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J J Steenbergen

Publications and source records attributed to J J Steenbergen.

7 recordsLinked to original sources

Cellular composition of milky spots in the human greater omentum: an immunochemical and ultrastructural study.

BACKGROUND: Milky spots in the greater omentum of some animals are well organized perivascular infiltrates of leucocytes, and are considered to have characteristics of secondary lymphoid tissue. To determine whether milky spots in the human greater omentum can also be regarded as secondary lymphoid tissue, we studied milky spots in an unstimulated state. METHODS: Patients were selected on the basis of absence of disease in the peritoneal cavity that might influence the state of the milky spots. Using monoclonal antibodies against macrophages, B-lymphocytes and T-lymphocytes, and immunoperoxidase labeling, the number of these cells and their location in milky spots were studied by light microscopy. However, the stromal components of the greater omentum, especially those within the milky spots, were studied by electron microscopy. RESULTS: Milky spots in the human greater omentum are relatively uniform vascularized accumulations of mononuclear cells comprising macrophages (67.9% +/- 9.4, mean +/- standard deviation), B-cells (10.1% +/- 3.4), T-cells (10.2% +/- 3.7), and mast cells. However, no special B-cell and T-cell areas could be distinguished. On the ultrastructural level it was demonstrated that macrophages are present in different stages of maturation and can enter or leave the milky spots. Furthermore, no cells characteristic of secondary lymphoid organs, such as interdigitating cells or follicular dendritic cells, were seen. CONCLUSIONS: These data indicate that unstimulated milky spots in the human greater omentum are to a great extent just a preformed specific accumulation of primarily macrophages within the stroma of the greater omentum, and therefore, cannot be regarded as true secondary lymphoid tissue. Milky spots could serve as a gateway for, as well as a provider of peritoneal macrophages when the intra-abdominal status so requires. Finally, the data from this study are compared with the data of other studies of human milky spots and those in animals.

Adolescent↗

Interleukin-8 production by human mesothelial cells after direct stimulation with staphylococci.

Mesothelial cells (MC) are able to produce interleukin-8 (IL-8) after stimulation with IL-1 beta or tumor necrosis factor alpha. The aim of our study was to investigate whether MC are able to produce IL-8 after direct stimulation with clinically relevant bacteria. We observed a significant IL-8 response by the MC which were directly stimulated with viable staphylococci.

Cells, Cultured↗

Isolation of cytotoxic Kupffer cells by a modified enzymatic assay: a methodological study.

Kupffer cell (KC)-mediated cytotoxicity against tumor cells is of interest, since the liver is a major site of metastatic growth of primary colorectal cancer. KC isolation methods from rat livers, to study the tumoricidal properties of these cells, are based on perfusion of the liver and are therefore not suitable for human KC isolation from liver biopsies. In view of application to isolate KC from small wedge human liver biopsies, we have developed an isolation procedure for rat KC that does not require perfusion techniques. Liver tissue fragments were incubated with pronase with continuous pH registration and neutralization. KC were subsequently separated from other non-parenchymal cells by Nycodenz gradient centrifugation and purified by counterflow centrifugal elutriation. KC and other non-parenchymal cells were identified by immunophenotyping with a cytoplasmic monoclonal antibody ED1 and by ultrastructural analysis. About 3 x 10(6) KC per gram liver were isolated with a final purity of > 95% without loss of viability. To ensure that functionally competent KC were isolated, we assayed cytotoxicity against CC531 tumor cells in a recent developed cell-mediated MTT assay. Maximum cytotoxicity of KC was approximately 40% at an effector to target ratio of 10. In conclusion our approach seems to be a useful and simple method to isolate KC with good functional properties from rat livers, without the need for perfusion techniques.

Animals↗

Milky spots in the mouse omentum may play an important role in the origin of peritoneal macrophages.

In the milky spots, precursors of cells belonging to the mononuclear phagocyte system (MPS), such as monoblasts, can be found on the basis of ultrastructural endogenous peroxidase cytochemistry. Therefore, in the present study, we investigated the milky spots using a panel of monoclonal antibodies, especially antibodies (ER-MP) that recognize macrophage precursor antigens. Early macrophage precursor antigens ER-MP12 and ER-MP58 were detected only on cells localized inside the milky spots. On the other hand, an antigen which disappears late in the course of macrophage differentiation, ER-MP20, was detected in high amounts on cells both inside and around the milky spots. This clearly indicates that macrophage precursors are centrally localized inside the milky spots, while more differentiated cells are found in peripheral areas. Moreover, long-term culture of milky spot tissue resulted in the forming of a monolayer of stromal cells which supported macrophage proliferation in vitro. In conclusion, both in situ and in vitro studies demonstrated that mouse milky spots have a microenvironment in which precursor cells of the MPS can home and proliferate, illustrating that milky spots play a role as a source of local macrophage generation, e.g. that of the free peritoneal macrophages.

Animals↗

Lymphoid tissue in rat oral mucosa: structure and function.

Lymphocyte and macrophage/dendritic cell populations in the oral cavity of the rat were studied by immunohistochemistry. Furthermore, the reactivity of the oral mucosa towards antigen and its position in the mucosal immune system was investigated by staining antibody-forming cells and comparing serum and saliva antibody titres. Although numerous lymphocytes and non-lymphoid cells were present in the oral mucosa, organized lymphoid tissue could not be found. The majority of the lymphocytes are T cells, particularly of the T-helper phenotype. Macrophages were found only in the connective tissue, whereas dendritic cells also occurred in the epithelium. The possible functions of the cells in the different sites of the oral mucosa are discussed. Antibody-forming cells were mainly detected in the draining superficial and posterior cervical lymph nodes and in the submandibular glands. The roles of the various compartments of the oral mucosal immune system are discussed with particular reference to the epithelium and connective tissue as induction sites, the draining lymph nodes as sites of proliferation and differentiation, and the submandibular glands as effector sites.

Animals↗

Immunoglobulin concentrations in nasal secretions differ between patients with an IgE-mediated rhinopathy and a non-IgE-mediated rhinopathy.

Nasal secretions from patients with an IgE-mediated rhinopathy, patients with a non-IgE-mediated rhinopathy, and healthy control subjects were collected with a newly developed direct aspiration system. Total protein, albumin, secretory IgA (sIgA), IgE, IgG, and IgM concentrations were measured in the nasal secretions to detect whether the nasal pathology is reflected in nasal secretion protein concentrations. It was found that protein and immunoglobulin concentrations in nasal secretions were inversely related to amount of secretion in the nasal cavity. Both patients' groups had a significantly higher sIgA to protein ratio than the healthy control subjects. Furthermore, patients with an IgE-mediated rhinopathy had significantly higher sIgA and IgM to total protein ratios in their nasal secretions than patients with a non-IgE-mediated rhinopathy. Probably these differences are due to changes in immunoregulation.

Adolescent↗