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A Traunecker

Publications and source records attributed to A Traunecker.

29 records · Page 2Linked to original sources
PubMed1986

Normal T cell development is possible without 'functional' gamma chain genes.

The T cell-specific gamma gene family is organized into four V, J and C gene segments containing clusters (gamma 1, gamma 2, gamma 3, gamma 4) in germline DNA. We found that the V, J and C elements of gamma 2 are physically linked on a stretch of 6 kb of DNA while those of gamma 3 are found within a 15-kb region. Rearrangements take place only within the clusters, explaining the rigid rearrangement patterns seen in T lymphocytes. New V gamma, J gamma and C gamma gene segments were discovered and characterized allowing the better understanding of the potential germline diversity of the gamma gene family. No correlation with T cell function, i.e. cytolytic or helper, and the type of the productive gamma rearrangement could be established. In contrast we found that functional T cell clones have been able to mature without any functional gamma chain genes.

Animals↗
PubMed1984

Somatic mutation creates diversity in the major group of mouse immunoglobulin kappa light chains.

Using a cloned cDNA of a mouse immunoglobulin kappa light chain synthesized in a myeloma MOPC321 (V kappa-21 subgroup C) as a probe we could detect 13 germ line V kappa gene segments. 11 of these were isolated. Using a set of overlapping cloned segments, we showed that nine of these germ line V kappa genes are arranged in two linkage clusters and that they all have the same transcriptional orientation (11, 12, 22). These two clusters occupy 90 and 30 kb of chromosomal DNA and contain six and three V kappa's, respectively. We determined the complete nucleotide sequences of five germ line V kappa's and showed that three of them encode the prototype sequence of V kappa-21 subgroups B, C, and E. None of these five germ line V kappa's encodes the variant amino acid sequences of known V kappa-21 subgroups. We thus conclude that, as in the lambda 1 light chains, the variant V regions are encoded by gene segments derived by a few somatic mutations from the corresponding germ line DNA. Such somatic mutations are not restricted to sequences encoding the hypervariable regions: they also occur in sequences encoding framework regions.

Amino Acid Sequence↗
PubMed1984

Different ways to modify monoclonal antibodies.

In this paper we summarize experiments which were undertaken to create altered antibody molecules. Three different approaches were used. Established hybridoma lines were re-hybridized to mouse spleen cells to generate arrays of secondary hybridomas which express one particular heavy chain and one specificity together with a multitude of different light chains. In such hybrids the influence of light chains to the antibody combining site and the influence of affinity to antibody effector functions can be studied. Another way to obtain altered antibodies was the selection of cells producing less lytic IgM. With this technique we obtained (among many other variants) a series of mu-deletion products which were used to map the fine specificity of rat anti-mouse mu monoclonal antibodies. Both the anti-mu antibodies and the deletion variants were used to assign the Clq binding to the fourth C mu-domain demonstrating the power of mutant IgM in the structure-function analysis. In a third series of experiments we show the feasibility of generating new antibody combining sites by the methods of molecular genetics. The variable region gene of a heavy chain was placed in front of a kappa-constant region gene. The plasmid construct was transferred into mouse myeloma lines which stably express a variable heavy-constant light chain protein. Upon fusion with a light chain producing line, chimaeric light chain dimers with a functional antibody combining site were secreted. These experiments demonstrate that new series of man-made antibody molecules can be made in the future.

Animals↗
PubMed1983

Functional immunoglobulin M production after transfection of cloned immunoglobulin heavy and light chain genes into lymphoid cells.

The rearranged immunoglobulin heavy (mu) and light (kappa) chain genes cloned from the Sp6 hybridoma cell line producing immunoglobulin M specific for the hapten 2,4, 6-trinitrophenyl were inserted into the transfer vector pSV2-neo and introduced into various plasmacytoma and hybridoma cell lines. The transfer of the mu and kappa genes resulted in the production of pentameric, hapten-specific, functional IgM.

Animals↗
PubMed1981

The role of DNA rearrangement and alternative RNA processing in the expression of immunoglobulin delta genes.

We have established the exon-intron structure of the gene coding for the constant (C) region of the mouse immunoglobulin delta heavy chain, using DNA clones isolated from BALB/c embryos and the delta mRNA extracted from two delta-producing hybridomas, B1-8. delta 1 and GCL2.8. At least three types of C delta gene structures are identified. A 2.7 kb delta mRNA reveals six exons. This delta mRNA may code for a membrane-bound delta chain. A second delta mRNA of 1.8 kb shares the first (5' side relative to direction of transcription) three exons with the 2.7 kb delta mRNA and in addition contains a fourth exon unique to this mRNA species. This delta mRNA most likely codes for a secreted delta chain. A third delta mRNA, also of 1.8 kb, shares the first four exons and a part of the fifth exon with the 2.7 kb mRNA. Its function, if any, remains unclear. We investigated the question of how a lymphocyte can produce the mu and delta heavy chains simultaneously, using the hybridoma GCL 2.8, which makes both IgM and IgD. Results of Southern gel blot analysis and gene cloning experiments indicate that this cell utilizes the same rearranged VH gene for the synthesis of the mu and delta chains, and yet maintains the embryonic configuration for the C mu and C delta genes and for the intervening region. Based on these results, we conclude that the VH sequence is spliced alternatively to the C mu or C delta sequence during processing of the primary RNA transcript. An alternative mechanism for the expression of the delta gene is found in hybridoma B1-8. delta 1, which actively secretes delta chains and synthesizes no mu chain. This mechanism involves deletion of the C mu gene, which brings the complete VH gene closer to the C delta gene.

Animals↗
PubMed1981

Linkage of the four gamma subclass heavy chain genes.

The genes for the heavy-chain constant regions of the four gamma subclass immunoglobulins were identified in a set of overlapping mouse DNA fragments representing about 100 kilobase pairs (kb) of the mouse genome that was cloned from bacteriophage lambda libraries of BALB/c mouse embryo DNA. R-loop mapping studies show that the genes are located 5'-C gamma 3-34 kb-C gamma 1-21 kb-C gamma 2b-15 kb-C gamma 2a-3' and lie in the same transcriptional orientation. Two DNA segments, one of 19 kb and another of 15 kb, that surround the C gamma 2b and C gamma 2a genes, respectively, show considerable homology and implicate a tandem duplication mechanism in the evolution of this gene cluster.

Animals↗
PubMed1981

Organization of four mouse lambda light chain immunoglobulin genes.

We have cloned four lambda light chain constant region (C) genes from mouse embryo DNA. Each carries its own joining (J) segment approximately 1.3 kilobases to its 5' side. The four C genes occur in two clusters, 5' J3C3J1C13' and 5' J2C2J4C43', with C4 being a new C lambda gene. We have also shown that V lambda 1 is joined productively with C lambda 3 in a lambda 3-producing myeloma, and it is most likely that V lambda 1 and V lambda 2 are the only V lambda genes. Based on the analysis of the germ line and rearranged variable region (V) lambda genes in myelomas we argue that the V lambda 1 and V lambda 2 genes are at the 5' side of the C3C1 and C2C4 clusters, respectively. We propose that the two clusters arose by duplication. We also speculate on the role of J-associated DNA sequences in regulation of expression of the lambda subtypes.

Animals↗
PubMed1980

Exon shuffling generates an immunoglobulin heavy chain gene.

From endonuclease EcoRI partial libraries of DNAs from mouse embryo and MOPC 141, a gamma 2b-producing myeloma, clones were isolated by using a DNA fragment carrying the gamma 2b constant (C) region gene as a hybridization probe. One clone from MOPC 141 contained a heavy chain variable (V) gene and the C gamma 2b gene, as demonstrated by R-loop mapping. The V gene and C gene in this clone were separated by a 3.9-kilobase intron. The characterization of this clone as well as the embryonic clones suggest that at least two recombination events occurred to create the gamma 2b gene in MOPC 141. One of the events is analogous to the V-J joining previously demonstrated in the light chain genes, which brings the major part of the V gene next to a short coding sequence (J). The other event we refer to as "C mu-C gamma 2b switch recombination" because a portion of the intron between the V gene and C gene of the rearranged gamma 2b gene is derived from the 5' flanking sequence of the embryonic C mu gene. A model suggesting how the phenomenon of switch seen in lymphocytes may occur is presented.

Animals↗
PubMedDate not supplied

[In vitro differentiation of B lymphocytes].

In order to find if lymphoid colony-forming cells in the mouse are precursors of lymphocytes or are more differentitaed cells with a high proliferative capacity, their distribution between spleen and lymph node was compared to that of mitogen-sensitive B lymphocytes and that of precursurs of B lymphocytes assayed in mass cultures. The results and the responses to culture conditions suggest that these cell populations are different from each other.

Animals↗