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C Milstein

Publications and source records attributed to C Milstein.

At least 145 records · Page 8Linked to original sources
PubMed1981

Complete sequence of an immunoglobulin mRNA using specific priming and the dideoxynucleotide method of RNA sequencing.

The complete sequence of the mouse immunoglobulin kappa light chain MOPC 21 messenger RNA has been determined using a chain termination method and chemically synthesised deoxyoligonucleotides to initiate the synthesis of a DNA molecule complementary to the mRNA template. Five such oligonucleotide primers have been used for the sequence analysis of this messenger RNA. The approach is excellent for comparative studies of mouse k-chain mRNAs because they can be made on impure mRNA preparations. The MOPC 21 light chain mRNA is 943 nucleotides in length excluding the poly(A) region. An unexpected finding was that there are only three bases in the 5' non-coding region and its significance in terms of ribosome binding is discussed; 87 code for the precursor or leader sequence of the protein, 642 for the mature protein and 211 for the 3' non-coding region. The codons for the precursor region allows the previously undetermined amino acid sequence to be predicted. In common with other precursor regions a high proportion of the predicted amino acids are hydrophobic.

Animals↗
PubMed1981

The Wellcome Foundation Lecture, 1980: monoclonal antibodies from hybrid myelomas.

When the lymphoid cells from immunized animals are fused with myeloma cells adapted to grow permanently in culture, hybrid cells can be isolated that are capable of permanent growth in culture, or as transplantable myeloma tumour in animals, and that at the same time express the antibodies of the immunized donor. Such hybrid cells can be cloned and the antibody produced by each clone is monoclonal. By this procedure therefore it is possible to dissect the heterogeneous immune response of an animal. The monoclonal antibodies can be permanently produced in unlimited quantities and the products are well defined chemical entities, unlike antibodies prepared in animals, which vary from animal to animal and even in different periods within a single animal. These properties have been of great importance in the use of antibodies as biochemical reagents in basic research in a variety of fields. They are also replacing conventional antibodies in standard laboratory practice.

Animals↗
PubMed1981

Expression of spleen cell immunoglobulin phenotype in hybrids with myeloma cell lines.

Fusions were performed between myeloma cell lines, of mouse and rat origin, and mouse or rat spleen cells. Two statistical methods have been used to measure the proportion of hybrids expressing a spleen cell-derived immunoglobulin phenotype, one of them applicable to cells growing under nonlimiting dilution conditions. The results indicate that there is strong preferential selection for hybrid cell growth with an immunoglobulin-secreting phenotype. The degree of preferential selection is dependent upon the myeloma cell line used and is most marked in the case of the rat myeloma lines. Surviving hybrids seem to originate from fusions of myeloma and spleen B(but not T) cells, but immunoglobulin production is lost more readily in certain combinations.

Animals↗
PubMed1981

Immunocytochemical detection of serotonin with monoclonal antibodies.

The development of the monoclonal antibody YC5/45 HLK (YC5/HLK) against a 5HT-bovine seroalbumin immunogen and its application for immunocytochemistry is described. The YC5/HLK antibody is the product of a rat x rat hybrid myeloma, producing a heavy chain and two light chains. In hemagglutination tests, the antibody cross-reacts to entirety with dopamine, serotonin, and tryptamine at high concentrations. The serotonin-albumin conjugate is 20,000 times more effective in displacing the binding antibody, while albumin itself goes unrecognized by the antibody. In fixed preparations of brain tissue, immunofluorescence is observed only in neurons known to contain serotonin, while no reaction is observed in dopamine-rich neurons. All immunofluorescence is extinguished by the use of agents that inhibit the biosynthesis of 5HT, but not of the catecholamines.

Animals↗
PubMed1981

Use of a monoclonal antibody specifically non-reactive with T cells to delineate lymphocyte subpopulations.

Rat monoclonal antibody M1/69.16 reacts with a heat stable antigen of mouse commonly expressed in the majority of cell types in blood, spleen, bone marrow and thymus, including cells of erythroid, myeloid and lymphoid series. However, subpopulations of cells in lymphoid tissues can be identified which are non-reactive with this antibody using the fluorescence-activated cell sorter. All surface Ig positive cells seem to react with M1/69.16 while more than 96% of Ig negative cells in spleen and lymph nodes are M1/69.16 negative. Most cells (80%-90%) in the M1/69.16 negative populations in spleen lymph nodes and bone marrow express Thy-l. Thus, peripheral T cells are specifically non-reactive with this antibody. In contrast, approximately 95% of thymocytes react with M1/69.16, leaving a minor population which is negative. The negative population (5%) is enriched in cells expressing high amounts of H-2 antigen and those bearing H9/25 antigen which is specific for lymphocyte subsets, indicating that M1/69.16 negative thymocytes represent a specific subpopulation, possibly "mature' thymocytes.

Animals↗
PubMed1980

A mouse immunoglobulin heavy chain deletion mutant: isolation of a cDNA clone and sequence analysis of the mRNA.

The mouse cell line IF2 secretes an immunoglobulin heavy chain lacking the CH1 domain. We have isolated and characterised a recombinant plasmid containing cDNA copies of the IF2 mutant mRNA. The cloned sequence extends from the nucleotides coding for amino acid 96 in the variable region through 100 nucleotides of untranslated region at the 3' end. The sequence of the cDNA insert reveals no discontinuity at the variable-hinge region junction, the site of the CH1 deletion. Experiments employing direct priming on the poly(A) tail of the IF2 heavy chain mRNA suggest that the 3' end of the cDNA clone (sequence C-C-C-T-G-C) is also the 3' end of the mRNA.

Animals↗
PubMed1980

H 9/25 monoclonal antibody recognizes a new allospecificity of mouse lymphocyte subpopulations: strain and tissue distribution.

C3 H/He-mg mice were immunized with C57BL/10 (B10) spleen cells and the immune spleen cells were fused with BALB/c myeloma cells (NS1). One of the monoclonal antibodies (H9/25 antibody) produced by the hybrid cells was studied. It reacts with subpopulations of B10 lymphocytes as well as some lymphoid tumor lines including some of the Abelson virus-induced leukemias. The antigen recognized by H9/25 antibody is expressed on lymphocytes from all the B10 congeneic mice tested as well as some other strains of mice. No linkage between genes coding for the antigen and H-2 loci was found as judged by its presence on cells of the B10 strains regardless of H-2 type and the distribution of the antigen on Bailey recombinant inbred mice. The antigen is expressed on subpopulations of lymph node cells, spleen cells, thymocytes and bone marrow cells. The strain distribution of the H9/25 antigen seems to be identical to that of Ly-6, Ly-8 and Ala-1 antigens. However, the tissue distribution of the antigen recognized by H9/25 antibody, while similar to these alloantigens, is unique and the antigen may be distinct from the other alloantigens.

Animals↗
PubMed1980

Monoclonal antibody H9/25 reacts with functional subsets of T and B cells: killer, killer precursor and plaque-forming cells.

Monoclonal antibody (McAb) H9/25 has previously been shown to react with an alloantigen expressed on subpopulations of mouse lymphocytes. We here investigated the expression of the antigen (H9/25 Ag) on functional subsets of T and B cells. Lymphocytes were depleted of H9/25 Ag-bearing cells by complement-dependent cytolysis or by the affinity to immobilized McAb H9/25, and the residual immunological functions were tested. In these tests, killer T cells generated in mixed lymphocyte cultures and their precursors were found to express H9/25 Ag. In contrast, the activity and frequency of helper T cells specific to keyhole limpet hemocyanin carrier were not decreased by the depletion of H9/25 Ag-bearing cells. On the other hand, IgG and IgM anti-TNP plaque-forming cells were found to express H9/25 Ag, while it was not detected on unprimed B cells, as tested by using TNP-Ficoll and TNP-lipopolysaccharide. Only a small proportion of TNP memory B cells seemed to express the antigen. These results revealed significant differences between H9/25 Ag and other known alloantigens in the distribution among functional subsets of lymphocytes.

Animals↗
PubMed1980

Chromosome segregation and expression of rat immunoglobulins in rat/mouse hybrid myelomas.

The chromosome segregation of hybrid myelomas from a fusion of rat immunized spleen cells and the mouse myeloma P3-X63-Ag8 has been analysed. Chromosome loss appears to be nonrandom. Most mouse chromosomes are retained. The rat chromosome are preferentially lost but a few--particularly 1-5 and 13--are rarely lost. The specific retention of some rat chromosomes explains the stability of rat Ig expression in mouse-rat hybrids. Correlation of chromosome loss and retention and loss of rat heavy and light chains leads us to propose chromosome 14 as coding for the heavy chain.

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