Biomedical subjects

M Wabl

Publications and source records attributed to M Wabl.

58 records · Page 4Linked to original sources

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↗

PubMedDate not supplied

HeLa-LAV, an epithelial cell line stably infected with HIV-1.

An HeLa-LAV cell line was established by infecting and subcloning previously described CD4-expressing HeLa cells with HIV-1. Cells of this line stably synthesize all major HIV proteins, release infectious particles of HIV-1, but do not die even after long term culture. More than 90% of the cells express the envelope protein gp120 on the surface. The cells can be easily and efficiently labeled with 51chromium, and exhibit a low spontaneous release. Because they are susceptible to killing by allogeneic cytotoxic T cells (CTL) when targeted to gp120, they ought to be a useful source of target cells in any kind of HIV-specific killing assays. The cells may also help studies on HIV replication in non-lymphatic/non-monocytic cells. The HeLa-LAV cell line will be freely available from the AIDS Research and Reference Reagent Program.

CD4-Positive T-Lymphocytes↗

PubMedDate not supplied

Immunoglobulin heavy chain binding protein.

Pre-B lymphocytes, and hybridomas derived from them, synthesize immunoglobulin heavy (IgH) chain in the absence of light (L) chain. In the Abelson virus transformed line 18-81, which is representative of the pre-B cell stage, we observed that at least some of the H-chains are bound to a protein other than L-chain. Here we show that the protein (which we term immunoglobulin heavy-chain binding protein, BiP) binds non-covalently to free IgH, but not to IgH associated with IgL.

Animals↗

PubMedDate not supplied

Critical test of a sister chromatid exchange model for the immunoglobulin heavy-chain class switch.

B lymphocytes may switch from producing an immunoglobulin heavy chain of the mu class to that of the gamma, epsilon or alpha class. To maintain the specificity, the new heavy chain must keep the original variable (V) region; this is achieved by deleting DNA sequences so that the V (consisting of joined VH, diversity (DH) and joining (JH) gene segments) and C (constant) gene segments coding for the new heavy chain are brought into close proximity (reviewed in ref. 5; we do not consider here the mu-delta situation). There are, in principle, three types of chromosomal rearrangements that yield a deletion: rearrangement within a chromatid; unequal sister chromatid exchange (as suggested by Obata et al.); and unequal recombination between chromosomal homologues. We have analysed the arrangement of C mu DNA in clones of the pre-B-cell line 18-81 that switches in vitro from mu to gamma 2b. The clones examined produce either mu, gamma 2b or no immunoglobulin chain. We report here that all the gamma 2b clones had lost at least one copy of C mu and no clones contained three copies of C mu. These findings formally exclude both unequal sister chromatid exchange and recombination between homologues as mechanisms for creating a gene encoding the gamma 2b chain.

Animals↗