Frozen sections of the labial biopsies from 18 patients with Sjögren's syndrome (SjS) were studied with a panel of monoclonal antibodies identifying T cells, B cells, macrophages, and their subsets in order to characterize the immunocompetent cells in the salivary glands. T cells were predominant cells in the specimens with mild lymphocytic infiltrations. However, both T cell-foci and B cell-foci surrounding the ducts or acini were observed in the specimens with marked lymphocytic infiltrations. Ratio of B cells to T cells increased in accordance with the degree of lymphocytic infiltration. CD4-positive T cells (inducer/helper T cells) were more predominant than CD8-positive T cells in almost of all cases. CD4-positive T cells frequently showed focal accumulations, while CD8-positive T cells (suppressor/cytotoxic T cell) were diffusely scattered. In almost of all cases, B cell foci were surrounded by or intermingled with CD4-positive T cells. Phenotypes of the cells in B cell foci varied among the cases. CD23 (B6)-positive B cells (activated B cells) were observed in a part of B cell foci in the cases with severe lymphocytic infiltration, while resting B cell foci (CD21-positive) were also observed in the same foci. CD23-positive B cells were close to ducts and a part of them weakly expressed B cell activation antigen (B5) simultaneously. On the other hand, CD14 (monocyte/macrophage antigen)-positive cells were scattered in the interstitium and also positive for HLA-DR antigen. Expression of HLA-DR antigen were also detected on the epithelial cells in the ducts and acini, and on the majority of infiltrating T cells. Intensity of the expression of HLA-DR antigen varied among the cases. These findings indicate that T cells and a part of B cells of the salivary gland in patients with SjS were activated, and might suggest that immune responses between T cells and HLA-DR-positive cells play an important role in the disordered immunoregulation in SjS.

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Oxygen-derived free radicals have been suggested to play an important roles in the pathogenesis of some pulmonary tissue damages such as adult respiratory distress syndrome (ARDS). In the case of ARDS, the production of superoxide from polymolphonuclear leukocyte has been reported to be increased. Alveolar macrophages play important roles in the pulmonary defense by participating in specific immunologic reactions, releasing chemotactic factors and releasing superoxide anions. However, oxygen-derived free radicals released from alveolar macrophages by the stimulation of endotoxin may cause pulmonary tissue damage.
In the present study, we could detect superoxide anions from alveolar macrophages by electron spin resonance using 5, 5-dimethyl-pyroline-N-oxide as a spin trap and chemiluminescence using 2-metyl-6 (p-methooxy-phenyl) 3, 7-dihydroimidazo (1, 2-a) pyrazin-3-one as a chemiluminescence probe. The administration of endotoxin to rats increases the production of superoxide radicals from alveolar macrophages stimmulated by phorbol myristate acetate (PMA). Moreover, by the preincubation of alveolar macrophages with endotoxin, the production of superoxide radicals was significantly increased. The administration of endotoxin to rats significantly increases total protein, albumin and angiotensin I converting enzyme in bronchoalveolar lavage fluid.
Although endotoxin did not cause superoxide production by alveolar macrophages, it enhanced superoxide production induced by PMA. It was suggested that superoxide from alveolar macrophages was one of the toxic factors which induced pulmonary injury by endotoxin.

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Responses of peripheral blood mononuclear cells (PBMC) that had been stimulated by various mitogens to human recombinant cytokines were examined. PBMC that had been stimulated by T-cell mitogen, PHA and Con A, responded to recombinant IL-2 remarkably. However, they did not respond to recombiant IL-3 and IL-4. On the other hand, PBMC that had been activated by T and B cell mitogen, pokeweed mitogen (PWM) and Staphylococcus aureus cowan I (SAC-1), responded to IL-3 and IL-4 significantly as well as to IL-2. Especially, PWM-stimulated mononuclear cells (PWM-blasts) responded well to human recombinant IL-3 and IL-4, and they showed marked DNA synthesis. PWM-blasts did not respond to IL-1, IFNγ, GM-CSF, G-CSF, M-CSF, IL-5 and IL-6. However, they showed significant response to TNF and lymphotoxin, as well as to IL-2, IL-3 and IL-4. PBMC that had been preincubated for 4 or 5 days incorporated ³H-thymidine in dose-dependent way, when they were cultured in the presence of rIL-3 or rIL-4. Responses of PWM-blasts to IL-3 were inhibited specifically, when anti-IL-3 monoclonal antiboby was added to the culture. IL-2 dependent cell line, CTLL-2, did not respond to human rIL-3 nor rIL-4, suggesting that combination of CTLL-2 assay and PWM-blast method enables us to discriminate between human IL-2 and IL-3/IL-4 activity in simple and convenient way. CD2⁺, CD4^-, CD8^- cells were shown to play an important role in IL-3 responsive cells of human peripheral blood.

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In the present study, the mechanism that explains failure of rejection of the fetus by the mother was immunologically analysed to be clarified and the immunocapacity of the regional lymph nodes draining the uterus (DLN) in pregnant women was attempted to be investigated.
Surface markers of mononuclear cells (MNC) of DLN in pregnant women were analyzed by FACS analyzer using monoclonal antibodies and the increased HLA-DR and CD11b positive cells were observed. Immunohistological study of DLN was also carried out and HLA-DR positive cells were predominantly found in T cell zone and the results indicated that the T cells were found to be activated due to the appearance of the HLA-DR positive cells. Furthermore, the immune responsiveness of MNC of DLN in pregnant women was tested by lymphoblastogenesis and the results were compared with normal controls. The mitogenic response of DLN-MNC was found to be higher than that of the controls. The response of DLN-MNC to cord MNC was also found to be higher than that of peripheral blood MNC. Therefore, the immunocapacity of DLN-MNC in pregnant women was thought to be increased by recognition of the fetal antigens.
Suppressor cell activity of DLN-MNC in pregnant women was shown to be not significantly elevated, while cytolytic activity was increased. Therefore, the suppressive effects of cytolytic activity is considered to be important role for the maintenance of pregnancy.

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A study was performed on the enterobacterial flora and secretory IgA concentration in feces of the cases, in which diarrhea occurred during the administration of antibiotics. In the cases, where clostridium difficile and its toxin were detected in feces, total number of bacteria in feces and secretory IgA concentration decreased and the enterobacterial flora was destroyed. In the enterobacterial flora of these cases, anaerobic bacteria such as Bifidobacterium, Bacteroides, Eubacterium, etc. decreased, while yeast increasd. In some of these cases, Streptococcus and Lactobacillus have also increased. In the cases, in which Cl. perfringens and Cl. butyricum were detected in feces, Cl. difficile was not found. In the cases, in which Cl. difficile was detected and to which Cl. butyricum preparation has been given perorally, the number of Cl. difficile was lower than the cases, to which Cl. butyricum preparation was not given, and the production of toxin was also low.

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The success in LAK therapy depends on obtaining a large amount of cells with high LAK activity. We considered that the absolute number of cells in culture is not the number of true LAK cells but that ³H-thymidine uptake by cells that react to rIL-2 is a mark of true LAK cells. We assumed that high LAK activity and the greatest ³H-thymidine uptake are the conditions to obtain a large amount of high LAK activity cells and examined this hypothesis. We also examined the enhancement of induction of the LAK activity by the addition of Nocardia rubra cell wall skeleton (N-CWS) and rIL-2 to the culture.
The optimal condition to obtain a large amount of high LAK activity cells was culturing 2.5×10⁶/ml lymphocytes for 5 days with 2 U/ml or more rIL-2 in the presence of human type AB serum. No LAK activity was induced by culturing with N-CWS alone, but the LAK activity was markedly enhanced by culturing with N-CWS and rIL-2. The LAK activity was enhanced by adding rIL-2 first and N-CWS next after 2 days, but no LAK activity was induced by adding N-CWS first and then rIL-2 2 days later.

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The plasma concentrations of Factor VIII related antigen (F VIII R: Ag), tissue plasminogen activator (tPA), thrombomodulin (TM), β-thromboglobulin (β-TG), platelet factor 4 (PF4) and fibrinopeptide A (FPA) in 35 patients with collagen diseases (17 systemic lupus erythematosus-SLE, 11 progressive systemic sclerosis-PSS, 7 rheumatoid arthritis-RA) were measured before and after the whole body exposure to cold at 4°C to compare the differences in reaction to cold, especially between those groups with and without Raynaud's phenomenon.
The mean concentration of F VIII R: Ag was revealed to be higher (p<0.01) in those patients than in healthy controls even before the cold exposure and regardless of Raynaud's phenomenon, and this alteration was more prominent in PSS. The mean basal levels of TM, β-TG, and PF-4, though ranged within normal, were also higher (p<0.01) in the patients compared with controls.
The mean level of F VIII R: Ag in the patients with Raynaud's phenomenon was significantly increased (p<0.05) after the cold exposure than in their counterparts, and this increment was greater in SLE.
From these results, it is conceivable that patients with collagen diseases, especially those presenting with Raynaud's phenomenon, are in a subclinical thrombotic state, so a consideration of prophylactic antithrombotic therapy would be suggested for them along with conventional one.

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Susceptibility of LAK cells to K562 cell line was investigated by HTCA (human tumor cell clonogenic assay), when target cells were pretreated with antineoplastic drugs. Percent inhibition of colony formation as cytotoxic effect of LAK cells and antineoplastic agents was significantly higher than that of LAK cells or antineoplastic agents alone. This susceptibility of combination of LAK cells with antineoplastic agents was additivism. The cytotoxicity of LAK cells was not affected by antineoplastic agents, enen when target cells were treated concurrently with antineoplastic agents. The susceptibility of LAK cells against intermittent drug-exposure K562 cells as a resistant cell line was investigated. Percent inhibition of colony formation of LAK cells against CDDP and MMC resistant K562 cell lines were significantly higher than that of parent K562 cell line. To investigate the difference of tumor-binding capacity and recognition of LAK cells against these cell lines, direct conjugate forming assay and cold target inhibition assay were examined. As a result of these assays, recognizing ability of LAK cells against resistant cell line was lower than that of parent cell line in opposition to inibitory effect of colony forming. Then we investigated the difference of membrane fragility of these cell lines. NP-40, a non-ionic detargent of membrane, was incubated with these cell lines. Consequently, the cell membrane of CDDP resistant cell line was more fragile than that of K562 cell, and it is suggested that the difference of susceptibility against these cell lines may be induced by membrane fragility.

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A monoclonal antibody, GC401, was established by fusing murine myeloma cells, SP2/O and the splenocytes of a. BALB/c mouse immunized with the human gastric cancer cell line, NUGC4. Immunohistochemically, GC401 reacted with glandular epithelium of various normal tissues and various cancers derived from glandular epithelium. Western blot analysis of the GC401 antigen indicated a molecular weight of ca. 30K, and the antigenic determinant was suggested to be a glycoprotein with disulfide bonds. In this study, we analyzed the change of phenotypic expression of differentiation antigens in human stomach by monoclonal antibodies GC302 (previously reported) and GC401. GC302 antigen was found not to be expressed in normal adult gastric mucosa, but to be expressed in the fetal gastric mucosa, intestinal metaplasia and almost all of the cancer of the stomach. Whereas GC401 antigen was expressed in all stage of differentiation of the stomach, GC302 reacted more strongly with intestinal metaplasia which was negative in high iron diamine staining and had less goblet cells. But GC401 reacted strongly with all types of intestinal metaplasia of human gastric mucosa.

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Long-term outcomes of sulphasalazine (SASP) therapy for rheumatoid arthritis (RA) were compared with those of auranofin (AF).
Eight of the 41 patients and 6 of the 39 patients who were treated with either SASP or AF stopped each treatment within 3 months because of adverse effects of the agents. Shortterm efficacy, therefore, was compared between the remaining 33 patients received SASP and 33 patients treated with AF: SASP or AF was effective in 79% or 58% of the patients, respectively. There was no significant difference in the incidences of short-term efficacy between SASP-and AF-treated groups. However, SASP more rapidly improved the activities of disease and laboratory parameters such as serum rheumatoid factor levels as compared with AF.
Cumulative continuation rates of treatment were compared between SASP-and AF-treated groups by using Kaplan-Meier method. The continuation rates were significantly higher in SASP-treated group within the range from 9 to 18 months after the beginning of treatments than AF-treated group. Thereafter, the rates of both treatments gradually lowered and became almost identical (about 20%).
Most of the adverse effects of SASP were mild and developed within one month after the beginning of treatment, although proteinuria developed in some of the AF-treated group several months later.
These results support the reported evidence that SASP is an effective and safe treatment for RA, and suggest that SASP is a candidate for the first-choice agent among several slow-acting anti-rheumatic drugs.

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A 67 year-old female was admitted with dyspnea, dry cough and fever. Idiopathic interstitial pneumonia was diagnosed by a history of illness, findings of chest X-ray and computed tomography. She was improved once by the administration of antibiotics and steroid therapy. But about 2 months later she worsened clinically and died of respiratory failure despite treatment with a high dose of corticosteroid and antibiotics. Autopsy proved that she died not of pneumonia but of respiratory and cardiac failure that was brought about by acute progressive interstitial pulmonaly fibrosis.
In the clinical course we examined complement fragments, iC3b, Bb, C4d, C3a and C5a, those were drastically up and down in accordance with the clinical course. Those results suggested that complement and its fragments through activation may take a some kind of role in the pathogenesis of idiopathic interstitial pneumonia.

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A case of systemic mast cell disease (SMCD) was reported. A 22-year-old woman who had been suffering from pruritus at the palms and legs since 17 years old, admitted to the hospital because of developing systemic pruritus and edema on the extremities.
On admission, she had swelling of left parotid gland and systemic peripheral lymphadenopathy. Laboratory findings showed eosinophilia (WBC 15, 000/mm³, eosinophil 51%) and elevated serum LDH (1, 309IU/l). These symptoms and abnormal laboratory findings fluctuated every 20 to 40 days. In addition, the elevation of serum IgE and mild hepatosplenomegaly were seen.
The increase of mast cells were seen in the biopsied specimens of bone marrow, lymph nodes, parotid gland, skin and liver. From these findings the diagnosis of SMCD was made.
Besides of these findings, peripheral blood lymphocytes revealed about 40% of CD3 (-) CD4 (+) cells among whole lymphocytes. Three-color staining was useful for the further examination of surface markers of these abnormal lymphocytes. Lymphocyte subset abnormality like this has not been reported in SMCD until now.
SMCD is a rare disease. But eosinophilia accompanies about 15% of SMCD. If we encounter the eosinophilia of unknown origin, we should take SMCD into account.

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A case of malignant rheumatoid arthritis (MRA) associated with fulminating sensorimotor neuropathy was reported. A 53-year-old female had been suffered from RA since 1969 and received long-term corticosteroid treatment. The motor and sensory nerve disturbance of upper and lower extremities was suddenly appeared on February 1, 1988, and she was transferred to Senami Hospital. Since she had severe sensorimotor neuropathy of forearms and legs, leukocytosis, thrombocytosis and increase of rheumatoid factor and acute phase reactants, the diagnosis of MRA was made. Because gangrene of both fingers and leg ulcers developed rapidly within a few days, she was treated with high dose corticosteroid combined with immunosuppressants and plasmapheresis. Despite of improvement of laboratory examinations the ulcers of both legs had been infected frequently and become worse, followed by bilateral below knee amputation. Vasa nervosum of the sural nerve and small arteries around the ulcers showed vasculitis.
This case was classified as Bywaters' type of MRA because of little internal organ damages. In addition to the conventional steroid and immunosuppressant treatment, more extensive therapy including intravenous cyclophosphamide therapy and long-term plasmapheresis should be considered in MRA with fulminating sensorimotor neuropathy.

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We report a case of Weber-Christian disease who showed hypothermia after treatment with prednisolone. A 12-year-old female was admitted to our hospital because of remittent fever and subcutaneous nodules with pain. She failed to respond to treatment with antibiotics and aspirin. The histological findings in skin biopsy revealed acute panniculits with vasculitis. The diagnosis of Weber-Christian disease suggested by clinical features was confirmed. Prednisolone at initial dosage of 40mg per day was used for therapy. Fifteen hours after prednisolone therapy was begun, her temperature dropped rapidly to 34.1°C. The temperature ranged between 34.2°C to 36.0°C for 40 hours and then rose slowly over the next 3 days. The other clinical symptoms and laboratory data improved by prednisolone therapy. This case was interesting in to consider the effect of adrenocorticosteroid to the thermo-regulation.

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A rare case of common variable immunodeficiency disease who developed idiopathic thrombocytopenic purpura was reported. In October 1981, a 29-year-old woman was admitted to the Keio University Hospital because of aseptic meningitis. On this occasion, she was found to have hypogammaglobulinemia (IgG 277 mg/dl, IgG 22 mg/dl, IgM 18 mg/dl). The in vitro culture experiments indicated that defective B cell function and activated suppressor T cells might account for hypogammaglobulinemia observed in this patient. A diagnosis of common variable immunodeficiency disease was made and monthly supplementation therapy by gammaglobulin was started. On July 1985, she was readmitted because of severe thrombocytopenia (10, 000/mm³). Physical examination revealed multiple purpura on her upper and lower extremities. Bone marrow examination showed normocellularity with normal number of megakaryocytes (70/mm³). Tests for circulating immune complexes were negative and platelet bound IgG (PBIgG) was within normal limits. Titers of antibodies to viral antigens (EB, cytomegalo, rubella, coxackie, measles, influenza) were not elevated. Treatment by prednisolone (60 mg/day) was ineffective and high dose intravenous immunoglobulin resulted in only transient increase of platelet counts. From the 17th hospital day, the vincristine slow infusion therapy was started and remarkable recovery of thrombocytopenia was obtained. Although it might be difficult to determine whether ITP in this case was of the acute or chronic type, we could not find an established case report of common variable immunodeficiency disease complicated with ITP. The mechanisms of action of slowly infused vincristine in ITP and pathogenetic mechanisms of ITP occurred in an immunodeficient patient were discussed.

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The influence of PSK on superoxide generation from polymorphonuclear leukocytes (PMN) was studied by Cypridana luciferin analog-dependent chemiluminescence and by electron spin resonance assay using 5, 5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trap. Superoxide generation from rat peritoneal PMN stimulated by OZ or PMA was augmented by the 8-day oral administration of PSK in vitro, but not by the incubation with PSK in vitro. PSK had scavenging effect of superoxide generated from the hypoxanthinexanthine oxidase system. These results suggest that the enhanced generation of oxygen radicals induced by PSK may play a beneficial role in the tumoricidal activity exhibited by PMN. The significance of the scavenging effect of superoxide by PSK remains unknown.

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