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Biomedical subjects

M J Truong

Publications and source records attributed to M J Truong.

6 recordsLinked to original sources

Decreased expression of eosinophil peroxidase and major basic protein messenger RNAs during eosinophil maturation.

We evaluated the levels of mRNAs encoding cationic proteins in peripheral blood eosinophils (PBE) purified from patients with eosinophilia and in eosinophils differentiated from cord blood cells (CBC) by culture with recombinant human interleukin-3 (rhIL-3), rhGM-CSF, and rhIL-5. Messenger RNAs encoding eosinophil peroxidase (EPO), major basic protein (MBP), eosinophil-derived neurotoxin (EDN), and eosinophil cationic protein (ECP) were detected by Northern blot hybridization with the respective specific oligonucleotide probes. In mature PBE, MBP mRNA appeared to be absent, whereas EPO mRNA was barely detectable in only 5 of the 19 patients. In contrast, EDN and ECP mRNAs were observed in the PBE of all patients. In CE, EPO, and MBP, mRNAs were abundant in immature eosinophils and their amounts decreased after differentiation toward eosinophils. ECP and EDN mRNAs followed the same patterns, but mRNAs were less abundant at all timepoints studied. Study of mRNA t1/2 during the time course of differentiation indicated that changes in the stability of the different mRNAs were not responsible for the variations observed in the steady-state levels. Together, these results suggest that regulation of expression differs among EPO, MBP, EDN, and ECP mRNAs during the time course of eosinophil differentiation.

Actins

Cloning and expression of a cDNA encoding a non-classical MHC class I antigen (HLA-E) in eosinophils from hypereosinophilic patients.

A cDNA library, constructed from purified blood eosinophils, was screened with the B cell CD23 cDNA probe. A clone designated EO15 has been isolated and found to encode a non-classical HLA class I gene transcript. EO15 was compared with HLA-E and found to be 99.9% similar at the nucleotide level and to extend further in the 3' untranslated region. The presence of an additional polyadenylation signal in the EO15 3' end suggests that EO15 clone represents a copy of the 3.3-kb mRNA species detected in Northern blot analyses. HLA-E transcripts of 1.9 and 3.3 kb have been described in a variety of cell types. The two EO15 mRNA species, similar in size to the previously defined HLA-E mRNA, were present at high levels in blood leukocyte populations and at variable levels in different cell lines. The EO15 transcripts were found at abundant levels in hypodense and normodense eosinophils from hypereosinophilic patients. In situ hybridization confirmed the expression of EO15 mRNA in eosinophils. Neutrophils and lymphocytes from normal donors of from patients with hypereosinophilia also strongly expressed EO15 mRNA. Among the cell lines studied, the highest levels of EO15 transcripts were detected in B and monocytic cell lines, whereas intermediate and lower levels were found in eosinophilic, NK-like, megakaryocytic, and T cell lines, respectively. Similar to its effect on classical HLA class I transcripts, IFN-gamma increased the levels of EO15 mRNA in eosinophils and neutrophils from hypereosinophilic patients. These results suggest that purified blood eosinophils as well as neutrophils express EO15/HLA-E mRNA; however, further experiments are needed to investigate the localization and the function of EO15 protein products.

Antigens, Differentiation, B-Lymphocyte

Eosinophil IgE receptor and CD23.

In the present review, eosinophil Fc epsilon RII was compared to CD23, a differentiation marker of B cells. Biochemical analysis revealed that molecules of similar molecular weight were immunoprecipitated from eosinophils and B cells by an anti-CD23 monoclonal antibody (mAb) or by BB10, and anti-eosinophil Fc epsilon RII. By flow cytometry, a correlation was found between the binding of anti-CD23 mAb and myeloma IgE. However, a low expression of different epitopes of CD23 was observed in various hypereosinophilic patients. Northern blot analysis of eosinophil RNA with the cDNA probe of CD23 revealed a weak message in only 3 of the 6 patients expressing membrane CD23. The inhibition by anti-CD23 mAbs of IgE-mediated cytotoxicity and IgE binding to eosinophils clearly indicated the participation of CD23 or a related molecule in IgE-dependent eosinophil functions. However, the differential effects of anti-CD23 mAbs on eosinophils and B cells suggest major differences in the characteristics of the molecule expressed by eosinophils and by B cells.

Antibodies, Monoclonal

Human eosinophils from hypereosinophilic patients spontaneously express the p55 but not the p75 interleukin 2 receptor subunit.

The expression of interleukin 2 receptor (IL2R) on eosinophils was investigated in patients with hypereosinophilia. Hypodense activated eosinophils have been described in various diseases such as parasitic or allergic diseases, hypereosinophilic syndrome (HES) associated in some cases to myeloproliferative markers, and more recently described in patients undergoing recombinant IL2 treatment. The presence of p55 alpha chain of IL2R (CD25) on purified eosinophils collected from blood of hypereosinophilic patients was detected by flow cytometry. In 10 out of 19 cases, more than 10% of eosinophils were CD25+. Cross-linking studies on enriched eosinophils showed one 64-75-kDa band, consisting of IL2 (15 kDa) cross-linked to the IL2R p55 subunit. In Northern blot analysis the two messenger mRNA (3.5 and 1.5 kb) encoding the IL2R p55 subunit were identified after hybridization with a CD25 cDNA probe. In contrast, the presence of the IL2R p75 subunit was not detected. These data provide preliminary evidence for the expression of a low-affinity receptor for IL2 on in vivo activated eosinophils and raise the question of the role played by this cytokine in eosinophil differentiation and activation.

Eosinophilia

Heterogeneous expression of CD23 epitopes by eosinophils from patients. Relationships with IgE-mediated functions.

The receptor for IgE (Fc epsilon RII) on human eosinophils presents some common characteristics with CD23, a differentiation marker of B cells. We have used flow cytometry for evaluating the expression of various epitopes of CD23 on purified eosinophils from patients with eosinophilia. A correlation was found between the binding of myeloma IgE protein and the binding of a monoclonal antibody (mAb 135), directed against the IgE-binding site of B cell CD23. Using two additional anti-CD23 mAb, directed (8-30) or not (3-5) against the IgE-binding site, a low expression of these CD23 epitopes was observed on eosinophils from different eosinophilic patients. Northern blot analysis of eosinophil RNA with the cDNA probe of CD23 revealed a low-abundance transcript in three of the six patients expressing membrane CD23. The inhibition by all anti-CD23 mAb of IgE-mediated cytotoxicity and IgE binding to eosinophils clearly indicated the participation of a CD23-related molecule in IgE-dependent eosinophil functions.

Antibodies, Monoclonal