Isolation and characterization of a cDNA clone encoding the murine homologue of the human 20K T3/T-cell receptor glycoprotein

The antigen receptor on the surface of human T lymphocytes, which consists of a heterodimer of relative molecular mass (Mr) 90,000 (90K) (alpha- and beta-chains), is associated with the T3 antigen (gamma = 25K, delta = 20K and epsilon = 20K). A working model for the mode of action of the T3/T-cell receptor complex is that the clonotypic alpha- and beta-chains are involved in the recognition and binding of antigen in the context of polymorphic major histocompatibility complex (MHC) gene products on the surface of target cells. Antigen binding by the clonotypic receptor probably results in conformational changes in this structure which are recognized by and subsequently trigger the associated T3 complex to transmit signals into the cell, resulting in a proliferative response. The similarity in structure between murine and human clonotypic antigen receptors suggests that such a mechanism of recognition and activation also exists in mouse T lymphocytes, but so far there has been no evidence for the existence of a murine T3 complex. Here we demonstrate the existence of a T3 delta-chain mRNA in murine T lymphocytes. Our sequence data strongly suggest that this mouse mRNA codes for a complete T3 delta polypeptide chain and reveal some interesting properties of the protein.     

Selective inhibition of the anchorage-independent growth of myc-transfected fibroblasts by retinoic acid

Fischer rat 3T3 (FR3T3) fibroblasts transfected with a cellular myc gene can be induced to grow and form colonies in soft agar by treatment either with epidermal growth factor (EGF) alone or with the combination of platelet-derived growth factor (PDGF) and type-beta transforming growth factor (TGF-beta). We now show that induction of anchorage-independent growth by each of these sets of growth factors involves different cellular pathways which can be distinguished by their sensitivity to retinoic acid. Colony formation induced by the combined action of PDGF and TGF-beta is 100-fold more sensitive to inhibition by retinoic acid than is colony formation induced by treatment of the myc-transfected cells with EGF. Moreover, retinoic acid (10(-8) M) is inhibitory for colony growth whenever TGF-beta is present, regardless of whether the effects of TGF-beta are stimulatory, as occurs in the presence of PDGF, or inhibitory, as found in the presence of EGF.     

Amino-acid sequence of a Ca2+ + Mg2+-dependent ATPase from rabbit muscle sarcoplasmic reticulum, deduced from its complementary DNA sequence

We have cloned and sequenced complementary DNA encoding a Ca2+-ATPase of rabbit muscle sarcoplasmic reticulum. We propose a model of the protein which has 3 cytoplasmic domains joined to a set of 10 transmembrane helices by a narrow, penta-helical stalk. In this model, ATP bound to one cytoplasmic domain would phosphorylate an aspartate in an adjoining cytoplasmic domain, inducing translocation of Ca2+ from binding sites on the stalk.     

Selective activation of Lyt 2+ precursor T cells by ligation of the antigen receptor

Resting T lymphocytes may be activated either physiologically, by the specific recognition of antigen in association with molecules encoded by the major histocompatibility complex (MHC), or non-physiologically using mitogens such as concanavalin A (Con A). The former activation process is difficult to analyse because resting precursor T cells specific for a particular antigen-MHC combination can only be isolated in the presence of a large excess of bystander cells of irrelevant specificity; clonal populations of uniform specificity are not useful for studying the activation of naive T cells because there is no reason to believe that such cloned cells ever return to the state of resting precursors. Mitogens may activate a large fraction of resting T cells, but analysis is again complicated because the target molecule(s) of most mitogens is unknown and the relationship of this kind of activation to physiological induction by antigen plus MHC molecules remains unclear. By using a monoclonal antibody specific for the antigen receptors on approximately 25% of all T cells of both Lyt 2+ and Lyt 2- subsets, we have studied the induction of lymphokine responsiveness in resting normal T cells. This antibody, immobilized on Sepharose beads, is sufficient to activate Lyt 2+ T cells, but not Lyt 2- T cells, to clonal expansion in the presence of a mixture of lymphokines (10% rat spleen Con A supernatant). We report here that clonal growth of the T cells obeys single-hit kinetics in limiting-dilution microcultures, suggesting that a single cell type is limiting. We conclude that cytotoxic T-lymphocyte (Tc) precursors require only ligation of the antigen receptor before they become responsive to lymphokines, whereas helper T-lymphocyte (Th) precursors require additional signals.     

Synaptic connectivity of a local circuit neurone in lateral geniculate nucleus of the cat

Although receptive fields of relay cells in the lateral geniculate nucleus of the cat nearly match those of their retinal afferents, only 10-20% of the synapses on these cells derive from the retina and are excitatory. Many more (30-40%) are inhibitory and largely control the gating of retinogeniculate transmission. These inhibitory synapses derive chiefly from two cell types: intrinsic local circuit neurones and cells in the adjacent perigeniculate nucleus. It has been difficult to study the functional organization of these inhibitory pathways; most efforts have relied on indirect approaches. Here we describe the use of direct techniques to study a local circuit neurone by iontophoresing horseradish peroxidase (HRP) into it, which completely labels the soma and processes of cells for subsequent light- and electron microscopic analysis. Although the response properties of the labelled cell are virtually indistinguishable from those of many relay cells, its morphology is typical of 'class 3' neurones (see Fig. 1 legend), which are widely believed to be interneurones (but see ref. 12). Here, we refer to the cell as a 'local circuit neurone', which allows for the possibility of a projection axon, rather than as an 'interneurone', a term that commonly excludes a projection axon. We find that the labelled cell has a myelinated axon, but that the axon loses its myelin within 50 microns of the soma and has not yet been traced further. The dendrites of the labelled cell possess presynaptic terminals that act as intrinsic sources of inhibition on geniculate relay cells. We also characterize other morphological aspects of this inhibitory circuitry.     

Osmotic stress mimics effects of vasopressin on learned behaviour

It has been suggested that arginine vasopressin (AVP) is involved in the retention of learned responses, in addition to its classical physiological functions of water retention and modulation of blood pressure. AVP administered subcutaneously (s.c.), intraventricularly or intracerebrally can prolong extinction of active avoidance behaviour and can enhance retention in inhibitory (passive) avoidance. These effects have been interpreted as a direct action of AVP on the central nervous system to facilitate memory consolidation. AVP also has facilitatory effects on cognitive function in humans, and marked deficits in AVP function have been associated with certain types of psychopathology. Alternative hypotheses for the behavioural actions of AVP have involved motivational constructs such as arousal, and our recent work has focused on the role of arousal resulting from the activation of peripheral visceral signals in the behavioural effects of peripherally administered AVP. The development of a specific antagonist for AVP, 1-deaminopenicillamine-2-O-methyl tyrosine arginine vasopressin (dPTyr(Me)AVP), which can reverse the behavioural effects of exogenously administered AVP, has provided a powerful tool for examining the role of AVP in the behavioural responses produced by physiological challenges known to release vasopressin. However, the relationship between the behavioural effects of exogenously administered AVP and the behavioural function of endogenously released AVP has not been evaluated. We report here that a potent peripheral osmotic stimulus, the intraperitoneal (i.p.) injection of hypertonic saline, at doses known to release AVP both centrally and peripherally, will produce behavioural effects similar to those of exogenously administered AVP. Furthermore, the prolongation of active avoidance induced by this osmotic stimulus is reversed by pretreatment with dPTyr(Me)AVP, suggesting that endogenously released AVP may also produce behavioural effects.     

Transposition of structural genes to an expression sequence on a linear plasmid causes antigenic variation in the bacterium Borrelia hermsii

In Borrelia hermsii, a spirochaete that causes relapsing fever, the switch between expression of two frequent variable major protein (VMP) types (7 and 21) is associated with a DNA rearrangement. Both cell types 7 and 21 contain untranscribed 7 and 21 VMP genes on linear plasmids. The serotype 7 cells contain an additional copy of the 7 VMP gene fused to an expression sequence on another linear plasmid. Switching to the 21 serotype involves removal of the transcribed 7 VMP gene and fusion of a copy of the 21 VMP gene to this same expression sequence. Thus recombination between linear plasmids can activate different VMP genes.     

Metallothionein gene cluster is split by chromosome 16 rearrangements in myelomonocytic leukaemia

The metallothioneins (MTs) are a family of proteins of low relative molecular mass which bind heavy-metal ions. MTs exist in several molecular forms (MT-I, MT-II) and are encoded by a multi-gene family containing at least 14 closely related genes and pseudogenes. These proteins function in the regulation of trace-metal metabolism, the storage of these ions in the liver, and as a protective mechanism against heavy-metal toxicity. Somatic cell hybridization has shown that most MT genes, including the functional MT genes (MT1A, MT1B, MT2A), lie on human chromosome 16. Using in situ hybridization, we have now localized the MT genes to band q22 of chromosome 16. This chromosomal band is also a breakpoint in two specific rearrangements, the inv(16)(p13q22) and t(16; 16)(p13;q22) rearrangements, found in a subgroup of patients with acute myelomonocytic leukaemia (AMML). Hybridization of a MT probe to malignant cells from two patients with an inv(16) showed labelled sites on both arms of the inverted chromosome, indicating that the breakpoint at 16q22 splits the MT gene cluster. Similar results were obtained when this probe was hybridized to metaphase cells from two patients with a t(16; 16). These results suggest that the MT genes or their regulatory regions may function as an 'activating' sequence for an as yet unidentified cellular gene located at 16p13.     

Structure, expression and divergence of T-cell receptor beta-chain variable regions

Analysis of three new T-cell receptor beta-chain variable regions together with those in the literature indicates that they have both remarkable similarities and differences with those of immunoglobulin. Less than 10 V regions appear to predominate in the thymus. V beta sequences are much more heterogeneous at the amino acid level than are immunoglobulin V regions and they appear to diverge between species much more quickly, apparently the result of additional hypervariable regions. Three of these putative new hypervariable regions lie outside of the classical immunoglobulin binding site, an indication that important interactions may be occurring in these regions with polymorphic MHC determinants.     

HLA-restricted T-cell recognition of Epstein-Barr virus-infected B cells

In mice the cytotoxic T-cell response to several types of virus is influenced by genes within the major histocompatibility complex; in particular, genetic control is exercised at the effector cell level through a requirement that virus-specific cytotoxic T cells recognise viral antigens in association with H-2K and H=2D region gene products on the surface of infected cells. In man the restriction which the analogous HLA-A, -B and -C-region gene products might place on virus-specific T-cell function is still in dispute. The earliest and most controversial evidence concerns the Epstein-Barr virus (EBV), a B lymphotropic agent which causes infectious mononucleosis (IM) and which induces an unusually vigorous T-cell response; cytotoxic T cells from IM patients' blood were shown to be EBV-specific yet, in contrast to mouse systems, apparently free of any obvious HLA restriction. Since then T-cell recognition of EBV-infected B cells has assumed particular significance as a model system for the study of cytotoxic T-cell function in man. This report describes the results of a new approach clearly indicating that HLA-A and -B region products do indeed have a role in this system.