[Human leukocyte differentiation antigens and CD classification]. / Lidské leukocytární antigeny z hlediska CD klasifikace.

The autor summarizes the information about human leukocyte antigens. Expression of membrane markers is included in accordance with CD (Cluster of Differentiation) classification. The CD molecules are used ubiquitously in human medical research, immunodiagnosis and treatment and also CD profiles are intended to be used by biologists, pathologists and clinicians generally. Profiles nearly 400 known leukocyte cell surface molecules are summarized in CD system (CD1-CD350). Most CD molecules are integral membrane proteins that have one or more pass through the plasma cell membranes. Knowing which regions of a protein are intracellular or extracellular is important in the selection of peptides for immunization, for the expression of domains of the protein, and to understand the interaction of the protein with other proteins. CD markers are great targets for diagnostic and research of different types of diseases as a potential treatment for a variety of tumors.

Lactosylceramide in lysosomal storage disorders: a comparative immunohistochemical and biochemical study.

Immunohistochemical studies of the presence of lactosylceramide (LacCer) in lysosomal storage disorders (LSDs) were done using anti-LacCer monoclonal antibody of the CDw 17 type (clone MG-2). No sign of an association between LacCer and the lysosomal system in normal cells was observed, except for histiocytes active in phagocytosis. A comparative study of a group of LSDs showed a general tendency for LacCer to increase in storage cells in Niemann-Pick disease type C (NPC), and types A and B, GM1 gangliosidosis, acid lipase deficiency, glycogen storage disease type II and mucopolysaccharidoses. LacCer accumulated in storage cells despite normal activity of relevant lysosomal degrading enzymes. The accumulation of LacCer displayed variability within storage cell populations, and was mostly expressed in neurons in NPC. An absence of the increase in LacCer in storage cells above control levels was seen in neuronal ceroid lipofuscinoses (neurons and cardiocytes) and in Fabry disease. Gaucher and Krabbe cells showed significantly lower levels, or even the absence, of LacCer compared with control macrophages. Results of immunohistochemistry were corroborated by semiquantitative lipid thin-layer chromatography (TLC). It is suggested that different associations of LacCer with the lysosomal storage process may reflect differences in glycosphingolipid turnover induced by the storage-compromised lysosomal/endosomal system.

The interaction and functional properties of leukocyte molecules of human leukemia/lymphoma cells.

The expression of 82 cell surface molecules was analysed on pathological cells from 52 patients with acute lymphoblastic leukemia (T and B), acute myeloid leukemia (FAB: M0, M1, M2, M3, M4, M5), chronic myeloid leukemia, chronic myelo-monocytic leukemia) and non Hodgkin's lymphoma (T and B types). We have selected patients with a high leukocyte count and percentage of blasts. Staining for membrane molecules was done by the immunofluorescence method and evaluated by flow cytometry. The findings indicate that expression of membrane molecules on pathological cells is quantitatively and qualitatively different in individual cases. The leukemia/lymphoma cells in their crude from represent the main phenotypes of normal haematopoietic cells, which reveal the great diversity of immunophenotypes within the main functional characterization of blood malignancies. The immunophenotype heterogeneity of leukemic cells has proved to be much greater than the match with existing classification criteria, this fact could raise the necessity for further functional evaluation and specification of immunophenotyping of the leukemia/lymphoma cells. The concept of explanation of pathogenesis and pathophysiology of different types of leukemia/lymphoma cells on the basis that they are derived from normal haematopoiesis must be accepted, because the number of membrane markers and their functional properties are correspondingly convincing.

[The CD nomenclature of leukocyte antigens]. / CD nomenklatura leukocytárních antigenu.

The author gives the basic information about human leucocyte antigens of haematopoietic cells. At present, the membrane markers (antigens) included into CD nomenclature are gradually detected, classified and ranged, many of them being well defined as to genetic determination, chemical structure and function.

[Hematopoiesis from the aspect of CD antigen expression]. / Hematopoéza z hlediska exprese CD antigenu.

The author summarized the expression of membrane markers (CD antigens) on haematopoietic cells. The proliferation and differentiation of cells in individual lineages from initial haematopoietic stem cell and their myeloid and lymphoid precursors is evaluated.

Occurrence of cytokine receptors on different lymphoid leukaemic cells.

We have studied the expression of cytokine receptors CD25 (IL-2R alpha, 55 kD), CD116 (hGM-CSFR, 145 kD), CD117 (CSFR, 145 kD), CD120a (TNFR, 55 kD), CD120b (TNFR, 75 kD), CD121a (IL-1R, type I, 80 kD), CDw123 (IL-3R), CD124 (IL-4R, 140 kD), CD126 (IL-6R, 80 kD), CD127 (IL-7R, 75 kD), CDw128 (IL-8R), CD130 (gp130 subunit), CDw131 (common beta), CD132 (IL-2Rgamma), CD134 (OX40) and also CD95 (Fas antigen) on the lymphoid leukaemic cells. Cells from peripheral blood or bone marrow of 24 patients with disorders in lymphoid lineage mostly included acute lymphoid leukaemias (with a high leukocyte count and percentage of blasts) were analysed for the expression of surface membrane molecules by the immunofluorescence method evaluated by flow cytometry. The findings indicate that some monoclonal antibodies have a reactivity against cytokine receptors of pathological cells in individual cases, but with very variable qualitative and quantitative expression (number copies/cell). The lymphoid leukaemic cells demonstrate unique cytokine receptor profiles, which reveal the great diversity of immunophenotypes within the main functional characterisation of T and B lymphoproliferative malignancies.

[Immunohistochemical detection of cytokine receptors on cryostat tissue sections]. / Imunohistochemická detekce cytokinových receptoru na kryostatových tkánových rezech.

We have studied tissue expression of the cytokine receptors using a high sensitivity biotin-streptavidin system on cryostat sections. We used a panel of monoclonal antibodies from the 6th International Workshop on Human Leukocyte Differentiation Antigens, namely CD25 (IL-2R alpha), CD95 (FAS antigen), CD116 (GM CSFR), CD117 (SCFR), CD120 alpha (TNFR I), CD120b (TNFR II), CD121a (IL-1R I), CDw123 (IL-3R), CD124 (IL-4R), CD126 (IL-6R), CD127 (IL-7R), CDw128 (IL-8R), CD130 (gpl130), CD131 (IL-3R), CD132 (IL-2R gamma), CD134 (OC-40), CD135 (FLT3/FLK2). Examined tissues (lymph nodes and spleens) were obtained from 12 patients with folicular non-Hodgkin's lymphoma, periferal T non-Hodgkin's lymphoma, B lymphoma, myeloma, Hodgkin's disease, two cases of T cell rich B-lymphoma, autoimmune haemolytic anemia and two cases of rudimentary trombocytopenic purpura. Our results indicate that immunohistological technology using native tissues on cryostat sections, monoclonal antibodies and the visualisation with biotin-streptavidin is a particularly suitable supplementary staining procedure for detection of the cytokine receptors in tissues.

[Chemokines and chemokine receptors. Review article]. / Chemokiny a chemokinové receptory. (Prehledový clánek).

Chemokines form a new superfamily of small glycoproteins. They are key molecules that activate and direct the migration of different types of leukocytes from the blood stream into sites of infection and inflammation. In addition to this role certain chemokines have been reported to act on different types of cells (e.g. hematopoietic progenitor cells, dendritic cells, fibroblasts, keratinocytes). Other of them also play a role in wound healing, in angiogenesis and in viral infections. These molecules have a high degree of amino acid sequence homology and they have four conserved cysteins forming two essential disulphide bonds. They are divided into four classes (families) depending on the position of the first two cystein residues. Chemokines mediate their proinflammatory effects by binding to a variety of specific receptors, belonging to the G protein-coupled superfamily of seven-transmembrane (serpentine) receptors. Some of this receptors serve as coreceptors for HIV-viruses, some of them could be expressed as markers preferentially in Th1 or Th2 subpopulations. This paper summarizes data on chemokines and their receptors, target cells and production in physiological and pathological conditions.

Aberrant markers expression in T- and B-lymphoid and myeloid leukemia cells of different differentiation stages.

The aim of the study was to ascertain if in T acute lymphoblastic leukemia (T-ALL), B acute lymphoblastic leukemia (B-ALL) and acute myeloid leukemia (AML) of different differentiation stages the coexistence of aberrant markers correlate with the degree of leukemic blasts maturation. We evaluated the results of surface and intracellular markers in 42 T-ALL, 86 B-ALL and 71 AML cases. A large panel of monoclonal antibodies (MoAbs) against T-cell, B-cell, myeloid cell and non-lineage specific structures has been used. Patients had dual-color flow cytometric immunophenotyping performed by FACStar flow cytometer. The correct immunological diagnosis of followed new cases before any treatment has been performed and simultaneously the presence of atypical/aberrant phenotypes has been studied and correlated with leukemia cells differentiation stage. A great deal of T-ALL and AML, in opposite to B-ALL cases, revealed a high proportion of atypical phenotypes (55, 75 and 36%, respectively), which are absent in nonleukemic cells. We found out that these atypical phenotypes were present in T-ALL, AML (not clearly in B-ALL) through all differentiation stages and so we obtained an evidence that they might represent an abnormal/atypical rather than an immature phenotype, as it was postulated till now by several authors.

[The Th1/Th2 paradigm. The role of the Th1 and Th2 lymphocyte subpopulations in the regulation of immune processes]. / Th1/Th2 paradigma. Role Th1 a Th2 lymfocytárních subpopulací v regulaci imunitních deju.

A short report on the interaction of Th1 and Th2 lymphocyte subpopulations in the regulation of immune processes is reviewed. Th1 and Th2 subsets have been characterized on the basis of cytokines they secrete and the immune functions they mediate and also on the basis of cytokine and chemokine receptor expression. The clinical and diagnostic influence of immune processes in pathological stages is also mentioned.

[Interactions of cytokines with their receptors in the regulation of hematopoiesis, clinical and diagnostic applications]. / Interakce cytokínu s jejich receptory v regulaci hematopoézy, klinické a diagnostické aplikace.

A short review on the interaction of hematopoietic growth factors with their receptors in the regulation of hematopoiesis. The introductory notes on the nature of cytokines and cytokine receptors, their biological and physicochemical properties as well as clinical and diagnostic consequences are mentioned.

Phenotypic heterogeneity and aberrant markers expression in T-cell leukemia.

For exact determination of lineage assessment there is a need of surface membrane and intracellular (cytoplasmic and nuclear) immunophenotyping performed by flow cytometry. We evaluated in detail the results of surface and intracellular immunophenotyping of 34 T-ALL cases. The great heterogeneity of T-cell differentiation markers has been observed which did not allow relevant subclassification of T-ALL according to the existing subclassification schemes and the proposed three-stage model of physiological T-cell differentiation. Therefore, a simplified classification based on the CD3 marker expression either on cell membrane or in cytoplasm has been created with allocation of T-ALL into two main phenotypic groups. From 34 in detail examined T-ALL cases a great deal-27 (79%) belonged to an immature phenotype (Stage I) and only 7 (21%) expressed more mature phenotype (Stage II). Simultaneously the presence of atypical/aberrant T-cell phenotypes has been studied. We showed that in T-ALL it was possible to specify some cases with leukemia-associated phenotype with coexistence of atypical markers which are absent in nonleukemic cells. In a majority of cases early B-lineage marker (CD10) and in a smaller proportion of them non-lineage associated marker (CD34) were observed. Myeloid marker CD13 was observed in one case of the immature T-ALL, together with CD10 and CD34. As these atypical markers were present through all differentiation stages of T-ALL we obtained a strong evidence that they might represent an abnormal rather than an immature phenotype. The prognostic significance of T-ALL subtypes and aberrant markers coexpression have been discussed. Simultaneously it was shown that quantitative immunofluorescence could provide an additional important diagnostic marker also in T-ALL cases.

[Progression of multiple myeloma during treatment with recombinant G-CSF and absence of G-CSF and IL-6 cell surface receptors on malignant cells]. / Progrese mnohocetného myelomu pri lécbe rekombinantním G-CSF, s neprítomností receptoru pro G-CSF a IL-6 na povrchu maligních bunek.

Recombinant human hematopoietic growth factors are widely used in the treatment of multiple myeloma (MM) especially due to the increasing role of autologous blood stem cell transplantation (ABSCT). We report a patient with MM in whom rapid extramedullary progression of disease was observed during stem cell mobilization with G-CSF. In 56-year-old man with relapsing IgG lambda MM myeloablative therapy with ABSCT was planned 2 years after diagnosis. G-CSF is increasing doses was used for mobilization. Ten days after the start of G-CSF therapy 2 extramedullary (subcutaneous) myeloma infiltrates appeared. For the second mobilization high dose cyclophosphamide and VP-16 with subsequent G-CSF was used. During the time of chemotherapy tumour infiltrates disappeared, however, after one week of G-CSF treatment rapid progression of disease with the formation of multiple extramedullary infiltrates occurred and the patient died in June 1996. Small pieces of subcutaneous tumour infiltrates were removed at autopsy and immediately frozen in liquid nitrogen. Using the panel of specific antibodies the expression of cytokine receptors (IL-1, 2, 3, 6, 7, 8, 10, SCF, gp130, G-CSF, GM-CSF, EPO) and Fas, Pgp and 24-34 kD multidrug resistance-associated protein were examined. However, no expression of cytokine receptors on tumor cells was found. On the contrary, high positivity of surface MDR associated proteins was observed.

Expression of cytokine receptors on different myeloid leukemic cells.

We have studied the expression of cytokine receptors CD25 (IL-2Ra,55kD), CD116 (hGM-CSRF,145kD), CD117 (CSFR,145kD), CD120a (TNFR,55kD), CD120b (TNFR,75kD), CD121a (IL-1R, type I, 80kD), CD123 (IL-3R), CD124 (IL-4R, 140kD), CD126 (IL-6R, 80kD), CDw127 (IL-7R, 75kD), CDw128 (IL-8R), CD130 (gp130 subunit), CD131 (common beta), CD134 (OX40) and also CD95 (Fas antigen) on the myeloid leukemic cells. Cells from peripheral blood or bone marrow of 30 patients with disorders in myeloid lineage included mostly acute myeloid leukemias (with high leukocyte count and percentage of blasts) were analyzed for the expression of surface membrane molecules by indirect immunofluorescence method evaluated by flow cytometry. The findings indicate that some monoclonal antibodies have a reactivity against cytokine receptors of pathological cells in individual cases, but with very variable qualitative and quantitative (number copies/cell) expression (preliminary results). The leukemic cells demonstrate unique cytokine receptor profiles, which reveal the great diversity of immunophenotypes within the main functional characterization of blood malignancies. The immunophenotype heterogeneity of leukemic cells has proved to be much greater than to match with existing classification criteria. This fact could raise the necessity of further evaluation and specification of cytokine markers of the myeloid acute leukemias. On the other hand, detection of cytokine receptors on the leukemia cells is important for cytokine therapy.

[Presence of CD markers on hematopoietic cells]. / Výskyt CD znaku na hematopoetických bunkách.

The author summarizes the expression of membrane markers of haematopoietic cells. The proliferation and differentiation of cells in individual lineages as well as immunophenotype mozaik of membrane molecules included into CD nomenclature is evaluated.

Characterization of platelet antigen for CD45RA monoclonal antibodies.

CD45RA monoclonal antibodies recognize the higher molecular weight isoforms (220 and 205 kDa) of leukocyte common antigen family (CD45), which are typically expressed on B cells and unstimulated T cells. We have found that there are at least three distinct CD45RA monoclonal antibodies which react with platelet 42 kDa (P42) intracellular protein antigen, which seems to be different from any to date described platelet proteins with similar molecular weight. This platelet antigen is a single chain protein, very likely not a glykoprotein, with isoelectrical point between 6.8 and 7.5. P42 does not seem to be a membrane protein and is not associated with platelet cytoskeleton. Results of immunofluorescence assay suggest that P42 may be redistributed to platelet surface after platelet aggregation.