Search

Borna disease virus (BDV) infection has been suspected to be a possible etiological factor in human psychiatric disorders and recently in chronic fatigue syndrome. Evidence of the correlation of BDV infection with these disorders remained unclear. Kagoshima is known to be one of the major areas in which human T-cell leukemia virus type 1 (HTLV-1) is endemic; this is the first isolated human retrovirus that causes adult T-cell leukemia with neurological symptoms. The present study aimed to clarify whether BDV and HTLV-1 infections are associated with psychiatric disorders among Japanese patients. Subjects were 346 patients with psychiatric disorders (schizophrenia, 179; mood disorder, 123; and others, 44) and 70 healthy controls. Anti-BDV antibodies from plasma samples were screened by the indirect immunofluorescence (IF) method using BDV-infected MDCK cells. Results revealed that only three samples were found to be weakly positive for BDV in the IF assay and seronegative by Western blot (immunoblot) assay. Furthermore, BDV-p24 related RNA in peripheral blood mononuclear cells from 106 of 346 psychiatric patients and 12 or 70 healthy controls by p24-reverse transcription PCR was examined. Two mood disorder patients were positive for BDV-p24 RNA but seronegative. To detect anti-HTLV-1 antibodies the plasma samples were screened by the particle agglutination method and no significant difference in seropositivity for anti-HTLV-1 antibody was found between the patients and healthy controls. These results also suggested that there is a lack of association between BDV and HTLV-1 infections with psychiatric disorders among Japanese patients.

Distribution of CD11b Ag, a member of the leukocyte integrin (beta 2 integrin) family, on human T cells is not well characterized. We investigated in this study the tissue distribution and age-related alteration of human CD11b+ T cells. Significant proportions of CD11b+ cells were found in T cells from adult peripheral blood (18-58 yr) (20%), liver (32%), and spleen (16%), but were rarely observed in those from thymus, lymph nodes, intraepithelial lymphocytes, lamina propria lymphocytes, and lung. In contrast, percentages of CD28+ T cells, a counter population of CD11b+ T cells, were highest in lymph nodes (91%), modest in intraepithelial lymphocytes (82%), lamina propria lymphocytes (77%), PBL (77%), spleen (73%) and lower in thymus (62%), lung (60%) and liver (56%). The CD11b+ T cells from peripheral blood gradually increased with age, whereas the CD28+ T cells showed an age-dependent decrease. Namely, the percentages of CD11b+ T cells were 6% in newborns (cord blood), 15% in young adults (18 to 22 yr), 25% in middleaged adults (45 to 58 yr), and 27% in aged adults (72 to 86 yr), whereas those of CD28+ T cells were 96%, 83%, 72%, and 71%, respectively. Proportions of CD11b+ T cells in TCR-gamma delta+ or CD8+ T cells were, respectively, higher than those in TCR-alpha beta+ or CD4+ T cells. Age-dependent increase of CD11b+ T cells in CD8+ T cells was more obvious than that of CD4+ T cells. Furthermore, CD11b+ CD28+ T cells increased in the peripheral blood only of aged adults. The percentage of CD11b+ CD28+ T cells in TCR-gamma delta+ T cells was highest in young adults (25%), whereas that in CD4+ T cells was highest in aged adults (15%). CD11b+ CD28+ T cells expressed much potent IFN-gamma, IL-2, and IL-4 at mRNA level than CD28+ CD11b- T cells or CD11b+ CD28- T cells, and had potential cytotoxic activity. Roles of CD11b+ T cells in vivo were discussed.

The lymphocyte function associated antigen 1 has been found only on leukocytes and lymphoid tissues; however, the expression of lymphocyte function associated antigen 1 on nonhematopoietic cells has not been reported previously. In this study, immunohistochemical expression of lymphocyte function associated antigen 1 was examined on various tissues from 35 patients with malignant diseases and 36 patients with benign diseases including benign tumors. The expression of lymphocyte function associated antigen 1 was found on various exocrine tissues (eg, gastric glands, bronchial epithelium, alveolar epithelium, duodenal glands, bile ducts, pancreatic acini, and salivary glands) uninvolved by tumor in patients with malignant diseases. Localization of lymphocyte function associated antigen 1 was limited to the exocrine glands and differed from tissue-infiltrating leukocytes. The expression of lymphocyte function associated antigen 1 on exocrine tissues was confirmed in all 35 cases of malignant diseases that were examined. These included a wide spectrum of carcinomas and hematopoietic tumors. In contrast, none of the 36 cases with benign diseases examined expressed lymphocyte function associated antigen 1 on their exocrine glands. These results indicate a strong correlation between lymphocyte function associated antigen 1 expression on exocrine glands and malignant disease. The expression of lymphocyte function associated antigen 1 on nonhematopoietic cells was further confirmed in nonhematopoietic cell lines. Two of 19 nonhematopoietic cell lines (MKN45 and PANC-1; exocrine gland cell lines) examined expressed lymphocyte function associated antigen 1 on both cell surface and cytoplasm. These results suggested that immunohistochemically defined lymphocyte function associated antigen 1 molecules on nontumorous exocrine gland cells are a potential marker for the presence of malignant diseases.

It has continuously been growing information on bidirectional communication between the neuronal and immune systems, while increasing evidence suggests that neuromodulators released from the neuronal systems influence cells of the immune system. On the other hand, activated cells of the immune system release an array of immunomodulators that influence the physiologic function of the nervous system. The evidence suggests that the nervous and immune systems are integrated, interdependent homeostatic system.

Recent evidence has provided information that the immune system can be activated by non-antigenic stimulus e.g. stress. Information on the bidirectional communication between the neuronal and immune systems has been growing, while increasing evidence suggests that neuromodulators, released from the neuronal systems, influence the immune cells. Conversely, the activated immune cells release an array of immunomodulators that influence the physiologic function of the nervous system. An attempt was made to restore the altered immune function following exposure to stress by the treatment of the olfactory system with various fragrances. From the data in experimentally designed mouse models, it was found that the individual sensitivity to the fragrances was restricted by the genetic background (H-2) of the mice. The adaptation to the fragrances with regard to the recovery of the immune response to stress, was also characterized. The ability of the adaptation to the fragrances was found to differ not only due to the genetic background of the mice but also different fragrances.

A simple and fast method to measure the natural killer (NK) cell cytotoxicity has been established. In our novel assay, following the incubation of K562 target and effector cells, the cells were stained with FITC-conjugated anti-transferrin receptor (CD71) mAb. The K562 targets alone were labeled with anti-CD71 mAb, since CD71 is highly expressed on K562 cells but not on mononuclear cells. The fluorescence intensity of CD71 on K562 cells were analyzed using flow cytometry, and the cytotoxicity was evaluated. In our assay, designated as a "CD71 assay", no spontaneous release of anti-CD71 mAb was observed from labeled K562 cells and living K562 cells were able to be divided from dead K562 cells. The data obtained by this assay was well correlated with that by the standard 51Cr-release assay as well as the C-FDA assay. In addition, the CD71 assay is a safe and simple analytic procedure since no isotope is required.

A method for measurement of hyaluronic acid (HA) level in serum was developed based on using "hyaluronic acid binding protein" (HABP)-coated polystyrene beads. After the beads and test serum being mixed, the mixture was incubated together with reaction buffer for 2 hours, and then the beads were washed. Subsequently, biotinylated HABP was added to the washed beads and incubated for 1 hour. Then peroxidase-conjugated avidin was added to the mixture and incubated again for 1 hour. After the beads being washed, a substrate solution was added to the washed beads and left for 1 hour. Then the reaction was stopped by adding 2N-H2SO4. The absorbance at 492 nm was recorded. The analytical range of HA in serum by this method was found to be between 10-800 micrograms/l, and the precision of the HA assay (CV%) was between 3.0-8.4 in the "with-in" assay (n = 10), and 4.8-8.9 in the "between" assay (n = 5). The analytical recovery of HA assay was between 92-115%. In this study, the results in screening of the serum HA level in RA patients (n = 107), OA patients (n = 16) and healthy subjects (n = 30) showed that the HA level of RA patients was demonstrated significantly higher than that of healthy subjects and OA patients.

Numerous reports have recently indicated that the DNA content of various malignant tumors can be of great value in predicting biological behavior and prognosis of the tumors. This study was undertaken to determine the nuclear DNA content in the primary and recurrent ovarian carcinoma of the same 20 consecutive patients by flow cytometry, and the results on clinical outcome was examined. The tissue samples of the recurrent tumors were obtained at second-look laparotomy. Of the primary tumors, 12 were diploid, 4 were pure aneuploid, and 4 were "mosaic", while of the recurrent tumors, 17 were diploid and 3 showed pure aneuploid. No significant difference of the DNA index at recurrence was observed. DNA ploidy was preserved in 12 out of 20 patients at recurrence. The time to recurrence after the initial treatment showed no significant difference versus DNA ploidy and change of ploidy. The sites and status of recurrence differed by DNA ploidy. At recurrence, the patients with DNA diploidy had a tendency to survive longer than those with DNA aneuploidy. The determination of DNA ploidy at recurrence may be useful as a variable parameter in predicting the survival of patients with ovarian carcinomas.

Peripheral mononuclear cells taken from rheumatoid arthritis (RA) patients, which had previously been treated in vitro with the neuropeptide Substance P (SP) followed by stimulation with the mitogen PHA, showed a significantly higher percent positivity of lymphocytes with the double-markers CD4+ CD2+, CD8+ CD25+, CD4+, HLA-DR+ and CD8+ HLA-DR+ than those of the cells untreated by SP. No effect of SP was detected in normal controls, except that the cells with the double-marker CD4+ CD25+ were slightly increased. Furthermore, in RA patients the percentage of CD4+ CD45R- cells was increased, while that of CD4+ CD45R+ cells was decreased, by pre-treatment with SP and stimulation with PHA. Monocytes of patients with rheumatoid arthritis after the treatment with SP released a significantly higher amount of oxygen-intermediate than those of normal controls with or without SP. From these results, it could be considered that SP plays an important role in RA inflammation.

Several methods were compared for quantitating the B lymphocytes in human peripheral blood. RBC rosette assays including sheep and bovine red blood cells, and immunobead rosette assays including beads coated with anti-kappalambda and trivalent anti-GAM antibodies respectively were investigated. The anti-kappalambda bead assay proved to be simpler, faster, higher in reproducibility, and appeared to be more accurate than any of the other assays.

The effect of fourteen plant lectins on human peripheral blood lymphocytes in rosette formation with sheep red blood cells (SRBC) was examined. Among the lectins used, phytohemagglutinin (PHA), lens culinalis (LCH), wheat germ agglutinin (WGA), and ricinus communis I (RCAI) showed a significant increase in active T rosette counts when the results were compared with the effect of other lectins. This enhancement in active T rosette forming cells is found within one hour after the treatment with lectin and is dependent upon the concentration of lectin. The results suggest that the changes in lymphocyte membrane dynamics via stimulation by certain lectins increases the capacity to form active T rosettes.

Autoantibodies including rheumatoid factor (RF), anti-DNA antibody (ADA), and anti-nuclear factor (ANF) in addition to cold agglutinin and heterophil antibody titers were tested for in a total of 219 patients with common viral diseases. The diseases included varicella, influenza, measles, mumps, herpes zoster, hand-foot-mouth syndrome, and exanthem subitum . A high incidence of RF (23%) was demonstrated in varicella patients, and ADA and ANF (16% and 12%, respectively) were most frequently detected in the influenza cases. Those autoantibodies were most frequently found in patients with influenza. Each serum complement component and total hemolytic complement (CH50) were also assayed. Elevated levels of the fourth (C4) and ninth (C9) components of complement, along with elevated CH50, were observed in most patients. Follow-up studies indicated that those autoantibodies as well as other antibodies disappeared 3 to 8 weeks after the onset of infection. The present study indicates that substantial but transient alterations in the immune system accompanied by autoimmune phenomena and elevated levels in the complement components can occur in viral infections.

An improved assay for the simultaneous assessment of phagocytic uptake (via Immunobeads) and metabolic integrity (via NBT dye reduction) was used to evaluate the neutrophil function in neonates, one year olds, and adults. The unique advantage of this assay is that it evaluates both phagocytic and killing function simultaneously which appears to make the assay more reliable in the detection of neutrophil dysfunction. This assay allowed us to establish a subtle but significant neutrophil dysfunction in full term unstressed neonates which proved to be more pronounced in both the stressed and the preterm neonates. The one year olds, however, were found to have neutrophil function comparable to that of an adult.

Paraformaldehyde-fixed platelets from normal donors were used for detection of antibody to platelet by in vitro sensitization (indirect method) utilizing rabbit anti-human IgG heavy chain specific antibody-coated polyacrylamide gels (Immunobeads). The sensitized platelets formed rosettes with Immunobeads and the positive rosette count was over 30%, while control showed less than 8% when normal sera were used. This method was also applicable for detecting antibody-sensitized platelets in vivo (direct method) in patients with autoimmune thrombocytopenia. This method is simple, rapid and reproducible for clinical use. Direct and indirect immunofluorescent antibody tests and a blocking test with anti-human serum also supported the results of Immunobead rosetting technique.

A human polymorphonuclear leukocyte (PMN) activating factor (AFH-S) was isolated from a tropical plant seed, Aleurites Fordii Hemsl. (AFH) in PBS. It is found that the AFH-S stimulate human PMNs and induced superoxide generation and chemotaxis. Superoxide generation was affected by the extracellular calcium ion or pretreatment with H-7 (PK-C inhibitor), but not by mepacrine (PLA2 inhibitor) or pertussis toxin (islet-activating protein: IAP). Furthermore, a lag time exists dose-dependently. In addition the cytosolic calcium was not increased by the stimulation with AFH-S. Thus, the receptor for AFH-S was suggested to be independent from Ni-like protein, PI-response, or PLA2-activation, and stimulate PMNs through activation of PK-C.

This paper describes a method for the simultaneous determination of human thymus-dependent lymphocyte (T cell) and thymus-independent lymphocyte (B cell) populations by mixing lymphocytes, sheep red blood cells, and immunobeads. This method also detects lymphocytes possessing both T and B cell markers. T cells are detected by rosette formation with AET-treated sheep red blood cells and B cells are identified by rosette formation with polyacrylamide gel coated with antibody specific to immunoglobulin classes. The major advantages of this technique is that it is simple, convenient, and reproducible. Three different types of rosettes are distinguishable under light microscopic observation. The detection of double marker lymphocytes, using this combined technique, may have further application in clinical investigation.

Serum levels of IgE, other immunoglobulins and C'3 were measured in 36 MS patients, and the results compared with those of 40 age- and sex-matched patients hospitalized on Neurology Service. Diagnoses among controls included cerebral infarction, cervical spondylosis, nonmigrainous headache, seizure disorders and peripheral neuropathy. Six patients in the MS group and seven in the non-MS group had a past history of allergy to food, drugs, dust or other substances. IgE levels were measured by double-antibody radioimmunoassay. Other immunoglobulins (G, M, D and A) and C'3 were quantitated by Mancini's method. Results indicate that the median IgE and C'3 levels of MS patients were slightly lower than those for non-MS subjects. Concentrations of other immunoglobulins were similar for the two groups. CSF samples from 12 MS and five non-MS patients were studied and none of these contained measureable (greater than 7.5 U/ml) IgE. In view of: 1) recent reports describing mast cells in MS plaques, 2) the suggested role of biogenic amines in the pathogenesis of demyelinating diseases, and 3) because IgE exists in free and cell bound state, it is suggested that tissue surveys of MS plaques for IgE deposits similar to those seen in glomeruli in nephrotic syndromes may provide a clue to the pathogenesis of MS.

An improved assay for the simultaneous assessment of phagocytic uptake (via Immunobeads) and metabolic integrity (via nitroblue tetrazolium (NBT) dye reduction) was used to evaluate the neutrophil function in neonates, one year olds, and adults. The unique advantage of this assay is that it offers greater standardization since the beads are commercially available with a known quantity of immunoglobulin bound to their surface which allows for considerably less variation than particles opsonized in vivo. Blood samples were collected from 20 full term healthy neonates, 20 healthy one year olds, and 20 healthy adults. The neutrophils were isolated, and phagocytic and killing function compared among the three groups. It was found that the neonates had a small but significant neutrophil dysfunction with respect to phagocytic and intracellular killing ability when compared to the one year olds and adults. Additionally, blood was collected and evaluated from five premature infants with varying degrees of stress. Their neutrophil dysfunction was much more pronounced. Although it was previously thought that full term healthy neonates have no demonstrable neutrophil dysfunction unless stressed in some manner, a subtle dysfunction was identified even in unstressed neonates. It was, however, found to be more pronounced if the neonate was either premature or stressed in some manner. It is hoped that this assay, through its greater sensitivity and ease of standardization, will uncover subtle neutrophil dysfunctions in various disease states that as yet remain undiagnosed. The clinical significance of such subtle neutrophil dysfunctions is not yet known.

One hundred and ninety-nine patients were divided into three groups and examined for their skin reactions to dinitrochlorobenzene (DNCB). The groups consisted of those patients with rubella, those with cancer and those with dermatoses receiving long terms, small dose glucocorticosteroids. Eleven patients who did not respond to the first 0.1% challenge of DNCB two weeks after the initial sensitization responded to subsequent 0.1% DNCB challenges. Seventeen patients who were insensitive to a 0.1% DNCB challenge showed a positive skin reaction when higher concentrations (greater than 0.6%) of DNCB were used. These results indicate that among DNCB-negative patients, as determined by the present method of testing, there are those who are actually sensitized with the first dose but do not show a positive reaction.

Membrane surface antigens on peripheral blood lymphocytes were analyzed by a laser flow cytometry system using monoclonal antibodies. Four normal subjects among 1,998 normal and diseased groups were found whose lymphocytes were not detected by OKT4 monoclonal antibody, while Leu3a and KOLT-1, which were also specific for helper/inducer T-lymphocytes as OKT4, were present in these subjects. The cases with the defect of the OKT4 antigen suggest that there is some modification of the antigen, lack of epitopes, or genetic polymorphism in the appearance of the OKT4+ cell surface antigen on normal human lymphocyte.

Humoral and cellular immunologic studies were carried out to define the effect of steroid treatment on the immune system of patients with non-immunologically-mediated dermatoses. The cellular immune response as measured by PHA and Con A induced lymphoblastogenesis was highly impaired, being reduced in 79% and 66%, respectively, of the cases. The peripheral and bone marrow lymphocyte subpopulation and PPD skin reaction were only slightly affected by steroid administration. Histologic studies of lymph node biopsies showed atrophy, fibrosis and fat degeneration. The immunoglobulin and complement levels and humoral antibody responses to typhoid vaccine, diphtheria toxoid and bacteriophage phi--x 174 were mildly affected. The impairment of immune systems induced by steroids was largely reversible, while the PHA induced blastogenesis and pathological changes in the lymph nodes were irreversible. In comparison with experimental animal studies using massive doses of steroids, the immune system in man was not significantly affected by administration of small doses on a long term basis in clinical practice.

The effect of various cellular metabolic inhibitors on the active T rosette formation of human peripheral blood lymphocytes with sheep red blood cells (SRBC) was studied. Cytochalasin B greatly inhibited SRBC rosette formation. However, colchicine, mytomycin C, cyclohexamide, and sodium azide had no effect. These results suggest that the ability of the lymphocyte to form active T rosettes is dependent on the lattice microfilament network, but not on the cellular metabolism. The effect of cytochalasin B on human peripheral blood lymphocytes activated by phytohemagglutinin (PHA), wheat germ agglutinin (WGA), lens culinalis (LCH), or ricinus communis agglutinin I (RCAI) was also examined. Cytochalasin B significantly inhibited the active T rosette formation induced by LCH or RCAI, but did not inhibit that induced by PHA or WGA. This suggests that different cellular mechanisms are involved in the activation of lymphocytes by either PHA and WGA or LCH and RCAI.

The neutrophil function of 12 patients with cystic fibrosis was evaluated via the new Immunobead/NBT dye reduction test. While the neutrophils appeared to be hyperfunctional when studied in the standard assay using fetal calf serum, we demonstrated a significant neutrophil dysfunction when studied in their own respective serum. The dysfunction involved both the phagocytic and intracellular killing ability of the cells, which we postulate is due to an inhibitory factor present in the patient's serum. The inhibitory factor did not appear to be related to the level of circulating immune complexes.

The cell membrane markers on peripheral blood lymphocytes of 10 patients with Vogt-Koyanagi-Harada disease (VKH) were analyzed by laser flow cytometry using monoclonal antibodies. Seven patients with active uveitis and 98 normal control subjects were also studied. The percentage of OKT3+, OKT4+ and OKT11+ cells were significantly lower while OKIa1+ cell were higher in VKH compared to controls. The percentage of cells reacting with monoclonal antibodies were slightly affected during steroid therapy, but returned to the pretreatment level after cessation of the therapy. OKIa1+ cells were shown to be increased in active uveitis while percentages of cells with other cell markers remained unchanged when the results were compared with that of the control. The findings suggest that the analysis of cell membrane markers on lymphocytes in VKH could be a significant parameter in clinical medicine.

In this study we were able to confirm the presence of double marker lymphocytes (D cells) in human peripheral blood from normal subjects in addition to patients with SLE and malignant lymphoma. Quantitation of D cells and Null lymphocytes (N cells) in peripheral blood was not feasible via conventional rosette methods. The present combination method, using two indicators, neuraminidase treated sheep red blood cells (nSRBC) and Immunobeads, allows for simultaneous demonstration of T, B, N and D lymphocytes in a simple reproducible manner. The mean percentage of the D cell counts with this method was 1.8% of total lymphocytes from normal donors, 7.0% in patients with SLE, and 29.2% in patients with malignant lymphoma. The simultaneous quantitation of T, B and N cells in these blood samples was compared to the results of conventional methods, i.e. E rosettes for T cells and EAC rosettes for B cells, which gave similar results. This combination method for simultaneous enumeration of human peripheral lymphocyte subpopulations may offer significant advances in the study and diagnosis of various clinical entities.

We studied cell surface antigens on fresh and cultured peripheral blood lymphocytes (PBL) of healthy HTLV-I carriers. OKT4, OKT8 and OKIa1-positive cells were found to be within the normal range as compared with controls both in fresh and in cultured PBL. The percentage of Tac positive cells in HTLV-I carriers was shown to be higher in fresh PBL than that of controls. Furthermore the value increased significantly during cell culture in vitro and large blastoid cells appeared. The results suggested that HTLV-I proliferation in infected lymphocytes is activated in a serum-free environment and that small resting lymphocytes are transformed into Tac-positive large blastoid cells.

A human T-cell line, designated as MKB-1, was established by cloning procedures in a suspension culture from a peripheral blood of a 17-year-old female patient with acute myeloblastic leukemia. The immunological marker profile of MKB-1 indicated that unlike a myeloid phenotype of the original leukemic cells, the cells were positive for CD3 (both cell surface and cytoplasm), T cell receptor (TcR) alpha/beta heterodimer, CD4, CD5, CD7, CD10, CD57 (Leu7), SN-1 and the cytoplasmic TcR beta chain. These findings indicate the T cell nature of the established cells. Terminal deoxynucleotidyl transferase (TdT) was also detected in 60%. We did not detect markers of human myeloid and B cell associated antigens, HLA-class II or immunoglobulin chains. Cytogenetic study revealed that the MKB-1 cells had a female hypo-tetraploid karyotype with chromosomal abnormalities including a translocation between chromosomes 10 and 14. The breakpoint of chromosome 14 of this translocation, 14q11.2, is known to be the location of TcR alpha and delta genes; t(10; 14) (q26; q11.2) is a variant type of a T cell neoplasm-associated translocation, t (10; 14) (q24; q11.2). The MKB-1 cell line is unusual in that its T cell characteristics are phenotypically and cytogenetically distinct from the original myeloid leukemia cells.

Nine patients had birdshot retinochoroidopathy. Associated ophthalmologic findings included rhegmatogenous retinal detachment, rubeosis iridis, glaucoma, and a high incidence of disciform macular degeneration. Four patterns of birdshot spot distribution were noted on fundus examination. Immunologic studies showed a significantly elevated percentage of EA rosettes and an elevated C4 complement level. Electroretinograms showed b-wave amplitude reduction, with a disproportionate implicit time prolongation. The etiology of this syndrome remains unknown.

By sensitizing human red blood cells with antisera specific for corresponding blood group antigens and subsequent rosette formation, we were able to distinguish homozygosity of the antigens from heterozygosity. Three types of rosette assays were utilized: a protein A rosette, an immunoglobulin-coated bead rosette, and an erythrocyte-antibody rosette with human lymphocytes. In the Rh blood group system, we were able to demonstrate the dosage effect of the Rho(D) antigen by rosette assay in addition to determining that the deletion of the C and/or E alleles increased the rosette forming cell count when red blood cells with different genotypes were sensitized with Rh immune globulin (anti-D). The assay proved to be reproducible in distinguishing between homozygous and heterozygous Rh red blood cell antigens, and may be adaptable to study many antigenic markers on cell surfaces.

In order to clarify the abnormalities of cell-mediated immunity in patients with mesangial proliferative glomerulonephritis [IgA nephropathy (IgA N) and Non-IgA nephropathy (Non-IgA N)], lymphocyte subsets were analysed by using monoclonal antibodies with flow cytometric two-color analysis and interleukin-2 (IL-2) production from lymphocytes of the patients was measured by ELISA system. A markedly decreased percentage (11.6 +/- 10.5%) of CD4+ 45R+ cells was found in the patients with IgA N when the results were compared with the normal controls (20 +/- 6%) (P less than 0.01). No difference was found between patients with IgA N and the controls as to the percentage of CD4+ 45R-, CD8+, 11+ and CD8+ 11- cells, respectively. Patients with Non-IgA N also showed a significantly decreased percentage of CD4+ 45R+ cells (10.9 +/- 6.5%), while CD8+ 11+ cells was parallely lowered (7.4 +/- 5%) in compared with the controls (10 +/- 4%). The percentage of HLA-DR positive cells was found to be increased in the patients with both IgA N and Non-IgA N, although the antigen bearing cells were reduced after stimulation with Concanavalin-A (Con-A). No difference in IL-2 production from lymphocyte of both patients groups and the controls cultured with Con-A was found. These results suggested that a deficiency of suppressor inducer T cells played a part of the pathogenesis of IgA N and Non-IgA N.

A hairy cell leukemia cell line designated "Hair-M" was established in a suspension culture derived from the peripheral blood of an 86-year-old Japanese male with a diagnosis of hairy cell leukemia. The Hair-M cells were identified as having prominent hair-like cytoplasmic projections by examination with phase-contrast and scanning electron microscopy. These cells displayed ruffled membranes and stublike microvilli similar to those observed on the surfaces of cells in the peripheral blood of the patient. Immunologic and cytochemical studies on the Hair-M cells confirmed derivation from the clone of the patient's leukemia cells. Although the cultured Hair-M cells had definite B-cell characteristics, such as IgG kappa-chains on the surface and in cytoplasm, they also demonstrated Tac antigen, which is usually expressed on activated T-cells, and myelomonocyte antigens determined by OKM-1 and MCS-1 monoclonal antibodies. Other cell surface markers, including E(-), IgGFc(-), IgMFc(-), C3R(+), Ia-like antigen(+), OKT9(+), OKT10(+), and terminal deoxynucleotidyl transferase(-), were detected; no Epstein-Barr virus-determined nuclear antigen was detected. The karyotype of the Hair-M cells was determined to be 46XY with -11, -14, and two marker chromosomes. The Hair-M cells also had phagocytic activity to rabbit anti-human IgG serum-coated polyacrylamide gel particles.

Lymphocytes from a patient with Wiskott-Aldrich syndrome (WAS) were employed for a study of the intracellular viscosity and fluorescein permeability through the cell membrane by a fluorescence polarization spectrofluorometer, which was designed to calculate polarization value and permeable fluorescein intensity automatically. Fluorescein diacetate (FDA) was used as the indicator probe. The fluorogenic substrate is taken up by viable cells and converted to a fluorescent molecule, fluorescein, by intracellular esterase, where upon the fluorescein easily effluxes through the cell membrane. The response to stimulation with phytohemagglutinin (PHA) for 45 min led to a decreased polarization value (p-value) as compared to lymphocytes of healthy donors, and the fluorescein efflux through the cell membrane was greater than that of healthy donors. Fluorescein efflux from lymphocytes in the patient during 48 and 72 h incubation with or without PHA was markedly increased. In healthy donors, the degree of fluorescein permeability was not increased during the culture. These results indicate that intracellular viscosity of lymphocytes is altered in initial mitogenic stimulation, but that there was some abnormality in the fluorescein permeability properties through the cell membrane of lymphocytes in a patient with WAS.