Common variable hypogammaglobulinemia (CVH) probably includes a variety of disorders, in that, B cell or helper T cell deficiency and suppressor T cell hyperfunction.
A 24-year-old male had had recurrent eczema since his tender age. He had suffered from idiopathic thrombocytopenic purpura(ITP) at 11-year-old, peptic ulcer and colitis at 17-year-old, pyelonephritis at 18-year-old. He suffered from liver damage and ITP at 21-year-old and was given glucocorticoid and thrombocyte infusion. Since Dec' 1980, he was taken pneumonia and low immunoglobulin level was found. The quantitative immunoglobulin levels were IgG 126, IgA, 3, IgM 29, IgD 1 (mg/dl), IgE 14 IU/ml. Peripheral lymphocyte counts were normal and T: B ratio was 72: 26 (control 68: 32). The patient had delayed hypersensitivity skin reactions for PHA and SK-SD. His lymphocytes responded to in vitro stimulation by PHA or Con A.
Ig production in vitro by PWM-stimulated lymphocytes from the patient was examined. Co-culture of B cells from the patient and T cells from normal resulted in reduced synthesis of all classes of Ig. Combination of the patient T cells and normal B cells also gave low Ig synthesis and X-ray treatment of the patient T cells did not enhance Ig synthesis. Moreover, coculture of normal lymphocytes and patient T cells resulted in reduced synthesis of Ig. Thus it was demonstrated that poor immunoglobulin synthesis was due to B cell insufficiency, and hypofunction of helper T cells, as well as hyperfunction of suppressor T cells.

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In order to investigate whether leukemia and lymphoma cells would be cytolysed by autochthonous and allogeneic peripheral blood mononuclear cells (PB-MNC) or mononuclear cells from lymphnode (LN-MNC), cytotoxic activity was measured by a 6-hr ⁵¹Cr-release assay by culturing fresh and/or cryopreserved malignant cells with PB-MNC or LN-MNC from healthy donors and from the patients. Significant cytolysis was recognized in 5 out of 10 materials from lymphoma patients (50%), and 1 out of 6 materials from leukemia patients (16.7%). These cytolysis were either due to direct cell-mediated cytotoxicity or due to humoral antibody (ADCC). However, the degree of these cytolysis was lower than natural killer (NK) activity against K-562 cells in all cases. It was noteworthy that LN-MNC from the accessory lymphnode of a patient showed cytotoxic activity as Killer T-cell, against autochthonous lymphoma cells and that this killer activity was inhibited by competitive inhibition test by Raji cells. Since these findings in vitro were coincident with clinical signs in some patients, specific and or nonspecific immunotherapy against these patients witg hematologic malignancy should be advanced in future.

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Cell surface antigens of normal human peripheral lymphocytes after activation with mitogenic lectins; phytohemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM) were analysed by dual laser flow cytometer.
Normal human peripheral blood mononuclear cells were isolated by Ficoll-Conray sedimentation method. The cells were washed three times with RPMI-1640 medium and were resuspended into RPMI-1640 medium supplemented 10% fetal calf serum. The cell suspension was adjusted to a concentration of 1×10⁶/ml and PHA, Con A and PWM were added to the cell suspension at an optimal concentration of the lectins respectively. The cells were then cultured at 37 C° humidified 5% CO₂ atmosphere for 72 hrs.
After 72 hrs, the cells were harvested, washed three times with phosphate buffered saline (PBS) and cell surface markers were analysed by direct, indirect and combined immunofluorescent staining methods. The combined staining method was carried out first, by indirect staining with tetramethyl rhodamine isothiocyanate (TRITC) conjugated goat anti-mouse IgG serum as the secondaly reagent and second, by direct staining with fluorescein isothiocyanate (FITC) conjugated T4 and T8 mouse monoclonal antibody.
Activated T cell antigen recognized with anti-Tac monoclonal antibody, other T cell antigens recognized with T4, T8, OKT-9, OKT-10 monoclonal antibodies and immune associated (Ia)-like antigen recognized with I2 monoclonal antibody were markedly expressed on the cell surface of lectin stimulated normal humal peripheral lymphocytes.
Through the dual laser flow cytometric analysis, all T4 positive cells exhibited the Tac antigen. The T8 positive cells also exhibited the Tac antigen but not on all cells. The Tac negative but the T8 positive and double marker cells representing both T4 and T8 positive were also demonstrated.
Acute lymphoblastic leukemia antigen (CALLA) with J5, B cell antigen with Bl and myelomonocyte antigen with Mo 2 monoclonal antibody were not recognized on the distribution of two membrane surface antigens on the cells with a single procedure.

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Recently it has been shown that the natural killer (NK)-target cell interaction results in animmediate burst of chemiluminescence. In this paper we investigated to apply chemiluminescence assay for the evaluation of human NK cell activities.
Degrees of chemiluminescence were dependent on the concentration of lymphocytes or target cells, but optimal concentrations of lymphocytes and target cell were 2×10⁶/ml, 3×10⁶/ml respectively. The responder cell that generates chemiluminescence after target-effector interaction is most likely a lymphocyte for a variety of reasons. First, monocyte-depleted fraction contained more than 95% of lymphocytes. Removal of NK cells with monoclonal anti HNK-1 antibody and complement abolished the chemiluminescence response. Lymphocytes or target cells (K 562) generated very little chemiluminescence response. Furthermore, cell to interaction between allogeneic or autologus lymphocytes did not generate any chemiluminescence. A survery of 10 normal peripheral lymphocytes revealed a well correlation (P<0.01) of NK cell activities between chemiluminescence and ⁵¹Cr release assays.
These results propose that chemiluminescence assay would be another assay system for human NK cell activities.

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Antinuclear antibodies in sera obtained from 183 psychiatric patients treated with psychotropic drugs were studied.
Antinuclear antibodies (ANA) were detected by the indirect immunofluorescent method using rat liver cells as substrate. Farr assay was employed to determined the binding capacity for DNA. The immunoglobulin classes and complement fixing ability (CF) of anti-dsDNA were detected by the indirect immunofluorescent method and the complement fixation method, respectively, using Crithidia luciliae kinetoplasts as the substrate. Anti-ENA was detected by the hemagglutination test (PHA) and double immunodiffusion. Anti-histone was detected by the histone reconstitution of the immunofluorescent test. HLA typing for antigens of the A, B and C was performed by the standard lymphocyte microcytotoxicity test provided by the Terasaki laboratory. Typing for HLA-DR was performed on B lymphocytes isolated on nylon wool column. As controls, 75 normal Japanese individuals were tissue typed with the same antisera.
ANA was detected in 37 patients (20.2%). Total doses of carbamazepine in the patients with ANA were significantly larger than in those without ANA. The patients with ANA hada photosensitivity and oral ulceration significantly more than those without ANA. The patients with ANA had a tendency to have an association with HLA-DRw8.
Anti-DNA antibodies by Farr assay were detected in 25 patients, but titer of anti-DNA was low. Anti-dsDNA by Crithidia immunofluorescent technique was detected in two patients. However, CF of anti-dsDNA was not detected in any patients. Anti-ENA by PHA technique was detected in 21 patients. The patients with anti-ENA had arthralgia and hypergammaglobulinemia significantly more than those without anti-ENA. Anti-histone antibodies were detected in three patients, who were treated with chlorpromazine.

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Islet cell surface antibody (ICSA) was examined on sera from the patients of Graves' disease, Hashimoto's thyroiditis and systemic lupus erythematosus (SLE). Five out of 22 patients (23%) of Graves' disease, 8 out of 32 patients (25%) of Hashimoto's thyroiditis and none of SLE patients and healthy subjects were proved to have ICSA. The incidence of ICSA was associated with the duration of the disease in Graves' disease patients (P<0.01), but had no correlation with sex, age of onset or microsome antibody (MCHA) titer. Antilymphocytotoxic antibody (ALA) was examined on sera from ICSA positive patients with high MCHA titer and SLE patients. The incidence of ALA of the former ((%) was significantly lower than that of the latter (55%) (P<0.05). These findings may suggest that plural antibodies against different endocrine organs are produced in a part of the thyroid disease and that the mechanisms of immunological disorders of such patients are different from those of SLE.

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Anti-DNA antibody synthesis by lymphocytes from normal individuals and systemic lupus erythematosus (SLE) patients was investigated. Anti-DNA antibody synthesis in response to DNA was demonstrated in SLE, but not in normal individuals. DNA was T cell-dependent antigen in anti-DNA antibody synthesis. Anti-DNA antibody synthesis by SLE lymphocytes was suppressed by T₈ cells and enhanced by T₄ cells in inactive stage. However, in active stage, SLE T₈ cells failed to suppress anti-DNA antibody synthesis, even in any T₄/T₈ ratio. These results indicated that DNA-specific suppressor T cell dysfunction was in part responsible for the elevated anti-DNA antibody synthesis in SLE lymphocytes in response to DNA.

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A method was described about normal human peripherial B-lymphocyte colony formation in vitro. Irradiated T lymphocytes, and non-T cell population were cultured with 30 μg of phytohaemagglutinin-p (PHA-P) into methylcellulose gel, after a day of preincubation with PHA-P, and these lymphocytes. Colonies more than 50 cells were counted under a dissecting microscope.
As a result, a linear relationship between the number of seeded cells and the growth of colonies was seen in methylcellulose gel. 570±333 colonies per 5.0×10⁵ seeded cells were detected at 4 days of culture. Cells in colonies had surface immunoglobulin by using FITC-labeled anti-human immunoglobulin F (ab)2 rabbit serum under a fluolescent microscope.
And cells in colonies had characters differentiated to plasma cells and lymphocytes in Giemsa stain, but no E rosette-forming ability.

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The responsiveness of human peripheral B cells when stimulated by Salmonella paratyphi B (SPB) was studied.
The human B cells showed nonspecific and polyclonal differentiation to immunoglobul-inproducing cells (ISC) when stimulated by SPB.
SPB-induced generation of ISC from human B cells was found to be T-independent and Mφ-independent and was not affected by DNA synthesis.
These results suggested that this bacteria does not stimulate the proliferation of B cells but does stimulate differentiation from B cell to ISC and the secretion of polyclonal Igs.
Therefore it was verified that SPB stimulates B cells differently from other mitogen. SPB made it possible for detailed analysis of the activating system of human B cells.

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A 63-year-old female is described with Sjögren's syndrome without any evidence of rheumatoid arthritis or connective tissue disorder, but partial lipodystrophy had been marked since 10 years old. There was no abnormal findings on 50 gGTT and serum lipids. Immunological and histological study revealed positive SS-A antibody, positive lip-biopsy findings, normal renal biopsy finding and normal levels of serum complement that was reported common with partial lipodystrophy.
Although, both of the Sjögren's syndrome and lipodystrophy are rareconditions, 3 cases of this association were reported already in the world. This case was understood 1st instance of lipodystrophy and Sjögren's syndrome in Japan. As there was good evidence of Sjögren's syndrome written in former report of lipodystrophy and immunological disorder on serum complement with high incidence in partial lipodystrophy, this intersting association of lipodystrophy and Sjögren's syndrome may not be coincidental.
It would be helpfull studying evidence of sicca syndrome in patients with lipodystrophy easily diagnosed from the characteristic appearance to understand the relation of lipodystrophy and Sjögren's syndrome.

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C3 nephritic factor (C3NeF) was found in patients with membranoproliferative glomerulone phritis (MPGN) and in some cases of partial lipodystrophy. C3NeF was supposed to be the auto-antibody to C3 convertase (C3bBb) of the alternative complement pathway and stabilize it, resulting in characteristic complement profile in their serum.
Recently C4 nephritic factor (C4NeF), which was thought to be the auto-antibody to C3 convertase of the classical complement pathway (C4b2a), was found in patients with systemic lupus erythematosus (SLE) or poststreptcoccal glomerulonephritis.
We found C4NeF activity both in serum and urinary IgG fraction of a patient with SLE, who presented nephrotic syndrome and MPGN at the time of renal biopsy. The C4NeF activity in serum and urine disappered after the treatment of steroid therapy.
We also found C3NeF activity only in serum IgG fraction, not in urinary IgG fraction, of a patient with ulcerative colitis, sclerosing cholangitis and chronic glomerulonephritis, of which biopsy specimen showed MPGN. However, the activity was atypical in comparison with C3NeF reported so far, because of its unstability and weakness in activating complement of normal human serum and it suggested heterogeniety of C3NeF.
While C4NeF activity was detected both in serum and in urine, C3NeF was found only in serum. Thinking into consideration that selectivity of the urinary protein from the two patients did not differ, we speculated C3NeF had high affinity to the glomerulus.

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RU 41740, a glycoprotein extracted from Klebsiella Pneumoniae, has an ability to activate human complement in a dose response fashion either through the classical or the alternative pathway in vitro, when measured by the lysis of either sensitized sheep erythrocytes or unsensitized rabbit erythrocytes, as well as by immunoelectrophoresis.
Some of bacterial or plant-derived polysaccharides have been reported to restore impaired host immune responses mainly through their cellular immunity, and partly through humoral immunity. The present data led us to speculate that the agent might exert its anti-infectious ability via the activation of complement system.

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