So-called “non-specific bindings” are troublesome problems in flow cytometric surface marker assay with monoclonal antibodies. Especially in the detection of weakly positive populations and in the fine evaluation of fluorescence intensity, it should be resolved for practical purpose.
We examined the reactivities of mouse serum IgG fractions and mouse monoclonal isotypic control antibodies to some cell lines bearing Fc receptor, and also referred these subclass specificities to other monoclonal antibodies against defined surface markers. Well controlled staining procedures and use of selected monoclonal antibodies and fluorochromic anti-mouse antibodies disclosed that most cause of the non-specific binding was thought to be deduced to the antibody binding activities of Fc receptors.
We attempted to eliminate Fc receptor derived non-specific binding by adding a step to the conventional staining procedures. The pretreatment of the cells with commercial human gamma globulins was quite effective in the neglection of false positivities of control antibodies. However this simple but useful technique might have some adverse possibilities that it decreases the fluorescence intensity of some special markers, for example Leu-11a (antibody directed against Fc receptor).
Negative control staining with control antibodies is essential for practical surface marker assay and non-specific binding can be prohibited by blocking of Fc receptor with human globulins in most cases.

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Pokeweed mitogen (PWM) is widely used to induce immunoglobulin production of PBL. Since PWM predominantly stimulates suppressor T cells, it is sometimes impossible to evaluate correctly the function of B cells or helper T cells. It is known that Staphylococcus aureus Cowan 1 (SAC) induces T cell-independent proliferation of B cells and T cell-dependent differentiation of B cells. To study the activation of T cells stimulated by SAC, two surface antigens (T4, T8 and Tac) were determined simultaneously by indirect immunofluorescence. By this method, SAC was demonstrated predominantly to activate the T4⁺ T cells, but not the T8⁺ T cell subset.
Suppressor T cells are believed to be involved in the pathogenesis of some cases of common variable immunodeficiency (CVID). Such cells suppress PWM induced Ig production in coculture of normal and patient lymphocytes. Peripheral blood lymphocytes (PBL) and T cells of two patients with CVID and a patient with agammaglobulinemia were shown to suppress PWM induced Ig production of control PBL. However T cells of these three patients showed normal helper activity in SAC induced Ig production. The data indicate that the parallel use of SAC and PWM is always needed for studying the mechanisms responsible for altered immunoregulation in patients with hypogammaglobulinemia.

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A case of bilateral pneumothoraces caused by subpleural rheumatoid nodules in malignant rheumatoid arthritis (RA) is reported. A 56-year-old man with a long history of RA was admitted to another hospital in February 1984 because of high fever, progressive arthritis, cough and sputa. Chest X-ray film disclosed bilateral pleural effusions and a coin lesion in the left lung field. He had no history of occupational exposure to dust. The white-cell count was 13, 600, the erythrocyte sedimentation rate 63mm per hour, CRP 9+, CH₅₀ 23U/ml and RAHA 1:160. He was initially treated with prednisolone 30mg/day and antituberculosis drugs, but experienced little improvement. Prednisolone was increased to 60mg/day and the patient became afebrile. He was transferred to this hospital in April 1984.
He had active polyarthritis (classical RA, stage IV, class III), but no subcutaneous nodules. Thoracentesis yielded a greenish yellow effusion with protein 17.6g/dl, glucose 0mg/d1 and cell count 970/mm³ (neutrophils 48%, histiocytes 16%, lymphocytes 35%). Cultures for tuberculosis were negative and cytologic examinations showed no atypical cells. The echocardiogram disclosed a pericardial effusion. Chest X-ray film revealed bilateral pneumothoraces and pleural effusions, and a coin lesion in the left lung. Computed tomography of the chest revealed bilateral pleural effusions and pleural thickening and a subpleural tumor in the left lung. In addition, he had episcleritis and peripheral neuropathy in the right leg. He was diagnosed as malignant RA. He was treated with chest tube drainage without improvement. In September, the right thoracotomy with decortication and abrasion was performed. Several yellow, subpleural nodules were observed. Histological examination of the nodule disclosed infiltration of histiocytes and lymphocytes, and giant cells around necrosis or cavity, but typical “palisading arrangement” was not found. The fluorescent staining for tubercle bacilli was negative. In this case, the pneumothorax was most probably caused by cavitation and rupture of subpleural rheumatoid nodules.
Pneumothorax is an unusual complication in the pleuropulmonary manifestations of RA and so far no case has been reported in Japan.

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A case of cytomegalovirus (CMV) infection following pulse therapy in systemic lupus erythematosus (SLE) is reported.
A 39-year-old woman developed polyarthralgia and fever during her second pregnancy at the age of 29. Alopecia, Raynaud's phenomenon and malar erythema developed later. She was admitted to another hospital in January 1982 because of polyarthralgia. Laboratory examination showed positive antinuclear antibody, high titers of anti-DNA antibody, leukopenia and telescoped urinary sediments. She was diagnosed as SLE and was treated with prednisolone 40mg/day. Polyarthralgia and malar erythema disappeared, but telescoped sediments remained unchanged.
She was transferred to this hospital in April 1982 for the purpose of renal biopsy. Laboratory findings on admission were as follows: the whitecell count 3, 800, CRP 1+, the erythrocyte sedimentation rate 97mm per hour, RA test negative, antinuclear antibody positive, anti-DNA antibody 805U/ml (RIA), anti-RNP, Sm, SS-B antibodies negative, anti-SS-A antibody 1:16, CH₅₀ 13.6U/ml, C3 33mg/dl, C4 9mg/dl, and circulating immune complex 20.0ug/ml (Clq binding). 24-hour urinary protein was 1.3 g/day, and urinalysis revealed 13-15 white blood cells, 12-13 red blood cells, 10-13 granular casts and 1-2 hyaline casts per field. The creatinine clearance was 91ml/min. Renal biopsy showed active diffuse proliferative lupus nephritis. She was treated with prednisolone 60mg/day, but telescoped sediments were unchanged. Pulse therapy was porformed twice with methylprednisolone 1g/day for three days in August. In September, anti-DNA antibody and serum complement were almost normal. Velcro rales became audible in both lung bases and arterial blood gas values worsened. Interstitial pneumonitis due to SLE was suspected and prednisolone was increased to 100mg/day. Immunosuppressive drugs and plasmapheresis were added. The titer of antibody to CMV (complement fixation) rose from 1:16 in April to 1:128 in September. Urine culture for CMV performed in September was reported as positive in October. Prednisolone was tapered. Blood gases improved. In February 1983, ulcers of stomach and tongue developed. The histological examination revealed cytomegalic inclusion bodies. Lung biopsy was not performed.
CMV infection is rare in adults and reported mostly in patients with organ transplants and neoplasms, who usually received corticosteroids or immunosuppressive drugs and were severely immunocompromised. CMV infection in adults is thought to be due to recrudescence of latent virus, triggered by an immunologic defect in the hosts. CMV infection in SLE was so far reported only in one case in Japan. CMV infection should be kept in mind in cases of SLE under severe immunosuppression such as pulse therapy.

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OK-432, one of the so-called biological response modifiers (BRMs), has been reported to restore cellular and humoral immune responses in tumor-bearing individuals. However, since host immune responses in tumor-bearing individuals are modified by development of disease, by application of various forms of anticancer therapy, surgical operation, and by radiation therapy, it is sometimes impossible to evaluate accurately the effect of these nonspecific BRM restoring impaired immune response in patients with malignancy. Avoiding this problem, we studied the effect of BRM on host immune responses in the aged individuals without malignancy, who had shown decreased cellular immunity detected by PPD and PHA skin tests and in vitro blastoid transformation test of lymphocytes by PHA. Although OK-432 is used widely by intradermal or intramuscular injections, it is more convenient for the patients to receive oral administration than intramuscular injection having the side effects such as chill and fever.
Five KE or 20 KE of OK-432 was orally given three times a week for 1 month to the aged patients without malignancy who showed decreased skin responses to PPD and PHA. Oral adminstration of 5 KE of OK-432 evoked an increased skin response to PPD except 2 patients. PPD skin response was much more increased by the oral administration of 20 KE of OK-432, and the increased responsiveness was detected even 2 months after withdrawal of OK-432. There was same tendency in the skin response to PHA. The increased skin reaction to SuPS (lipopolysaccharide fraction of Streptococcus pyogenes Su strain) was observed in the one group of patients treated with 5 KE of OK-432, while there was no significant increased response by oral administration of 20 KE of OK-432.
It was concluded that the activation of non-specific host immune responses assessed by the skin tests used by us could be evoked by the oral administration of OK-432 as well as intramuscular injection.

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