Studies were conducted on 192 homosexual Japanese males who visited our department of internal medicine for the determination of human immunodeficiency virus (HIV) antibody in serum. Besides the history taking for their daily sexual lifestyle (the number of the partners of their homosexual intercourse, nationality of their partners, and the presence or absence of the experience of pederasty), various immunological tests were performed to determine lymphocyte subsets, capacity of interleukin 2 production, phagocytic capacity of neutrophils and PPD skin reaction, and the serologic test for syphilis and antibody determination of various viruses (hepatitis B virus, herpes simplex, varicella zoster virus, cytomegalovirus and EB virus) were also performed. Although the serologic test for HIV antibody was negative in all 192 cases in the present series, abnormalities in T lymphocyte subsets such as decreased CD4⁺ lymphocyte, elevated CD8⁺ lymphocyte, decreased CD4⁺/CD8⁺ ratio were detected in about 1/3 of the cases. Moderate to marked elevations in CD11⁺ lymphocyte count were noted in 70% or more of the cases. As compared with the normal healthy population, the positive rates of syphilis and hepatitis B virus antibodies were significantly higher in our subjects. The antibody titers against other viruses were also higher in these patients, and, particularly in cases presenting high antibody titers against cytomegalovirus, CD11⁺ lymphocytes tended to increase, suggesting their involvement in reactivation of viruses. However, no correlation was found between these abnormalities and the patterns of the sexual lifestyle of our subjects. The function of the neutrophiles, capacity of interleukin 2 production and PPD skin reaction were normal in all cases. From these, it was suggested that a considerable number of homosexual Japanese males have suppressed immunological functions even if their test for HIV antibody is negative and they are, therefore, susceptible to HIV infections.

View full abstract

Sheep erythrocytes are uniform electrophoretically and those treated with tannic acid (electrophoretic mobility test indicator cells: EIC) are able to adsorb a minute quantity of a biologically active substance, causing a change in the electrical charge on the surface. This change can be precisely detected with a tanned erythrocyte electrophoretic mobility test (TEEM) by using a newly designed automated cell electrophoresis analyzer with high reproducibility developed by the Shimazu Corporation.
For the purpose of the study on biologically active substance produced through a immunological response, EIC were incubated in the sera of human renal transplants, cancer patients and blood transfused recipient mice and in the lymphocyte supernatants stimulated with mixed lymphocyte culture (MLC) and some mitogens and in IL-2 and IAP, and each changes in the electrical charge on the surface of EIC were observed using this analyzer.
On the study of the sera of human renal transplants, the electrophoretic mobility of EIC (TEEM) was unchanged in the non-rejection group. In the rejection group, 3_??_7 days before the rejections occurred, the TEEM tended to be accelerated (P<0.005), and when the rejections recovered, it tended to be delayed.
On the sera of cancer patients, no significant differences of TEEM were seen compared to the sera of healthy volunteers.
The TEEM was also unchanged by the sera of blood transfused recipient mice compared to non blood transfused mice and by the lymphocyte supernatants stimulated with MLC, PHA and PPD and by IL-2 and IAP.
The results of this study suggest that among the sera obtained from some immunological responses, only the sera of renal transplanted patients showed the change of electrical charge of EIC and this certain TEEM accelerating biologically active factor which is neither IL-2 nor IAP has already been released into the serum before the rejection is clinically confirmed, and it is expected that TEEM could be effectively used in the future to detect a rejection at an earlier stage and predict the prognosis more accurately in human renal transplants.

View full abstract

Human peripheral blood monocytes are the reticuloendotherial cells, which are known to act on the elimination of soluble immune complexes in vivo. Phagocytosis of soluble immune complexes by phagocytes has been detected by radioimmunoassay. We described here new method of the measurement using enzyme-linked immunosorbent assay (ELISA). Peroxidase-anti-peroxidase soluble complex (PAP) was used as a soluble immune complex in this method. Monocytes were isolated from peripheral blood mononuclear cells by their adherent activity to plastic culture dishes. Monocytes (5×10⁴ per well in 200μl of medium) were placed in the wells of a 96-well culture microplate and mixed with 100μl of PAP after culturing for 24 hs. The cells were then incubated in a humidified atmosphere of 5% CO₂ at 37°C for 60min. After washing four times with cold phosphate buffered saline, 100μl of 1% Nonidet P 40 was added to each well to destroy the monocytes. Fluid (50μl) from each well was placed in the wells of a flat-bottom microplate for ELISA and then 150μl ABTS at 0.2mg/ml in substrate buffer was added to each well. After 60min at room temperature, the plate was scanned in a microplate ELISA reader at a wavelength of 405nm, with correction being made for medium controls.
The phagocytic activity of monocytes was not observed at 4°C, and was suppressed in the presence of a specific inhibitor of glycolysis (2-deoxyglucose) and blockers of oxidative metabolism (NaN₃, KCN). Aggregated IgG also suppressed the phagocytosis. As a result, the phagocytic activity of monocytes detected by this method was thought to be dependent on temperature, cellular metabolism or Fc receptors.
Moreover, we examined the effect of lymphokines on this monocyte activity. The phagocytic activity was enhanced by the addition of 48-h culture supernatant from tonsil lymphocytes stimulated with concanavalin A, and by gamma- and beta-interferon, but not by alpha-interferon or interleukin 2.

View full abstract

Prostaglandin E₁ (PGE₁), prednisolone (PSL) or both of them were administered to rats subcutaneously. Thereafter, numbers of polymorphonuclear leucocytes (PMN) in the peritoneal cavity of the rats stimulated with oyster-glycogen were counted and further O^-_i production of the peritoneal PMN was measured after stimulation of (PMNs) with the opsonized zymosan (OZ).
PGE₁ inhibited the accumulation of PMN into the peritoneal cavity at doses ranging from 0.02 to 2mg/kg, and also suppressed the O^-_i production of PMN at doses ranging from 0.2 to 2mg/kg. PSL inhibited both accumulation and O^-_i production of PMN at a dose of 4 mg/kg.
The combined administration of PGE₁ and PSL inhibited more extensively both accumulation and O^-_i production of PMN than that of each drug alone. These results suggest that the anti-inflammatory effect of PSL could be accelerated by a combined use with PGE₁.

View full abstract

The cholestatic factor, a lymphokine, has been shown to be produced from the lymph node cells of sensitized guinea pigs in vitro by stimulation with a specific antigen, PPD (purified protein derivatives). Two active fractions can be prepared from the supernatant of the medium by Sephadex G-75 gel filtration and DEAE cellulose column chromatography. When these fractions are injected into the mesenteric vein of normal rats, a marked reduction in bile flow is induced. Since dimeric immunoglobulin A (IgA) is richly contained in the bile and it is rapidly and actively transported from the blood to bile by hepatocytes, the authors investigated the effect of the cholestatic factor on dimeric IgA transport. As a result, this lymphokine induced a fall in dimeric IgA secretion, suggesting that it may also influence the vesicular transport system of the hepatocyte.

View full abstract

The authors reported and discussed the clinical and immunological features of 80 children registered as sufferers of common variable immunodeficiency to the All Japan Immunodeficiency Registry up to April 16, 1986.
12.3% of the patients reported some cases suspected of common variable immunodeficiency in their family members.
Mean age of the diagnosis of the patients was 65.0±49.0 months old. Mean age of the 60 survivors at the time of registration was 112.1±66.4 months old, and mean age of death of the 19 expired patients was 76.3±84.4 months old.
Respiratory tract infection was the most common initial symptom among the patients, and 92.5% of the all patients reported susceptibility to infection. Pneumonia (55.0%), otitis media (37.4%), pyodermia (30.0%), gastroenteritis (17.5%) and sepsis (10.0%) were common infection among the reported patients.
Eczema (15.0%) and bronchial asthma (8.8%) were occasionally seen and there were two cases with rheumatoid arthritis, one with SLE and one with Hashimoto's thyroiditis. One patient was suffered from B cell type malignant lymphoma.
48 of all 80 patients were also reported the levels of serum immunoglobulins, and 87.5% of the patients showed significantly low levels of IgG for their age. Among these patients with hypo-IgG-emia, 43.8% also showed significantly low levels of both IgA and IgM and 29.2% also showed significantly low levels of IgA. 54.2% of 24 patients with hypo-IgG-emia were reported to have decreased numbers of B cells in the peripheral blood.
Replacement of IgG with human serum imunoglobulin preparations was reported to be the most common effective therapy applied to these patients with common variable immunodeficiency.

View full abstract

A new cytotoxicity assay was developed to measure Natural Killer cell activity. This assay does not need ⁵¹Cr labelled target cells. FITC (Fluorescein Isothiocyanate) was used for labelling of target cells, K 562, in acidic conditioned medium. Target cells were coincubated with peripheral blood mononuclear cells in plastic tubes. After 4 hours incubation, these cells were resuspended in a Fuchs-Rosenthal chamber.
Then these mononuclear-cells and target cells were counted by NIKON epifluorescent microscope.
Only alived target cells glittered and it was easy to distinguish alived target cells from mononuclear cells. NK cell activities measured by this new assay had very high correlation (r=0.887 p<0.001) with standard ⁵¹Cr release assay.

View full abstract

Systemic Lupus Erythematosus (SLE) is one of the chronic autoimmune diseases with various inflammatory manifestations involving multiple organ systems, accompanied by multiple immunological abnormalities. The Preliminary Criteria for the Classification of SLE by American Rheumatism Association (ARA) had been widely used for the diagnosis of patients with SLE since 1971.
The criteria was newly revised by ARA in 1982. The sensitivity and the specificity of the 1982 Revised Criteria was evaluated in our patients with connective tissue diseases. The sensitivity for SLE (78 cases) was 94.9%, and the specificities against PSS (17 cases), PM (10 cases) or RA (19 cases) were all 100%.
The sensitivity for the diagnosis at initial visits in patients with SLE was also 78.2% and the 1982 Criteria was considered to be quite useful for early diagnosis of SLE.
The results indicated that the 1982 Criteria could be more useful in diagnosis of SLE than the 1971 Criteria.

View full abstract

The distribution of binding sites for three lectins labeled with rhodamine was studied by using immunofluorescence microscopy in the colonic mucosa in ulcerative colitis.
Binding sites for Dolichos biflorus agglutinin (DBA), a lectin specific to terminal α-N-acetylgalactosamine residue, and soy bean agglutinin (SBA), specific to teminal α or β-N-acetylgalactosamine residue of surface glycoprotein in the endoscopically uninvolved mucosa in ulcerative colitis were almost equal in distribution to those of normal human colon. These two lectins were markedly decreased in distribution in the endoscopically involved mucosa, whereas they were also decreased in the boundary zone which was considered as locating between the involved and the uninvolved portion. On the other hand, there was no definite difference in binding sites for concanavalin A (Con A), a lectin specific to terminal α-mannopyranoside, α-glucopyranoside, between normal colon and ulcerative colitis.
These results suggest that a quantitative and/or qualitative alteration in the colonic mucin has an important role in the colonic mucosa in ulcerative colitis. And lectins may be developed into a powerful probe for the detection of biochemical change in the colonic mucosa in ulcerative colitis.

View full abstract

We pointed previously two problems in treating hepatocellular carcinoma (HCC) with lymphokine activated killer (LAK) cells: one is the difficulty in inducing LAK activity and the other is the existence of serum inhibitory factor in LAK generation. In this paper, 14 HCC patients were injected with recombinant interleukin 2 (rIL-2) 15 μg/day continuously for 5 to 27 days, and were assessed for its immunological influence and clinical effect. Results indicated that LAK activity which was lower estimated before the treatment, was remarkably increased in 9 of 12 patients and more than 25 percent reduction of serum α-fetoprotein (AFP) concentration was observed in 5 to 13 patients. Therefore, it could be suggested that continuous intravenous-administration of rIL-2 seemed possible to overcome a problem in adoptive immunotherapy in HCC.

View full abstract

Clinical studies and typing for HLA-A, HLA-B, HLA-C and HLA-DR loci on five cases with Sjögren's syndrome with aseptic meningoencephalitis were performed. In our cases, four of the five patients had sicca syndrome alone without other connective tissue disease. Only one patient had Sjögren's syndrome with adult Still's disease Three of the five patients had clinical manifestation of Raynaud's phenomenon and non-deforming arthritis. Two of the five patients had renal tubular acidosis. Four of the five patients were subclinical Sjögren's syndrome. No HLA antigens specific for Sjögren's syndrome were found out in all patients. The CSF pressure was normal. There was mild to moderate pleocytosis in each case with mononuclear cell predominance. The CSF protein concentration was elevated in five of the eight episodes. The initial CSF: blood glucose ratios was low in four of the five episodes. Reccurent episodes of aseptic meningoencephalitis were documented in two of the four patients. Viral, fungal, and bacterial cultures were uniformly negative.
The association of Sjögren's syndrome with aseptic meningoencephalitis is discussed.

View full abstract

A 45-year-old female developed fever, erythema over the whole body, and swelling of general lymph nodes from mid-May 1984. At that time, the LDH was 929IU/l, each fraction was clear, and no anomaly was detected by immunoprecipitation reaction in free liquid media. The WBC in peripheral blood was 20, 900/μl, 18% of which were mature eosinophils. After administration of prednisolone at a daily dose of 30mg, the patient recovered and left the hospital. In February of the following year, fever and general erythema recurred. The WBC in peripheral blood was 31, 800/μl, of which mature eosinophils accounted for 42%. In sternal myelograph, mature eosinophils accounted for 9.2%. No whipworm eggs were found. A slight enlarged heart shadow and pericardial cavity fluid were noted, and a skin biopsy disclosed acanthosis and an infiltration of eosinophils around the blood vessels, but no vasculitis. The LDH in mid-March was 1, 182IU/l, and isozymes in each fraction were indistinct and LDH-linked immunoglobulins LDH-Ig G, A, M (κ, λ) were detected by immunoprecipitation reaction in free liquid media (molecular weight 520, 000). She was diagnosed of hypereosinophilic syndrome. From mid-March she was administered prednisolone at a daily dose of 40mg. The temperature declined gradually, the general erythema and pericardial cavity fluid subsuided, and the peripheral blood eosinophils also disappeared. The LDH became normalized in about October. At the end of March, the anomaly pattern was type 3-5, and LDH-Ig G, A (κ, λ) was detected. In mid-April, the anomaly pattern was 3-5, and LDH-Ig G (κ, λ) was observed. At the end of May, the anomary disappeared and not detected by immunoprecipitation reaction in free liquid media, but in the same serum after 2 weeks in cold storage, anomaly of type 3-5 appeared, and LDH-Ig G (κ, λ) was detected by immunoprecipitation reaction in free media. At the end of May 1986, the LDH was 231IU/l, the isozyme fractions were clear, and no anomaly was detected by immunoprecipitation reaction in free liquid media, nor was it detected in the same serum after 2 weeks in cold storage. This case presents three interesting points in considering the onset mechanism of LDH-Ig. 1) At the time of the first admission, the LDH was high, but no anomaly was noted. At the time of the second admission, when the LDH increased again, an anomaly occurred. 2) As a results of treatment, LDH-IgM, IgA, IgG disappeared in this sequence. 3) No anomaly was noted in the serum sampled during the process, but an anomaly was found in same serum after 2 weeks in cold storage.

View full abstract