Phenotypic and functional heterogeneity of the murine alveolar macrophage-derived cell line MH-S

Abstract

We have previously reported that MH-S, an established murine alveolar macrophage-derived cell line, mediated profound inhibition of in vitro antibody production, as did their freshly isolated alveolar macrophage (AM) counterparts. In this communication we show that like freshly recovered AMs, the MH-S cell line also displays phenotypic and functional heterogeneity. Sorting of parental MH-S cells by flow cytometry based on reactivity with anti-Mac-1 antibody yielded two subsets. Further analysis by staining with monoclonal antibodies against well-characterized murine macrophage cell surface markers revealed that both Mac-1+ and Mac-1- subsets expressed the mature murine macrophage antigen (F4/80) and class II major histocompatibility complex molecules, but with different intensity. In contrast, the two subsets stained equivalently with antibody against the Fc gamma II receptor, whereas neither subset stained with anti-CD4 antibody. Examination by light microscopy revealed plemorphism in the Mac-1+ population with many of the cells appearing spindle shaped and having elongated processes, whereas a majority of the cells in the Mac-1- population were spherical in shape. Functionally, cells from the Mac-1+ population were less inhibitory of in vitro antibody production and produced significantly more nitric oxide in response to stimulation with lipopolysaccharide than were cells in the Mac-1- population. Essentially similar results were obtained using cloned Mac-1+ and Mac-1- MH-S cells. The finding of heterogeneity in an established cell line that displays functions similar to those of freshly recovered AMs suggests that distinct subsets of AMs may be involved in the pathogenesis of disease processes in the lung.

Noninflammatory phagocytosis of monosodium urate monohydrate crystals by mouse macrophages: Implications for the control of joint inflammation in gout

“…The cell lines, which were of mouse origin since a comparable panel of human cells was not available, were placed in order of differentiation on the basis of F4/80 and BM 8 antigen expression, as well as phagocytic capacity. Although to our knowledge, this is the first time these cell lines have been arranged in this way, the cells individually displayed phenotypes consistent with published information (15,(18)(19)(20)(21)(22)(23)(24)(25). Our finding that the partially differentiated macrophage cell line RAW264.7 synthesized TNF␣ in response to MSU crystals is in agreement with findings of a previous study (26).…”

“…The monocytoid cell line WEHI-265 (21) exhibited moderate F4/80 expression and phagocytic ability. The remaining 4 cell lines tested, RAW264.7, J774A.1, IC-21, and MH-S, exhibited increasingly high levels of F4/80 and BM 8 antigen, as well as phagocytic ability, consistent with a known macrophage phenotype (22)(23)(24). These data, together with an increasingly stellate morphology and plastic-adherent phenotype (data not shown), allowed us to establish a clear order of monocyte to macrophage differentiation, as follows: M1, WEHI-3, WEHI-274.1, WEHI-265, RAW264.7, J774A.1, IC-21, and MH-S.…”

Clinical response and immunomodulation following experimental challenge of calves with type 2 noncytopathogenic bovine viral diarrhea virus

“…the enhanced cell phagocytic capability) of immune cell activities. Because MHC II molecules are primarily associated with antigen-presenting functions [1], and various subpopulations of macrophages are present in mammals [5,27], one may suggest that BVDV exerts these apparently dual immunoregularory effects on different subsets of monocytic cells that might be present in the bovine PBMC suspension. Whatever the mechanism could be, similar dual effects on the functional activity of immune cells have been shown in other animal virus systems [20,31].…”

The Vsa Shield of Mycoplasma pulmonis Is Antiphagocytic

“…The cell line consists of a heterogeneous population of Mac-1 Ϫ and Mac-1 ϩ cells (35). MH-S cells were grown in sodium bicarbonate-buffered Dulbecco's minimum essential medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS).…”