Different accessory function for T_H1 cells of bone marrow derived macrophages cultured in granulocyte macrophage colony stimulating factor or macrophage colony stimulating factor

Abstract

The ability of macrophages to stimulate immune responses is heterogeneous and may have influence on the type of the developing immune response. Therefore, in an attempt to define different functional states of mouse macrophages, we made use of the two macrophage growth factors: macrophage colony stimulating factor (M-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF). Generation of macrophages from freshly isolated bone marrow cells in the presence of GM-CSF results in a population expressing profound antigen presenting function for mouse TH1 cells, resulting in strong lymphokine production and proliferation of the T cells. Furthermore, high amounts of a novel soluble cytokine active on mouse TH1 cells are generated during the interaction of TH1 cells with macrophages elicited with GM-CSF. In contrast, macrophages grown from bone marrow cells for at least 14 days in the presence of M-CSF express only minimal antigen-presenting function for TH1 cells. Treatment of such macrophages for 24 h with either IFN-gamma or GM-CSF allows the distinction between two further functional states. Those treated with IFN-gamma efficiently presented antigen towards TH1 cells. The T cells produced large amounts of lymphokines and proliferate well. However, synthesis of the novel soluble cytokine (active on TH1 cells) was not detectable. The generation of this mediator requires a short-term treatment with GM-CSF of macrophages developed in the presence of M-CSF prior to their interaction with TH1 cells.

Interleukin‐12/T cell stimulating factor, a cytokine with multiple effects on T helper type 1 (Th1) but not on Th2 cells

“…with M@ as APC which can present antigen (TcRKD3-signal) and deliver co-stimulators such as IL-12 [40]. The co-stimulatory effect of IL-12 on the synthesis of IFN-y by T h l clones activated via theTcR/CD3-complex with an anti-CD3 mAb is demonstrated in Fig.…”

“…Supernatants containing TSF were produced by stimulation of 25 X lo6 Tcells of the Th1 line LNC.2 [24] which were seeded onto 10 x lo6 syngeneic GM-CSF-Ma adhering on petri culture dishes in 5 ml medium containing 50 pg/ml PPD as antigen [40]. After 20 h supernatants were collected, concentrated and dialyzed against PBS by ultrafiltration.…”

“…TSF-containing supernatants were obtained by stimulation of Thl cells of line LNC.2 with PPD as antigen presented by syngeneic M@ that had been developed from bone marrow cells in the presence of GM-CSF as described previously [40]. After concentration of the supernatants by ultrafiltration TSF was partially purified by a three-step procedure involving adsorption on a dye (Procion Red) column followed by cation ion exchange chromatography and HPLC on a reversed phase C4-column (Fig.…”

Immunologic Protection against Canine Heartworm Infection.

“…The authors have proved that cell-mediated immunity rather than antibody [9], especially macrophages, is important for mice to protect against L3 of Brugia malayi. Macrophages are activated also by T helper 1 (Th1) cytokines [5,15] produced by T lymphocytes and itself. In the present study, adding FCA is thought to stimulate Th1, even though those dogs showed eosinophilia that is induced by T helper 2 cytokine such as interleukin 5.…”

Downregulation of Mitogen-Activated Protein Kinases by theBordetella bronchisepticaType III Secretion System Leads to Attenuated Nonclassical Macrophage Activation

“…All media were supplemented with 10% heat-inactivated fetal calf serum (HyClone), penicillin at 100 IU/ml, and streptomycin at 100 g/ml. At day 9 BMM⌽ were trypsinized and transferred into new medium containing 20 ng of GM-CSF per ml (13,37) and incubated for another 24 h. For MAPK analysis, BMM⌽ and 10-day-old BMDC were serum and growth factor deprived in RPMI for 1.5 h prior to infection to reduce basal levels of phosphorylated MAPK (32). Serum deprivation of RAW 264.7 cells was performed for 16 h. For all other assays, cells were transferred to new medium containing 5 ng of GM-CSF per ml and serum.…”