The proliferation of CD3-LAK cells could be induced from normal peripheral blood mononuclear cells (PBMC) after in vitro treatment with CD2 monoclonal antibody (MoAb) plus rabbit complement and culture with IL-1 plus IL-2. This presentation exhibited the results of the cloning of the LAK cells.
PBMC from healthy donors were incubated with CD2 MoAb plus rabbit complement for 1 hr, then washed with HBSS and cultured with rIL-1β (0.5μg/dl) plus rIL-2 (2μg/dl) in RPMI-1640 culture medium. Two to ten days later, the LAK cells were cloned under limiting conditions in 96-well microtiter plates. Ten to 14 days later the cloned cells were pipetted into a culture flask and continued to culture with the addition of rIL-1 plus rIL-2. After the cloned cells increased to sufficient numbers of cells, their surface markers were analyzed using various MoAb by FCM and histostaining techniques, and the non-major histocompatibility-restricted cytotoxicity (NMRC) was tested against target K562 cells.
The results showed 112 clones were WT31+γ/δ-1-, 62 clones were WT31-γ/δ-1+ and 101 clones were WT31-γ/δ-. Most of the WT31-γ/δ-1- clones showed CD2+, CD7+ and CD56+, but no CD3, βF 1 and TCRδ-1 on their cell surfaces. Some of the WT31-γ/δ-1- clones showed CD2- and CD56-. The NMRC showed 13 clones out of 27 WT31-γ/δ-1- clones tested had a strong cytotoxicity to K562 target cells but none of the WT31+ nor γ/β+ clones had.
The surface markers of 10 out of 13 NMRC positive clones showed both CD2 and CD56 were positive. From these results, CD2 and CD56 antigens may have some important effect on CD3- LAK cells regarding to the NMRC activity.

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House dust mite Dermatophagoides farinae (Df) is an important aeroallergen in bronchial asthma of childhood. We have studied herein whether peripheral blood mononuclear cells (PBMC), obtained from children with stable asthma and strongly positive RAST results for Df, can produce colony-stimulating activities (CSAs) by stimulation with crude Df antigens in vitro. Levels of CSAs in culture supernatant were assessed by an in vitro clonal culture. PBMC of the patients, unlike those of non-allergic controls, released significantly higher levels of eosinophil (Eo)-CSA and granulocyte-macrophage (GM)-CSA when stimulated with Df antigens than unstimulated PBMC. The addition of polymyxin B, substance that inactivates endotoxin, did not suppress the Df-antigen-induced generation of the CSAs. The supernatant of PBMC stimulated with Df antigens contained significantly higher levels of both the CSAs in the patients than in the controls. These results indicate that PBMC stimulated with Df antigens can produce elevated levels of Eo-CSA and GM-CSA in children with Df-allergic asthma in comparison to those in non-allergic children.

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We measured anti-nuclear antibody (ANA) by the method of enzyme-linked immunosorbent assay (ELISA) in 150 cases with collargen disease and evaluated its clinical usefulness in comparison to the indirect immunofluorescence antibody method (IF). ELISA-ANAs with three different types of antigens were especially examined. When the cut off values were set at 3.0 units ELISA-ANA and 20 dils IF-ANA, the sensitivity, specificity and coincidence were 82.1%, 80.0%, 82.0% (IF vs ANA-I ), 88.6%, 70.0%, 87.3% (IF vs ANA-II), 97.9%, 66.7%, 96.0% (IF vs ANA-III), respectively. The measured values by ANA-III were significantly higher than those by the other types (p<0.01). The correlation cofficients between ELISA-ANA-III and IF-ANA in SLE, Primery SjS and PSS; NCTD; PM/DM were r=0.618, 0.591 and 0.682, respectively.
The case with anticentromere or anti PCNA antibody positive by IF-ANA could be detected only by ELISA-ANA-III among ELISA.
We conclude that ELISA-ANA-Ill is a useful screening test for total ANA detection.

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The proportion of CD4/CD8 T-cell subsets and HLA-DR and IL2 receptor (IL2R)-expression on peripheral T-lymphocytes from patients with infectious mononuleosis (IM) were examined.
There was a significant increase in both the proportion and the absolute number of CD8 positive (CD8⁺) cells in comparison with those in healthy adults; thus the ratio of CD4⁺ cells to CD8⁺ cells was significantly decreased. HLA-DR expression on T-cells, particularly on CD8⁺ cells, was increased. The expression of IL2Rα (CD25) was not increased, but that of IL2Rβ was increased. The number of CD7⁺ cells decreased and that of CD45RO⁺ CD8⁺ cells increased. These findings were thought to represent the activation of CD8⁺ T cells and the increase of memory T cells.
We found a significant increase of TdR-incorporation of the lymphocytes in IL2 stimulation in contrast to PHA stimulation. By IL2 stimulation for 48h, the number of HLA-DR⁺ CD8⁺ cells was increased as compared with medium alone and the proliferating cells were CD7⁺ CD8⁺, though most of the fresh cells from patients were CD7^- CD8⁺. And after PHA stimulation for 48h, the fluorescence intensity of CD7 increased and HLA-DR^- cells were changed into HLA-DR⁺ cells. After incubation for 48h, the number of viable cells from patients with IM was lower than that from healthy adult volunteers.

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Association of age and serum IgE levels with pathophysiology of asthma was examined in 64 patients with asthma. Positive RAST scores of 2+ or more to inhaled allergens were significantly higher in patients with high serum IgE than in those with low serum IgE in 4059 year-olds. The proportion of BAL lymphocytes was significantly higher in patients over age 60 with high serum IgE compared with those between 20 and 39 and between 40 and 59 with either high or low serum IgE. The release of histamine or leukotriene C₄, from leucocytes was significantly higher in patients with high serum IgE than in those with low serum IgE in 20-39 (both histamine and LTC₄) and over 60 year-olds (histamine).

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The very low incidence of male patient in systemic lupus erythematosus (SLE) indicates that genetic factors and sex hormones may be importantly associated with the disease. Seventeen male SLE patients were examined to clarify the difference in clinical features, laboratory findings, and endocrinological data from female SLE. Seventeen female SLE patients matched in age of onset and duration of disease were studied as controls. Photosensitivity (p=0.04), lymphadenopathy, and neurological involvement were more frequent in males, with arthritis and skin rash being seen less frequently. The incidence of renal involvement was same in both sexes, however the nephrotic syndrome was noted more often in males. Leukopenia occurred more commonly in the females (p=0.07). Other laboratory findings revealed no significant difference. All patients except one female received corticosteroid. The mean dose of maximum prednisolone was 90.1mg for males and 44.1mg for females.
Sex hormones were measured in 10 male patients. No male patient showed increased estrogen levels. Three male patients who received high dose of prednisolone or immunosuppressive drugs showed decreased plasma testosterone. Elevated plasma FSH was found in 4 patients, and 3 of whom received immunosuppressive drugs. Thus, male SLE would appear severer than female SLE and the abnormality of hormonal status in male SLE was possibly due to the therapeutical agents.

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A 55-year-old woman with pustulosis palmaris et plantaris (PPP) presented with sudden hearing loss (SHL). After steroid therapy for SHL, she had lumbago, pain of sterno-cost clavicular joints, sweating, and palpitation. The bone scan showed accumulation of 99 mTc in sterno-cost-clavicular joints. On the basis of physical and endocrinological examinations, she was diagnosed as pustulotic arthro-osteitis (PAO) with Basedow's disease. Antithyroid drug therapy for Basedow's disease could control both thyroid function and arthralgia. She showed hypersensitivity to antimony chloride. After patch test, thyroid function, arthralgia and PPP were worsen. Removal of antigen from oral cavity resulted in one year remission of PAO.

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A patient of chronic fatigue syndrome (CFS) with low CD4/CD8 was reported. A 23-year-old male was admitted to our hospital because of low grade fever and general fatigue. Besides, the patient complained of sore throat, myalgias, muscle weakness, headaches which lasted for a long time. Infectious disease, connective tissue disease, and malignancy were ruled out. Based on the clinical criteria proposed by CDC criteria, he was diagnosed CFS. In addition, low CD4/CD8 ratio and lymulus lysate test positivity were persisted for a long time. These findings suggested that CFS was based on a kind of immunological disorder.

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A 43-year-old female was first diagnosed as having rheumatoid arthritis (RA) because of polyarthritis. She was treated with non-steroid anti-inflammatory drugs, Salazosulphapyridin and Thiopronin. Subsequently lymphnode swelling and muscular weakness in the neck appeared, accompanied by high fever. On admission to Ida Kawasaki Metropolitan Hospital, the most likely diagnosis was thought to be polymyositis or MCTD, because of the muscular weakness, elevation of myogenetic enzyme and positive anti-n-RNP antibody. Later, a diagnosis of systemic lupus erythematosus (SLE) was strongly suspected because of positive antinuclear antibodies, positive LE cell phenomenon and high level of anti-DNA antibody. Finally, the diagnosis of SLE was confirmed by cervical lymphnode biopsy showing necrotizing lymphadenitis with a hematoxylin bodies. This case suggested that lymphnode biopsy may be a useful tool to diagnose SLE in such patients with lymphadenopathy.

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Splenectomy is indicated for patients with hypersplenism, and it is well known that various kinds of immunological problems may develop following this procedure. An IgG subclass abnormality, however, has not been described before. We present a 15-year-old Japanese boy whose serum level of IgG 2 was higher than that of IgG 1 one month after splenectomy, and both of them returned to the normal range in six months.
This patient had an autotransplantation of bone for his multiple osteocystoma at the age of 10. He never experienced a severe form of infection. His platelet count started to decrease around the age of 10. From twelve to fifteen years of age, when the platelet level fell to 60, 000/μl, a bleeding tendency such as nasal bleeding and purpura became obvious. At 15, this boy was admitted to our hospital for splenectomy because his spleen was getting significantly larger, and the platelet count was further decreasing. The serum IgG concentration was low, 559 mg/dl, with subclasses IgG 1 350 mg/dl and IgG 2 181 mg/dl before the splenectomy. The serum level of IgG 1 and IgG 2 elevated to 637 mg/dl and 895 mg/dl, respectively one month following the splenectomy. It required six months that these IgG subclasses returned to the normal level. This may be due to the different distribution of B cells and plasma cells producing IgG subclasses in various organs.

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A 52-year-old woman was admitted to our hospital in December 1989 because of polyarthritis which involved her bilateral wrists, finger joints, shoulders, right knee and ankle. In 1982, she developed myasthenia gravis and hyperthyroidism. She received thymectomy and partial thyroidectomy in 1985, which made her free of symptoms without any medication. She began to suffer from polyarthralgia with morning stiffness in May 1989. Her rheumatoid factor was positive and X rays of the joints showed bony erosion. Furthermore, leukocytepenia (1, 200-2, 200/mm³) and splenomegaly was also noted, therefore, she was diagnosed as Felty's syndrome. Although titer of the anti-acetylcholine receptor antibody was high, she showed no symptoms of myasthenia gravis. Oral methotrexate (7.5 mg/week) and prednisolone (10mg/day) improved her polyarthritis, but the leukocyte count still remained under 3, 000/mm³. A combination of these diseases was rare and this case seemed to be of interest in considering the variety of the overlap of autoimmune diseases.

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