Foreign MHC Research Articles

Safety and Short-Term Toxicity of a Novel Cationic Lipid Formulation for Human Gene Therapy

Among the potential nonviral vectors for human gene therapy are DNA-liposome complexes. In a recent clinical study, this delivery system has been utilized. In this report, a novel cationic lipid, dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium (DMRIE), has been substituted into the DNA-liposome complex with dioleoyl phosphatidylethanolamine (DOPE), which both improves transfection efficiencies and allows increased doses of DNA to be delivered in vivo. The safety and toxicity of this DNA-liposome complex has been evaluated in two species, mice and pigs. The efficacy of DMRIE/DOPE in inducing an antitumor response in mice after transfer of a foreign MHC has been confirmed. No abnormalities were detected after administration of up to 1,000-fold higher concentrations of DNA and lipid than could be tolerated in vivo previously. Examination of serum biochemical enzymes, pathologic examination of tissue, and analysis of cardiac function in mice and pigs revealed no toxicities related to this treatment. This improved cationic lipid formulation is well-tolerated in vivo and could therefore allow higher dose administration and potentially greater efficiency of gene transfer for gene therapy.

Does the HIV envelope induce a chronic graft-versus-host-like disease?

Evidence is accumulating that susceptibility to disease following HIV infection is genetically restricted. In addition, activation of the immune response appears to play an important role in disease progression. Here, John Habeshaw and colleagues propose that HIV envelope glycoproteins mimic foreign MHC molecules and, in doing so, stimulate alloreactive lymphocytes. This activation may explain the marked clinical and immunological similarities between chronic GVHD and AIDS.

Non‐Deletional Mechanisms of Peripheral and Central Tolerance: Studies with Transgenic Mice with Tissue‐Specific Expression of a Foreign MHC Class I Antigen

The studies described here reveal a surprising variety of non-deletional manifestations of tolerance (anergy and unresponsiveness) in both the thymus and peripheral lymphoid organs. This can be observed in anti-Kb TCR transgenic mice crossed with normal Kb positive mice, and in TCR mice crossed with transgenic mice expressing Kb in restricted tissues, such as liver, epithelial cells, cells of neuroectodermal origin etc. Thymic induction of unresponsiveness: Transgenic mice were prepared with the a, beta TCR genes from a CD8-dependent and Kb-specific CTL clone. In homozygous H-2b mice clonotype+ cells were found in the thymus but none or few in the periphery, suggesting that deletion occurs in the thymus although lack of migration to the periphery has not been ruled out. In H-2 heterozygous mice (TCR.H-2kxb) deletion was incomplete in the thymus and clonotype+ cells accumulated substantially in the periphery. Most of them had downregulated their CD8 molecules. The clonotype+ cells in both the thymus and periphery were unresponsive to the Kb antigen in vitro, suggesting the induction of anergy in the thymus. The inefficient negative selection in H-hkxb heterozygous mice is probably due to the lower Kb expression in comparison to H-2b homozygous mice. We then further reduced the amount of Kb expression in the thymus by constructing Kb transgenic mice using the 0.8 Kb fragment of the keratin IV promoter. In the thymus expression was only observed on a subset of medullary epithelial cells. When these mice were crossed with the TCR transgenic mice than no deletion was observed in the thymus. However, the clonotype+ CD8+ thymocytes could not respond to Kb in vitro, indicating that they had been rendered anergic in the thymus. These data show that unresponsiveness can be induced in the thymus, that the extent of clonal deletion can vary greatly owing to a change in antigen expression by a factor of two as in H-2 heterozygous versus homozygous mice, and that expression of Kb on a few medullary thymic epithelial cells is sufficient to induce anergy. Peripheral induction of unresponsiveness: To study the consequence of tissue-specific tolerogen expression we have generated transgenic mice expressing Kb exclusively in cells of neuroectodermal origin (GFAP.Kb mice) in the liver (alumin.Kb mice), or in certain epithelial cells outside the thymus (2.4 keratin IV.Kb mice). Consistent with the absence of Kb expression in the thymus there was no deletion of clonotype+ CD8+ cells in the thymus, and the thymocytes were fully functional.(ABSTRACT TRUNCATED AT 400 WORDS)

Regulation of MHC expression

The level of expression of class I and class II MHC antigens in any particular cell is not a fixed characteristic, but is subject to wide fluctuation. The "resting" level of expression varies widely in different cell types, e.g., the cells of the exocrine pancreas and the myocardial cell express neither class I nor class II MHC antigens, whereas B lymphocytes express high levels of both class I and class II. The factors regulating these resting levels are beginning to be elucidated at the DNA level. The resting levels of class I and class II MHC antigen expression can be very greatly increased, primarily as a consequence of exposure to various cytokines released during inflammatory responses, but also in response to other non-specific stimuli. This probably represents an evolutionary adaptation to increase the capacity of infected tissues to present foreign peptides to the immune system, and thereby favour the survival of the host. However, when this phenomenon occurs in a transplanted organ, the increased expression of the now foreign MHC antigens in the context of the new host very likely plays a role in the graft's own destruction. The increased expression of class I MHC antigens, especially if the cell does not express these antigens at all at the time of grafting, will very likely render the cell susceptible to CD8+ cytotoxic T lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Superantigens: Mechanism of T-Cell Stimulation and Role in Immune Responses

Superantigens combine with MHC class-II molecules to form the ligands that stimulate T cells via the V beta element of the T-cell receptor. Two groups of superantigens have been described so far: first, endogenous murine products that include the Mls determinants, and second, bacterial products such as the Staphylococcal enterotoxins. Here, we review studies that address the interactions between the foreign superantigens and MHC class-II molecules, the mechanism of T-cell stimulation, and the role that tolerance to self-superantigens plays in shaping the T-cell repertoire. We speculate on the possible evolutionary significance of superantigens.

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.

Superantigens: Mechanism Of T Cell Stimulation And Role In Immune-Responses

Superantigens combine with MHC class-II molecules to form the ligands that stimulate T cells via the Vβ element of the T-cell receptor. Two groups of superantigens have been described so far: first, endogenous murine products that include the MIs determinants, and second, bacterial products such as the Staphylococcal enterotoxins. Here, we review studies that address the interactions between the foreign superantigens and MHC class-II molecules, the mechanism of T-cell stimulation, and the role that tolerance to self -superantigens plays in shaping the T-cell repertoire. We speculate on the possible evolutionary significance of superantigens.

Antigen Processing and Presentation by Epidermal Langerhans Cells: Induction of Immunity or Unresponsiveness

The seminal observation made 30 years ago that T cells do not discriminate between native and denatured proteins, whereas B cells generally do, can now be explained by the fact that T cells never see antigens in their native conformation and that intact proteins cannot associate simultaneously with MHC molecules and the TCR. This difference in the ability to recognize antigen based on conformational specificity appears to be a consequence of the fact that the T cell sees antigen not free in solution, but on the surface of an APC in association with MHC molecules. The metabolic events that protein antigens undergo within APC, prior to their presentation in an appropriately processed form to T cells, are called antigen processing. The end-product of antigen processing for CD4+ T cells is a relatively short peptide fragment bound to class II MHC molecules on the surface of an APC that can be recognized by the TCR on the T cells. Because this event is difficult to monitor directly, antigen processing can only be assayed in conjunction with the temporally distal event of T-cell activation, manifested ultimately as proliferative responses or lymphokine secretion. In addition to occupancy of the TCR by the peptide/class II complex, several other antigen-nonspecific receptor-ligand interactions between APC and T cells are required for optimal T-cell activation. M phi, B cells, and LC/DC comprise the principal APC for CD4+ T cells. M phi and B cells have been studied extensively with respect to their antigen processing and presenting capacities. Only recently, however, have such capacities been investigated in LC and DC; these studies now indicate freshly isolated LC (but not cultured LC and DC) to possess efficient antigen processing capabilities. In this respect, LC have been proposed to represent evolving (or "maturing") forms of DC: Freshly isolated LC (which retain morphologic and functional properties of epidermal LC in situ) are the equivalent of tissue forms of DC, whereas cultured LC resemble lymphoid or circulating DC. Cultured LC and DC appear to be the sole effective APC for inducing primary T-cell responses in vitro. Possibly underlying this property is the ability of cultured LC and DC (but not M phi, B cells, or freshly isolated LC) to induce formation of T-cell clusters during the course of such responses. The capacity of accessory cells to function as APC varies depending upon the type of APC and T cells examined.(ABSTRACT TRUNCATED AT 400 WORDS)

Genetics of the spondarthropathies

Recent investigation of the possible role of bacteria in the pathogenesis of AS has provided very interesting data. What is at present lacking is a clear demonstration that the findings point to the actual mechanisms involved in the initiation of the disorder. Rapid progress in three related areas of research gives hope that, in the relatively near future, the genetic basis for susceptibility to AS will be elucidated. These are the demonstration of the detailed structure of an HLA class I molecule, of the primary amino acid structure of B27 heavy chain with its subtypes, and of the nature of the interaction between foreign proteins and MHC molecules which leads to antibody and cytotoxic cell responses. It is just possible that the B27 molecules have a disease-promoting capability because of some structural characteristic independent of their antigen binding site. However, it may perhaps be considered more likely that it is the propensity of the specific antigen-binding site itself to bind to a particular group of antigenic peptides that will explain the susceptibility of B27-positive individuals to several clinical disorders. The ability to study the properties of antigenic epitopes which preferentially bind to the very variable binding site of different MHC molecules raises the possibility of revealing the antigenic structures which bind to B27 molecules in patients with AS. This could in turn lead to the source of these antigens in the environment. There has been a tendency to assume that one simple model will explain all the B27-associated disorders but it may be preferable to keep an open mind about the possibility that the mechanisms involved in AS, in the bacteria-induced acute arthropathies and in acute anterior uveitis may not be identical. At the same time, there is a need to continue further direct investigation of the role of microbiological agents in AS both in vitro and in vivo, as ultimately it is most likely that, by blocking the effects of such agents as may be shown to be involved, progress in our ability to influence the progress of the disease in a fundamental way will be achieved. There is still little information as to how the tissues involved in AS come to be the particular targets of the pathological process and currently proposed theories of pathogenesis have not yet provided very satisfactory answers to this problem.

High determinant density may explain the phenomenon of alloreactivity

When two members of the same species differfrom each other in their MHC-encoded antigens, mixing their lymphocytes leads to a vigorous T-cell response. This is the phenomenon of alloreactivity, exempl here as a mixed lymphocyte response. In this article Michael Bevan presents a hypothesis to account for alloreactivity, based on the notion that the alloreactive target determinant is a sequence or conformational determinant contributed entirely by the foreign MHC molecule in isolation, which is expressed at high density on the antigen-presenting cell.

Analysis of early events occurring during neonatal induction of tolerance to histoincompatible cells in mice

Mice have been analyzed at different times post neonatal inoculation of F 1 hybrid lymphoid cells for their ability to respond to foreign MHC antigens expressed on the tolerizing cell population, and to impart that response phenotype to normal adult spleen cells. At early times following neonatal challenge with F 1 spleen cells all mice produced serum immunoglobulins of F 1 donor origin which inhibited the response of normal adult cells. The antigenic specificity of the inhibitory factors suggests a role for an antireceptor antibody in the inhibition seen, and the presence of such serum-mediated inhibition is indicative of (and may be causally related to) the late appearance of a cell-mediated suppressor mechanism in these mice.

Analysis of lymphocytes reactive to histocompatibility antigens: II. Exponential alloantigen-dependent lymphocyte growth in vitro

Rat thoracic duct lymphocytes were maintained in continual blast transformation and cell division by repeated in vitro stimulation with allogeneic cells. This resulted in increases in responder cell numbers of up to 10,000-fold in 10-day periods. Growth of responder lymphocyte populations was dependent upon cell density, culture medium nutrients, and the presence of antigen in the form of allogeneic cells. A titration assay for mixed lymphocyte interactions (MLI) was used to relate absolute growth of cells in preparative cultures to [ 3H]thymidine incorporation in analytical MLI. Growth of lymphocyte populations derived by repeated stimulation with cells bearing a single foreign MHC haplotype was supported to lesser, variable degrees by stimulation with unrelated “third party” stimulator cells. The extent of this operational cross-reactivity was assessed by parallel line analysis of MLI titrations of responder lymphocytes enriched for specific alloreactivity.

Cell Interactions Between Histoincompatible T and B Lymphocytes

Abstract Two possible explanations for the failure of primed histoincompatible T and B lymphocytes to cooperate in secondary responses of the IgG antibody class have been investigated in the present study: 1) The possible existence of subtle suppressive mechanisms developing as a consequence of mixing two histoincompatible cell populations; and 2) The possible inability of histoincompatible carrierprimed T cells to recognize and/or be induced to function by carrier determinants presented to them in association with foreign MHC antigens (i.e., the “altered-self” recognition hypothesis). Absence of suppression has been verified by two different approaches, including: 1) the failure of histoincompatible T cells, even in great excess, to interfere with physiologic cooperation between syngeneic T and B lymphocytes; and 2) the failure of histoincompatible B cells to interfere with physiologic cooperation between semi-syngeneic F1 hybrid T cells and parental B cells. The unlikelihood of the “altered-self” explanation for failure of histoincompatible T and B cells to cooperate has been indicated by an inability to circumvent the requirement for I-region identity by priming T cells with allogeneic macrophagebound antigen even when the latter cells are of identical haploytpe with the allogeneic B cells employed in the final cooperation assay. These results strongly substantiate the existence of true genetic restrictions in T-B cell intractions in secondary responses to hapten-protein conjugates. The validity of other observations indicating an absence of genetic restrictions in certain circumstances, considered in the context of our own data, has suggested the possibility that virgin T and B lymphocytes reciprocally influence one another during the course of cell interactions in response to antigenic and/or other signals, a process we term “adaptive differentiation.”