In vitro characterization of rat bone marrow-derived dendritic cells and their precursors

Abstract

Although the rat is commonly used for basic immunology and transplantation research, phenotypic and functional characterization of rat dendritic cells (DCs) lags behind similar studies in the human and mouse. Therefore, these features were examined using DCs propagated from cultures of rat bone marrow maintained in a medium supplemented with granulocyte-monocyte colony-stimulating factor. Analysis of cytospin preparations of cultured cells showd that DCs arise from OX7+ myelomonocytic precursors. Typical mature rat DCs were morphologically similar to their mouse and human counterparts and expressed major histocompatibility complex (MHC) class II (common part determinant of Ia), OX62 (integrin molecule), OX7 (CD90), ICAM-1 (CD54), and CTLA4 counterreceptor, but were negative for OX8 (CD8), OX19 (CD5), W3/25 (CD4), and ED2, a rat macrophage marker. Functional analysis of OX62+ sorted DCs showed that they could effectively present the soluble antigen ovalbumin to naive T cells in vitro. A combination of anti-MHC class II monoclonal antibody and CTLA4-immunoglobulin inhibited allostimulatory ability more effectively than either reagent alone. Implications for studying the role of DCs in immune responses in the rat are discussed.

Cytokine gene therapy of gliomas: effective induction of therapeutic immunity to intracranial tumors by peripheral immunization with interleukin-4 transduced glioma cells

“…It has been reported that OX62 represent a unique marker for DC. 38 However, other investigators have reported OX62 is negative on matured rat DCs, 39 and our unpublished data from rat bone marrow culture coincide with their results (data not shown). Therefore, we did not use this to evaluate DC/monocyte type TILs.…”

Identification and Characterization of the Antigen Presenting Cell in Rat Autoimmune Myocarditis: Evidence of Bone Marrow Derivation and Non-requirement for MHC Class I Compatibility with Pathogenic T Cells

“…Our findings differ from those of Spencer and Fabre [21] in that we could not define a second population of APC in the normal rat heart with increased MHC class II expression, but distinct from the population of cells which stain with the ED2 antibody. Furthermore, in contrast to prior morphological observations in normal rat heart [28], we were unable, using immunohistochemistry, to demonstrate a separate dendritic cell population using either OX62 for veiled dendritic cells [29,30] or ED5, a marker of follicular dendritic cells in the lymph node [27]. Negative staining for veiled dendritic cells in normal heart agrees with the findings of Brenan and Puklavec who failed to identify OX62 positive cells in rat heart and skeletal muscle [30].…”

Allergen-Induced Changes in Bone-Marrow Progenitor and Airway Dendritic Cells in Sensitized Rats

“…An increase of DC in the airways can be the result of an increased influx, a local proliferation, or enhanced survival, or a decreased efflux of DC via the lymph vessels. We found that a 3-d exposure to antigen in sensitized rats induced a 60% increase in the yield of DC grown from bone-marrow precursors in the DC growth factor GM-CSF (52). This is, to our knowledge, the first description of an effect of inflammatory conditions on bone-marrow precursors for DC.…”