As reported by Brenchley et al. src inhibitor dasatinib and Inhibitors,Modulators,Libraries confirmed by others, plasma levels of lipopolysaccharide, a Gram negative bacterial endotoxin, are higher in chronic HIV infected patients with HAART than in the unin fected. Bacterial infection in HIV patients Inhibitors,Modulators,Libraries influ ences the severity and rate of disease Inhibitors,Modulators,Libraries progression. Peripheral LPS induces various inflammatory and immu nological reactions including the production of cyto kineschemokines, such as tumor necrosis factor a 1, and IL 6. TNF a enhances HIV 1 transport across the BBB and LPS induces an increase in HIV 1 infected monocyte trans port across the BBB. In our previous in vivo study, we found that the peripheral injection of LPS enhanced gp120 uptake by brain. These studies suggest that elevated levels of inflammatory mediators, including cytokineschemokines and LPS, regulate the permeabil ity of the BBB to HIV 1.

BECs express LPS receptors, such as Toll like receptor 2, TLR 4, and CD14 and are targets of LPS. The barrier function of the BBB is affected by various Inhibitors,Modulators,Libraries cytokineschemokines in the blood compartment. Several studies using in vitro BBB models have shown that LPS increases the paracel lular permeability of the BBB. LPS induces or enhances the secretion of Inhibitors,Modulators,Libraries several cytokines by BECs. Thus, bacterial infection and the accompanying inflammatory state could be involved in the enhance ment of HIV 1 entry into the brain. We recently reported that LPS increased transcellular transport of HIV 1 across the BBB through p38 mito gen activated protein kinase.

Here, we examined whether LPS enhanced release of cytokines by BMECs mediated the transcellular transport of HIV 1 and was regulated by MAPK signaling pathways. Materials and methods Radioactive labeling HIV 1 CL4CEMX174 prepared and ren dered noninfective by aldrithiol 2 treatment as pre viously described was a kind gift of the National Cancer Institute, inhibitor purchase NIH. The virus was radioactively labeled by the chloramine T method, a method which preserves vial coat glycoprotein activity. Two mCi of 131I Na, 10 ug of chloramine T and 5. 0 ug of the virus were incu bated together for 60 sec. The radioactively labeled virus was purified on a column of Sephadex G 10. Primary culture of mouse brain microvascular endothelial cells BMECs were isolated by a modified method of Szab�� et al. and Nakagawa et al. The animals were housed in clean cages in the laboratory with free access to food and water and were maintained on a 12 h dark, 12 h light cycle in a room with controlled temperature and humidity. All procedures involving experimental animals were approved by the local Animal Care and Use Committee and were per formed in a facility approved by Association for Assess ment and Accreditation of Laboratory Animal Care.