Reduced inflammatory potential of peritoneal macrophages recruited in mice pretreated with a glycolipid synthesis inhibitor.

Integral components of mammalian cell membranes, glycosphingolipids (GSL) reside in specialized plasma membrane microdomains critical for cell signaling. N-alkylated nojirimycins are compounds developed for GSL substrate deprivation therapy, blocking GSL synthesis by specifically inhibiting an essential enzyme, ceramide glucosyltransferase. Peritoneal macrophages recruited in mice pretreated with an inhibitory N-alkylnojirimycin displayed a reduced capacity to release either TNFalpha or interleukin-6 when re-exposed to whole killed E. coli in vitro. Cell viability and protein content were not affected. A nojirimycin analogue without GSL inhibitory capacity had no effect. The results show inhibition of GSL synthesis in vivo by an N-alkylnojirimycin can reduce the response to an inflammatory stimulus and indicate N-alkylnojirimycins have experimental and potential clinical value for modulating innate immune responses in vivo.

Enhanced metabolism of LDL aggregates mediated by specific human monocyte IgG Fc receptors.

Macrophage-derived foam cells are important constituents of atheromatous lesions. In addition to the scavenger receptor pathway, uptake of immune complexed lipoproteins through IgG Fc receptors (Fc gamma receptors) represents an additional pathway of macrophage foam cell development that may be important during atherogenesis. The importance of this mechanism is suggested by studies showing that the titer of autoantibodies to modified lipoproteins correlated with the extent of occlusive disease in patients, and that those antibodies exist in human lesions. Human mononuclear phagocytes possess three structurally and functionally distinct classes of Fc gamma receptors, each of which could be associated with a unique pathway of lipoprotein metabolism. In order to determine whether uptake of an acute lipid load through each type of Fc gamma receptor was associated with foam cell development, we used bispecific antibodies consisting of anti-LDL monoclonal antibodies conjugated to anti-Fc gamma receptor monoclonal antibodies to study the effects of targeting LDL aggregates to each specific type of Fc gamma receptor on freshly isolated adherent human monocytes. Relative to appropriate controls, LDL degradation, cellular sterol mass, and foam cell development of monocytes were enhanced by targeting LDL aggregates to Fc gamma RI or Fc gamma RII, and this was accompanied by an apparent impairment of LDL degradation. Uptake was specific to the Fc gamma receptors and was not influenced by the presence of scavenger receptor ligands. Thus, with the bispecific approach, the functions of each class of Fc gamma receptor can be studied on an individual basis with respect to several aspects of cellular cholesterol metabolism. This will be critical for determining which of these receptors are potentially most important in the clearance of lipoprotein immune complexes during atherogenesis.

Redirected targeting of LDL to human monocyte Fc gamma receptors with bispecific antibodies.

Recent studies indicate that low density lipoprotein (LDL)-immune complexes consisting of anti-LDL antibodies bound to LDL may contribute to macrophage foam cell development by uptake through immunoglobulin G (IgG) Fc receptors. As human mononuclear phagocytes possess three structurally and functionally distinct classes of IgG Fc receptors, we developed a system whereby the effects of LDL-immune complexes could be studied with respect to each type of IgG Fc receptor. Novel bispecific antibodies consisting of anti-Fc gamma receptor antibodies linked to anti-LDL antibodies were used to prepare bispecific LDL-immune complexes for targeting to specific Fc gamma receptors. In this report, the effects of bispecific LDL-immune complexes directed to Fc gamma receptor types I, II, and III were studied primarily with monocytes and were compared with the effects of similarly prepared bispecific complexes that targeted LDL to human leukocyte antigen (HLA) class I antigens. Each type of bispecific antibody was effective in targeting 125I-LDL to its respective site on the cell surface. Using fluorophore-labeled LDL and flow cytometry, bispecific complexes directed to Fc gamma receptor types I or II but not to HLA class I antigens caused a two- to sevenfold increase in cell-associated fluorescence relative to control cells treated with LDL in the absence of bispecific antibody. Uptake occurred in the presence of excess unlabeled LDL, acetylated LDL, and antioxidants. That the bispecific complexes triggered metabolic uptake was supported by studies of kinetics and temperature dependence. Using 125I-labeled complexes, metabolic degradation of LDL was demonstrated in association with each of the three types of Fc gamma receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

High-frequency ventilation does not affect pulmonary surfactant, liquid, or morphologic features in normal cats.

Effects on pulmonary surfactant, liquid, and morphologic features were compared in normal cats ventilated with conventional and high-frequency ventilators for 4 h. Normal blood gases and comparable mean airway pressures (approximately 7 cm H2O) were maintained with the two techniques. Peak tracheal pressure during conventional ventilation (16.5 +/- 0.9 cm H2O) was significantly greater than that during high-frequency ventilation (8.5 +/- 1.1 cm H2O, p less than 0.00001). Pressure-volume curves were identical in control, conventional, and high-frequency ventilation groups. There was no effect on pulmonary lavage surfactant, whether measured functionally or biochemically. Likewise, lung liquid and morphologic aspects examined by both light and electron microscopy were not affected. These results provide evidence that high-frequency ventilation under these conditions does not cause pulmonary damage.