THE manufacture of predefined specific antibodies by means of permanent tissue culture cell lines is of general interest. There are at present a considerable number of permanent cultures of myeloma cells1,2 and screening procedures have been used to reveal antibody activity in some of them. This, however, is not a satisfactory source of monoclonal antibodies of predefined specificity. We describe here the derivation of a number of tissue culture cell lines which secrete anti-sheep red blood cell (SRBC) antibodies. The cell lines are made by fusion of a mouse myeloma and mouse spleen cells from an immunised donor. To understand the expression and interactions of the Ig chains from the parental lines, fusion experiments between two known mouse myeloma lines were carried out.

Continuous cultures of fused cells secreting antibody of predefined specificity

The Human Plasma Proteome History, Character, and Diagnostic Prospects

A human IgGI antibody has been reshaped for serotherapy in humans by introducing the six hypervariable regions from the heavy- and light-chain variable domains of a rat antibody directed against human lymphocytes. The reshaped human antibody is as effective as the rat antibody in complement and is more effective in cell-mediated lysis of human lymphocytes.

Reshaping human antibodies for therapy.

The title of the Nobel Lecture of George Palade (1 August, p. 347) should have been "Intracellular aspects of the process of protein secretion."

https://science.sciencemag.org/content/sci/189/4200/347.full.pdf

Intracellular aspects of the process of protein synthesis

In this contribution we put forward a novel hypothesis concerning the aetiology of Type 2 (non-insulin dependent) diabetes mellitus. The concept underlying our hypothesis is that poor foetal and early post-natal nutrition imposes mechanisms of nutritional thrift upon the growing individual. We propose that one of the major long-term consequences of inadequate early nutrition is impaired development of the endocrine pancreas and a greatly increased susceptibility to the development of Type 2 diabetes. In the first section we outline our research which has led to this hypothesis. We will then review the relevant literature. Finally we show that the hypothesis suggests a reinterpretation of some findings and an explanation of others which are at present not easy to understand.

/pdf/type-2-non-insulin-dependent-diabetes-mellitus-the-thrifty-wkzyogfl8p.pdf

Type 2 (non-insulin-dependent) diabetes mellitus: the thrifty phenotype hypothesis.

IMMUNOGLOBULIN light chains are synthesized in heterologous cell-free systems from reticulocytes1 and Krebs II ascites cells2 and in reconstituted homologous systems from lymph nodes3,4 and myeloma tumours5,6. Although the synthesis appeared to be essentially faithful, absolute identity with authentic light chains was not demonstrated and some small differences were observed. We report here a clear discrepancy between the product of certain cell-free reactions and authentic light chain. With the product of other cell-free reactions no such discrepancy is seen. We propose that light chains are initially synthesized as a precursor of slightly higher molecular weight and subsequently converted into the authentic product.

A possible precursor of immunoglobulin light chains

MONOCLONAL antibodies can be produced by cultures of permanent cell lines derived by fusion of suitable myelomas with spleen cells from immunised animals1. So far, mainly two mouse myelomas, X63-Ag8 and NSI/1-Ag4.1, have been used for this purpose1,2. When cells from immunised mice are used in the fusion, the mouse × mouse hybrid myelomas can be grown as tumours in appropriate mouse strains. We describe here the preparation and use of a new clone of a rat myeloma, which is suitable for the derivation of rat × rat hybrid myelomas producing specific rat antibodies. The spent medium of hybrid myeloma cultures usually contains 1–20 μg ml−1 antibody. The serum and ascites of the tumour-bearing mice, with few exceptions, yield 1–20 mg ml−1 of antibody, which is 1,000 times more concentrated and very convenient for larger preparations.

Rat × rat hybrid myelomas and a monoclonal anti-Fd portion of mouse IgG

Thymocyte antigens CD1 [Thy,gp45,12] are thought to be the human counterparts of mouse thymus leukaemia (TL) antigens1,2. Serological and biochemical analyses indicate that at least three subsets exist3–6, the first of which (HTA 1/T6) was initially identified by the monoclonal antibody NA1/34 (refs 7,8). Like TL, CD1 are expressed on cortical thymocytes as well as on some lymphoid neoplasias, and resemble in structure major histocompatabiliry complex (MHC) class I antigens. However HTA 1/T6 is loosely associated with β2-microglobulin9–10 and is also found linked by a disulphide bridge to CD8 (T8)11,12. A molecular genetic approach is needed to investigate the CD1 system, to clarify its relationship to TL antigens and to understand its regulation. We report the isolation of complementary DNA (cDNA) clones encoding a CD1 antigen. These clones reveal a novel family of genes which are MHC-related but are neither equivalent to mouse TL antigens nor linked to the MHC.

A novel family of human major histocompatibility complex-related genes not mapping to chromosome 6

EACH immunoglobulin chain is the integrated expression of one of several V and C genes, coding respectively for their variable and constant sections1. The restricted expression of these genes leads to molecules of a single class and type with identical combining sites at both halves of the antibody molecule2–4. This symmetry is essential for the formation of the antigen-antibody lattice. Its importance is emphasized by allelic exclusion, whereby each cell expresses only one of two possible alleles4. To understand this problem further we have studied the expression of these genes in hybrid cells obtained by fusion of two cell lines producing different immunoglobulins. Successful fusion of this type has not yet been demonstrated but fusion between immunoglobulin-producing cells and non-immunoglobulin-producing cells has been reported. Thus myeloma cells have been fused to fibroblasts5–7 and also to a non-immunoglobulin-producing lymphoma line8. Substantial immunoglobulin production in the hybrids was observed only in the latter case. Here, we demonstrate that fusion between two immunoglobulin-secreting cell lines produces hybrid cells which secrete immunoglobulin of both parental types.

C. Milstein

Papers

Continuous cultures of fused cells secreting antibody of predefined specificity

A possible precursor of immunoglobulin light chains

Rat × rat hybrid myelomas and a monoclonal anti-Fd portion of mouse IgG

A novel family of human major histocompatibility complex-related genes not mapping to chromosome 6

Fusion of Two Immunoglobulin-producing Myeloma Cells