Peripheral blood and synovial fluid mononuclear cells (MNC) were cultured with pokeweed mitogen for 7 days, and the media which contained secreted IgM rheumatoid factor (IgM RF) were assayed by double antibody radio-immunoassay technique, which was specific and sensitive enough to assay them.
Peripheral blood and synovial fluid MNC from seropositive RA patients synthesized high levels of IgM RF, whereas those from seronegative RA patients, patients of osteoarthritis and normal subjects synthesized low levels of it. The IgM RF titers in media were correlated well with the level present in the sera of the donors which were assayed by haemagglutination methods. Synthesis of IgM RF by synovial fluid MNC but not by peripheral blood MNC led to a concomitant synthesis of total IgM. Moreover B lymphocytes demanded helper activity of T lymphocytes for the synthesis of IgM. These data suggested that peripheral blood and synovial fluid MNC from patients with RA play an important role in the synthesis of IgM RF in vivo.

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Chemiluminescence (CL) in whole blood has been analyzed to assess the phagocytic function of granulocytes and the serum opsonic activity simultaneously. Granulocytes are well known to show CL during phagocytosis, which is derived from the interaction between activated oxygen species generated in cells and substances phagocytosed. The total activity of activated oxygen species generated from phagocytes can be measured in luminol-dependent CL. This method, namely luminol-dependent CL, has become an elegant and simple method to estimate the phagocytic function of granulocytes or monocytes. The method described here is an improved one, in which whole blood can be directly used and serum opsonic activity can be measured simultaneously. The methods are as follows; a 0.1ml of freshly drawn whole blood is put into a CL measuring vial, in which a 0.4ml of medium (Eagle's medium buffered with 50mM HEPES containing 10U/ml of heparin) has been put. After 20min incubation at 37°C, 10μl of luminol at the final concentration of 40μg/ml is added to a specimen. Following equilibration for 10min, a background CL is measured. Then a 10μl of stimulant is added to the specimen. CL is continuously recorded by a recorder connected to the CL machine (BLR-102, Aloka). The results obtained in this study are as follows;
1) The peak CL in whole blood is about 1/30 of that of granulocytes alone, however, the CPM is high enough for the estimation of phagocytic function of granulocytes.
2) The peak CPM correlates to the number of granulocytes present in specimen, and/or function of granulocytes.
3) The time showing the peak CL depends on the activity of serum opsonin.
4) If aggregated gamma globulin is used as the stimulant, it can quickly activate granulocytes without opsonin.
5) Sheep red blood cells can be a stimulant, if complement and specific antibody are present in blood. These results seem to confirm that the measurement of CL in whole blood offers much information concerning functions of phagocytic system in whole blood.

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In a thought that hypothalamus, pituitary, and adrenal system may be acting as a factor to control immunization, studies were conducted with pituiterectomized rats and adrenectomized rats for histological examination of thymus and spleen and for evaluation of the number of peripheral lymphocytes, amount of immunoglobulin, and DNCB test.
The pituiterectomized rats demonstrated reduced T-cell area of thymus and spleen, reduced number of peripheral lymphocytes, and negativization of results of DNCB test, while both IgM and IgG of immunoglobulin showing liquid immunization gradually increased after the extirpation.
Especially, IgG increased about doubly at 30 days. On the other hand, the adrenorectomized rats showed no change in thymus, spleen, or the number of peripheral lymphocytes, but the immunoglobulin IgG was transiently reduced into half immediately after the extirpation, but gradually increased thereafter into approx. 1.5 times number at 2 weeks. The increase in the amount of immunoglobulin may be attributed to the reduction in T-cells, but details will have to be further studied. The above-mentioned findings may be deemed to be the evidence that the pituitary and adrenal system exert important influences upon the immunization abilities.

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Human lymphocyte subpopulations were studied histochemically for acid alpha-naphthyl acetate esterase (ANAE) and 5'-nucleotidase (5'-N).
The percentage of ANAE positive T lymphocyte of normal subject was 24.0±9.7% in cord blood, 41.4±9.8% in infancy, and 55.9±10.9% in adult, respectively.
The lymphocytes from patients with X-linked agammaglobulinemia had normal ANAE activity. In aplastic anemia, Down syndrome and Lesch-Nyhan syndrome, ANAE activity in their T lymphocytes was significantly reduced. And, the lymphoblasts isolated from patients with acute T-lymphoblastic leukemia (T-ALL) had lacked ANAE activity.
The percentage of 5'-N positive B lymphocyte of normal subject was 13.8±7.3% in cord blood, 57.9±12.7% in adult, respectively. In chronic B-lymphocytic leukemia (B-CLL), Burkitt lymphoma and aplastic anemia, B lymphocytes had markedly decreased 5'-N activity.
These enzyme defects may reflect a immature lymphocytes in at least some of the patients. The presence of these enzyme abnormalities seemed to play an important role in human lymphocyte differentiation and in a certain immuno-deficient condition. Also, these enzyme may be utilized as a cell marker for different stages of maturation. The significance of the lymphocyte enzyme abnormalities and their relation to the immune disorders are discussed.

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We studied transudation of i. v. immunoglobulin preparations in secretions from the normal lungs and salivary gland by measuring anti-tetanus toxoid activity. Time course was studied by serially obtaining alveolar washings, saliva and sera after the administration of commercially available i. v. immunoglobulin preparations, i.e., pepsin digested immunoglobulin (F(ab')₂), sulfonized immunoglobulin (whole molecule) and PEG treated immunoglobulin (whole molecule) preparations. We found that the F(ab')₂ preparation disappeared from the sera more rapidly, but transudated more efficiently in alveolar washing and saliva than the whole molecule preparations. Since mucous membranes are the site of primary infection, these findings are of particular importance in preventing and treating various infectious diseases in mucous membranes.

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Endotoxin protein (EP) was isolated from the TCA-extracted lipopolysaccharide (LPS) of Salmonella minnesota R595 by the use of aquous phenol. The EP stimulated human B lymphocytes to synthesize DNA, even though T lymphocytes were depleted with the nylon column method, the E-rosetting method, or the mouse anti-human monoclonal antibody and complement. However, it is not concluded whether the EP is a human T-independent B cell mitogen or not, because it was not denied that a few T lymphocytes remained in the B lymphocyte fraction. The maximum uptake of ³H-TdR to lymphocytes was at 3 days after culture. The EP stimulated the lymphocytes from every human donors, and its stimulation was greater than that of LPS. Further, the EP produced the increase of immunoglobulin synthesis in human B lymphocytes.
Sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis revealed that the EP was heterogeneous protein, and its molecular weight was from approximately 10, 000 to 40, 000. Lipid A was detected in the EP by the limulus test, and the lipid A portion of the EP could be responsible for the mitogenic activity as well as LPS, since its mitogenic activity was inhibited by polymyxin B. It seems unlikely that the mitogenic activity of EP was due to the contamination by LPS, because the EP scarcely contained 2-keto-3-deoxyoctonate (KDO) which was reported to bridge between the polysaccharide and the lipid A in LPS.

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A case of allergic granulomatous angitis (Churg and Strauss) was reported and the role of circulating immune complexes (CIC) and serum complement in pathogenesis of the disease was discussed.
A 30-year-old woman with history of bronchial asthma for 2 years duration developed abdominal pain, diarrhea, melena, fever, marked emaciation and weakness of the extremities. The clinical investigation revealed pulmonary infiltrates, multiple gastric ulcers, marked leucocytosis with eosinophilia, thrombocytosis, proteinuria, microcopic hematuria and mononeuritis multiplex. The histlogical examination of the specimens obtained by musclar and renal biopsy demonstrated vasculitis with eosinophilic infiltration. Corticosteroid therapy (prednisolone 40 mg/day orally) induced rapid clinical improvement.
CIC estimated by Raji cell assay and immunofluorescent study on polymorphonuclear leucocytes (PMN-IF) were positive in the sample of the serum obtained at the time of admission. In the clinical course of the patient, CIC detected with PMN-IF method were decreased in level during the period of initial corticosteroid therapy but reincreased as diminishing the dose of prednisolone without clinical exacervation.
The components of the serum complement, C3, C4, C5 and C9 showed rather high value in the initial period of the disease and decreased gradually as administration of prednisolone, but never showed abnormal low level. Serum Clq level, which was slightly lower than normal in the initial period, showed no significant change in the clinical course of the case.
These findings suggest that CIC play some role in pathogenesis of allergic granulomatous angitis, although their corelation with serum complement is not yet clear.

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Soft-tissue calcification, a well known roentogen manifestation of connective tissue disease, is commonly associated with PSS and DM-PM. However the occurrence of soft-tissue calcification in SLE has been rarely emphasized. We observed an SLE patient with widespread softtissue calcification. A 15 years old girl developed SLE in 1974 with recurrent symptoms of arthralgia, Raynaud's phenomenon, fever, hair loss and positive ANA (1:>1280 speckled pattern), hypergammaglobulinemia. She was treated with prednisolone 15 mg every day and responded it. In May 1975, she had a mild proteinuria, granular cast and slight hypocomplementemia, but she well responded to 30 mg prednisolone every day. In Feb. 1977, she had a coin sized, hard and painful subcutaneous mass at left buttock. And then, she noticed similar masses at right buttock, right shoulder, bilateral popliteal fossa and thighs. X-ray films of the extremities and the pelvis showed extensive soft-tissue calcifications. In May 1978, she was admitted for painful, enlarged subcutaneous masses at bilateral buttocks. At the admisson, LE cell, ANA and DNA-Ab. were negative, but RNP, SSA-Ab. and unknown ENA-Ab. were detected. Serum enzyme levels (GOT, LDH, CPK, aldolase, amylase), serum calcium concentration, inorganic phosphorus values, ionized calcium concentration, parathyroid function (P. T. H., %TRP) and renal function were all within normal limits from 1974 to 1978. Subcutaneous masses at buttocks were excised. Histological examination revealed a subcutaneous soft-tissue calcification without evidence of fat necrosis, myositis, and vasculitis. Excised material was a compounds of hydroxy appatite and tricalcium phosphate by the infrared spectroscopic analysis.
The occurrence of soft-tissue calcification in SLE was rare and only 25 case reports have been described. Analysis of SLE cases revealed that soft-tissue calcification occurred in patients with a long standing illness, sometimes associated with skin ulcer, myositis and fat necrosis. But the pathogenesis of soft-tissue calcification in this case was unclarified. It was a matter of interest to us in this case that ^99mTe-MDP compounds accumulated at the site of soft-tissue calcification. We suggest that ^99mTe-phosphate compounds are capable of detecting soft-tissue calcification and useful for its diagnosis.

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