Expression of the fibronectin receptor VLA-5 is regulated during human B cell differentiation and activation

Abstract

We examined the expression of VLA-5, a fibronectin receptor, during human B cell development and activation. VLA-5 is a member of the integrin supergene family; VLAs are heterodimers of at least six unique alpha chains sharing a common beta chain; most are involved in cell attachment to extracellular matrix (ECM). A hypothesis of haematopoietic development is that maturing cells leave the bone marrow because of the loss of VLA-5 during differentiation. However, mature B cells are not primarily circulating cells, and the role of ECM receptors in homing to peripheral lymphoid tissue and inflammatory sites is unknown. To examine the expression of VLA-5 during B cell development, cell lines blocked at specific stages of differentiation were evaluated for their synthesis and surface expression of VLA-5 using VLA-5-specific antibody and cDNA probes. VLA-5 mRNA and surface expression were found in the pre-B cell lines, REH and Nall 1, but not in more differentiated Raji cells or in several EBV-transformed peripheral B cell lines. Circulating peripheral B lymphocytes and resting tonsillar and splenic B lymphocytes expressed no VLA-5 by FACS analysis. Interestingly, mRNA and surface expression of VLA-5 were found in SKW, a highly differentiated, IgM-secreting line. In addition, low levels of staining for VLA-5 expression could be demonstrated when tonsillar or peripheral blood B lymphocytes were stimulated by Staphylococcus aureus Cowan (SAC). All cell lines expressed VLA-3 and VLA-4, two other receptors reported to mediate fibronectin binding in some cell types. Thus, our studies provided no evidence for developmental or inflammatory regulation of these receptors. Binding studies, however, demonstrated that adherence of both pre-B REH cells and SKW cells to fibronectin was almost completely inhibited by a monoclonal antibody to VLA-5 alpha. In addition, Raji cells, which lack VLA-5 but express VLA-3 and VLA-4, showed very low level binding to fibronectin. This demonstrates that for some B lymphocytes VLA-5, rather than other possible fibronectin receptors, primarily mediates attachment to fibronectin. These data also suggest that human VLA-5 expression is regulated during B cell development, with expression at a very early stage and then again after activation. This pattern of loss and reacquisition of an ECM receptor may be relevant to normal B cell maturation and to function during immunologic injury.