Inducible gene targeting in mice (2007)
Kuhn, R., Schwenk, F., Aguet, M., Rajewsky, K.
A method of gene targeting that allows the inducible inactivation of a target gene in mice is presented. The method uses an interferon-responsive promoter to control the expression of Cre...
Kanters, E., Pasparakis, M., Gijbels, M.J.J., Vergouwe, M.N., Partouns-Hendriks, I., Fijneman, R.J.A., ...
Lymphoid development in mice with a targeted deletion of the interleukin 2 receptor gamma chain.
DiSanto, J P, Müller, W, Guy-Grand, D, Fischer, A, Rajewsky, K
The interleukin 2 receptor gamma chain (IL-2R gamma) is a component of the receptors for IL-2, IL-4, IL-7, and IL-15. Mutations in IL-2R gamma in man appear responsible for the X chromosome-linked...
Küppers, R, Rajewsky, K, Zhao, M, Simons, G, Laumann, R, Fischer, R, ...
Hodgkin disease (HD) is characterized by a small number of putative malignant cells [Hodgkin and Reed-Sternberg (HRS) cells] among a background of lymphocytes and histiocytes. The lineage of HRS...
We have reevaluated the in vivo lifespan of B lymphocytes based on incorporation of the thymidine analogue 5-bromo-2'-deoxyuridine into the DNA of dividing cells. To exclude potential alterations in...
B lymphocyte-specific, Cre-mediated mutagenesis in mice.
Rickert, R C, Roes, J, Rajewsky, K
Adaptation of the P1 phage-derived Cre /loxP site- specific recombination system to the gene targeting technique allows for the conditional deletion of genes in mice. To selectively modify genes in B...
Temporally and spatially regulated somatic mutagenesis in mice.
Schwenk, F, Kuhn, R, Angrand, P O, Rajewsky, K, Stewart, A F
In mice transgenesis through oocyte injection or DNA recombination in embryonal stem (ES) cells allows mutations to be introduced into the germline. However, the earliest phenotype of the introduced...
Berg, D J, Kühn, R, Rajewsky, K, Müller, W, Menon, S, Davidson, N, ...
Previous studies in vivo have shown that IL-10 infusion can prevent lethal endotoxic shock. Mice deficient in the production of IL-10 (IL10T) were used to investigate the regulatory role of IL-10 in...
Stepwise intraclonal maturation of antibody affinity through somatic hypermutation.
Using recombinant DNA techniques, we reconstructed a genealogical tree (Sablitzky, F., Wildner, G. & Rajewsky, K. (1985) EMBO J. 4, 345-350) that connects three clonally related B cells producing...
Antibodies generated from human immunoglobulin miniloci in transgenic mice.
Wagner, S D, Williams, G T, Larson, T, Neuberger, M S, Kitamura, D, Rajewsky, K, ...
One approach to the production of human monoclonal antibodies focusses on the creation of transgenic mice bearing human immunoglobulin gene miniloci. Whilst such loci undergo lymphoid-specific gene...
PCR-based cDNA library construction: general cDNA libraries at the level of a few cells.
Belyavsky, A, Vinogradova, T, Rajewsky, K
A procedure for the construction of general cDNA libraries is described which is based on the amplification of total cDNA in vitro. The first cDNA strand is synthesized from total RNA using an...
From a hybrid mouse cell line (B1-8) that secreted an IgM, lambda 1 anti-(4-hydroxy-3-nitrophenyl)acetyl antibody but that had no detectable surface IgM, selection for a variant with lambda 1 chains...
Radbruch, A, Müller, W, Rajewsky, K
Mouse B lymphocytes can be activated polyclonally by bacterial lipopolysaccharide (LPS) to secrete Ig and perform Ig class switch. In the presence of the T-cell lymphokine B-cell differentiation...
Isolation of variants of mouse myeloma X63 that express changed immunoglobulin class.
Radbruch, A, Liesegang, B, Rajewsky, K
We have used fluorescence-activated cell sorting with class-specific antisera to isolate spontaneous variants in the expression of immunoglobulin heavy chain class from the mouse myeloma cell line...
Recombination between antibody heavy chain variable-region genes: evidence for gene conversion.
Krawinkel, U, Zoebelein, G, Brüggemann, M, Radbruch, A, Rajewsky, K
The murine hybridoma line B1-8.delta 1 secretes monoclonal IgD lambda 1 antibodies specific for the hapten (4-hydroxy-3-nitrophenyl)acetyl (NP). The variable (V) region of these antibodies is defined...
Steplewski, Z, Spira, G, Blaszczyk, M, Lubeck, M D, Radbruch, A, Illges, H, ...
Three different monoclonal antibody isotypes, IgG1, IgG2b, and IgG2a, were derived by selection of isotype-switch variants from the CO-19-9 hybridoma. All three antibodies retained their binding...
Isolation of myeloma variants with predefined variant surface immunoglobulin by cell sorting.
Liesegang, B, Radbruch, A, Rajewsky, K
We describe a procedure for the isolation of somatic cell variants in which gene products are expressed on the cell surface that are not expressed in the wild type. Cloned cells of the myeloma line...
Antigen induces chronic idiotype suppression.
Hapten coupled to isologous gamma globulin can substitute for anti-idiotypic antibody in the induction of neonatal chronic idiotype suppression. The immunologically suppressed animals fail to express...
Zou, Y R, Takeda, S, Rajewsky, K
The production of lambda chain-expressing B cells was studied in mice in which either the gene encoding the constant region of the kappa chain (C kappa) or the intron enhancer in the Ig kappa locus...
A novel gene product associated with mu chains in immature B cells.
Shirasawa, T, Ohnishi, K, Hagiwara, S, Shigemoto, K, Takebe, Y, Rajewsky, K, ...
A previously unreported B cell specific gene, which we have named 8HS-20, was isolated from the cDNA library of a pre-B cell clone by subtraction and differential hybridization. This gene is...
Takeda, S, Zou, Y R, Bluethmann, H, Kitamura, D, Muller, U, Rajewsky, K
Immunoglobulins (Ig) secreted from a plasma cell contain either kappa or lambda light chains, but not both. This phenomenon is termed isotypic kappa-lambda exclusion. While kappa-producing cells have...
Developmental and functional impairment of T cells in mice lacking CD3 zeta chains.
Ohno, H, Aoe, T, Taki, S, Kitamura, D, Ishida, Y, Rajewsky, K, ...
CD3 zeta is a component of the T cell antigen receptor (TCR) complex and is important for signal transduction. We have established mice selectively lacking CD3 zeta but able to express CD3 eta, a...
Küppers, R, Zhao, M, Hansmann, M L, Rajewsky, K
Germinal centres are areas of intense B lymphocyte proliferation inside primary B cell follicles in spleen and lymph nodes. Rearranged V genes from single human B cells, isolated from histological...
Antibody engineering for the analysis of affinity maturation of an anti-hapten response.
Allen, D, Simon, T, Sablitzky, F, Rajewsky, K, Cumano, A
The influence of structural variation, previously observed in a panel of V186.2 VH/V lambda 1-expressing anti-NP antibodies from the secondary response, on the affinity of these antibodies was...
A novel 34-kd protein co-isolated with the IgM molecule in surface IgM-expressing cells.
Hombach, J, Leclercq, L, Radbruch, A, Rajewsky, K, Reth, M
Plasmacytoma cells, transfected with a vector encoding a membrane-bound IgM molecule, do not show cell surface IgM expression, although complete IgM molecules are assembled intracellularly. The...
Förster, I, Gu, H, Rajewsky, K
Antibody V gene expression was studied in a subpopulation of murine B cells (Ly1 B) which was enriched by cell transfer and had earlier been shown to persist in the immune system over long periods of...
Gu, H, Förster, I, Rajewsky, K
Sequence analysis of rearranged VHDJH genes of B lineage cells from various stages of ontogeny indicates that short sequence homologies at the breakpoints of recombination contribute to V region gene...
Brüggemann, M, Radbruch, A, Rajewsky, K
The hybridoma cell line B1-8.delta 1 expresses an IgD antibody with specificity for the hapten 4-hydroxy-3-nitro-phenylacetyl (NP). Two monoclonal antibodies, each recognizing an idiotypic...
Immunoglobulin V region variants in hybridoma cells. II. Recombination between V genes.
Dildrop, R, Brüggemann, M, Radbruch, A, Rajewsky, K, Beyreuther, K
The mouse hybridoma line B1-8.delta 1 secretes a monoclonal IgD, lambda 1 anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) antibody with defined idiotypic determinants. Two spontaneous V-region variants...
Yoshida, N, Radbruch, A, Rajewsky, K
In cultures of murine bone marrow cells colonies of 10(3)-10(4) cells were identified which consisted to a large part of pre-B and B cells. Cell mixing experiments with genetically marked cells...
Somatic mutation and clonal expansion of B cells in an antigen-driven immune response.
Sablitzky, F, Wildner, G, Rajewsky, K
The variable (V) regions of three closely related monoclonal antibodies produced by hybridomas which had been isolated from a single mouse were sequenced at the level of the mRNA. The sequences and...
Sablitzky, F, Weisbaum, D, Rajewsky, K
The non-expressed, rearranged JH loci of two hybridoma cells, thought to be derived from a single B cell precursor were cloned and partially sequenced. Identical DJ rearrangements including N...
Holtkamp, B, Cramer, M, Rajewsky, K
In the heterozygous T lymphoma line LDHB, variants which have lost the expression of individual H-2 class I genes are spontaneously generated in vitro at a frequency of 10(-1)-10(-2). A cell line...
Dildrop, R, Bovens, J, Siekevitz, M, Beyreuther, K, Rajewsky, K
Idiotope Ac38, a V region determinant of the lambda 1 chain-bearing, germ line encoded antibody B1-8, is expressed at high frequency (approximately 1/40) in lambda 1 chain-bearing B cells. Here, we...
Molecular basis of an isogeneic anti-idiotypic response.
The nucleotide sequences of the variable region genes expressed in the heavy and light chains of six isogeneic anti-idiotope antibodies recognizing idiotopes on two closely related antibodies with...
Brüggemann, M, Müller, H J, Burger, C, Rajewsky, K
Somatic mutation occurs at a low rate in the rearranged antibody V region genes of the hybridoma line B1-8. delta 1 which expresses an antibody with specificity for the hapten...
Clonal recruitment and somatic mutation in the generation of immunological memory to the hapten NP.
The nucleotide sequences of the variable regions of lambda 1 chain bearing anti-NP antibodies from the secondary response of C57BL/6 mice were determined. The data indicate that the V186.2 VH gene...
Ku80 is required for immunoglobulin isotype switching.
Casellas, R, Nussenzweig, A, Wuerffel, R, Pelanda, R, Reichlin, A, Suh, H, ...
Isotype switching is the DNA recombination mechanism by which antibody genes diversify immunoglobulin effector functions. In contrast to V(D)J recombination, which is mediated by RAG1, RAG2 and DNA...
Antibody domain mutants demonstrate autonomy of the antigen binding site.
We have constructed derivatives of a lambda I light chain-bearing anti-(4-hydroxy,3-nitrophenyl)acetyl (NP) antibody which have the V regions exchanged between heavy chains and light chains of the...
Lymphoid development in mice with a targeted deletion of the interleukin 2 receptor gamma chain.
DiSanto, J P, Müller, W, Guy-Grand, D, Fischer, A, Rajewsky, K
The interleukin 2 receptor gamma chain (IL-2R gamma) is a component of the receptors for IL-2, IL-4, IL-7, and IL-15. Mutations in IL-2R gamma in man appear responsible for the X chromosome-linked...
Küppers, R, Rajewsky, K, Zhao, M, Simons, G, Laumann, R, Fischer, R, ...
Hodgkin disease (HD) is characterized by a small number of putative malignant cells [Hodgkin and Reed-Sternberg (HRS) cells] among a background of lymphocytes and histiocytes. The lineage of HRS...
We have reevaluated the in vivo lifespan of B lymphocytes based on incorporation of the thymidine analogue 5-bromo-2'-deoxyuridine into the DNA of dividing cells. To exclude potential alterations in...
B lymphocyte-specific, Cre-mediated mutagenesis in mice.
Rickert, R C, Roes, J, Rajewsky, K
Adaptation of the P1 phage-derived Cre /loxP site- specific recombination system to the gene targeting technique allows for the conditional deletion of genes in mice. To selectively modify genes in B...
Temporally and spatially regulated somatic mutagenesis in mice.
Schwenk, F, Kuhn, R, Angrand, P O, Rajewsky, K, Stewart, A F
In mice transgenesis through oocyte injection or DNA recombination in embryonal stem (ES) cells allows mutations to be introduced into the germline. However, the earliest phenotype of the introduced...
Berg, D J, Kühn, R, Rajewsky, K, Müller, W, Menon, S, Davidson, N, ...
Previous studies in vivo have shown that IL-10 infusion can prevent lethal endotoxic shock. Mice deficient in the production of IL-10 (IL10T) were used to investigate the regulatory role of IL-10 in...
Stepwise intraclonal maturation of antibody affinity through somatic hypermutation.
Using recombinant DNA techniques, we reconstructed a genealogical tree (Sablitzky, F., Wildner, G. & Rajewsky, K. (1985) EMBO J. 4, 345-350) that connects three clonally related B cells producing...
Antibodies generated from human immunoglobulin miniloci in transgenic mice.
Wagner, S D, Williams, G T, Larson, T, Neuberger, M S, Kitamura, D, Rajewsky, K, ...
One approach to the production of human monoclonal antibodies focusses on the creation of transgenic mice bearing human immunoglobulin gene miniloci. Whilst such loci undergo lymphoid-specific gene...
PCR-based cDNA library construction: general cDNA libraries at the level of a few cells.
Belyavsky, A, Vinogradova, T, Rajewsky, K
A procedure for the construction of general cDNA libraries is described which is based on the amplification of total cDNA in vitro. The first cDNA strand is synthesized from total RNA using an...
From a hybrid mouse cell line (B1-8) that secreted an IgM, lambda 1 anti-(4-hydroxy-3-nitrophenyl)acetyl antibody but that had no detectable surface IgM, selection for a variant with lambda 1 chains...
Radbruch, A, Müller, W, Rajewsky, K
Mouse B lymphocytes can be activated polyclonally by bacterial lipopolysaccharide (LPS) to secrete Ig and perform Ig class switch. In the presence of the T-cell lymphokine B-cell differentiation...
Isolation of variants of mouse myeloma X63 that express changed immunoglobulin class.
Radbruch, A, Liesegang, B, Rajewsky, K
We have used fluorescence-activated cell sorting with class-specific antisera to isolate spontaneous variants in the expression of immunoglobulin heavy chain class from the mouse myeloma cell line...
Recombination between antibody heavy chain variable-region genes: evidence for gene conversion.
Krawinkel, U, Zoebelein, G, Brüggemann, M, Radbruch, A, Rajewsky, K
The murine hybridoma line B1-8.delta 1 secretes monoclonal IgD lambda 1 antibodies specific for the hapten (4-hydroxy-3-nitrophenyl)acetyl (NP). The variable (V) region of these antibodies is defined...
Steplewski, Z, Spira, G, Blaszczyk, M, Lubeck, M D, Radbruch, A, Illges, H, ...
Three different monoclonal antibody isotypes, IgG1, IgG2b, and IgG2a, were derived by selection of isotype-switch variants from the CO-19-9 hybridoma. All three antibodies retained their binding...
Isolation of myeloma variants with predefined variant surface immunoglobulin by cell sorting.
Liesegang, B, Radbruch, A, Rajewsky, K
We describe a procedure for the isolation of somatic cell variants in which gene products are expressed on the cell surface that are not expressed in the wild type. Cloned cells of the myeloma line...
Antigen induces chronic idiotype suppression.
Hapten coupled to isologous gamma globulin can substitute for anti-idiotypic antibody in the induction of neonatal chronic idiotype suppression. The immunologically suppressed animals fail to express...
Zou, Y R, Takeda, S, Rajewsky, K
The production of lambda chain-expressing B cells was studied in mice in which either the gene encoding the constant region of the kappa chain (C kappa) or the intron enhancer in the Ig kappa locus...
A novel gene product associated with mu chains in immature B cells.
Shirasawa, T, Ohnishi, K, Hagiwara, S, Shigemoto, K, Takebe, Y, Rajewsky, K, ...
A previously unreported B cell specific gene, which we have named 8HS-20, was isolated from the cDNA library of a pre-B cell clone by subtraction and differential hybridization. This gene is...
Takeda, S, Zou, Y R, Bluethmann, H, Kitamura, D, Muller, U, Rajewsky, K
Immunoglobulins (Ig) secreted from a plasma cell contain either kappa or lambda light chains, but not both. This phenomenon is termed isotypic kappa-lambda exclusion. While kappa-producing cells have...
Developmental and functional impairment of T cells in mice lacking CD3 zeta chains.
Ohno, H, Aoe, T, Taki, S, Kitamura, D, Ishida, Y, Rajewsky, K, ...
CD3 zeta is a component of the T cell antigen receptor (TCR) complex and is important for signal transduction. We have established mice selectively lacking CD3 zeta but able to express CD3 eta, a...
Küppers, R, Zhao, M, Hansmann, M L, Rajewsky, K
Germinal centres are areas of intense B lymphocyte proliferation inside primary B cell follicles in spleen and lymph nodes. Rearranged V genes from single human B cells, isolated from histological...
Antibody engineering for the analysis of affinity maturation of an anti-hapten response.
Allen, D, Simon, T, Sablitzky, F, Rajewsky, K, Cumano, A
The influence of structural variation, previously observed in a panel of V186.2 VH/V lambda 1-expressing anti-NP antibodies from the secondary response, on the affinity of these antibodies was...
A novel 34-kd protein co-isolated with the IgM molecule in surface IgM-expressing cells.
Hombach, J, Leclercq, L, Radbruch, A, Rajewsky, K, Reth, M
Plasmacytoma cells, transfected with a vector encoding a membrane-bound IgM molecule, do not show cell surface IgM expression, although complete IgM molecules are assembled intracellularly. The...
Förster, I, Gu, H, Rajewsky, K
Antibody V gene expression was studied in a subpopulation of murine B cells (Ly1 B) which was enriched by cell transfer and had earlier been shown to persist in the immune system over long periods of...
Antibody domain mutants demonstrate autonomy of the antigen binding site.
We have constructed derivatives of a lambda I light chain-bearing anti-(4-hydroxy,3-nitrophenyl)acetyl (NP) antibody which have the V regions exchanged between heavy chains and light chains of the...
Gu, H, Förster, I, Rajewsky, K
Sequence analysis of rearranged VHDJH genes of B lineage cells from various stages of ontogeny indicates that short sequence homologies at the breakpoints of recombination contribute to V region gene...