Anti-CD18 Antibody [MEM-48] (A86019)

Bst-2/Tetherin inhibits the release of HIV by tethering newly formed virus particles to the plasma membrane of infected cells. Although the mechanisms of Tetherin-mediated restriction are increasingly well understood, the biological relevance of this restriction in the natural target cells of HIV is unclear. Moreover, whether Tetherin exerts any restriction on the direct cell-cell spread of HIV across intercellular contacts remains controversial. Here we analyse the restriction endogenous Tetherin imposes on HIV transmission from primary human macrophages, one of the main targets of HIV in vivo. We find that the mRNA and protein levels of Tetherin in macrophages are comparable to those in T cells from the same donors, and are highly upregulated by type I interferons. Improved immunocytochemistry protocols enable us to demonstrate that Tetherin localises to the cell surface, the trans-Golgi network, and the macrophage HIV assembly compartments. Tetherin retains budded virions in the assembly compartments, thereby impeding the release and cell-free spread of HIV, but it is not required for the maintenance of these compartments per se. Notably, using a novel assay to quantify cell-cell spread, we show that Tetherin promotes the transfer of virus clusters from macrophages to T cells and thereby restricts the direct transmission of a dual-tropic HIV-1. Kinetic analyses provide support for the notion that this direct macrophage-T cell spread is mediated, at least in part, by so-called virological synapses. Finally, we demonstrate that the viral Vpu protein efficiently downregulates the cell surface and overall levels of Tetherin, and thereby abrogates this HIV restriction in macrophages. Together, our study shows that Tetherin, one of the most potent HIV restriction factors identified to date, can inhibit virus spread from primary macrophages, regardless of the mode of transmission.

Accumulated research has suggested the importance of the adhesion molecules modulation as therapeutic approach for bronchial asthma. Adhesion molecules expression alteration contributes to the pathogenesis of asthma. In order to probe the roles of expression imbalance of adhesion molecules in asthma pathogenesis, expression profiling of adhesion molecules was performed using cDNA microarray assay. The results showed that the expression pattern of adhesion molecules was altered in peripheral blood leucocytes of asthma patients. In this study, we focused on one of the abnormally expressed molecule, integrin β4, which was down-regulated in all asthma patients, to analyze the relevance of asthma susceptibility with the alteration of integrin β4 expressions. Real time PCR was used to verify the down-regulation of integrin β4 in additional 38 asthma patients. Next, the 5'flanking region of integrin β4 DNA were amplified, sequenced and site-directed mutagenesis technology in correspondent variation sites were carried out. Among 4 variation sites found in 5' flanking region of integrin β4, 3 were related to asthma susceptibility: -nt1029 G/A, -nt 1051 G/A, and -nt 1164 G/C. A reduction of human integrin β4 promoter activity was observed at mutants of these sites. This study demonstrates that various adhesion molecules in asthma patients are abnormally expressed. Mutations in 5' flanking region result in reduced integrin β4 expression, which is related to increased risk of asthma.

Monoclonal antibodies to CD18, the common chain of leukocyte integrins, recognize in various types of human lymphoid and myeloid cells under the conditions of nonreducing Western blotting three species of CD18 of mol. wt. 96, 87, and 78 kDa, respectively. Using a unique monoclonal antibody MEM-148 reacts exclusively with free CD18 molecules, but not with leukocyte integrin heterodimers. We demonstrate that only the upper one (96 kDa) is present on the cell surface within the CD11/CD18 integrin heterodimers, while the lower ones (87 and 78 kDa) are found intracellularly as free molecules unassociated with CD11 chains or other molecules. These intracellular free CD18 chains may in part represent biosynthetic precursors; alternatively, these species may represent an intracellular source of the recently observed free, proteolytically truncated CD18 chains expressed on the surface of activated myeloid cells.

Six monoclonal antibodies were prepared that recognize a broadly expressed antigen composed of noncovalently associated 95 kDa and 180 kDa subunits. Antibodies MEM-25, MEM-83 and MEM-95 are shown to react with a determinant(s) present on the isolated larger subunit identified as the CD11a glycoprotein, MEM-48 reacts with the isolated smaller subunit which is identical to the CD18 glycoprotein. Antibodies MEM-30 and MEM-94 recognize probably a determinant in the CD11a/CD18 complex (i.e., LFA-1 antigen) dependent on association of the constituent chains. Cytotoxic activity of NK cells in inhibited by antibodies MEM-25, MEM-95 and less strongly by MEM-30 and MEM-94.