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Biomedical subjects

J Allison

Publications and source records attributed to J Allison.

At least 91 records · Page 5Linked to original sources
PubMed1991

Dysmyelination in transgenic mice resulting from expression of class I histocompatibility molecules in oligodendrocytes.

Major histocompatibility complex (MHC) molecules are not normally expressed in the central nervous system (CNS). However, aberrant expression has been observed in multiple sclerosis lesions and could contribute to the destruction of myelin or the myelinating cells known as oligodendrocytes. The mechanism of cell damage associated with aberrant MHC molecule expression is unclear: for example, overexpression of class I and class II MHC molecules in pancreatic beta cells in transgenic mice leads to nonimmune destruction of the cells and insulin-dependent diabetes mellitus. We have generated transgenic mice that express class I H-2Kb MHC molecules, under the control of the myelin basic protein promoter, specifically in oligodendrocytes. Homozygous transgenic mice have a shivering phenotype, develop tonic seizures and die at 15-22 days. This phenotype, which we term 'wonky', is due to hypomyelination in the CNS, and not to involvement of the immune system. The primary defect appears to be a shortage of myelinating oligodendrocytes resulting from overexpression of the class I MHC molecules.

Animals↗
PubMed1991

Overexpression of beta 2-microglobulin in transgenic mouse islet beta cells results in defective insulin secretion.

Overexpression of heavy chains of the class I major histocompatibility complex in islet beta cells of transgenic mice is known to induce nonimmune diabetes. We have now overexpressed the secretory protein beta 2-microglobulin in beta cells. Transgenic mice of one lineage had normal islets. Mice of another lineage did not become overtly diabetic but showed significant depletion of beta-cell insulin. When mice were made homozygous for the transgene locus, they developed diabetes. Introduction of the beta 2-microglobulin chain into class I heavy chain transgenic mice resulted in a significant improvement in their islet morphology and insulin content, and the female mice remained normoglycemic. These results suggest that different transgene molecules overexpressed in beta cells can cause islet dysfunction, though not necessarily overt diabetes, and that this effect is mediated by the level of transgene expression. Evidence is provided to show that beta-cell disruption by transgene overexpression occurs at the level of protein and involves a defect in insulin secretion.

Animals↗
PubMed1991

A transgenic approach to the study of peripheral T-cell tolerance.

There is now convincing evidence for the imposition of self tolerance by means of the clonal deletion of self-reactive T cells operating within the thymus. Since not all self components may be encountered there, the question must be asked whether tolerance can occur post-thymically. To test this, we and other investigators have used transgenic technology to direct expression of a known "nonself" gene to a given extrathymic tissue. No lymphocytic infiltration was ever seen in transgene-expressing tissues, even if the mice were given normal syngeneic (nontransgenic) spleen cells intravenously or were stimulated with H-2Kb spleen cells. Infiltration did, however, occur in irradiated transgenic recipients of H-2Kb immune spleen cells. In MET-Kb mice, this infiltrate diminished with time, raising the possibility that peripheral tolerance may even have been induced in immune cells. H-2Kb-bearing skin was accepted in young RIP-Kb mice but rejected in older mice, which had lost more than 75% of their beta cells as a result of the overexpression of H-2Kb. This loss of tolerance thus occurred when the concentration of the tolerogen, H-2Kb, fell below some critical threshold. Following in vitro stimulation, spleen cells from young RIP-Kb mice could not kill H-2Kb-bearing targets, but could respond to third party targets. Thymus cells, on the other hand, could be stimulated to kill both targets, clearly indicating that tolerance was not imposed intrathymically. Spleen cells from older RIP-Kb mice (those that had lost most of their beta cells) killed both targets, which is in agreement with the in vivo data. Reactivity to H-2Kb was restored to young spleen cells by providing them with IL-2. Two hypotheses were proposed to account for the above findings: tolerance results either from the deletion or functional silencing of high-affinity effector cells or of regulatory, IL-2-producing helper T cells. As it is difficult to distinguish between these, we have produced a second series of transgenic mice (F3+) with rearranged TCR genes encoding an anti-H-2Kb TCR and derived "double-transgenic" (F3+RIP+) offspring by mating these mice with RIP-Kb mice. The transgenic TCR utilized the V beta 11 segment which can be detected by a monoclonal antibody. There were in the thymus very few CD4+ and very few CD4+8+ cells in both F3+ and F3+RIP+ mice and, in the double-transgenic mice, there was no evidence of deletion of CD8+V beta 11+ cells in the periphery although they showed tolerance to H-2Kb-bearing skin.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1991

Clonal deletion of autospecific B lymphocytes.

Using mice transgenic for functional, rearranged immunoglobulin heavy and light chain genes, it can be demonstrated that B lymphocytes reactive with cell surface-bound class I MHC antigen can be controlled by clonal elimination. Even low-affinity cell-bound ligands can induce deletion. Deletion can occur in the pre-B to B cell transitional stage or after the B cells exist the bone marrow, depending on where the cells first encounter autoantigen. IgD appears to play no role in protecting cells from deletion. It is argued that defects in B-cell tolerance alone may be sufficient to lead to systemic autoimmunity.

Animals↗
PubMed1991

Inherent beta-cell dysfunction induced by transgenic expression of allogeneic major histocompatibility complex class I antigen in islet cells.

Insulin-dependent diabetes mellitus (IDDM) is generally believed to be an autoimmune disease resulting from T-cell dysfunction that produces beta-cell damage, but it is conceivable that some forms of IDDM are not immunologically mediated. The effect of the expression of a foreign transgenic MHC class I antigen (H-2Kb), restricted to pancreatic islet beta-cells, was tested in vitro and in nude (athymic) mice to determine whether beta-cell dysfunction was due to non-immune mechanisms. The models used clearly excluded immune involvement in beta-cell damage. Fetal pancreas from transgenic and littermate control mice was maintained in organ culture for up to 18 days and insulin secretion into the medium assessed. For the initial 3-4 days in vitro, fetal control and transgenic pancreas secreted similar amounts of insulin, but thereafter insulin secretion by the transgenic tissue decreased in comparison with the controls. When the cultured pancreas was transplanted into nude mice, the transgenic issue produced smaller grafts than the control pancreas, but there was wide variation in graft size. Expression of H-2Kb antigens in beta-cells of nude transgenic mice also resulted in early-onset diabetes. The insulin content in the pancreas of young H-2Kb transgenic euthymic mice, (previously shown not to have insulitis), was reduced but glucagon content was normal. The reduction in in vivo insulin production was similar chronologically to the reduced insulin production by transgenic islets in vitro. These data confirm the non-immune loss of beta-cell function in MHC-transgenic mice and they may be a model for atypical Type I diabetes.

Animals↗
PubMed1990

Renaissance of gas chromatography-time-of-flight mass spectrometry. Meeting the challenge of capillary columns with a beam deflection instrument and time array detection.

This report describes the use of a unique beam deflection time-of-flight mass spectrometer to address some of the demands made on mass spectrometry by new developments in high-resolution capillary column gas chromatography. An integrating transient recorder is used in combination with this beam deflection time-of-flight instrument to apply the concept of time array detection in capturing all of the mass spectral information available from the ion source, thereby greatly enhancing the signal-to-noise ratio quality of the mass spectral data. The applicability of the time array detection approach to gas chromatography-mass spectrometry is demonstrated in the context of an analysis of the standard Grob mixture for assessing performance of capillary column chromatography. During analysis of the Grob mixture by gas chromatography-mass spectrometry, mass spectra were recorded at a rate of 20 scan files per second. The data indicate that this rate of mass spectral scan file generation is adequate to provide a suitable data base for reconstruction of the chromatographic profile. In addition, the effective scan rate is high enough that there is no distortion in the relative peak intensities throughout the individual mass spectra of components regardless of the relatively high dynamic changes in partial pressure of the analyte as reflected by the sharp peaks in the chromatographic profile. The experimental results indicate that the beam deflection time-of-flight mass spectrometer can provide mass spectra at a scan file generation rate much higher than that possible with the conventional quadrupole or magnetic sector mass spectrometer, but at comparable detection limits.

Chemical Phenomena↗
PubMed1990

Selection of the T-cell repertoire in transgenic mice expressing a transplantation antigen in distinct thymus subsets.

Transgenic mice that expressed a transplantation antigen, H-2Kb, in an unusual tissue distribution have been developed. Gene-regulatory elements from the immunoglobulin heavy-chain locus (Emu enhancer and heavy chain promoter) were linked to the class I Kb gene and the construct microinjected into fertilized mouse eggs of a different haplotype. It was expected that such gene-regulatory elements would direct expression of the foreign class I molecules only to B and T lymphocytes. However, expression was also detected in a subset of thymus medullary epithelium. The Kb molecules expressed on this thymic subset were unable to positively select T cells for passage to the periphery. The mice were, however, tolerant of the cell types expressing the foreign Kb molecules and were also tolerant of Kb presented as skin grafts. These results suggest that not all components of thymic epithelium are involved in positive selection of T cells and that transplantation antigens expressed on non-dendritic cells can induce tolerance.

Animals↗
PubMed1990

Prevention of diabetes in non-obese diabetic I-Ak transgenic mice.

The non-obese diabetic (NOD) mouse develops insulin-dependent diabetes mellitus (IDDM) with mononuclear cell infiltration of the islets of Langerhans and selective destruction of the insulin-producing beta-cells, as in humans. Most infiltrating cells are T lymphocytes, and most of these carry the CD4 antigen. Adoptive transfer of T cells from diabetic NOD mice into irradiated NOD or athymic nude NOD mice induces diabetes. Susceptibility to IDDM in NOD mice is polygenic, with one gene linked to the major histocompatibility complex class II locus, which in NOD mice expresses a unique I-A molecule but no I-E. Speculation exists as to the role of the I-A molecule in the diabetes susceptibility of NOD mice, especially regarding the significance of specific unique residues. To examine the role of the NOD I-A molecule in IDDM pathogenesis, we made NOD/Lt mice transgenic for I-Ak by microinjecting I-Ak alpha- and beta-genes into fertilized NOD/Lt eggs. Insulitis was markedly reduced and diabetes prevented in NOD/Lt mice expressing I-Ak.

Animals↗
PubMed1990

Efficiency and persistence of licking in rats.

Thirsty rats tested in 60-min sessions licked an empty spout for access to a water spout. We raised the behavioral price of the water lick by requiring more licks at the empty spout per lick at the water spout. At relatively high prices the rats made relatively few water licks but licked more efficiently, getting more water per lick. Controls showed that the rise in efficiency with price was not attributable to two variables previously confounded with price: the total number of water licks, and the temporal pattern of access. At a higher price the rats also licked more persistently, making more extra water licks before the shutter could occlude the spout. However, yoked controls showed that the greater persistence resulted not from price, but from the relatively stringent pattern of access that accompanied the higher price. Extra water licks diminished as the session progressed, apparently from satiety, not fatigue. The results have implications for behavioral ecology, behavioral economics, motivation, and the methodology of reward.

Animals↗
PubMed1990

Tolerance and diabetes in transgenic mice over-expressing class I histocompatibility molecules in pancreatic beta cells.

The Class I gene, H-2Kb, was linked to the rat insulin promoter and the construct inoculated into fertilized mouse eggs to produce lines of transgenic mice. Mice which expressed the Class I molecule in the beta cells of the pancreas developed diabetes and progressive loss of their pancreatic beta cells. This occurred whether the transgene product was syngeneic or allogeneic with respect to its host. No lymphocytic infiltration was ever seen in transgene expressing mice, even in those deliberately immunized with H-2Kb-bearing cells. When the transgene product was allogeneic, spleen cells from the transgenic mice stimulated in vitro with irradiated B10.A(5R) cells (KbDd), could kill H-2d targets in vitro, but not targets bearing H-2Kb. Responsiveness of spleen cells to H-2Kb targets returned with advancing age, as the severity of diabetes increased. The results indicate that diabetes in this model occurs independently of the immune system, and point to an extra-thymic mechanism of tolerance induction dependent on the continuous presence of antigen.

Animals↗
PubMed1990

An electromagnetic cervical range-of-motion system and its sensitivity to environmental metal.

A three-dimensional cervical spine range-of-motion system was developed using an electromagnetic tracking system for data collection and a personal computer for analysis and graphic representation of the data. A test was designed to investigate the sensitivity of the electromagnetic device to the proximity of metal. It was found that position errors could be kept below 0.5 cm, and rotation errors could be kept below 1.3 degrees , if all metal was at least 33 cm from the source and sensor. The sensitivity of the system to metal should not cause serious problems in the typical clinical environment if simple precautions are taken.

Cervical Vertebrae↗
PubMed1990

Transgenic models of T-cell self tolerance and autoimmunity.

Self-tolerance is generally induced by intrathymic clonal deletion of T cells with reactivity directed to antigens synthesized within the thymus (Kappler et al. 1987, Kisielow et al. 1988). It may also be induced in peripheral T cells when these encounter antigens unique to extra-thymic tissues. Two transgenic models have been particularly useful in the study of peripheral self tolerance: in one model, a known antigen is expressed in a particular extra-thymic site; in the other, the T-cell repertoire is predominantly reactive to this antigen. We, and others, have shown that expression of class I or II MHC molecules in defined extra-thymic sites leads to a state of T-cell tolerance. To account for this, we have proposed two hypotheses which have different implications for autoimmune disease. According to one, tolerance is imposed by deletion or functional silencing of specific high-affinity cytolytic T cells; alternatively, the target cell for tolerance induction may be a regulatory IL-2-producing T-cell, rather than the effector cell itself. To distinguish between these hypotheses it is essential to examine the fate of T cells which have the potential to react to the transgene product. Since the frequency of such T cells is low and there is no dominant clonotype for H-2Kb, which is the class I molecule we used, it was necessary to create double transgenic mice by mating class I transgenic mice with transgenic mice whose T-cell pool was compared of cells reactive to H-2Kb and could be detected by an antibody directed to the TCR. Initial studies showed that such T cells did persist despite the presence of antigen to which they may be reactive. If these double transgenic mice can be shown to be tolerant, they will offer a rich source of tolerant T cells for detailed investigation of their phenotype and fate, and they will be most useful in enabling us to probe the mechanisms responsible for the induction of peripheral self tolerance. Transgenic mouse technology has also been used successfully to unravel the genetic influences which may lead to or prevent autoimmunity. In particular, we have prevented autoimmune diabetes in the nonobese diabetic mouse by introducing a non-NOD MHC class II gene and further work is implicating the failure of intrathymic positive selection of a protective cell as one step in the pathogenesis of diabetes in NOD mice.

Animals↗
PubMed1989

Tolerance of class I histocompatibility antigens expressed extrathymically.

Although convincing evidence has been obtained for the imposition of self-tolerance by the intrathymic deletion of self-reactive T cells, the development of tolerance to antigens which are expressed only in the periphery is not so well understood. We have approached this question by creating transgenic mice which carry a class I major histocompatibility complex (MHC) gene (H-2Kb) linked to the rat insulin promoter. Mice expressing the transgene develop diabetes, but do not appear to mount an immune response against the transgene-expressing pancreatic beta-cells, even when the transgene is allogeneic with respect to the endogenous host H-2 antigens. We have now explored the mechanism of this tolerance further. We find that spleen cells from pre-diabetic transgenic (RIP-Kb) mice do not kill targets bearing H-2Kb, whereas thymus cells from the same mice do. The unresponsiveness of these spleen cells can be reversed in vitro by providing recombinant interleukin-2 (rIL-2). In older, diabetic mice, responsiveness develops as the pancreatic beta-cells are lost. Our results point to an extrathymic mechanism of tolerance induction, dependent on the continuous presence of antigen and the lack of IL-2 in the local environment of potentially reactive T cells.

Animals↗
PubMed1989

Mass spectrometric analysis of cardiac glycosides by the desorption/ionization technique potassium ion ionization of desorbed species.

The analysis of cardiac glycosides by the desorption/ionization (D/I) mass spectrometric technique potassium ion ionization of desorbed species (K+IDS) is presented. K+IDS mass spectra of digitonin, digoxin, digoxigenin, digitoxin and ouabain are discussed to demonstrate the capabilities of this D/I method. The K+IDS analysis consists of two steps: thermal desorption of neutral molecules representative of the analyte, followed by gas-phase addition of K+ ions to these species. Structural and molecular weight information of the cardiac glycosides is obtained with the K+IDS technique. The most intense peak in the K+IDS mass spectrum of an analyte, M, is frequently the [M]K+ ion. Interpretation of the K+IDS mass spectra is simple, since one thermal degradation mechanism dominates. This mechanism is a 1,2-elimination process. A variation of the original K+IDS technique, performed by changing the ionizing metal from K+ to Na+ (i.e. Na+IDS), is presented for the analysis of digoxin. The Na+IDS mass spectrum of digoxin contains more structural information than the K+IDS mass spectrum of that compound. This may lead to a means of controlling the types of information obtainable with this D/I technique by varying the cation that is thermionically generated. K+IDS analyses can be performed rapidly, no sample derivatization is necessary, no matrix is required and little instrument modification is necessary.

Cardiac Glycosides↗