CONTENTS
Mini-Review
- Alterations in Glycoconjugates Related to the Transformation Grade of Human Uroepithelial Cell Lines
MACIEJ UGORSKI and DANUTA DUŚAbstract. To study the changes in carbohydrate expression associated with the progression of malignant phenotype, we have used the series of human urinary bladder epithelial cell lines. Non-tumorigenic and non-invasive cell lines, with infinite life span, were defined as transformation grade II (TGr II): Cell lines producing tumors in nude mice and invading fragments of embryonic chick hearts in vitro were defined as TGr III. In the present review the results obtained from our immunochemical, biochemical and structural studies on glycosphingolipids and glycoproteins of 10 human urothelial cell lines representing both grades of transfor- mation are summarized and discussed in relation to the literature data. TGr III cell lines are characterized by decreased level of the GMZ ganglioside, presence of sialyl-Lea ganglioside, increased amount of more highly branched, tri- and tetraantennary N-acetyllactosamine-type glycans and higher number of binding sites for peanut lectin and wheat germ agglutinin lectin, as compared to non-tumorigenic and non-invasive TGr II cell lines.
Keywords: human uroepithelial cell lines; transformation grade; glycoconjugates.ugates.
- Mouse Monoclonal IgA Antibodies Lack Interchain Disulfide Bonds
HUBERT KROTKIEWSKI, ANNA LASKOWSKA and BOŻENA KROTKIEWSKAAbstract. A mouse monoclonal IgA antibody (A008), anti-blood group A antigen, was shown to dissociate into heavy (H) and light (L) polypeptide chains under non-reducing conditions. The dissociation occurred in the presence of 02% sodium dodecyl sulphate (2 h at room temperature) or at elevated temperature (1 h at 70C). The free H and L polypeptide chains could be separated in SDS-polyacrylamide gel electrophoresis or by gel filtration in the presence of SDS. The dissociation of IgA (A008) antibody into heavy and light polypeptide chains was reversible, since the fractions containing gel filtration-isolated chains, after removing the detergent, pooling together and incubation at 4oC created again the immunoglobulin molecules with activity close to the native value. Similarly, the same IgA antibody heated at 70oC for 1 h recovered its antibody activity after keeping at 4″C. Three other mouse monoclonal IgA antibodies showed the same ability to dissociate into heavy and light polypeptide chains after exposure to SDS or elevated temperature, which suggests that this outstanding property is a chracteristic feature of the mouse monoclonal IgA antibodies and it comes out from the lack of H-H, L-L and H-L interchain disulfide bonds in these molecules.
Keywords: immunoglobulin polypeptide chains; non-reducing conditions; H-H dimers; disulfide bonds; re-association.iation.
- Human Serum Immunoglobulins Binding to Synthetic Fragments of Human Chorionic Gonadotropin
IWONA E. KOCHANOWSKA and APOLINARY SZEWCZUKAbstract. Human sera obtained from healthy individuals, patients with cancer diseases, infertile and pregnant women were investigated for the presence of immunoglobulins (Ig) binding to synthetic fragments correspon- ding to sequence of human chorionic gonadotropin (hCG). Four synthetic peptides corresponding to some regions of (hCG and ßhCG subunits, coupled to polystyrene beads and 16 peptides overlapping sequences of (- and ßhCG subunits, synthesized on polyethylene pins, were used. IgG and IgM bound to hCG-synthetic peptides were assayed using goat or mouse monoclonal antibodies anti-human IgG labelled with horseradish peroxidase. Most of the investigated sera contained IgG which bound to examined synthetic peptides, but obtained results were dependent on serum donors. Some relationships between results obtained with peptides synthesized on modified pins according to Geysen et al. method and corresponding peptides synthesized by Merrifield method and coupled to beads were noted.
Keywords: ELISA; human chorionic gonadotropin (hCG); IgG and IgM binding to hCG fragments; synthetic peptides of hCG sequence.quence.
- Stability of Human Lutropin Studied with Immunoenzymatic Technique
EWA KUROWSKA, IWONA E. KOCHANOWSKA, MARIANNA KUROPATWA, .IANUSZ BORATYŃSKI and APOLINARY SZEWCZUKAbstract. Characteristics of two monoclonal antibodies against human lutropin and their use in enzyme immunoassays of the hormone are presented. Diluted solution of pituitary lutropin was unstable in buffered saline and could be stabilized with some proteins. Concentration of the lutropin in woman urine, collected during „lutropin-peak”, was markedly increased during long time storage at 5` C or at room temperature. Using HPLC it was demonstrated that pituitary lutropin, normal urine lutropin and „increased” urinary lutropin were eluted from ion exchange column almost with the same retention time.
Keywords: human lutropin; human luteinizing hormone (hLH); pituitary hLH, urine hLH, stability of hLH.of hLH.
Clinical Immunology
- Comparison of the Influence of Stress Exercise Test and Transesophageal Cardiac Pacing on Polymorphonuclear Neutrophils Functions
HENRYK WYSOCKI, ANDRZEJ MINCZYKOWSKI, ANDRZEJ WYKRETOWICZ, GRZEGORZ JUNICZAK and JULIUSZ SMIELECKIAbstract. Peripheral polymorphonuclear neutrophils (PMN) number, percent of PMN bearing IgG Fc recep- tors as well as PMN adherence were evaluated in 29 patients submitted to an exercise test. The peripheral PMN count significantly increased at the maximal work load. The increase in number of neutrophils bearing IgG Fc receptors was also noticed at that point, while PMN adherence to nylon wool columns did not change significantly. Nine subjects were additionally submitted to diagnostic transesophageal atrial pacing with a rate similar to maximal heart rate observed during the exercise test. It was revealed that atrial pacing in these patients had no influence on the peripheral PMN count, PMN adherence as well as number of neutrophils bearing IgG Fc receptors. We conclude, that granulocytosis observed during exercise can not be sollely attributed to the increased heart rate and increased cardiac output but other mechanisms like muscle work play a significant role in this process.
Keywords: stress; neutrophils; exercise test; cardiac pacing.pacing.
- Autologous Peripheral Blood Stem and Progenitor Cells Transplantation as a Valid Treatment Approach in Recurrent, Refractory Lymphoma
JERZY HOŁOWIECKI, JERZY WOJNAR, MIROSŁAW MARKIEWICZ, ANNA WACŁAWIK, KRYSTYNA JAGODA, HANNA KAMINSKA, ŁUCJA KACHEL, MARIA WOJCIECHOWSKA and DARIUSZ KATAAbstract. Autologous peripheral blood stem cell transplantation (APBSCT) is used similarly to autologous bone marrow transplantation (ABMT) to reconstitute bone marrow following myeloablative therapy in pa- tients with proliferative diseases of the blood. Eight patients with recurrent and refractory lymphoma (3 HD, 4 NH L) and multiple myeloma aged 17 – 55 were included into the study. Peripheral blood stem cells following their prior mobilisation with cyclophosphamide 4-7 g/m2 and/or G-CFS or Dexa- -BEAM + G-CSF were collected by subsequent leukaphereses on Fenwal CS3000. Nucleated cells were sepa- rated by sedimentation, cryopreserved in a programmed freezer and then stored at -196oC. Bone marrow has been additionally collected in one patient. Conditioning treatment prior to transplantation consisted of BCNU, etoposide and cyclophosphamide (CBV) in lymphomas and melphalan in multiple myeloma. Collec- ted material with mean cellularity 5.52 x l Os/kg and mean CD34+ contents 627 x 10(6/kg was reinfused by central line. G-CSF was given in 5 patients to hasten the bone marrow recovery. All patients fully recovered and left hospital on average 35.5 days following transplantation. No signs of relapse were seen throughout the observation period (mean 349.5 days). Neutrophils > 0.5 G/I were obtained on day +20, > 1.0 G/1 on day 30, platelets > 50 G/1 on day 29, > 100 G/1 on day 53, reticulocytes ) 0.015 on day 30, erythrocytes transfusions were needed up to day 39. Presented outcomes together with other reports indicate, that APBSCT is a highly efficient way to rescue repeatedly relapsing patients with proliferative diseases of the lymphatic systems, even those presenting with changes in the bone marrow (neoplasmatic infiltrate, hypoplasia or fibrosis).
Keywords: autologous peripheral blood stem cell transplantation; cytokines; malignant lymphomasmphomas
- Influence of Gold Salts Treatment on the Serum Concentration of IL-1 and IL-2 in Patients with Rheumatoid Arthritis
JOLANTA LEWANDOWICZ, BOŻENA DĄBKOWSKA and DARIUSZ NOWAKAbstract. Eighteen subjects (2 males, 16 females) with rheumatoid arthritis were treated with gold salts for 6 months. The statistically significant decrease of IL-1 and IL-2 levels during the treatment was observed. The mean serum concentrations of IL-1 and IL-2 decreased from 41.0± 11.5 to 28.6±7.3 (p < 0.001) and 98.3 ± 57.6 to 29.3 ± 20.5 fmol/ml (p < 0.001 ), respectively. This decrease was accompanied by a decline of serum concentration of acute phase proteins and immunoglobulins. These results can explain a strong an- ti-inflammatory action of gold salts in patients with rheumatoid arthritis.
Keywords: interleukins; gold salts; rheumatoid arthritis; non-steroid anti-inflammatory drugs. drugs.
Immunotherapy
- Synthesis and Immunological Activity of Mono- and Disubstituted Derivatives of 2,5-Dimercapto-1,3,4-Thiodiazoles
STEFANIA H. KOWALCZYK-BRONISZ, JULIAN POMORSKI, BARBARA BUBAK, MACIEJ PREGIEL and BARBARA BŁASZCZYKAbstract. It is presented that the newly synthesized mono- and disubstituted 2,5-dimercapto-1,3,4-thiodiazole derivatives exert immunotropic activity. This effect was evaluated by measuring plaque forming cell number, circulating immunoglobulins level, autologous rosette number, mitogenic activity and by performing popliteal lymph node assay. It is shown that intensity of immunotropic activity depends on chemical structure: mono- substituted derivatives had a strong suppressive effect, disubstituted compounds showed the variable activity. One compound of this group exerted a stimulatory effect. The tested compounds were active in popliteal lymph node assay and had no mitogenic activity.
Keywords: plaque forming cells; immunoglobulins; autologous rosettes; mitogenic activity; popliteal lymph node assay. assay.
- Lactoferrin Lowers the Incidence of Positive Coombs’ Test in New Zealand Black Mice
MICHAŁ ZIMECKI, ZBIGNIEW WIECZOEK, JOEL MAZURIER and GENËVIEVE SPIKAbstract. The results presented in this communication show that New Zealand Black (NZB) mice treated for a prolonged period with bovine lactoferrin (BLF) exhibit a decreased frequency of positive Coombs’ reaction. This effect was dose dependent and best pronounced at a dose of 50 µg/dose. We found, in addition, that incubation of peritoneal cells with BLF resulted in a decreased number of cells recognising Hb antigen on autologous erythrocytes. The data indicated that lactoferrin may be of therapeutical value in treatment of autoimmune disorders.
Keywords: lactoferrin; New Zealand Black mice; autoimmune hemolytic anemia.anemia.
- Influence of Lys-Residue Deletion on the Immunomodulatory Activity of Peptides Related to Immunosuppressive Region of Lactoferrin
JACEK I. SŁOŃ, IGNACY Z. SIEMION, MICHAŁ ZIMECKI and ZBIGNIEW WIECZOREKAbstract. The effects of Lys residue deletion from peptides, related to immunosuppressive regions of lactoferrin (LF), were studied in the humoral and cellular immune responses to sheep erythrocytes (SRBC) in mice. We found that the deletion of Lys residue, vicinal to Pro, influences mostly the immunomodulatory activity of short fragments (tetra-, and pentapeptides) of immunosuppressive 231-245 mini-domain of LF. The activity of longer peptides, related to this mini-domain, is influenced to much lower extent by Lys removal. In particular, the tetrapeptide derived from Arg-Lys-Pro-Val-Asp sequence by deletion of Lys, exhibits strongly reduced immunosuppressive activity. On the contrary, the similar change within Arg-Lys-Pro-Val-Thr sequen- ce, derived of 575-589 loop of LF, produces a tetrapeptide Arg-Pro-Val-Thr, that is active as immunosupp- ressor both in humoral and cellular immune response. A quite strong immunosuppressive activity of tripeptide Lys-Arg-Pro is also worth consideration, especially when compared with the loss of activity observed for Thr-Arg-Pro tripeptide. The data presented herein indicate that exclusion of Lys residue from the LF-derived immunomodulatory peptides differentially alters their effects on the immune response to SRBC in mice.
Keywords: lactoferrin; immunoregulatory fragments of; lysine.lysine.
Various
- Lymphocyte Mitogen-Induced Proliferation in Patients with Allergic Rhinitis
JOANNA CHOROSTOWSKA-WYNIMKO, DANUTA KLENIEWSKA, IRENA SOKOLNICKA, EWA ROGALA and EWA SKOPIŃSKA-RÓŻEWSKAAbstract. Lymphocytes play a central regulatory role in mechanisms contributing to impairied function of immune system in atopy. The aim of our study was evaluate the mitogen-induced proliferation of lymphocytes in a group of asymptomatic, seasonal allergic rhinitis patients. A highly significant lower mitogen-induced proliferation and, in contrast to other studies, significantly lower background proliferative activity of lym- phocytes were found in the atopic persons, comparing to the controls. We concluded that the decreased mitogen-induced proliferation of lymphocytes observed in allergic patients reflects abnormal T cell function, which is due to the atopic status, and not only as it was believed to the antigen-induced lymphocyte activation.
Keywords: lymphocyte mitogen-induced proliferation; allergic rhinitis.initis.
- Effect of Domperidone-Induced Hyperprolactinemia on Selected Immune Parameters in Healthy Women
JOZEF ROVENSKÝ, MILAN BUC, ZDENEK LOJDA, MARIE RUZICKOVÁ, STANISLAVA BLAZÍCKOVA, LŮBICA RAUOVÁ, TIMOTEJ MIŚTINA, MILAN VIGAŚ and VLADIMÍR LAĆKOVIĆAbstract. Domperidone, anti-emetic drug, given to healthy female volunteers, induced an elevation of plasma prolactin (PRL) concentration with the peak in 1-4 h. The release of prolactin had a transient stimulating effect on theophylline sensitive T lymphocytes and on concanavalin A induced mitogenic activity, suggesting an enhanced activity of T suppressor lymphocytes. The relative number of CD4+ lymphocytes decreased markedly one hour after domperidone administration and returned to normal values within 2 h (that means 3 h after taking the drug). The number of lymphocytes positive for dipeptidyl peptidase IV exhibited similar transient increase and normalization of activity. No change was observed in the number of CD8+ lym- phocytes. The production of interferon by leukocytes treated with Newcastle disease virus was found to be significantly increased 2 h after domperidone administration. The results suggest that prolactin can selectively stimulate some functions of cellular immunity as well as the release of cytokines (IFN). The present study may contribute to the understanding of the role of the immune system in endogenous hyperprolactinemia.
Keywords: domperidone; hyperprolactinemia; cellular immunity.munity.
- Cell Migration between Graft and Host – an Analysis with Monoclonal Antibodies after Allogeneic Rat Kidney Transplantation
MACIEJ KRZYMAŃSKI, ANDRZEJ OKO, ANA MARIA WAAGA,JAN ŻEROMSKI, KARIN ULRICHS and WOLFGANG MÜLLER-RUCHHOLTZ
Abstract. In order to analyse migration patterns of donor MHC class II cells out of transplanted kidney and accumulation of host cells within the graft, immunomorphological studies were performed using monoclonal antibodies in rat allogeneic kidney transplantation model. To answer the question of how many donor cells migrate out of the renal cortex MRC 0 x 3 monoclonal antibody (MoAb) against LEW MHC class II antigens was used. In the grafts explanted after 4, 24 48 and 73 h, a slow reduction of donor class II cells was observed and some areas in cortex showed only very few, if any, donor cells. At the same time, starting from day 2 after transplantation accumulation of donor cells was found in perivascular spaces. Spleen sections stained at 24, 48, 72 and 96 h after transplantation revealed donor cells present in recipient’s spleen. They were detected up to day 3 after surgery. Their numbers, however, decreased after day 2. After 2 and 3 days, accumulations of recipient’s cells between tubules were detected. It was found that many cells in infiltrations were stained with anti-T lymphocyte MoAb. Expression of class II antigen on rat kidney cells increases significantly from the day 4 after transplantation.
Keywords: kidney transplantation; immunomorphology; monoclonal antibodies; cell migration; experimental model. model.