When mice were intravenously injected with heat-killed Propionibacterium acnes followed by an intravenous injection of a small amount of gram-negative lipopolysaccharide (LPS) 7 days later, most of the mice died of massive hepatic cell necrosis. However, when K76, an anti-complementary agent which acts on C5, was administered prior to LPS injection, remarkable improvements in the survival rate and in the histological changes of the liver were observed. How K76 suppressed the induction of massive hepatic cell necrosis was further studied. Since our previous report indicated that the liver adherent cells caused hepatocyte injury, whether adherent cells or hepatocytes were affected by K76 was investigated. As a result, K76 not only inhibited the activation of liver adherent cells and suppressed the release of the cytotoxic factor, but it directly affected the hepatocytes and protected them from the cytotoxic factor.

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An uncharacterized antibody was found in a patient afflicted with primary biliary cirrhosis or chronic active hepatitis. This antibody (which we tentatively called the MM antibody) could be detected by means of double immunodiffusion using the microsomal fraction of rat liver as the antigen. However, cytoplasmic staining in both rat liver and kidney could not be obtained when viewed by indirect immunofluorescent microscopy. The MM antibody did not react with mitochondrial fraction of rat liver. The antibody showed nonidentity with anti-liver kidney microsome antibodies when tested by means of double immunodiffusion. It was found in 5 of 5 PBC-CAH overlap patients who were diagnosed histopathologically, 3 of 19 PBC patient sera and in 2 of 14 CAH patient sera. However, it was not found in non-autoimmune hepatitis patients, nor in the healthy subjects. In conclusion, the detection of MM antibody might be useful for the diagnosis of autoimmune liver diseases particularly, PBC-CAH overlap patient.

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Epstein-Barr (EB) virus specific cytotoxic T lymphocyte (EBV-CTL) activity and the effects of various lymphokines, OK-432 and indomethacin on EBV-CTL activity were examined in 10 children with chronic active EB-virus infection. And the results were as follows.
EBV-CTL activity of the patients was 10.7%; 5.6-20.4% (mean and the range from mean-1 SD to mean +1 SD) and was significantly lower than the EBV-CTL activity of 55 control group (22.9%; 12.6-41.7%), though EBV-CTL activity in 9 patients was included in the normal value (mean±2 SD of control group). Interleukin 2 (IL-2) and interferon α (IFN-α) activated EBV-CTL activity of the patients was 21.4% (mean) and 14.1%, and were also significantly lower than those of control group (38.0% and 35.5%). Though the grade of the augmentation of EBV-CTL by IL-2 and IFN-α were also low, more than 10% augmentation by IL-2 or IFN-α was achieved in 2 cases.
EBV-CTL activity was augmented by IFN-γ in 2 cases and OK-432 in one case, but indomethacin did not enhance EBV-CTL activity of any patients.
The killer cell activity of EB-virus non-primed effector cells of the patients was same as control group, and was not augmented by IL-2 or IFN-α either.
These results indicate that though EBV-CTL activity of the patients with chronic active EB-virus infection was hardly enhanced by IL-2 or IFN-α, these lymphokines may be useful in some patients because EBV-CTL activity was enhanced by IL-2, IFN-α, IFN-γ or OK-432 in a few patients.

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Humoral and cellular immunity of patients with collagen diseases was compared between young and aged groups along with the young and aged controls.
Similar abnormalities in several immunological parameters were observed in both young and aged patients. However, decrease in C4 levels and lymphocyte counts and increase in monocyte counts observed in the young patients improved to those of the controls in the aged patients.
Decrease in CD4 cells and increase in CD8 cells, subsequently decrease in CD4/CD8 ratio, found in the young patients turned back to the value of the control group in the aged patients. These improvements were considered to be due to decrease in disease activity through the treatment. In contrast, increase in IgG and IgM levels and decrease in IgA levels observed in the young patients, compared with those of controls, were enhanced in the aged patients. These immunoglobulin abnormalities found in the aged patients might be due to hypersecretion of BSF from their lymphocytes than those from young patients since increased secretion of BSF in the aged people has been reported.

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Ten patients with selective IgM deficiency (SIgMD) were studied for cell surface immunoglobulin, immunophenotype of T lymphocytes and immunoglobulin synthesis in vitro by peripheral blood lymphocytes. Serum IgM levels were less than 25mg/dl, while IgA, IgG, and IgD were within normal levels. Ten patients had respiratory or urinary tract infections, three were diagnosed as having systemic lupus erythematosus (SLE) and one as rheumatoid arthritis (RA). Surface immunoglobulins were almost within normal range except two cases who had very low surface immunoglobulins. However, normal values were detected after 7 days of culture in the presence of normal T lymphocytes with Pokeweed mitogen. E rosett counts were normal in all patients. As for immunophenotype of T lymphocytes, OKT4/OKT8 ratios were low in all the cases. Analysis of lymphocyte function for immunoglobulin synthesis in vitro, using a co-culture of counterpart T and B cells from healthy individuals and patients with SIgMD, revealed the excessive function of IgM specific suppressor T lymphocytes. These results suggest that the increased function of IgM specific suppressor T cells was responsible for the IgM deficiency in all ten cases.

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To assess the role of decidual cells in direct contact with paternal antigens in maintenance of pregnancy, immunosuppressive action of the culture suprenatant of decidual cells were investigated. Desidual cells were obtained from artificially aborted healthy pregnants on 8 to 10 weeks of gestational age. Dispersed cell suspensions were prepared by enzyme digestion method (collagenase 1mg/ml and DNase 0.05mg/ml). Desidual cells were enriched by Ficoll-Hypaque^® density gradient, following dish adherent method. 1×10⁶/ml of decidual cells were cultured for 48 hours in RPMI-1640 medium supplemented with 10% fetal calf serum, and the supernatants were harvested.
The supernatant added at a concentration of 50% inhibited the reactions of human peripheral blood lymphocytes (PBL) by polyclonal mitotic activities of phytohemagglutinin (PHA), mixed lymphocytes culture (MLC), and killer T cell induction of PBL against alloantigen by 40%, 35% and 50%, respectively. To diminish the contamination of macrophages (spectific gravity: 1.045_??_1.075), discontinuous density gradient method using Percoll^® was performed. Desidual cells were enriched over 90% in the fraction of specific gravity of 1.033 to 1.044 (fraction 2; Fr 2). The culture supernatant of the desidual cells (Fr 2) suppressed not only the mitotic activities of PHA to PBL by 29.6% at a concentration of 50%, but also the MLC by 40%, 19% and 8.75% at concentrations of 50%, 25%, 12.5%, respectively. The culture supernatant also suppressed the killer T cell induction by 53.3% and 13.8% at concentrations of 50% and 25%, respectively.
To dissect the mechanism of immunosuppressive action of the culture supernatant, the suppressive effect of the supernatant on IL-2 production or IL-2 receptor expression of PBL stimulated by PHA, were investigated. The supernatant suppressed not only IL-2 production (0u/ml) but also IL-2 receptor expression (32.2%) compared with controlls (9.4u/ml, 48.1%). The supernatant also suppressed the immunoglobulin (IgG and IgA) production from PWM stimulated B cells separated from PBL. Those results indicate that desidual cells play a role in the maintenance of pregnancy by producing immunosuppressive substance which inhibit both T cell and B cell immunity to fetus.

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In order to study the mechanisms involved in the decreased LAK activity in HCC patients, factors involved in LAK activity induction were investigated on peripheral blood mononuclear cells (PMNC) of HCC patients with normal and low LAK activity and healthy volunteers.
1) In the experiments using PMNC of healthy volunteers and HCC patients with normal LAK activity, the addition of anti-γ-interferon (γ-IFN) to the LAK induction system at the start of incubation resulted in the suppression of LAK activity. In HCC patients with low LAK activity, γ-IFN production in LAK activity induction was significantly lower than that in healthy volunteers, HCC patients with normal LAK activity and liver cirrhosis patients. The addition of recombinant γ-IFN (r-γIFN) to the LAK induction system in HCC patients with low LAK activity enhanced the decrease in LAK activity.
From these results, it was suggested that γ-IFN played an important role in the induction of LAK activity.
2) PMNC subpopulations of HCC patients with low LAK activity were not significantly different from those of healthy volunteers. In healthy volunteers and HCC patients with normal LAK activity, LAK activities and γ-IFN concentrations in culture supernatants were decreased by the deletion of the Leu 7 positive or the Leu 11 positive cell populations, however the level of decreased LAK activities were still higher than that of HCC patients with low LAK activity. These were not enhanced significantly by the addition or r-γIFN.
In conclusion, it was suggested that the decrease in LAK activity in HCC patients was due to not only the impaired γ-IFN production but also the abnormalities of various LAK precursor cells.

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The presence of neutralizing antibody activity (NAb) against HIV-1 and HIV-1 reverse transcriptase inhibitory antibody (RTIAb) were tested in the sera of 72 HTLV-1 carriers and 61 healthy controls living in Nichinan District, Miyazaki. One of 72 HTLV-1 carriers, a patient with hemophilia, was found to be HIV-1 Ab positive, while all 61 control individuals were HIV-1 Ab (-). Two out of 71 HIV-1 (-) but HTLV-1 carriers and the HIV-1 (+) and HTLV-1 (+) hemophiliac patient had strong RTIAb, while all controls were RTIAb (-). Although the HTLV-1/HIV-1 co-infected hemophiliac patient had strong NAb, while all of 71 HIV-1 (-) but HTLV-1 carriers had no NAb. Overall results suggested that some antigenic determinants of HIV-1 RT which effect the enzyme activity may be weakly cross-reactive to some anti-HTLV-1 antibody. However, the RTIAb (+) sera had no NAb capacity against HIV-1, implying that HIV-RTIAb may not support neutralization against HIV-1 in vivo. The results also suggest that the exposure of HTLV-1 may not contribute to NAb against HIV-1 in HIV-1 infected individuals.

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Nine monoclonal anti-platelet autoantibody producing human cell lines were established from patients with idiopathic thrombocytopenic purpura (ITP) by Epstein-Barr virus (EBV) transformation and their characteristics were studied. Three of the antibodies showed cross-reactivity with single-stranded DNA (ssDNA), as determined by enzyme-linked immunosorbent assay (ELISA) and one of them was also found to bind to the synthetic polynucleotides, poly (dT) and poly (I), as well as to ssDNA.
One of these EBV-transformed human cell lines was fused with the mouse myeloma P3X63 Ag8.653. The resulting hybridoma could be grown in nude mouse ascites by which large quantities of human monoclonal antibodies were obtained.

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A new enzyme linked immunosorbent assay (ELISA) was developed for detecting anti-red blood cell (RBC) antibodies. RBC ghost protein was immobilized on the microtiter plates with 0.05M carbonate buffer, pH 9.6. One percent bovine serum albumin (BSA) in phosphate buffered saline (PBS) was used as a blocking solution. Indirect Coombs' test positive sera and antibody eluates from patients' RBC were added to the wells, and then optimally diluted horseradish peroxidase-conjugated goat anti-human IgG was added. Absorbed peroxidase activity was detected by o-phenylenediamine, and color reactions were terminated by addition of 1 N HCl. Absorbance measured at 490nm.
The reactivities of indirect Coombs' test positive sera were statistically higher (P<0.01) than that of normal sera (0.295±0.355, 0.024±0.030, respectively), and antibody eluates from patients' RBC also responded. But a few samples did not respond. This method is suitable for handling many samples and more sensitive than Coombs' test. But, because some samples positive in Coombs' test did not respond in this method, it is necessary to investigate the difference of the antigenicity between the RBC and the RBC ghost.

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Dipeptidyl aminopeptidase IV (DAP IV) has been demonstrated localizing in certain human T cell. In vitro mitogen stimulations (PHA, Con A and PWM) was employed to assess possible participation of lymphocyte DAP IV in immune responses.
In healthy human peripheral lymphocytes, mitogen stimulations produced a marked increase in DAP IV activities; and similar increase was observed also in culture supernatant. In addition, those phenomena paralleled with increase in released soluble IL-2 receptor.
The principal action of DAP IV remained unknown, but it might, with its connection to production of IL-2 receptor, be part of a correlating mechanism having to do with an immune regulatory system.
Clinical data from the patients with Behcet's disease, rheumatoid arthritis and systemic lupus erythematosus were discussed in a comparable manner.

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Rat mast cells were challenged with compound 48/80 or calcium ionophore A23187 and protein kinase C activity in the cell pellets and amount of histamine release in the supernatant was measured after stopping the reaction. Histamine was significantly released from compound 48/80-activated mast cells at 30 sec after stimulation. In parallel with it, protein kinase C activity in the cell pellets increased at 30 sec and 1 min, and returned to basal value at 3 min after the stimulation. When mast cells were incubated with various concentration of 48/80 for 30 sec, the amount of histamine release and the protein kinase C activity increased dependently on the concentration of 48/80. Significant histamine release from A23187-acti-vated mast cells was found at 1 min after stimulation. Also, protein kinase C activity in the cell pellets showed a tendency to increase at 1 min and returned to basal value at 5 min after stimulation.
These findings suggest that transient increase of protein kinase C activity may be involved in mast cell activation process.

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A 65-year old Japanese woman was admitted to our hospital because of mental confusion and hemorrhagic tendency. She had a 10 years history of mild anemia and Raynaud's phenomenon, and also had xerostomia, xerophthalmia and oral aphthae since 1982. On admission, fever, delirious state, suggillations on the face, chest, arms, legs and abdomen, anemia and mild jaundice were observed. In addition, definite keratoconjunctivitis sicca and oral aphthae were seen. No lymphadenopathy nor hepatosplenomegaly was palpable. Laboratory examinations revealed microhematuria, anemia with fragmented red cells, increased indirect bilirubin and LDH and low level of haptoglobin indicating the presence of microangiopathic hemolytic anemia. Increased FDP, prolonged prothrombin time, low fibrinogen and severe thrombocytopenia clearly indicated the complication of DIC. Autoantibodies; antinuclear antibody, anti-DNA antibody, anti-ENA antibody, anti-RNP antibody, anti-microsoma antibody, antithyroglobulin antibody and elevation of the platelet associated IgG (Pa IgG) were detected, but LE-cell, anti-Sm antibody and coombs tests were negative. Histological examinations showed the deposition of IgG in dermoepidermal junction and fibrinogen in epidermis perivascular lesion of finger, mild lymphocyte infiltration in the microsalivery gland and vascular thrombas in the populus ulcer.
She was diagnosed as having autoimmune disease (SLE and Sjögren's Syndrome) and TTP-like disorder with DIC. High does of prednisolone, gabexate mesilate, fresh whole blood, and platelet consentrates were administered, and her symptomes were improved. The possible relationship between TTP with DIC and the autoimmune disorders was discussed.

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A 75-year-old female was admitted to the Hospital of Yamanashi Medical School on April 22, 1985, because of rheumatoid arthritis (RA) and hemorrhagic diathesis.
Laboratory examinations, on admission, revealed an acceleration of erythrocyte sedimentation rate (124 mm/hr), positive RA test and thrombocyte count of 5×10⁴/μl. The white blood cell count and the red blood cell count were within normal range. Examination of bone marrow disclosed no decrease of megakaryocyte counts. Shortened life span of thrombocytes, increased uptake of ⁵¹Cr-labeled thrombocytes in spleen and elevated levels of platelet-as-sociated IgG (PAIgG) were compatible with autoimmune thrombocytopenia. Treatments with large doses of γ-globulin or steroid hormone failed to restore her thrombocyte count. It decreased progressively to 1×10⁴/μl within one year and platelet transfusions were occasionally required. She died of sepsis from suppurative arthritis on July 17, 1986.
In contrast to the frequent association of RA with thrombocytosis, RA is rarely associated with thrombocytopenia. Recently, with the measurement of PAIgG, autoimmune thrombocytopenia in patient with RA has been reported. Although thrombocytopenia in this case appears to be compatible with such an autoimmune mechanism, no case with such a severe thrombocytopenia and hemorrhagic diathesis has been reported.

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