|  In preliminary experiments (data not shown), an increased LPS-proliferative effect was noted in CBMCs when compared with regular adult volunteer-derived blood. In previous reports, TLR4 may be activated by minimal amount of endotoxin in the preparations of HMGB1 rather than HMGB1 itself. As a result, synergistic effects of HMGB1-LPS on upregulation of RAGE and TLR4 expression were found. T cells from alymphoplasia (aly) mice that contain mutant NIK showed impaired proliferation and IL-2 production in response to anti-CD3ϵ stimulation (12). We demonstrate here that while inducing only low levels of TNF-alpha alone, DNA from both Gram-negative and Gram-positive bacteria synergizes with subthreshold concentrations of LPS (0.3 ng/ml) to induce TNF-alpha in the RAW 264.7 macrophage-like cell line. RASF were lysed in a buffer containing 1.0% (vol/vol) Nonidet-P40, 150 mM NaCl, 20 mM Tris-HCl (pH 7.5), 1 mM EDTA, and a protease inhibitor mixture (Roche, Basel, Switzerland). Important differences exist between the adult and neonatal human immune systems despite the apparent presence in the neonate of a full repertoire of Th1, Th2, cytotoxic T cells, B cells and dendritic cells (1). group box chromosomal protein 1 (HMGB1) or lipopolysaccharide Shanghai 200433, China, 2Department of Rheumatology & Immunology, Changhai Hospital, Second Military Medical University, 168 Changhai Road, Shanghai 200433, China, 3Department of Orthopedics, Changhai Hospital, Second Military Medical University, Shanghai 200433, China. Finally, the inability to mount a T cell-independent B cell response to polysaccharide antigens until 2 years of age leads to susceptibility to infection by encapsulated bacteria (4). Crit Care Med. We have demonstrated that TLR4 is not involved in the synergistic effects of HMGB1-IL-1β complex in macrophages [12]. Receptor for advanced glycation endproducts, B. Bartok and G. S. 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