The fluorescence polarization technique to measure cytoplasmic microviscosity of human peripheral lymphocytes was established. The previous method, which was proposed by Cercek, has been corrected because a serious mistake was revealed in the process of cell filtration. The mean value of fluorescence polarization in lymphocytes from 20 healthy donors was 0.190±0.005. The mean value of fluorescence polarization as % of control after stimulation of lymphocytes with phytohemagglutinin (PHA) was 83.6±3.8. The optimal conditions in the measurement were discussed.

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The suppressive mechanism of Interleukin 2 (IL-2) production was investigated in cancer patients.
IL-2 production of peripheral blood mononuclear cells (PBMC) derived from control subjects and patients with non-treated advanced carcinoma were 524.1±250.4 nano Jurkat Units (nJ. U.)/cell and 139.3±85.4 nJ. U./cell, respectively. The mean value of IL-2 production in cancer patients was significantly lower than that of normal subjects. IL-1 release from bacterial lipopolysaccharide-stimulated monocytes in cancer patients was significantly higher than that of normal subjects and the supplementation of IL-1 did not restore the IL-2 production, suggesting that the impairment of IL-1 release was not responsible for the decrease of IL-2 production in cancer bearers.
On the other hand, monocyte depletion increased the IL-2 release in PBMC of cancer patients. Moreover, the monocytes derived from cancer patients possessed a higher suppressive activity on IL-2 production than those of normal subjects. These results suggest that the increase of the suppressive monocytes is responsible for the impairment of IL-2 production. However, this suppression was not abolished by treatment with indomethacin, a inhibitor of prostaglandin synthesis, but was restored by treatment with phorbol myristate acetate (PMA).
Our experiments suggest that the IL-2 decrease in cancer patients is chiefly mediated through a suppressive monokine which is diminished by treatment with PMA rather than indomethacin.

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Human peripheral dendritic cell was successfully obtained with little contamination of macrophage. Dendritic cell showed a cluster formation with autologous T cells. These T cells were mainly CD-4 positive and partially blastoid. Clusters were isolated and then recultured. CD-4 positive cells were grown from isolated clusters confirmed by phase contrast microscopy and ³H-thymidine incorporation.
This cell growth was augmented by pretreatment of dendritic cell with α-interleukin 1. Cluster supernatant contained some factor to drive SAC-stimulated B cell into antibody secreting cell.
Our experiment using peripheral dendritic cell would be able to apply the clinical investigation on dendritic cell function in some patients.

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The tumor cytostatic activity of neurophils induced by intraperitoneal administration of PSK in mice was evaluated in terms of inhibition of DNA and RNA synthesis as respectively determined by inhibition of ³H-thymidine and ³H-uridine uptake.
CDF₁ mice were intraperitoneally injected 50mg/kg of PSK. A large number of neutrophils were migrated in the peritoneal cavity 12 hours after the injection. Since PSK could activate the complement system, promoting the generation of a complement-derived chemotactic factor C5a, 1.0ml of normal human serum (NHS)was administered as a sourse of complement to another group of mice along with 50mg/kg of PSK. Numerous number of neutrophils were also detected 6 hours after the administration of PSK with NHS. C5a in the peritoneal cavity elevated l hour after the injection in both groups intraperitoneally injected with PSK or PSK and NHS, but there was no significant difference between two groups.
Neutrophils at 6 hours after the injection of PSK showed 1.5 to 2 times higher inhibitory effect of ³H-thymidine uptake than those of casein-induced neutrophils as a control. They also revealed inhibitory effect of ³H-uridine uptake. But these neutrophils showed no inhibitory effects on NIH3T3 fibroblast cells. Effect of neutrophils induced by PSK with NHS was not higher than that of PSK alone in ³H-thymidine or ³H-uridine uptake.
These findings suggested that intraperitoneal injection of PSK could induced the tumor cytostatic neutrophils in the peritoneal cavity in mice.

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Circulating immune complexes (CIC) in eleven patients with mixed connective tissue disease (MCTD) were determined by C1q solid phase radioimmunoassay. A significant increase in the CIC level was observed in sera from patients with MCTD, as compared with those of progressive systemic sclerosis, polymyositis and normal controls. The CIC level in patients with MCTD, however, was much lower than that of systemic lupus erythematosus. It was associated with the disease activity in the most cases and a significant inverse correlation was observed between the CIC level and the serum C3 level. There was, however, no correlation between the CIC level and the titer of anti-RNP antibody.
By using the Western blot method, no RNP antigen was detected in the CIC obtained from all MCTD patients tested. On the other hand, substantial amounts of the anti-RNP antibody were detected in the CIC. The titer of the anti-RNP antibody in the CIC seemed to be associated with the CIC level, suggesting that the CIC may mainly be composed of the anti-RNP antibody.

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The production of interleukin 1 (IL 1) in cultured peripheral monocytes from patients with Kawasaki disease was examined by co-mitogenic assay using mouse thymocytes, compared with that from acute febrile diseases. The supernatants of the cells treated with lipopoysaccharide (LPS) at 1-20 hr of incubation (early phase) and 20-72 hr of incubation (late phase) were used as IL 1 specimens. The cells showed the high production of IL 1. The marked production was recognized in the cells not treated with LPS during the late phase when compared with that from healthy adults. This indicated that the cells were in vivo activated and the production of IL 1 was induced. The patients with Kawasaki disease showed the high production of IL 1 at hospital discharge although no symptoms appeared on them. The production of IL 1 alpha and IL 1 beta was determined by using neutralizing antibodies (anti-IL 1 alpha or/and anti-IL 1 beta antibodies) in the supernatants. In the patients with high production of IL 1 the supernatants of the early phase and the late phase revealed the high activity of IL 1 beta and IL 1 alpha, respectively.

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B cell abnormality in 13 patients with primary a/hypo-gammaglobulinemia were studied. Number of peripheral B cells was normal in 7 of 13 patients but very small in the remaining six patients.
In the first step, we established lymphoblastoid cell lines (LCL) by EB virus and evaluated immunoglobulin production by LCL. LCLs were established in 7 patients, whose number of peripheral B cells was normal. IgM dominant pattern of immunoglobulin production was seen in 6 of the 7 LCLs as LCLs from cord blood cells. These results suggested that the B cells of these patients were immature, like the B cells of cord blood. Only three LCLs could be established from the remaining 6 patients, whose number of peripheral B cells was very small. These three LCLs produced dominantly IgM, dominantly IgG and no immunoglobulin. Various B cell abnormalities were sugested, even when the number of peripheral B cells was very small.
Next, we purified B cells from 5 patients and evaluated B cell functions, such as activation, proliferation and differentiation by using SAC and IL 2. In this assay system, B cell abnormality was devided into three groups. We concluded that the abnormality was in the step of activation/proliferation by SAC in 1 patient, in the step of proliferation response by IL 2 in 1 patient and in the step of differentiation to Ig secreting cell by IL 2 in 2 patients. B cells from 1 patient, whose LCL secreted IgG and IgM equally, showed no abnormality even in this assay.
Finally, we examined DNA and mRNA of immunoglobulin of the LCL, which produced no immunoglobulin. It was confirmed that the LCL was pre B cell line, and suspected that μm-RNA of the LCL had an abnormality because the number of M. W. was small compared to that of control LCLs secreting IgM.

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The concentrations of adenosine deaminase (ADA) and neopterin (NP), known to be released from activated lymphocytes and monocytes/macrophages, were measured in the sera of hemophiliacs with human immunodeficiency virus (HIV) infection to study whether these measurements were useful or not as early predictors for development to ARC and full blown AIDS.
The mean concentration of ADA in sera of 38 hemophiliacs with anti HIV antibody (54.0±17.7IU/l) was higher as compared with that of 30 patients without the antibody (38.6±13.9IU/l) with statistical significance (p<0.01) and this tendency was observed more clearly in those with a reversed CD 4/CD 8 lymphocyte ratio (p<0.01).
The concentration of NP was also significantly higher in anti HIV antibody positive group (24.9±18.3nmol/l) than that of negative one (7.9±4.2nmol/l) (p<0.01), and this tendency was more prominent in patients with CD 4/CD 8 lymphocyte ratio less than 0.2 (p<0.01).
Moreover the concentrations of ADA and NP, measured serially in sera of 4 patients who developed AIDS, gradually increased in accordance with and/or ahead of the decreases in anti p24 antibody titer and CD 4/CD 8 lymphocyte ratio and reappearance of HIV, but turned ot decrease at their terminal stages.
From these, it was suggested that in patients with HIV infection, serial measurement of serum ADA and NP levels, combined with check-ups on anti p24 antibody, HIV and CD4 and CD8 lymphocyte counts was clinically useful and important to predict and find their development to ARC and AIDS, and manage them in advance.

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Human cord blood lymphocytes respond well to concanavalin A or phytohemagglutinin to proliferate, and human cord blood B cells produce immunoglobulins, especially IgM, after the stimulation by Epstein-Barr virus which is known to be polyclonal B cell activator. Pokeweed mitogen (PWM) which is a T cell-dependent polyclonal B cell activator poorly induces immunoglobulin from cord blood lymphocytes, while adult peripheral blood mononuclear cells (PBMC) well produce polyclonal immunoglobulins by PWM stimulation. However, antigen-specific IgM antibody is produced in the intrauterine infection by foetus. Hence, we tried to examine whether cord blood lymphocytes can respond to sheep red blood cells (SRBC) to produce SRBC-specific antibody utilizing the in vitro culture system consisting of cord blood lymphocytes and SRBC as antigen. In this culture system, SRBC-specific immunoglobulin secreting cells (IgSC) are enumerated after the 8 day culture, and SRBC-specific immunoglobulins secreted by B cells are considered to be IgM. In our experiments, cord blood lymphocytes produced no IgSC specific for SRBC, while adult PBMC produced 310_??_3.87 (geometric mean _??_ SD) SRBC-specific IgSC/10⁷ cells. PBMC derived from 1 to 15-year-old children responded to SRBC in vitro to produce 24_??_3.75 SRBC-specific IgSC/10⁷ cells intermediately. SRBC-specific IgSC were apparently present in the peripheral blood in childhood compared to cord blood, but the number of IgSC in childhood was significantly lower than that in adult (p<0.001). It was reported in the accompanying paper that interleukin 2 (IL 2) augmented SRBC-specific IgM antibody production and the number of SRBC-specific IgSC when exogenous IL 2 was added to the culture consisting of adult PBMC and SRBC as antigen. It was also clarified in this paper that human recombinant interleukin 1 (IL 1) augmented the number of SRBC-specific IgSC in this assay system. Hence, exogenous IL 1 or IL 2 was added to the in vitro culture consisting of cord blood lymphocytes or PBMC obtained from 1 to 15-year-old children or adult individuals with SRBC as antigen. Cord blood lymphocytes produced no SRBC-specific IgSC in the presence of exogenous IL 1 or IL 2, while PBMC in childhood remarkably responded to SRBC in the presence of IL 1, IL 2, and produced large number of SRBC-specific IgSC which were almost equal to the number of SRBC-specific IgSC of adult PBMC cultured with SRBC in vitro in the absence of exogenous IL 1 or IL 2. Childhood PBMC were essentially similar to adult PBMC in the response to SRBC in the presence of IL 1 or IL 2, while childhood PBMC less effectively produced SRBC-specific IgSC in the absence of IL 1 or IL 2 than adult PBMC.

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Dialyzable leukocyte extracts (DLE) have been shown to contain Transfer Factor (TF), which is able to transfer DTH (delayed-type hypersensitivity) activity from a positive donor to a negative recipient with antigen specificity. DLE have been used for the treatment of patients with various types of immunodeficiencies. Many reports on DLE have dealt with not only transfer activity but also other biological activities, and therapeutic effectiveness of DLE is regarded as the integration of those activities.
DLE are produced from human leukocyte lysate and in some cases the lysate is treated with DNase. Japanese Red Cross Blood Center has been supplying RCTF-1 as DLE which has been produced from DNase treated lysate. It had been reported that DLE from DNase treated Iysate (DNase (+) • DLE) and from DNase non-treated lysate (DNase (-) •DLE) both have the same ability of DTH transfer. In many foreign countries DNase(-) •DLE have been adopted for clinical use. Therefor, we studied the effects of DNase (-) and DNase (-) •DLE for human leukocyte blastformation and IgG production in vitro.
DNase (+) •DLE decreased PHA stimulated lymphocyte blastformation and IgG production, but DNase(-) •DLE did not. These inhibitory activities were resistant to promase treatment, and partially purified fraction of DEAE-Sephacel chromatography showed similar UV (ultraviolet) adsorpted patterns to nucleotide's. On the other hand, DNase (-) •DLE did not show a similar UV spectram pattern. Accordingly, it is possible that these inhibitors were unspecified oligonucleotides, appearing from the step of DNase treatment of lysate.
Improvement by DLE therapy has been reported for a variety of diseases, regardless of whether DNase (+) •DLE or DNase (-) •DLE were used. They are almost similar in clinical effectiveness. Thus, the inhibitory activities of DNase (+) •DLE in vitro, shown in this report, may be irrelevant to the activity in vivo. So, it seems to be better to use DNase (-) •DLE for studying DLE function in vitro.

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We report a SLE patient with cardiac tamponade as a presenting symptom, who was successfully treated with steroid pulse therapy.
A 19-year-old girl was admitted to a hospital because of severe chest pain and dyspnea. She was well until one month before entry, when fever, anterior chest pain and polyarthritis developed. Antibiotics and predonisolon (40mg/day) were administered without benefit. She was refered to our hospital on Feb 17, 1987. On admission, active arthritis and malar rash were observed and large amount of pericardial effusion compressing cardiac output was demonstrated by echocardiography. Diagnosis of active SLE complicated with cardiac tamponade was made, and steroid pulse therapy was started. All of the above symptoms subsided within a week after pulse therapy, followed by amelioration of pericardial effusion as well as immunological paramaters. Although cardiac tamponade as a presenting symptom in SLE is rare, it may be fatal without correct diagnosis and prompt treatment. However, it has been reported that oral predonisolone alone sometimes fails to improve cardiac tamponade. Here, we describe a first case successfully treated with steroid therapy and discuss the therapeutic approach to SLE complicated by cardiac tamponade.

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Trigeminal neuropathy has been known as a most common neuropsychiatric complication in mixed connective tissue disease (MCTD). In 1978, Bennett reported four cases of aseptic meingitis observed in MCTD. We report here three cases of aseptic meningitis in patients with MCTD.
All three patients were female and had Raynaud's phenomenon, swollen hands, lymphadenopathy, muscle involvement and arthralgia. They had anti-nRNP antibodies in high titers, but not anti-Sm nor ant-DNA antibodies.
There meningitis was accompanied by fever and severe headache but not by nuchal rigidity. In addition, one of the patients showed nausea, vomiting and dullness of consciousness. The cerebrospinal fluid (CSF) analysis revealed the increased levels of initial pressure, cell counts and protein amounts in all three cases. Mononuclear cells but not neutrophils were dominant in CSF in all three cases. No microorganisms were found in CSF and no specific increase of antibodies to viruses were found in both serum and CSF of three patients indicating an aseptic nature of their meningitis. The meningitis in all three MCTD patients responded to the moderate to high doses of corticosteroid but not to low doses of steroid nor antibiotics.

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A 32-year-old woman with 16 years duration of rheumatoid arthritis was admitted to the Juntendo University hospital in December 1985, and was diagnosed as Felty's syndrome be-cause of the simultaneous occurence of rheumatoid arthritis, splenomegaly, and the marked leukopenia. In this case, anti-DNA antibody and anti nuclear antibody have shown positive, and erythema, thrombocytopenia, and polyarthritis were recognized. These incidence are of definite SLE (Systemic Lupus Erythematosus). This indicates Felty's syndrome may be a connecting link lying somewhere between rheumatoid arthritis and SLE.
Felty's syndrome is very rare in Japan: only 30 cases have been reported since 1972. This case is characterized by the marked thrombocytopenia, which can be found in 55% of the above 30 cases. Thrombocytopenia was hypothesized by Thorne to be the fourth criteria of Felty's syndrome in 1982. Thrombocytopenia is not essential criterion, but, as a relatively frequent occurrence, it is valuable to illuminate its mechanism.

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