release from the sensitized guinea pig lung with the specific antigen challenge in vitro was significantly enhanced by the preexposure of the animal to sulfuric acid mist. 2. The contents of histamine in the lungs were also significantly increased by sulfuric acid exposure. 3. The amount of histamine release from the lung with the antigen challenge in vitro was well correlated with the degree of respiratory difficulty expressed in the breathing curve patterns during the inhalation challenge with the antigen in vivo. These results suggest that sulfuric acid mist exposure might accelerate the allergic respiratory disorder by increasing the histamine contents in the lung.

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