CD63 tetraspanin slows down cell migration and translocates to the endosomal-lysosomal-MIICs route after extracellular stimuli in human immature dendritic cells

Abstract

We analyzed herein whether members of the tetraspanin superfamily are involved in human immature dendritic cell (DC) functions such as foreign antigen internalization, phagocytosis, and cell migration. We show that CD63, CD9, CD81, CD82, and CD151 are present in immature DCs. Whereas CD9 and CD81 are mostly expressed at the cell surface, CD63 and CD82 are also located in intracellular organelles. Complexes of monoclonal antibody (Mab) FC-5.01-CD63 or Fab-5.01-CD63 were rapidly translocated "outside-in" and followed the endocytic pathway through early endosomes and lysosomes, reaching major histocompatibility complex (MHC) class IIenriched compartments (MIICs) in less than one hour. Internalization of CD63 was also observed during Saccharomyces cerevisiae phagocytosis. Moreover, an association of CD63 with the ␤-glycan receptor dectin-1 was observed. Mabs against CD9, CD63, CD81, and CD82 enhanced by 50% the migration induced by the chemokines macrophage inflammatory protein-5 (MIP- 5 IntroductionDendritic cells (DCs) comprise a family of professional antigenpresenting cells (APCs) that are sentinels of the immune system. 1 In this regard, DCs have been shown to efficiently stimulate both naive B and T cells and to elicit primary immune responses. 2,3 Their remarkable effectiveness is due to their ability to capture, process and present antigens along with costimulatory signals, and to migrate to secondary lymphoid tissues. 4 Different stages of maturation are responsible for the different functions of DCs. Immature DCs, widely present in peripheral tissues, efficiently uptake antigens but express moderate levels of major histocompatibility complex (MHC) class II and costimulatory molecules. In contrast, mature DCs poorly acquire antigens but express higher levels of MHC class II and costimulatory molecules, are able to migrate into lymph nodes, and become potent activators of resting T cells. [2][3][4][5] Since antigen internalization and processing, as well as cell migration, are essential properties of DCs, the study of the possible involvement of tetraspanins in these processes is a challenging question, as it could bring new insight into the physiologic role of these molecules in DCs and other cells. In recent years, considerable interest has arisen in the expanding tetraspanins family, which are integral membrane proteins with 2 extracellular domains (EC1 and EC2) that are variably glycosylated. 6 The most conspicuous members of this family are CD9, 7 CD63/lamp-3, 8 CD81/TAPA-1, 9 CD82/KAI1, 10 and CD151. 11 Tetraspanins form several specific complexes implicated in a variety of cellular processes such as migration, adhesion, proliferation, and signal transduction, initially leading to the idea that these proteins could play a "molecular facilitator" role. 6 There is growing evidence that points to a possible role of tetraspanins in antigen processing and presentation. In fact, it has been recently found that CD63 is modified after translation during maturation of DCs, and this event is accompanied by morph...

The tetraspanin CD63 is involved in granule targeting of neutrophil elastase

“…21 Tetraspanin complexes act as molecular facilitators in many cellular processes, 35,36 and different partner proteins can become clustered together with CD63. 37 It has been proposed that membrane proteins such as the H,K-ATPase 25 and membrane type 1-matrix metalloproteinase 23,38 are transported by CD63.…”

“…31,32 Therefore, the possible presence of proteoglycans was examined, as previously described by Lemansky et al 32 COS cells coexpressing NE and GFP-CD63 were incubated with radioactive sulfate, immunoprecipitated with anti-cproNE, and subjected to SDS-PAGE followed by fluorography. ProNE did not pull down any 35 S-labeled macromolecules (data not shown), indicating that proteoglycan was not present.…”

Recruitment of CD63 toCryptococcus neoformansphagosomes requires acidification

“…Here we have addressed the behavior of CD63 in the maturation of phagosomes using live cell imaging. To date, characterization of CD63 has been accomplished in human cell lines, largely through biochemical analysis (12,16,27). The data presented here focus on the behavior of CD63 and class II MHC in live primary cultures of mouse APCs in the context of antigen capture by phagocytosis.…”

“…More recently, the focus has shifted to its interaction with class II MHC and its role in antigen presentation. In professional APCs such as dendritic cells (DCs), subcellular fractionation and biochemical analysis show that human CD63 segregates into class II MHC-rich domains within the endocytic pathway and cell surface (10)(11)(12). Furthermore, this tetraspanin protein preferentially resides in membrane patches enriched for particular class II MHC-peptide complexes (13).…”

Regulation of NK cell trafficking by CD81

“…Moreover, growing evidence indicates tetraspanins to be critically involved in the regulation of cell motility [18][19][20]. In fact, Mantegazza and colleagues recently demonstrated the tetraspanin CD63 to slow down migration of DC possibly via interfering with surface expression of integrins.…”