Granulocyte‐macrophage colony‐stimulating factor‐cultured bone marrow‐derived macrophages reveal accessory cell function and synthesis of MHC class II determinants in the absence of external stimuli

Abstract

The antigen-mediated activation of a number of T cell clones by bone marrow (BM) cells cultivated in the presence of various colony-stimulating factor (CSF) preparations was investigated. BM macrophages (BMM phi) grown in L929 cell supernatant as a crude source of macrophage colony-stimulating factor (M-CSF) as well as BM cells propagated in the presence of recombinant M-CSF exhibited transient antigen presentation potential to some T cell clones, being maximal on day 7 and having declined to a low level by day 19 of in vitro culture. Treatment of these long-term-cultivated BMM phi populations with recombinant interferon-gamma (IFN-gamma) resulted in predominant antigen presentation capacity. In contrast, BM cells differentiated in the presence of recombinant granulocyte (G)M-CSF developed highly efficient accessory cell function to all T cell clones examined. This function became apparent earlier, was retained during the time period tested (up to day 19 of continuous culture) and did not require prior stimulation by IFN-gamma. The functionally competent cells were shown to belong to the monocyte/macrophage lineage. These findings are consistent with the demonstration of substantial levels of major histocompatibility complex (MHC) class II molecules synthesized by GM-CSF-cultured BM cells in the absence of exogenous IFN-gamma. In contrast, M-CSF grown BM cells synthesized only minute amounts of Ia antigens unless they were stimulated by IFN-gamma. Because GM-CSF-cultivated BM cells proved clearly superior to M-CSF-grown and IFN-gamma-activated BM cells with respect to antigen-presenting capacity but exhibited lower levels of MHC class II molecules, other properties acting in addition to surface Ia antigens might be responsible for their pronounced T cell accessory function.

Differential Processing of CD4 T-cell Epitopes from the Protective Antigen of Bacillus anthracis

“…Macrophages were grown from femoral bone marrow cells of BALB/c and BALB.K mice by culture for 6 days in culture medium supplemented with 5% of a supernatant from the L929 cell line as a source of macrophage colony stimulating factor, as described previously (41). Macrophages were activated by treatment overnight with 1 ng/ml interferon-␥ (R&D Systems, Abingdon, UK) before use in antigen presentation assays.…”

Presentation of the candidate rheumatoid arthritis autoantigen aggrecan by antigen‐specific B cells induces enhanced CD4+ T helper type 1 subset differentiation

“…Bone marrow cells from femurs of BALB/c mice were differentiated into macrophages [7 days with media supplemented with 5% horse serum (Sigma-Aldrich) and 10% L929 supernatant, 34 (refreshed on days 3 and 6) or DC (10 days with media supplemented with 20 ng/ ml granulocyte-macrophage-colony-stimulating factor 35 (PreproTech, Inc. Rocky Hill, NJ) (refreshed on days 3, 6 and 8)]. Macrophages were activated with 100 ng/ml interferon-c (IFN-c) …”

Class II (HLA-DR, HLA-DP, and HLA-DQ) Antigens on Colonic Epithelia in Ulcerative Colitis: A Comparison between Uneventful and Intractable Cases.

“…Although the mechanisms of class II antigens expression on colonic epithelia are unclear, it has been shown that cytokines, such as IFN-y (Carrel et (Crawford et al 1987), and granulocyte/ macrophage-colony stimulating factor (Falk et al 1988 ;Fischer et al 1988), have the ability to induce expression of class II antigens on various human cells . Therefore, these cytokines might also induce expression of class II antigens on colonic epithelia.…”