CONTENTS
Mini-Review
- Retrograde Transport of Acidic Fibroblast Growth Factor as a Part of the Growth Factor Signaling
ANTONI WIĘDŁOCHAAbstract. The plasma membrane represents an impermeable barrier to proteins and other macromolecules. However, certain exogenous proteins are able to cross cellular membranes and gain access to the cytosol. The best examples are bacterial and plant protein toxins, acting on intracellular targets. During last few years the number of known proteins possessing the capability to cross cellular membranes in the reverse direction and reach the nucleus has increased (acidic and basic fibroblast growth factor, interleukin 1, angiogenin, Schwannoma derived growth factor, homeoprotein Antennapedia, HIV-1 Tat protein are some examples). Here, the role of transport of exogenous acidic fibroblast growth factor to the nuclear location as a part of the growth factor signaling is discussed, and the current knowledge on this issue is reviewed.
Keywords: acidic fibroblast growth factor (aFGF); cellular membranes; transport of aFGF; tyrosine kinase.kinase.
Cytokines
- Immunomodulatory Activity of Conformationally Restricted Peptides Related to TGF-ß2 90 – 99 Sequence
ZBIGNIEW WIECZOREK, JACEK J. SŁOŃ and IGNACY Z. SIEMIONAbstract. Continuing our research on the immunomodulatory activity of the peptides related to the proteins of TGF-ß family, we have synthesized and investigated by Jerne’s plaque forming cell (PFC) test and delayed type hypersensitivity (DTH) test the following linear peptides and their cyclic (disulphide-bridged) analogues : Lys-Thr-Pro-Lys-Ile (Ia) and Cys-Lys-Thr-Pro-Lys-Ile-Cys (Ib), Tyr-Ile-Gly-Lys-Thr-Pro (IIIa) and Cys-Tyr-Ile-Gly-Lys-Thr-Pro-Cys (IIIb), Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile (IVa) and Cys-Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile-Cys (IVb), Tyr-Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile-Glu (Va) and Cys-Tyr-Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile-Glu-Cys (Vb). It has been found that the insertion of the sequence of Ia into the disulphide bridged Ib generates a product with enhanced immunosuppressive activity. The similar procedure applied to IIIa, IVa and Va produces cyclic peptides devoided of any activity or of a diminished activity in DTH test. The exchange of Pro residue of Ia D-Pro resulted in the analogue IIa with a preserved immunosuppressive activity. The disulphide-bridged peptide IIb, a cyclic analogue of IIa, exhibited the same activity as IIa in both: PFC and DTH tests.
Keywords: immunosuppression; immunopeptides; TGF-ß2 immunosuppressive sequences.uences.
- Colostrinine: a Proline-Rich Polypeptide from Ovine Colostrum Is a Modest Cytokine Inducer in Human Leukocytes
ANNA D. INGLOT, MARIA JANUSZ and JÓZEF LISOWSKIAbstract. A proline-rich polypeptide (PRP), now named colostrinine, molecular weight 18 000, was isolated from ovine colostrum and characterized by Janusz Lisowski et al. The nonapeptide (NP) which is an active fragment of PRP was obtained by chemical synthesis. In mice, PRP has many regulatory effects on the humoral and cellular immune response. The present paper describes PRP as a cytokine inducer. PRP at concentration of 1-100 m g/ml was found to induce production of interferon (IFN) and tumor necrosis factor (TNF) in human peripheral blood leukocytes and in whole blood cultures. The effects were dose related. The identified till now cytokines induced by PRP were IFN-g and TNF-a but many other cytokines may be stimulated also. NP was considerably less active as the cytokine inducer than the natural PRP. Two volunteers given orally once daily for two to three weeks 100 or 200 m g PRP in tablets were found to develop the tolerance to IFN induction and had the modified TNF response. Furthermore, the PRP-treated volunteers showed signs of psycho-stimulation. Taken together our observations suggest that ovine PRP is active in humans and may have therapeutic value as an immunostimulant and/or neurotropic cytokine.
Keywords: colostrinine; proline-rich polypeptide; interferon g ; tumor necrosis factor; human leukocytes; cytokine inducers; ovine colostrum.
- Immunological Status of Patients Subjected to Cardiac Surgery: Serum Levels of Interleukin 6 and Tumor Necrosis Factor a and the Ability of Peripheral Blood Mononuclear Cells to Proliferate and Produce these Cytokines in vitro
ANDRZEJ WŁASZCZYK, BARBARA ADAMIK, GRAŻYNA DUREK, ANDRZEJ KÜBLER and MICHAŁ ZIMECKIAbstract. The aim of this study was to monitor plasma interleukin 6 (IL-6) and tumor necrosis factor a (TNF-a ) levels in patients subjected to cardiac surgery under general anesthesia. In addition, proliferation of peripheral blood mononuclear cells (PBMC) to phytohemagglutinin (PHA) and the ability of these cells to secrete lipopolysaccharide-induced IL-6 and TNF-a during the observation period was investigated. IL-6 and TNF-a levels were measured using bioassays. We found that despite high variability in the postoperative response of the patients, characteristic kinetics in the appearance of the cytokines in plasma could be demonstrated. Most significant phenomenon was an increase of IL-6 level 1 day after operation associated with an inhibition of TNF-a concentration. Relatively high, preoperative concentrations of these cytokines were probably elicited by stress. Control, healthy donors, did not exhibit measurable levels of these cytokines. In terms of proliferative response of PBMC to PHA and of cytokine production in vitro, the patients could be classified into low and high responding. The proliferative response of PBMC from low responders was progressively increasing and the response of high responders did not exhibit meaningful changes throughout the observation period. PBMC from low producers of IL-6 showed also an increased ability to secrete this cytokine during the monitoring time while the cells from high producers yielded less IL-6 on the last day of the follow up. Regarding TNF-a production, PBMC from low responders reacted strongly by secretion of this cytokine on day 1 after surgery in contrast to high responders whose cells yielded less TNF-a after surgery. In majority of cases TNF-a production had a tendency to diminish on the last day of observation. We conclude that the changes in cytokine levels as well as an altered reactivity of PBMC in vitro reflect the immune response to the postoperative trauma and are desirable for a normal course of the immune response and wound healing.
Keywords: cardiac surgery; peripheral blood mononuclear cells; proliferation; interleukin 6; tumor necrosis factor a.
Immunochemistry
- Comparison of Serological Reactions of Rickettsiae-Infected Patients and Rabbit Anti-Proteus OX Antibodies with Proteus 0X2, 0X19 and OXK Lipopolysaccharides
KEN-ICHI AMANO, MACIEJ CEDZYŃSKI, ANNA S. SWIERZKO, KAORI KYOHNO and WIESŁAW KACAAbstract. The reactivity of anti-Rickettsiae human antibodies with Proteus OX cells is used as a presumptive rickettsial diseases diagnostic test Weil-Felix reaction. In presented studies we compare the reactivity of human anti-Rickettsiae and rabbit anti-Proteus antibodies with series of Proteus OX2, OX 19 and OXK lipopolysaccharides (LPS). Polyclonal rabbit anti-OX2, anti-OX19 and anti-OXK sera reacted only with the homologous LPS – OX2, OX19 and OXK, respectively. The antibodies of Japanese spotted fever patients were less specific and reacted with OX2 as well as OX19 LPS. The antibodies of scrub typhus patients recognized Proteus OXK LPS, only. The serological reactions of O-antigen of P. mirabilis S1959 indicated that this, previously serologically not classified, strain may belong to the OXK group. Bacteria, used in the studies, came from the American, Japanese and European strain collections. The series of OX2, OX19 and OXK LPS, isolated from these Proteus strains, presented pattern of electrophoretic mobility and serological reactivities specific for each of OX – group types.
Keywords: Proteus O-antigens serology; lipopolysaccharide, Weil-Felix test.il-Felix test.
- Alterations of Tamm-Horsfall Protein Immunoreactivity after Partial Desialylation and Deglycosylation
TERESA GRABSKA, TOMASZ BAGIŃSKI, ALEKSANDRA KUBICZ, MICHAŁ KOKOT, FRANCISZEK KOKOT, and JAN DUŁAWAAbstract. The role of the carbohydrate moiety of Tamm-Horsfall protein (THP) was investigated by studying the effect of partial enzymatic desialylation and deglycosylation on its immunoreactivity. In our experiments primarily sialic acid a 2,6 linked to galactose was split off, resulting in a 14% increase of immunoreactivity. It was increased up to 21 % by further removal of the sugar residues. We also found that complexes composed of THP and human serum albumin, immunoglobulin G or transferrin reacted with anti-THP antibodies. When these complexes were formed with desialylated or deglycosylated THP an essential increase of this reactivity (about 25%) occurred. Our studies indicate that modifications of the carbohydrate moiety of THP (which might occur in vivo by changes in the glycosylation process e.g. in pathologic conditions) lead to increased exposure of THP to the immune system.
Keywords: deglycosylation; desialylation; enzyme-linked immunoassay; immunoreactivity; Tamm-Horsfall protein.
- Selective Spectrophotometric Determination of Glucose and Fructose in the Presence of Aldoses Using Phenol-Acetone Reagent and Cerium(III) Chloride
ANDRZEJ GAMIAN, JANUSZ BORATYŃSKI, M. ANNA ŻAL and RENÉ ROYAbstract. Aqueous mixtures of glucose and fructose produce red solutions when treated with 2%(w/v) phenol in 5% (v/v) aqueous acetone in the presence of concentrated sulfuric acid. The color is stable for days, and the red chromophore has an absorbance maximum at 568 nm. When the concentration of phenol is raised to 25%, fructose, but not glucose, produces red solutions, allowing for the selective detection of ketoses. Two complementary methods have been developed to remove the interference of ketoses in solutions containing glucose. The first one relies on the selective reduction of ketoses with sodium borohydride in the presence of cerium(III) chloride prior to the addition of the phenol-acetone reagents. The second method is based on the differential specific determination of glucose using 2% versus 25% levels of phenol. The relative sensitivities of different sugars are also presented as well as the applicability of the methods using bacterial polysaccharides for immunochemical analyses. The quantitative determination of glucose or ketoses in the polysaccharides does not require hydrolysis prior to the estimation.
Keywords: glucose; fructose; sugar analysis; bioassay; glycoconjugate structure; spectrophotometry.trophotometry.
Experimental Therapy
- Treatment of Drug-Induced Agranulocytosis with Colony Stimulating Factors (G-CSF or GM-CSF)
JOANNA GÓRA-TYBOR, EUZEBIUSZ KRYKOWSKI and TADEUSZ ROBAKAbstract. The application of granulocyte-macrophage and granulocyte colony stimulating factors (GM-CSF and G-CSF has been progressively increased in the treatment of patients with agranulocytosis. The aim of our study was to compare the time of neutrophil recovery in patients with severe agranulocytosis treated with G-CSF or GM-CSF and the historical control group. We have studied 6 patients with agranulocytosis treated with stimulating factors and 7 patients in historical control group. Most of the patients have been exposed to thiamazole or non-steroid antiinflammatory drugs. Our results demonstrate that patients receiving colony stimulating factor have a significantly shorter period of recovery (the mean time 8.7 ± 1.98 days) than the historical control group (the mean time 11.0±2.24 days). We observed also a shorter time of antibioticotherapy and hospitalization in the group of patients treated with colony stimulating factor.
Keywords: drug-induced agranulocytosis; granulocyte colony-stimulating factor; granulocyte-macrophage colony-stimulating factor.factor.
- Interaction of 2-Chlorodeoxyadenosine in Combination either With Interferons or Recombinant Human Tumor Necrosis Factor a on Myeloid Progenitor Cells in vitro
ANNA KORYCKA and TADEUSZ ROBAKAbstract. The objective of the present study was to investigate the interactions of 2-chlorodeoxyadenosine (2-CdA) with interferon a or g (IFN-a , IFN-g ), as well as between 2-CdA and recombinant human tumor necrosis factor a (rhTNF-a ),0n the clonal growth of granulocyte-macrophage progenitor cells (CFU-GM) from patients with chronic myeloid leukemia (CML) and on clonogenic leukemic blasts (CFU-L) from acute myeloid leukemia (AML) patients. Progenitor cell culture in semisolid medium in vitro was applied and the percentage of colony growth inhibition was evaluated. The use of 2-CdA either with IFN-a or IFN-g and 2-CdA with TNF-a was found to inhibit, in a dose dependent manner, the growth of colonies formed by hematopoietic precursor cells from CML and AML patients as well as from normal individuals, with the greatest effect being observed after the use of 2-CdA and IFN-a at their highest concentrations.
Keywords: 2-chlorodeoxyadenosine; myeloid progenitor cells; recombinant human tumor necrosis factor a ; interferon a ; interferon g .
Various
- Psoriasis Vulgaris with the Early and Late Onset- HLA Phenotype Correlations
ANETA SZCZERKOWSKA-DOBOSZ, WALDEMAR PLACEK, ZOFIA SZCZERKOWSKA and JADWIGA ROSZKIEWICZAbstract. The purpose of the study was to classify psoriasis vulgaris basing on HLA phenotype and the age of onset of the disease. One hundred fifteen psoriatic patients were included into the study and divided into two groups: group I – with early onset ( < 40 years) and group II – with late onset ( 40 years). Each group was subclassified according to Cw6 antigen expression: IA-Cw6+ – early onset, IB-Cw6 – early onset, IIA-Cw6+ – late onset, IIB-Cw6 – late onset. HLA class I antigen typing was performed in each of 115 subjects using the microlymphocytotoxicity test. HLA class II antigens typing was also performed in 20 randomly selected patients by means of the two-step microlymphocytotoxicity test. The occurence Cw6, B 13, B 17 antigens was significantly increased in psoriatic groups in comparison with a control population. No difference between the groups was found with respect to clas II antigen frequency. Cw6, B 13 and B 17 were the most specific markers for psoriasis with the early onset, whereas B27, Cw2, B44 and Cw5 seemed to be associated with the late type of the disease.
Keywords: HLA and disease; psoriasis vulgaris; relative risk.e risk.
- Immunohistochemical Study in Breast Carcinoma. S100-Protein Positive Cells and Neuroendocrine Differentiation
ALBINA ZÓŁTOWSKA, GRAŻYNA MOSZKOWSKA, BARBARA ZAMORSKA and ANDRZEJ ROSZKIEWICZ
Abstract. In this study S100-protein positive cells were found in all 12 cases of benign dysplastic changes and in all of the investigated 53 cases. of breast carcinomas. These cells belong to interdigitating cells (IDC) and were located in a regular pattern in the basal cell layer of the ducts and alveolar nodules in benign lesions. However, in breast carcinomas S100-protein positive IDC were in various pattern of distribution; only in the stroma, in basal cell layer, between cancer cells and S100-protein positivity of IDC and epithelial cancer cells. This phenomenon suggests the antigenic modulation of cancer cells transferring them the property of S100-protein positive cells or it may be the consequence of fusion. Immunoreactivity for chromogranin A (ChgA) and synaptophysin (Syn) was found in epithelial cells in some cases of benign dysplastic changes. However, in some carcinoma cases ChgA and Syn were revealed in carcinoma cells, in myoepithelial cells, in some stromal mesenchymal cells and endothelial cells. Immunoreactivity to smooth muscle actin was shown in the endothelial and smooth muscle cells of the vessel’s wall, in myoepithelial cells and was also sporadic in carcinoma cells.
Keywords: S100-protein; interdigitating cells; chromogranin A; synaptophysin.synaptophysin.