CONTENTS
Reviews
- Stem Cell Transplantation: Past, Present and Future.
E. DONNALL THOMASAbstract. The early attempts at human allogeneic marrow transplants in the 1950’s and 1960’s were largely unsuccessful. The probability of success has improved steadily in the past two decades. Cure rates now range from 90% for non-malignant diseases transplanted early to 15% for patients with advanced leukemia. Most marrow transplants have involved an HLA matched sibling donor but, more recently, a matched unrelated volunteer marrow donor can be found for many patients without a family donor. Current research is focused on new preparative regimens for elimination of malignant cells, better prevention of graft-versus-host disease, and the use of hematopoietic growth factors and cytokines. Autologous transplants, which use the patient s own marrow, are increasing, particularly for breast cancer. The hematopoietic stem cells are responsible for marrow regeneration after a transplant. Suff cient numbers of stem cells for transplantation can now be obtained from the peripheral blood after mobilization of these cells by chemotherapy or hematopoietic growth factors. Transplants can also be achieved using stem cells obtained from cord blood at the time of delivery, tissue typed, and cryopreserved for later use. A variety of technological advances has reduced the hospitaliza- tion time for transplant patients with a corresponding saving in cost.
Keywords: bone marrow transplantation; stem cells; cord blood. blood.
- Role of Extracellular Matrix Proteins in Organ Transplantation.
JERZY W. KUPIEC-WEGLINSKI and ANDRZEJ GÓRSKIAbstract. Recent data indicate that the extracellular matrix (ECM) proteins can regulate the process of T cell activation.Lymphocytes express an array of surface integrin and non-integrin receptors (adhesion molecules) mediating those phenomena.Since ECM proteins accumulate in situ during allograft rejection,it is likely that such T cell :ECM interactions are relevant in the immunopathology of rejection.Adhesion molecules are also thought to affect the very early events between host leukocytes and vascular endothelium.Therefore,immuno- modulation of T cell interactions with the ECM proteins and endothelium may lead to the development of nov- el therapeutic strategies in clinical organ transplantation.
Keywords: organ transplantation; extracellular matrix; adhesion molecules; vascular endothelium.helium.
- Elastin Receptor and Cell-Matrix Interactions in Heart Transplant-Associated Arteriosclerosis.
ALEKSANDER HINEKAbstract. Vascular cells, as well as monocytes, neutrophils, and lymphocytes which may infiltrate vascular walls and tissues express a multifunctional 67 kD protein which also serves as a subunit of the cell surface „elastin receptor”. This protein differs structurally and functionally from other matrix adhesion molecules. Unlike the integrins or cadherins, it is not a transmembrane molecule, but can be immobilized on the cell surface by association with two other membrane-anchored proteins. Once expressed on the cell surface, it may mediate cell-matrix interaction in a calcium-independent manner. Unlike most integrins, which recognize the linear sequence on the matrix ligands (RGD), it recognizes the secondary structure of the matrix macromole- cules and binds to several non identical domains on ditferent matrix components, as long as they form the appropriate hydrophobic conformation. Similarly to the transmembrane selectins, the 67 kD protein has lectin-like properties with the galactosugars’ binding specificity. However, binding of galactosugar-bearing ligands interrupts its contacts with matrix proteins and displaces the 67 kD protein from the cell surface. Moreover, the 67 kD protein also serves as an intracellular chaperone which facilitates secretion of tropoelas- tin and asserrbly of elastic fibers. In this review I will address the role of this 67 kD protein in mechanisms of mutual interaction between vascular smooth muscle cells, infiltrating leukocytes, and several components of extracellular matrix during the development of heart-transplant associated arteriosclerosis.
Keywords: arteriosclerosis; growth factors; elastin; extracellular matrix; fibronectin; interleukins; MHC.s; MHC.
- Leukocyte Adhesion Defects.
AMOS ETZIONIAbstract. Studies of genetic deficiency syndromes have provided important insights into the molecular basis and biology of leukocyte emigration. Both animal and human deficiency syndromes have been described.
Keywords: genetic deficiency syndromes; leukocyte adhesion deficiency; adhesion molecules;animal models ; gene therapy.herapy.
Cellular Immunology
- Modified Cytotoxic T Lymphocyte Precursor Frequency Assay by Measuring Released Europium in a Time Resolved Fluorometer.
KHAWAJA HAQUE 1, CAROL TRUMAN 1, IAN DITTMER 2, GODFREY LAUNDY 1, PATRICIA DENNING-KENDALL 1,JILL HOWS 1, TERRY FEEST 2 and BENJAMIN BRADLEY 1Abstract. The frequency of cytotoxic T lymphocyte precursors (CTLpf) can be quantified by using the principle of limiting dilution analysis (LDA). Chromium 51 (SlCr) and europium (Eu) release assays are based on the measurement of marker release after lysis of targets by the effector cells. Although, slCr release has been widely used to quantify cell lysis since its introduction, it has several disadvantages such as handling and disposal of radioisotopes as well as health risk to personnel involved performing the assay. This situation has led us to adopt a non-radioactive cytotoxicity assay. After 7 days culture the PHA-stimulated targets are labeled with europium DTPA chelate. Lysis of labeled targets by effectors releases the Eu-DTPA complex in culture medium – a highly fluorescent substance. The amount of fluorescence can be measured in a time resolved fluorometer. We describe here some modifcations of the original protocol which include optimising IL-2 requirements, reduction of incubation times, addition of an extra spin before 37oC incubation, readjust- ment of target cells per volume of labeling buffer and other crucial parameters increasing the specificity and sensitivity of CTLpf assay. We are in agreement with others that the Eu-release assay is specific and re- producible. It can be used for the CTLpf estimation as well as other T cell and non-T cell cytotoxicity assays.
Keywords: europium-release; cytotoxicity; transplantation; fluorometry; recombinant interleukin 2; T lym- phocytes.ocytes.
- Stimulatory Effect of Ovine Colostrinine (a Proline-Rich Polypeptide) on Interferons and Tumor Necrosis Factor Production by Murine Resident Peritoneal Cells (Short Communication).
ZOFIA BLACH-OLSZEWSKA 1 and MARIA JANUSZ 2Abstract. We describe effects of ovine colostrinine (proline-rich polypeptide – PRP) isolated from ovine colostrum and nonapeptide fragment of PRP on interferon (IFN) and tumor necrosis factor (TNF) production by murine resident peritoneal cells (RPC). The cells from several mouse strains have been found to produce small amounts of IFN-ß and TNF-a constitutively. The colostrinine at concentrations of 1-100 ixg per one ml of cell suspension containing 1 x 106 RPC isolated from BALB/c mice, enhanced the IFN and TNF production by 3 – 30 folds. Upregulation of TNF and IFN production has been observed in the RPC cultures that produced spontaneously less than 16 units of the cytokines only. Synthetic nonapeptide fragment of the colostrinine (Val-Glu-Ser-Tyr-Val-Pro-Leu-Phe-Pro) at concentration of 1-100 gg/ml stimulated TNF syn- thesis but not IFN production. In 1996 Inglot et al. suggested that the colostrinines may be classifed as cytokines produced by the mammary gland of mammals. In this paper we have found that the ovine colost- rinine at low concentrations modulate the production of other cytokines (IFN-ß and TNF-a) in mouse cells that means that it may function in the cytokine network.
Keywords: ovine colostrinine; murine resident peritoneal cells; interferons; tumor necrosis factors.actors.
- Interleukin 12 and Direct Cytotoxicity of Spleen Lymphocytes to Listeria innocua-Phagocyting Syngeneic Macrophages in C57BL/6 and BALB/c Mice.
JACEK SZELIGA, DANUTA FIUK, MARIA WALENCKA and TERESA GOŚCICKAAbstract. Production of interleukin 12 (IL-12) during cytotoxic reaction of CS7BL/6 and BALB/c spleen lymphocytes or their subpopulations: T+B, T, CD4+ and CD8+ cells to L. innocua-phagocyting syngeneic macrophages was examined. The effector cell donors were untreated or L. innocua-infected. The number of surviving bacteria in phagocytes was tested and IL-12 level in culture supernatants of reacting cells was determined. CS7BL/6 mice, resistant to Listeria infection, were found to develop stronger cell cytotoxicity to bacteria-phagocyting syngeneic macrophages than BALB/c mice. The lymphocytes responsible for that pheno- menon were of CD8+ phenotype. IL-12 was produced only during nonspecific cytotoxic reaction of CD4+ and CD8+ T cells. It is suggested that the innate resistance of mice to Listeria is dependent on their ability to develop a specific cytotoxic reaction and IL-12 production.
Keywords: interleukin 12; cytotoxicity; phagocyting macrophages; Listeria.ges; Listeria.
- Sensitivity of Human Cutaneous Mast Cells to Anaphylactic and Nonimmunological Stimuli.
EWA BRZEZIIVSKA-BLASZCZYK1 and ANNA ZALEWSKA2Abstract. . Mast cells represent significant cellular element of the skin. It is now postulated that they play an important role in cutaneous homeostasis and are engaged in some pathological processes as well. The aim of our study was to examine sensitivity of human cutaneous mast cells to anaphylactic and anaphylactoid stimuli. The studies were performed on human cutaneous mast cells obtained from healthy skin by enzymatic dispersion technique. The mast cells were activated in vitro with anti-IgE, concanavalin A (Con A), compound 48/80, substance P (SP) or tumor necrosis factor a (TNF-a). We have noticed that skin mast cells were susceptible to Presentation the challenge with anti-IgE and Con A, and histamine release was dose- and time-dependent. In both cases histamine release was high (44.0±4.1% with anti-IgE at dilution I:500 and 20.1 ±2.4% with Con A in concent- ration 500 qg/ml). Cutaneous mast cells were challenged in a dose- and time-related fashion with compound 48/80, however histamine release was low (9.8±2.4%, at concentration of compound 48/80 – 100 ltg/ml). SP and TNF-a also activated mast cells but the magnitude of histamine release was not high (up to 7.1 ±0.9%, SP in concentration 10 4 M and 17.4 ± l.1 %, TNF-a in concentration 10 6 M) and maximal after 20 min reaction.
Keywords: cutaneous mast cells; anaphylactic histamine release; anaphylactoid histamine release; substance P; tumor necrosis factor aactor a
Experimental and Clinical Therapy
- Interferon-Inhibiting Activities in Sera of Patients with Lung Cancer (Short Communication).
KONSTANTINOS KARMANIOLAS, GEORGE TSANTAKIS and MARIA HAVREDAKIAbstract. A differential pattern of sera exhibiting interferon-inhibiting activity in cases of histologically confirmed lung cancer patients have been observed. The sera distribution was dependent on the clinical entity and the cell line specifďcity used measuring IFN-inhibition. Although either small cell lung cancer (SCLC) or non small cell lung cancer (NSCLC) sera exhibiting in a high percentage (80 and 75% respectively) such an activity, in any of the 3 lines tested, the ratio of sera exerting inhibition in each cell line was different. Such cell specific systems could be of prognostic/predicting value for effective therapeutic schedules of specific clinical entities.
Keywords: interferon-inhibiting activity; serum inhibitors; lung cancer patients.tients.
- Fffect of Bacterial Extract, IRS-I9, on the Concentration of Hydrogen Peroxide and Myeloperoxidase Activity in Nasal Washings of Patients with Chronic Bronchitis.
DARIUSZ NOWAK 1, MICHAŁ, PROZYIVSKI Z, TADEUSZ PIETRAS 1, ROBERT STOLAREK 1, PIOTR MAZERANT 3 and MICHAL LEDER 3Abstract. Twenty eight adult patients of both sexes with chronic bronchitis participated in an open study to determine the eh’ect of intranasal treatment with IRS-19, an immunomodulating agent, on the number of polymorphonuclear leukocytes (PMNL), HzOz concentration and myeloperoxidase (MPO) activity in nasal washings. The number of PMNL recovered from nasal spaces increased from 4460±3960 to 10490± 10950 cells/ml (p < 0.02) after two month administration of IRS-19. It was accompanied by 2.6- and 1.4-fold increase (p < 0.001) in MPO activity and HzOz concentration, respectively. However, no correlation was found betwe- en increments in these three variables. Since PMNL and MPO – HzOz – CI system are involved in the first line of defense against invading pathogens it is suggested that above mentioned changes may represent one among mechanisms leading to enhancement of antibacterial defence in the airways in response to treatment with IRS-19.
Keywords: chronic bronchitis; bacterial extract; IRS-19; nasal washings; myeloperoxidase; hydrogen peroxi- de; polymorphonuclear leukocytes.ar leukocytes.
- Urinary Gamma-Glutamyl Transferase Activity in Rats with Nonsteroidal Anti-Inflammatory Drug-Induced Nephrotoxicity (Short Communication).
SÜKRAN KOCAOGLU 1, AYSEN KARAN 1, TAYFUN BERKAN 2, GÜLÇIN BASDEMIR 3 and RIYAT AKPINAR 2Abstract. Excretion of urinary gamma-glutamyl transferase (GGT) was studied in rats following p.o. ap- plication of high doses (10 mg/kg/day) of indomethacin, diclofenac sodium or piroxicam for 28 days. Measure- ments of 24 h urinary GGT activity and urinary creatinine were carried out on 29th day. Histological examinations of kidneys were performed on day 29. The mean value for urinary GGT was found to be 0.77±0.05 U/mg creatinine (n =16) in the control group. The mean activities in the treated groups were as follows: 1.30 ± 0.15 U/mg creatinine (n =17, indomethacin); 1.22 ± 0.25 U/mg creatinine (n = 4, diclofenac); 1.54±0.39 U/mg creatinine (n = 5, piroxicam). The mean enzyme activities in indomethacin and piroxicam treated groups were significantly higher than in the control group (p < 0.02 and p < 0.03, respectively), while no significant dill’erence has been found between the group treated with diclofenac and control group (p 0.05). Histological examinations of renal tissues of indomethacin, piroxicam and diclofenac treated groups showed minimal glomerular abnormalities. Thus, determination of urinary GGT may be useful to investigate the nonsteroidal anti-inflammatory drugs (NSAIDs) related renal toxicity in rats.
Keywords: nephrotoxicity; gamma-glutamyl transferase; indomethacin; diclofenac; piroxicam.oxicam.
- Influence of Mesna on Urotoxic Effects of Selected Bromosubstituted Analogs of Ifosfamide.
HALINA KUŚNIERCZYK 1, LESZEK KONARSKI 1, PRZEMYSŁAW KOWALSKI2 and CZESŁAW RADZIKOWSKI 1Abstract. Bromofosfamides, the group of novel compounds closely related to ifosfamide, are currently in the stage of advanced preclinical evaluation. Ifosfamide, although itself the effective antineoplastic drug useful in situations which have proved refractory to cyclophosphamide therapy, has the side-effect toxicities caused by its metabolites that pose clinically a very real problem. One of their manifestations is the severe urinary tract toxicity which now could be adequately managed by conjunctive administration of mesna (sodium 2-mercap- toethane sulphonate). In this study we have compared the magnitude of urotoxic effects elicited by ifosfamide and two bromofosfamide compounds – racemate and S(-) isomer of chlorobromofosfamide (ClBrs) – selec- ted previously on the base of their superior antitumor activity in advanced animal tumor models. The urotoxic effects, expressed by the increase of urinary bladder weight and histopathologically defined organ wall edema, were estimated in healthy mice 24 h following single intraperitoneal or oral administrations of tested com- pounds which were applied in amounts equal to curative, sublethal or lethal doses. It was found that the expression of toxic effects revealed by both ClBrs was statistically significantly lower as compared to ifos- famide. Mesna coadministration prevented urotoxic effects almost completely in mice treated with ifosfamide or racemic ClBr. Somewhat lower efflcacy of uroprotection with mesna was observed in the case of S( – ) isomer of ClBr.
Keywords: ifosfamide; bromofosfamides; urotoxicity; mesna uroprotection.uroprotection.
- Vitamin C Inhibits Random Migration of Malignant Pleural Effusion Mononuclear Cells.
LJUBICA PROKIC 1, IVANKA VILIĆ l, LJILJANA VUCKOVIĆ-DEKIĆ 2, ZORA NESKOVIĆ-KONSTATINOVIĆ 2 and SNEZANA SUSNJAR 2Abstract. Although many epidemiological studies indicate protective effect of vitamin C against a variety of human malignancies its mechanism(s) of action is questionable. The presented results show that the part of its effect may be accomplished by mononuclear cells, as necessary participants in body defence. Namely, in a long-term in vitro assay we tested vitamin C influence on random migration ability of malignant pleural effusion mononuclears (PEM) obtained from breast cancer patients. Vitamin C in a dose- (50-500 g) and time-dependent (4-44 h) manner inhibited PEM motility, suggesting that immobilization of cells in situ may contribute to its beneficial effect in human cancers.
Keywords: malignant pleural effusion mononuclears; random migration; vitamin C.on; vitamin C.
Various
- Effect of Suramin on Human Interferon alpha Binding to Cell Receptors.
MARTYNA KANDEFER-SZERSZEŃ 1, KRISTINA RUUTH 2 and ERIK LUNDGREN 2Abstract. Radioiodinated human recombinant interferon a88 (l2sl-HuIFN-a88) binds to high affinity recep- tors of IFN sensitive Daudi cells (Burkitt lymphoma cell line). Suramin, a low molecular weight (1429), polyanionic compound at concentrations 105 -175 ItM completely abolished 125I-IFN-a88 binding to Daudi cells at low temperature (4oC). At 37oC, however, its effect was only partial and depended on incubation time. Suramin also dissociated IFN-a88-receptor complexes but, upon incubation of cells with IFN at 37C IFN-receptor complexes, became gradually less sensitive tu suramin action. Dissociation by suramin IFN-a88-receptor complexes prevented induction of (2-5)A synthetase activity and inhibited down-regula- tion of IFN receptors on Daudi cells. We suppose that the first reaction which represents IFN binding to the surface receptors is inhibited and dissociated by suramin, but when IFN is transferred to a tight activation complexes on the cell membrane or internalized, such complexes cannot be dissociated by suramin.
Keywords: interferon a receptors; suramin; (2-5)A synthetase.)A synthetase.
- Pathogenesis of Breast Carcinoma. Immunohistochemical Study.
ALBINA ŻÓŁTOWSKAAbstract. Synaptophysin and/or chromogranin A may be expressed in epithelial cells of normal and dysplastic mammary gland and in some cancer of the breast. This work indicates that some stromal cells form thin walled vascular channels and from their perivascular mesenchyma arise myoid-like cells, some with cross-like strations. These myoid-like cells have been stained with antibody to smooth muscle actin, rarely to desmin and sarcomeric actin as well as are strongly positive for synaptophysin and/or chromogranin A. From those cells arise cancer cells. Some cancer cells also expressed desmin, sarcomeric actin or smooth muscle actin. Because synaptophysin positive are neuromuscular endplates, it is a question whether the presence of synaptophysin in the progenitor of neoplastic cells of myogenic origin is due to the synaptic dysfunction in molecular mecha- nism of the epithelial-parenchyma and mesenchymal stroma interaction. The location of S100-protein positive dendritic cells distributed in regular pattern in suprabasal layer of the mammary gland indicate that these cells may arise from preadipocytes which take part in morphogenesis of the breast. All cases of the breast cancer demonstrated thymosin al, some of them also showed Hassall’s-like bodies, mucin secretion and minute calcification. Whether the interaction of epithelial cells and S100-protein positive dendritic cells in tissue other than the thymus has similar dynamic activity to that of Hassall’s bodies of the thymus remains to be determined. Both, epithelial cells of Hassall’s bodies and epithelial cells of the investigated breast are of myogenic origin. The above consideration suggests that the cells forming vascular channels are with a differen- tiation defect as they are created in altered stromal mesenchyma and the presence of thymic growth factors might induce tumor growth.
Keywords: breast cancer cells; synaptophysin; S100-protein; dendritic cells; preadipocytes; thymosin al.; thymosin al.
- Coincidence between Spontaneous Release of Interferon and Tumor Necrosis Factor by Colostral Leukocytes and the Production of a Colostrinine by Human Mammary Gland after Normal Delivery.
EGBERT PIASECKI l, ANNA D. INGLOT l, MAŁGORZATA WINIARSKA 3, KATARZYNA KRUKOWSKA l, MARIA,IANUSZ 2 and JÓZEF LISOWSKI 2
Abstract. Synaptophysin and/or chromogranin A may be expressed in epithelial cells of normal and dysplastic mammary gland and in some cancer of the breast. This work indicates that some stromal cells form thin walled vascular channels and from their perivascular mesenchyma arise myoid-like cells, some with cross-like strations. These myoid-like cells have been stained with antibody to smooth muscle actin, rarely to desmin and sarcomeric actin as well as are strongly positive for synaptophysin and/or chromogranin A. From those cells arise cancer cells. Some cancer cells also expressed desmin, sarcomeric actin or smooth muscle actin. Because synaptophysin positive are neuromuscular endplates, it is a question whether the presence of synaptophysin in the progenitor of neoplastic cells of myogenic origin is due to the synaptic dysfunction in molecular mecha- nism of the epithelial-parenchyma and mesenchymal stroma interaction. The location of S100-protein positive dendritic cells distributed in regular pattern in suprabasal layer of the mammary gland indicate that these cells may arise from preadipocytes which take part in morphogenesis of the breast. All cases of the breast cancer demonstrated thymosin al, some of them also showed Hassall’s-like bodies, mucin secretion and minute calcification. Whether the interaction of epithelial cells and S100-protein positive dendritic cells in tissue other than the thymus has similar dynamic activity to that of Hassall’s bodies of the thymus remains to be determined. Both, epithelial cells of Hassall’s bodies and epithelial cells of the investigated breast are of myogenic origin. The above consideration suggests that the cells forming vascular channels are with a differen- tiation defect as they are created in altered stromal mesenchyma and the presence of thymic growth factors might induce tumor growth.
Keywords: breast cancer cells; synaptophysin; S100-protein; dendritic cells; preadipocytes; thymosin al.; thymosin al.