Functional immunoglobulin transgenes guide ordered B-cell differentiation in Rag-1-deficient mice
We have examined the regulatory role of the individual components of the immunoglobulin antigen receptor in B-cell development by transgenic complementation of Rag-1 deficient (Rag-1⁻) mice. Complementation with a membrane µ heavy chain (µHC) gene allows progression of developmentally arrested Rag-1⁻ pro-B-cells to the small pre-B cell stage, whereas the introduction of independently integrated µHC and κ light chain (κLC) transgenes promotes the appearance of peripheral lymphocytes which, however, remain unresponsive to external stimuli. Complete reconstitution of the B-cell lineage and the emergence of functionally nature Rag-1⁻ peripheral B cells is achieved by the introduction of cointegrated heavy and light chain transgenes encoding an anti-H-2^k antibody. This experimental system demonstrates the competence of the µHC and κLC to direct and regulate the sequential stages of B-cell differentiation, defines the time at which negative selection of self-reactive B cells occurs, and shows that elimination of these cells occurs equally well in the absence of Rag-1 as in its presence. These data also support the hypothesis that Rag-1 directly participates in the V(D)J recombination process.
© 1994 by Cold Spring Harbor Laboratory Press. Received January 19, 1994; revised version accepted February 25, 1994. E.S. is grateful to Dr. En Li for providing the J1 ES cells and guidance into the ES cell technology. Dr. Ursula Storb generously provided the MOPC 21 κ transgenic line. We thank Dr. Tyler Jacks for advice. E.S. acknowledges the help of Thais Costa and Mercedes Sanchez on the Ca²⁺ release assays and Dr. Yongwon Choi for computer aid. E.S. was initially supported by an EMBO long-term fellowship, and is currently supported by Cancer Research Institute. C.R. is the recipient of a Damon Runyon-Walter Winchell Cancer Research Fund Fellowship. L.M.C. and M.S.S. acknowledge the support of Cancer Research Institute Investigator Awards. This work was supported by National Institutes of Health grant CA54162 to D.B. The publication costs of this article were defrayed in part by payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 USC section 1734 solely to indicate this fact.
Published - Genes_Dev.-1994-Spanopoulou-1030-42.pdf