NADPH-cytochrome c reductase and cytochrome b559 were purified from the membrane fraction of phorbol myristate acetate-stimulated porcine polymorphonuclear leukocytes. The highly purified reductase oxidized NADPH and generated superoxide when combined with partially purified cytochrome b559 in the presence of phosphatidylcholine. In the same system, but under the anaerobic condition, the reductase was found to reduce cytochrome b559.

Animals↗

PubMed1987

Healing length of superfluid 3He.

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Journal Article↗

PubMed1987

Predefined gene transfer for expression of a glycosphingolipid antigen by transfection with a cosmid genomic library prepared from a cell line in which the specific glycosphingolipid is highly expressed.

The deliberate transfer of globotriaosylceramide (Gb3) expression in mouse lymphoma L5178 cells was achieved by transfection with a cosmid DNA library prepared from human Burkitt lymphoma Ramos cells in which Gb3 was highly expressed. The recipient mouse lymphoma cells did not contain Gb3 but did contain its direct precursor, lactosylceramide. The transfected cells expressed Gb3, detected both chemically and immunologically, and contained human DNA detected by an Alu sequence probe. This model demonstrates a general method for studying glycosyltransferase genes and other factors necessary for the expression of glycosphingolipid antigens.

Animals↗

PubMed1987

Studies on transmission of human non-A, non-B hepatitis to marmosets.

Two sera obtained from four healthy blood donors, which caused non-A, non-B post-transfusion hepatitis in two recipients, were experimentally inoculated into nine marmosets. Three of seven marmosets developed acute hepatitis characterized by the elevation of serum concentrations of glutamic pyruvic transaminase (GPT) and/or isocitric dehydrogenase (ICD) 8-11 weeks after inoculation. Four of seven showed histopathological changes of acute hepatitis in liver biopsy specimens during the biochemically acute phase. In electron microscopic examination, attached membrane-like structures, which consisted of two-unit membranes of two neighboring endoplasmic reticula with electron-dense material between them, were noted in cytoplasm of hepatocytes during the acute phase of hepatitis. Furthermore, acute-phase sera obtained from two animals were inoculated into four additional marmosets, and non-A, non-B hepatitis was successfully passaged in two of them. The results of this study indicate that certain species of marmoset monkeys are susceptible to human non-A, non-B hepatitis agents and provide a useful animal model for non-A, non-B hepatitis.

Alanine Transaminase↗

PubMed1987

Detection of a novel HLA-DQ specificity: serological and immunochemical analyses by a monoclonal antibody.

A monoclonal antibody (mAb) with a novel human B-cell allospecificity was produced by immunizing a C3H/He mouse with the human B lymphoblastoid cell line EBV-Wa (HLA-DR4/Dw15/DQblank homozygous). The mAb, termed HU-46, reacted with B cells from not only DR4/Dw15-positive individuals but also certain DRw8/Dw8-positive ones whose DQ phenotypes had not yet been defined. Two-dimensional gel analyses indicated that the mAb recognized class II antigens which were encoded by the HLA-DQ locus. Furthermore, in genetic analysis, the gene encoding the class II antigen detected by HU-46 met the Hardy-Weinberg condition as a fourth allele of the DQ locus. We provisionally labeled this novel DQ specificity DQWa.

Alleles↗

PubMed1987

Congenital glioblastoma of the cerebellum with cytofluorometric deoxyribonucleic acid analysis.

A case of congenital glioblastoma arising from the cerebellum is presented with special reference to microspectrophotometric deoxyribonucleic acid (DNA) analysis of the tumor cells. Seven cases of this condition are already in the literature. The prognosis is poor and most of the reported cases survive no more than 2 months. Cytofluorophotometric DNA studies were helpful in elucidating the extreme heterogeneous distribution of DNA contents of the tumor cells, suggesting a biological aggressiveness probably related to the rate of proliferation.

Brain↗

PubMed1987

A cytotoxic monoclonal antibody (HU-39) that detects DRw8 + DRw12.

A monoclonal antibody, HU-39, was produced by immunizing BALB/c mice with a cultured human B lymphoblastoid cell line, Shi-C3 (HLA-A24, A31, B51, Bw52, DR2, DRw12, DQw1, DQw3). Utilizing the complement-dependent cytotoxicity test, HU-39 was found to detect a polymorphic determinant common to HLA-DRw8 and HLA-DRw12, a split antigen of HLA-DR5. Although HU-39 reacted with the cells from all of nine DRw12 positive individuals, the cells from only 18 out of 21 DRw8 positive individuals reacted with HU-39 and the remaining three were negative for HU-39. The cytotoxicity of the antibody was reduced after the surface HLA-DR molecules of two cell lines, GI and EBV-Sh, typed as DRw8 and DRw12, respectively, were masked with F(ab')2, of anti-HLA-DR monoclonal antibody. The results of the sequential coprecipitation test and the two-dimensional gel electrophoresis by using EBV-Sh also indicated that HU-39 preferentially recognizes an epitope borne on the DR molecules, but not on the DQ molecules. Thus, HU-39 appeared to be of great value as a tissue typing reagent to define DRw8 and DRw12, the latter of which had been difficult to assign because of the lack of monospecific alloantisera.

Antibodies, Monoclonal↗

PubMed1987

A novel human class II specificity, DQ "Wa," resides on DQ molecules of DR4,Dw15 and DRw8,Dw8 B-cell lines identified as DQ "blank".

The molecular localization of a novel human class II specificity, DQ "Wa," was investigated. A monoclonal antibody, HU46, which has previously been shown to react with DR4, Dw15 and DRw8, Dw8 B cells that type as DQ "blank," was used for the isolation and structural characterization of class II molecules bearing the DQ "Wa" determinant. The partial N-terminal sequence analysis of class II molecules bearing the DQ "Wa" determinant, purified from two B-cell lines, EBV-Wa (DR4, Dw15, DQ "blank") and GI (DRw8, Dw8, DQ "blank"), shows that the alpha and beta chain sequences are homologous to HLA-DQ. Within the limits of our analysis, the alpha and beta chains from both cell lines are identical. Both beta chains possess a phenylalanine residue at position 9 that differs from the tyrosine residue present at this position in beta chains of DQ alleles. These studies indicate that a novel human class II specificity, DQ "Wa," resides on a new allelic form of DQ molecules found in DR4, Dw15 and DRw8, Dw8 cells that are DQ "blank."

Alleles↗

PubMed1987

Purification and characterization of NADPH-cytochrome c reductase from porcine polymorphonuclear leukocytes.

NADPH-cytochrome c reductase [NADPH: ferricytochrome oxidoreductase, EC 1.6.2.4] was highly purified from the membrane fraction of porcine polymorphonuclear leukocytes by column chromatographies on DEAE cellulose DE-52, 2',5'-ADP-agarose, Sephacryl S-300, and Bio-gel HTP. Upon sodium dodecyl sulfate polyacrylamide gel electrophoresis, the purified preparation gave a main band with a molecular weight of 80,000. The enzyme contained 0.79 mol of FAD and 0.88 mol of FMN per mol, and was capable of exhibiting a benzphetamine N-demethylation activity in the presence of cytochrome P-450 purified from rabbit liver microsomes and dilauroylphosphatidylcholine, as is the case with liver NADPH-cytochrome P-450 reductase. The cytochrome c reductase activity of the polymorphonuclear leukocytes (PMN) enzyme was precipitated with rabbit anti-guinea pig liver NADPH-cytochrome P-450 reductase IgG followed by addition of guinea pig anti-rabbit IgG antibody. The biochemical and immunological properties of the PMN enzyme so far examined were similar to those of the liver enzyme, although its function in leukocytes has not yet been determined.

Animals↗

PubMed1987

New Dw8 associated class II specificities defined by cytotoxic T lymphocyte clones.

Two Leu2(-), Leu3(+), Leu4(+) human cytotoxic T lymphocyte (CTL) clones, BE-11 and AF-3, were generated against Epstein-Barr virus (EBV)-transformed cell line GI (Dw8/DRw8/DQWa homozygous). Blocking experiments with various monoclonal antibodies (MoAbs) revealed that the former recognized the DR molecule and the latter recognized the DQ molecule, respectively. Panel studies showed that CTL clone BE-11 lysed not only DRw8-positive cells but also DR1-positive ones. CTL clone AF-3 exhibited cytotoxicity against only Dw8/DRw8/DQWa typed cells. Until now, such specificities have not been defined serologically or biochemically. These results demonstrated that the previously unknown DR and DQ specificities could be defined by CTL clones, suggesting that CTL clones might be especially valuable tools for investigating the structural polymorphism of HLA antigens.

Antibodies, Monoclonal↗

PubMed1987

Genetic control of the immune responsiveness to Streptococcus mutans by the major histocompatibility complex of the rat (RT1).

The lymph node cells from 11 strains of rats, differing in the genotype of the major histocompatibility complex of the rat (RT1), were examined on the basis of their proliferative response to the cell wall antigen of Streptococcus mutans. The 11 rat strains fell into three groups: high, intermediate, and low responders. To demonstrate the influence of the major histocompatibility complex on immune responsiveness to S. mutans, further experiments were performed using the RT1-congenic rat strains WKAH.1L(LEW), WKAH. 1AV1(ACI), and WKAH.1J(LEJ), which differ only in the genotype of the RT1 region. Although the background genes of each strain were of WKAH origin, WKAH.1L(LEW) and WKAH.1AV1(ACI) rats showed a low response whereas WKAH.1J(LEJ) rats showed a moderate response to the S. mutans cell wall antigen. The results indicate that the immune response is controlled by the class II gene(s) in RT1. Furthermore, the RT1.D locus products were shown to play an important role in the restriction molecule, since a monoclonal antibody, HOK7, directed to the RT1.Dk locus products reduced the proliferative response of lymph node cells.

Animals↗

PubMed1987

Computerized glucose clamp method for the determination of insulin sensitivity in diabetic subjects.

A new, simplified computerized glucose clamp method was performed for estimation of glucose utilization at the insulin level of postprandial level. The program for the method is based on the mathematical algorithm using values of blood glucose, changes of its concentration and the desired glucose level. The coefficients of variation of the clamped blood glucose values during the last 60 minutes was 6.3 +/- 1.7(%) in normal, 3.7 +/- 0.3(%) in NIDDM, which was within satisfactory limit, and also indicated an attainment of steady state. Glucose infusion curves showed some bumps for the initial 60 min, which did not seriously affect the glucose utilization rate at steady state. In some cases, whose insulin sensitivity was high, CV was not low enough and occasional manual adjustment of K values was required. Average glucose infusion rate was 7.59 +/- 0.85 (mg/kg/min) in normal, and this was significantly lowered in NIDDM (42.3 +/- 3.4) at steady state, indicating a decreased insulin sensitivity for glucose utilization in NIDDM.

Adult↗

PubMed1987

[Detection of Chlamydia trachomatis (CT) in the smears taken from the infected sites using FITC labelled CT species-specific monoclonal antibody].

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Antibodies, Monoclonal↗

PubMed1987

[Lymphocyte cytotoxicity against autologous tumor cells and the effect of interferon-beta on their activities. (2) Evaluation of large granular lymphocytes (LGL)].

Large granular lymphocytes (LGL) were obtained by Percoll density gradient centrifugation of peripheral mononuclear cells from 15 patients with hematological malignancies (10 acute leukemias and 5 non-Hodgkin's lymphomas). Five-hour 51Cr-release cytotoxic assay by LGL was performed against frozen autologous tumor cells in these patients. Mean percentage cytotoxicity by LGL was 5.6%, and this was enhanced to 15.0% by the addition of IFN-beta to the culture medium. A decrease of cytotoxicity was observed when LGL was treated with anti-Leu 11b antibody plus complement, or when LGL was pretreated with the unlabelled K562 as a competitive inhibition assay. The addition of monocytes also induced a decrease of cytotoxicity, suggesting that monocytes may act as a suppressive agent in autologous tumor cell killing by LGL.

Antigens, Differentiation, T-Lymphocyte↗