Biomedical subjects

M Bodger

Publications and source records attributed to M Bodger.

5 recordsLinked to original sources

PubMed2001

Thrombopoietin induces histidine decarboxylase gene expression in c-mpl transfected UT7 cells.

The leukemic cell line UT7 is endowed with both megakaryocyte and basophil differentiation potential, as judged by its capacity to respond to PMA by displaying megakaryocytic and basophilic markers and to produce histamine by neosynthesis. Herein, we addressed the question whether the biological activities characteristic of basophil differentiation were still induced when c-mpl-transfected UT7 cells received a specific megakaryocytic differentiation signal delivered by thrombopoietin (TPO). Surprisingly, we found that histamine synthesis did effectively occur in response to the growth factor. This activity was not associated with megakaryopoiesis since it was not detected in megakaryocytes generated from CD34(+) cells cultured in the presence of TPO. Comparing different c-mpl-transfected cell lines, we found that the amount of histamine generated in response to TPO correlated with their responsiveness to PMA, but not with their level of c-mpl expression, thus revealing an intrinsic basophil differentiation potential. Both PMA- and TPO-induced histamine synthesis was reduced by PKC and MEKs inhibitors, indicating that the induction occurred through a common signalling pathway.

Basophils↗

PubMed1999

Modulation of histidine decarboxylase activity and cytokine synthesis in human leukemic cell lines: relationship with basophilic and/or megakaryocytic differentiation.

In the present study, we show that UT7D1 cells, derived from the pluripotent cell line UT7, express high levels of histidine decarboxylase (HDC) mRNA spontaneously. These cells conserve the ability to differentiate into megakaryocytes upon stimulation with PMA, while greatly increasing their HDC activity. We provide evidence that enhanced HDC activity reflects the basophil rather than the megakaryocytic differentiation potential of UT7DI cells. Indeed, in addition to HDC mRNA, they express spontaneously several other mRNA coding for molecules present in basophils (FcepsilonRI, CCR3, IL-4Ralpha, IL-5Ralpha). Furthermore, the basophil antigen Bsp-1 is displayed on the surface of some UT7D1 cells in response to PMA concomitantly with increased histamine synthesis and mRNA expression of typical basophil-derived cytokines (IL-6, IL-4, and IL-13). Nevertheless, PMA cannot sustain the differentiation of this lineage, because mRNAs for basophil markers gradually diminish during long-term culture, whereas molecules associated with the megakaryocytic lineage remain prominent. In support of the notion that HDC activity is not related with megakaryopoiesis, we show that PMA-induced CD41 expression and PDGF transcription occurs in the K562 cells, though neither HDC mRNA nor any known basophil marker are expressed in these conditions. In contrast, all these markers are expressed in the basophilic leukemia cell line KU812F. Interestingly, the megakaryocytic cell line HEL produces also substantial amounts of histamine and expresses FcepsilonRI, thus revealing its basophil differentiation potential. HEL as well as KU812F need not be stimulated with PMA to react with Bsp-1 mAb, suggesting that they are more engaged into the basophil differentiation scheme than UT7D1. Other leukemic cell lines unrelated to the megakaryocyte or basophil lineage, like HL60 and U937 do neither synthesize histamine nor express basophil markers before or after PMA stimulation. To our knowledge, this is the first evidence for a factor-dependent cell line with megakaryocyte/basophil bipotentiality with which early stages of basophil commitment can be analyzed.

Basophils↗

PubMed1990

Further characterization of surface membrane structures expressed on human basophils and mast cells.

Recently, we were able to establish the immunologic surface marker profile of human basophils and mast cells. In the present study, the characterization of these cell types was extended by the use of monoclonal antibodies (mAbs) to hemopoietic differentiation antigens. Basophils and mast cells were enriched by mAbs and complement from chronic myeloid leukemia blood (n = 5) and dispersed lung tissue (n = 4), respectively. A panel of 80 mAbs was tested for being reactive with purified cell populations using flow cytometry and/or a combined toluidine blue-immunofluorescence staining procedure. In addition to previous findings, basophils were found to react with mAbs directed against the 126-kilodalton dipeptidylpeptidase IV (CD26), platelet glycoprotein IIa (CD31), CD40 antigen known to share sequence homology with nerve growth factor receptor, leukosialin (CD43), CD44 antigen, the ICAM-1 antigen (CD54) and VIM2-reactive gangliosides involving the sialofucooligosaccharide sequence (CDw65L). Bsp-1 was found to be a specific marker for human basophils, whereas mast cells were not stained by this reagent. Basophils apparently lack CD22 antigen, gangliosides detected by CDw65 mAbs (except CDw65L) and CD71 antigen (transferrin receptor). Mast cells were found to express CD43 and CD44 antigen. In contrast, mast cells lack CD22, CD26, CD31, CD40 and CDw65 antigen. These results provide further evidence that both blood basophils and mast cells express a unique immunologic surface marker profile including binding sites for a variety of immunomodulating ligands and adhesion molecules.

Antibodies, Monoclonal↗

PubMed1984

Reactivity of the monoclonal antibody RFB1 in chronic B-cell leukaemias.

The monoclonal antibody RFB1, which reacts with early haemopoietic precursors in human bone marrow and peripheral T-lymphocytes, but not with pre-B cells and mature peripheral blood and marrow B lymphocytes, was tested in blood from 60 patients with chronic B cell leukaemias. All 37 cases of B chronic lymphocytic leukaemia (B-CLL) and 9 of hairy cell leukaemia (HCL) were positive. The antibody showed particularly strong reaction in HCL. On the other hand, RFB1 did not react with peripheral blood and B lymphocytes from 9 patients with non-Hodgkin lymphoma (NHL) in leukaemic phase and was negative or weakly expressed in 5 with prolymphocytic leukaemia (B-PLL). These findings may be significant for investigations on the cell origin of B-CLL and HCL. The reactivity of RFB1 was different from that of two anti-T1 monoclonal antibodies. The combined use of these reagents gave distinct patterns in B-CLL (RFB1+, T1+), HCL (RFB1++, T1-) and NHL and B-PLL (RFB1-/+/-, Tl-/+), suggesting they may be of value in the differential diagnosis of these B-lymphoproliferative disorders.

Antibodies, Monoclonal↗

PubMed1981

Delineation of human thymocyte differentiation pathways utilizing double-staining techniques with monoclonal antibodies.

Three new techniques have been introduced for the phenotypic analysis of human thymocytes and T cells. Monoclonal antibodies of different subclasses (i.e. IgG1 and IgG2) were used in various combinations and labelled with subclass-specific second layers conjugated to different fluorochromes. Tissue sections were studied with combinations of monoclonal and conventional reagents, and anti-mouse Ig-coated macrobead columns were used to separate antibody-tagged cells. With the help of these simple techniques, four stage of thymocyte differentiation (prothymocyte, cortical, intermediate and medullary cell types) can be described in detail and the microanatomical relation of T cell subsets with other cell types studied.

Antibodies, Monoclonal↗