Secretion of soluble pre-B cell receptors by pre-B cells.
Pre-B cells can express secretory mu (mu(s))- as well as membrane mu (mu(m))-chains. We evaluated the ability of mu(s)-chains to associate with surrogate light chains and assemble into a pre-B cell receptor (BCR) complex in pre-B cells, and explored whether mu(s)-chains could be exploited to generate a secreted soluble pre-BCR. We demonstrate that mu(s)-chains can associate with SLC internally. The mu(s)-containing complexes form higher order polymeric structures, but these are never assembled into completed covalent structures. Instead, the complexes are efficiently retained and rapidly degraded. Alteration of the intracellular redox state by incubation with 2-ME resulted in the secretion of mu(s)-chains, suggesting that they are retained by a thiol-mediated retention mechanism. To identify the sequences on mu(s)-chains responsible for their retention, we generated stable transfectants of a mu-negative pre-B cell line expressing either wild-type or mutant mu(s) constructs. Mutation of a single cysteine (Cys575) in the mu(s) tailpiece resulted in the release and secretion of the mu(s) H chains. These were associated with the surrogate light chain proteins lambda5 and VpreB, and thus appear to constitute an authentic secreted soluble pre-BCR. The soluble pre-BCR has a specificity distinct from Ab consisting of the same heavy chain V region paired with conventional light chains.
Bornemann
K D
KD
Department of Microbiology, Boston University School of Medicine, MA 02118, USA.
Brewer
J W
JW
Perez
E
E
Doerre
S
S
Sita
R
R
Corley
R B
RB
D000818:Animals; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D001704:Biopolymers / Q000378:metabolism; D002460:Cell Line; D003545:Cysteine / Q000235:genetics / Q000502:physiology; D007147:Immunoglobulin Light Chains / Q000378:metabolism; D007075:Immunoglobulin M / Q000378:metabolism; D007148:Immunoglobulin mu-Chains / Q000378:metabolism; D051379:Mice; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D012995:Solubility; D013234:Stem Cells / Q000276:immunology / Q000378:metabolism*
D001704:Biopolymers; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M; D007148:Immunoglobulin mu-Chains; D011947:Receptors, Antigen, B-Cell; C033027:secretory IgM; D003545:Cysteine