摘要:

AbstractWe investigated the immunomodulatory effect of cyclophosphamide on the induction of suppressor cell activities in peripheral blood lymphocytes. The sera from advanced breast cancer patients as well as concanavalin‐A (Con‐A) induced suppressor cell activities in lymphocytes from healthy volunteers. Pretreatment of these lymphocytes, including CD4+T cells with 4‐hydroperoxycyclophosphamide (4‐HC), an active form of cyclophosphamide, abrogated the induction of suppressor cell activities by either cancer sera or Con‐A. A decrease in CD4+CD45RA+suppressor/inducer T cells and reduction of Con‐A induced suppressor cell activities were observed in breast cancer patients with metastases that were treated with cyclophosphamide (CPM). These results suggest that cyclophosphamide may modulate the immune responses of breast cancer patients by interference with suppressor/inducer T cells. © 1994 Wil

ISSN:0887-8013    

DOI:10.1002/jcla.1860080302

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractWe evaluated four newly introduced assays for determination of glycated hemoglobin allowing the processing of large amounts of samples in a clinical routine laboratory. These methods were compared to the Bio‐Rad Diamat system. The investigated methods were the Merck Hitachi L‐9100, a fully automated HPLC analyser, the Abbott IMx glycated hemoglobin ion capture assay, the DAKO HbA1c enzyme linked immunosorbent assay (ELISA), and the Boehringer‐Mannheim HbA1c Tinaquant turbidimetric assay. All methods showed generally acceptable precision and good accordance with the Diamat system. Interference study showed influence of anaemia, polycythemia, rheumatoid factor, and chronic hemodialysis on the values of the DAKO ELISA and of anaemia and polycythemia on the values of the Boehringer‐Mannheim Tinaquant assay. All of the investigated methods allow referring of results either as measured or standardised HbA1c values, the latter obtained after calibration with reference to an ion exchange high‐pressure liquid chromatography method. Our data confirm the feasibility of this kind of standardisation of glycated hemoglobin assays, allowing direct comparison of results from various determination methods. © 1994 Wiley

ISSN:0887-8013    

DOI:10.1002/jcla.1860080303

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractIn this study, the effect of the synthetic glucocorticoid hormone dexamethasone (DXM) on HSV replication was studied in a DXM receptor‐positive mouse neuroblastoma (NB) cell line. In cells treated with 10−7M DXM and then infected with HSV, there was a statistically significant 9–18‐fold increase in the amount of virus produced in these cells compared to untreated controls. Adsorption kinetic studies with HSV were performed in DXM‐treated NB cells and untreated controls. It was found that there was a significant increase in the adsorption rate of HSV in the DXM‐treated cells as compared with the controls. During the course of these studies, a strain of NB cells was noted to have lost its ability to stimulate HSV replication following DXM treatment. Receptor binding assays were performed on cytosols prepared from NB cells that responded with an increase in HSV titers to DXM treatment and the new strain of NB cells that was DXM refractile. These latter cells were found to have lost their DXM receptors. These results indicate that the modulation of HSV replication of DXM treated cells was regulated by the presence of DXM receptors in these cells. Once lost, the cells do not respond to DXM treatment with increased HSV replication. These observations may lead to a clinical assay to determine patients with high glucocorticoid levels who may be at risk of recurrent herpes infections. © 1994 Wil

ISSN:0887-8013    

DOI:10.1002/jcla.1860080304

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractAdenylate kinase activity (AK) originating from erythrocytes, present in hemolyzed serum behaves like creatine kinase MM isoenzyme (CK‐MM) in some CK electrophoresis assays that employ, in their visualization reagent kits, adenosine monophosphate (AMP) as the sole inhibitor of AK, rather than a combination of AMP and a more potent inhibitor of erythrocyte AK, diadenosine pentaphosphate (Ap5A), to inhibit all contaminating‐AK activities in serum and quantify only the CK isoenzyme activities in serum following electrophoretic fractionation on agarose gel. This can spuriously overestimate the CK‐MM fraction and thereby result in underestimation of CK‐MM or CK‐BB isoenzymes if present. A hemolyzed serum sample obtained from an elderly patient was erroneously reported as containing low CK‐MB due to such overestimation of CK‐MM fraction in the sample. Supplementing the AMP already present in the visualization reagent formulation, used to estimate CK isoenzyme concentration in serum, with Ap5A can eliminate or effectively minimize AK interference, especially that caused by hemolysis, and thereby prevent reporting false‐negative CK‐MB result obtained with CK isoenzyme electrophoresis assays. © 19

ISSN:0887-8013    

DOI:10.1002/jcla.1860080305

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractWe studied the alteration of aldolase isozymes in the serum and tissues of patients with cancer and other diseases using radioimmunoassays specific for aldolaseA, B, and C subunits. Aldolase B was predominantly found in adult liver, where aldolase A and C were distinctly low. Aldolase A and B showed almost the same concentration in fetal liver, while in neonatal liver aldolase B protein concentrations were much higher than aldolase A. In contrast, aldolase A was the predominant isozyme found in hepatoma and gastric cancer tissues, whereas aldolase B was distinctly low in hepatoma tissues, and extremely low in gastric cancer tissues. These results suggest that the aldolase A is a more fetal type of liver isozyme than the aldolase B and C, and aldolase B is a more differentiated type of liver isozyme than aldolase A and C. Serum FDP aldolase activities were elevated in half of patients with liver diseases, all patients with muscle diseases and a few patients with cancer. Serum aldolase A levels were elevated in patients with muscle diseases and cancer, but not elevated in patients with liver diseases. In contrast, serum aldolase B levels were elevated in patients with liver disease, but not elevated in patients with muscle diseases and other diseases without liver injury. Serum aldolase B levels showed a trend to decrease in cancer patients with normal GPT levels. Serum aldolase A/B ratios were significantly increased in cancer patients with normal GPT levels, whereas they showed the decreased levels in patients with liver diseases. These results suggest that measurement of aldolase isozymes by subunit specific radioimmunoassays may be useful in the diagnosis of certain cancers, myopathies and liver diseases. © 1994 Wiley‐Liss, I

ISSN:0887-8013    

DOI:10.1002/jcla.1860080306

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractAntibody IgG to human T‐cell leukemia virus type I (HTLV‐I) in urine was detected by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys‐envgp46(188–224), as antigen, the sensitivity and specificity of which were 100 and 98.5%, respectively, using serum samples. Anti‐HTLV‐I IgG in urine was reacted simultaneously with 2,4‐dinitrophenyl‐bovine serum albumin–Cys‐envgp46(188–224) conjugate and Cys‐envgp46(188–224)‐β‐D‐galactosidase (Escherichia coli) conjugate. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG, eluted with ϵN‐2,4‐dinitrophenyl‐L‐lysine and transferred to polystyrene balls coated with affinity‐purified (anti‐human IgG γ‐chain) IgG. Finally, bound β‐D‐galactosidase activity was assayed by fluorometry. Thirty‐one urine samples from seropositive subjects and 100 urine samples from seronegative subjects were tested. The sensitivity and specificity were 87 and 100%, respectively, with unconcentrated urine samples and 94 and 100%, respectively, with approximately 10‐fold concentrated urine samples. These results were superior to those by the conventional ELISA

ISSN:0887-8013    

DOI:10.1002/jcla.1860080307

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractAllotransplant rejection is a T‐cell‐dependent reaction. Functional in vitro T‐cell assays are being used widely for donorrecipient matching in bone marrow transplantation and have recently been used in some centres for transplant monitoring. In order to assess tolerance induction after clinical transplantation, we measured the T‐cell response of the host against donor spleen cells of 33 kidney transplant patients before and every 3 months after transplantation over a period of 18 months. The T‐cell reactivity before transplantation was not significantly different in any of the assays in rejecting and nonrejecting patients. In the classical mixed lymphocyte culture (MLC), a donor‐specific loss of reactivity was seen only in a patient with a CMV‐associated irreversible transplant rejection. One patient with chronic rejection acquired a very high MLC response against donor spleen cells and a high response against third‐party cells. Little or nonspecific changes were seen in the MLCs of all other patients. Using the method of limiting dilution analysis (LDA), we found a significant reduction of donor‐specific cytotoxic T‐cell precursors (CTL‐p) within the first 3 months after transplantation in most patients with high antidonor CTL‐p frequencies before transplantation. The reduction of donor‐specific CTL‐p was seen in patients with rejection episodes as well as in patients without. Thus we conclude, in contrast to others, that MLC and CTL‐p LDA have no predictive value on the outcome of clinical transplant

ISSN:0887-8013    

DOI:10.1002/jcla.1860080308

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractThe human insulin domains, signal peptide, B‐chain, C‐peptide, and A‐chain, were highly expressed inEscherichia colias recombinant proteins N‐terminally fused to glutathione‐S‐transferase and a histidinehexapeptide. The recombinant proteins were purified from insoluble cell fraction by affinity chromatography using metal chelating matrix, which was charged with Ni+2iones. ELISA screening for autoantibodies directed to preproinsulin were performed with sera from patients with recently diagnosed insulindependent diabetes mellitus in order to localize the antigenic epitopes within the human preproinsulin. Of the patients, 14% had developed autoantibodies that recognized either the recombinant C‐peptide or the signal peptide. No reaction was observed with the A‐chain or B‐chain. © 19

ISSN:0887-8013    

DOI:10.1002/jcla.1860080309

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractA plasma protein cofactor, β2‐glycoprotein I (β2GPI), also known as apolipoprotein H, is necessary to detect certain antiphospholipid antibodies (aPA) to negatively charged phospholipids (PL) in the ELISA. Inasmuch as sera are diluted 1:100 before testing, the concentration of native β2GPI may be insufficient to provide an optimal aPA ELISA signal. Therefore, many laboratories add adult bovine serum (ABS) to the diluent buffer to provide a consistent level of cofactor for optimal aPA binding. To determine if other animal sera can provide the cofactor, cat, chicken, dog, horse, goat, guinea pig, mouse, pig, rat, and sheep were tested as diluent supplements in the aPA ELISA. To measure cofactor activity in these animal sera, ELISA for aPA to anionic phospholipids were performed. Two aPA positive patient plasmas were selected for study; one with cofactordependent and one with cofactor‐independent aPA. Only four of the animal sera tested (bovine, pig, sheep, and cat) supported the cofactor‐dependent aPA in ELISA. The cofactor‐independent aPA was positive in the presence of each animal serum except bovine and rat. In order to determine whether these animal sera contain a β2GPI‐like molecule, Western blot analyses were performed. By using a polyclonal antiserum produced to human β2GPI, specific β2GPI‐like cross‐reactivity was observed with all animal sera except the chicken. In summary, cofactor activity in animal sera varied significantly; however, bovine and pig sera appear to allow optimal binding of cofactor dependent aPA. ©

ISSN:0887-8013    

DOI:10.1002/jcla.1860080310

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY

摘要:

AbstractThis paper is a study to identify the clinical significance of high‐molecular‐mass alkaline phosphatase (ALP:E:C.3.1.3.1.), ALP–lipoprotein–X complex (LP‐X) and intestinal variant ALP. We used cellulose acetate and agarose gels and techniques including wheat germ lectin, cetavlon‐diethyl ether, thermostatability, neuraminidase andL‐phenylalanine to improve the electrophoretic separation of the alkaline phosphatase isoenzymes. Patients' serum samples were electrophoresed from a diverse group of individuals ill with cholestasis, neoplastic disease metastatic to the liver, hepatocellular carcinoma, cirrhosis, diabetes mellitus, and chronic renal disease. Agarose gels provided better separation of ALP isoenzymes than cellulose acetate gels. The results also indicated that high‐molecular mass ALP is present in patient's serum in conditions associated with cholestasis especially caused by hepatic malignancy. High‐molecular mass ALP was frequently found to co‐exist with the liver isoenzyme and LP–XALP complex. The intestinal variant was identified in patients with malignancy, cirrhosis, chronic renal disease and diabetes mellitus. Intestinal ALP coexisted concomitantly with a variant intestinal ALP. Intestinal variant ALP is most likely composed of intestinal ALP attached to a cellular membrane‐binding domain, or may be an artifact produced by neuraminidase incubation

ISSN:0887-8013    

DOI:10.1002/jcla.1860080311

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1994

数据来源: WILEY