CD1d favors MHC neighborhood, GM1 ganglioside proximity and low detergent sensitive membrane regions on the surface of B lymphocytes

Background: Cluster of differentiation 1 (CD1) represents a family of proteins which is involved in lipid-based antigen presentation. Primarily, antigen presenting cells, like B cells, express CD1 proteins. Here, we examined the cell-surface distribution of CD1d, a subtype of CD1 receptors, on B lymphocytes. Methods: Fluorescence labeling methods, including fluorescence resonance energy transfer (FRET), were employed to investigate plasma membrane features of CD1d receptors. Results: High FRET efficiency was observed between CD1d and MHC I heavy chain (MHC I-HC), β2-microglobulin (β2m) and MHC II proteins in the plasma membrane. In addition, overexpression of CD1d reduced the expression of MHC II and increased the expression of MHC I-HC and β2m proteins on the cell-surface. Surprisingly, β2m dependent CD1d isoform constituted only ~ 15% of the total membrane CD1d proteins. Treatment of B cells with methyl-β-cyclodextrin (MβCD) / simvastatin caused protein rearrangement; however, FRET demonstrated only minimal effect of these chemicals on the association between CD1d and GM1 ganglioside on cell-surface. Likewise, a modest effect was only observed in a co-culture assay between MβCD/simvastatin treated C1R-CD1d cells and invariant natural killer T cells on measuring secreted cytokines (IFNγ and IL4). Furthermore, CD1d rich regions were highly sensitive to low concentration of Triton X-100. Physical proximity between CD1d, MHC and GM1 molecules was also detected in the plasma membrane. Conclusions: An intricate relationship between CD1d, MHC, and lipid species was found on the membrane of human B cells. General significance: Organization of CD1d on the plasma membrane might be critical for its biological functions. © 2013 Elsevier B.V. All rights reserved.