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PubMed1986

Purification of DNA ligase II from calf thymus and preparation of rabbit antibody against calf thymus DNA ligase II.

DNA ligase II has been purified about 4,000-fold to apparent homogeneity from a calf thymus extract. The ligase consists of a single polypeptide with a molecular weight of 68,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. On fluorography after electrophoresis, a DNA ligase-[3H]AMP complex gave a single band corresponding to a molecular weight of 68,000. The Km values of the ligase for ATP and nicked DNA (5'-phosphoryl ends) were obtained to be 40 and 0.04 microM, respectively. Antibody against calf thymus DNA ligase II was prepared by injecting the purified enzyme into a rabbit. The antibody cross-reacted with DNA ligase II but not with calf thymus DNA ligase I. DNA ligase II was not affected by antibody against calf thymus DNA ligase I with a molecular weight of 130,000 (Teraoka, H. and Tsukada, K. (1982) J. Biol. Chem. 257, 4758-4763). These results indicate that DNA ligase II (Mr = 68,000) is immunologically distinct from DNA ligase I (Mr = 130,000).

Animals↗
PubMed1984

Characterization of thymus cells in hyperplastic thymuses in patients with myasthenia gravis and ulcerative colitis with monoclonal antibodies.

Recently, increasing attention has been paid to thymic relevance to pathogenesis in some autoimmune diseases. In this report, the thymus cells from 7 patients with myasthenia gravis (MG) and 6 patients with ulcerative colitis (UC), who had undergone thymectomy for complication of thymic hyperplasia, were studied. The thymus cells were characterized with monoclonal antibodies (Anti-Leu-2a and Anti-Leu-3a) which define human T-cell surface antigens. Although the control thymus consisted of 82-94% of thymocytes which were reactive with Anti-Leu-2a and 90-95% of cells reactive with Anti-Leu-3a, in UC patients both Leu-2a positive thymus cells (38-56%) and Leu-3a positive cells (68-82%) were decreased. Concerning MG thymocytes, Leu-2a positive cells were also decreased (66-81%), but the percentage of Leu-3a positive cells did not show a remarkable change (86-90%) compared with control thymocytes. Considering the above results and many reports telling functional and populational abnormalities of peripheral immunocompetent cells, the process of intra-thymic T-cell maturation may be impaired in these autoimmune diseases.

Adult↗
PubMed1982

The development of IgM- and IgG-containing plasmablasts in the white pulp of the spleen after stimulation with a thymus-independent antigen (LPS) and a thymus-dependent antigen (SRBC).

This study describes the development of IgM and IgG containing plasmablasts in splenic white pulp after a single intravenous injection of the thymus-independent antigen lipopolysaccharide (LPS) or the thymus-dependent antigen sheep erythrocytes (SRBC) using immunohistoperoxidase techniques. Attention has been paid especially to the sites where IgM and IgG blasts develop in the white pulp and their migration route, from the white pulp towards the red pulp. The distribution of IgM and IgG blasts in the different white pulp compartments, i.e. outer periarteriolar lymphocytic sheath (PALS), inner PALS, follicles and the area along the terminal arteriolar branches, has been studied. Our findings indicate that both the thymus-independent IgM response to LPS and the thymus-dependent IgM response to SRBC start in the outer PALS. During the course of the immune response against SRBC the early localization of IgG plasmablasts in the white pulp was dispersed through the whole PALS. Later in the immune response the IgG blasts in the white pulp were localized especially in and at the border of follicle centres. No significant development of IgG blasts was found after LPS administration. The results of the present study suggest that during the immune response the bulk of IgM blasts migrates via the outer PALS and along the terminal arteriolar branches into the red pulp.

Animals↗
PubMed1974

Induction of immunological tolerance to a thymus-dependent antigen in the absence of thymus-derived cells.

Specific immunological unresponsiveness was induced using thymus-dependent antigens in congenitally athymic (nu/nu) mice, in which no T-cell function has been demonstrated. The tolerance was induced in vivo by the injection of 5-10 mg of either FGG or DNP-HGG. Spleen cells from treated mice were tested in vitro for the ability to mount thymus-independent immune responses against FGG in the presence of polymerized flagellin POL, and the DNP determinant conjugated to POL. A specific deficiency in either the in vitro anti-FGG or anti-DNP response was demonstrated, depending on the antigen used for treatment of the spleen cell donor. Athymic mice treated with FGG were also tested by in vivo challenge with FGG given with POL as an adjuvant and were found to be hyporesponsive. Unresponsiveness to in vitro challenge was established by 24 h after the in vivo injection of FGG. It was found that the injection of POL with the FGG prevented the development of unresponsiveness, but not if the POL was given 24 h or more after the FGG. The unresponsiveness could not be overcome by confrontation with allogeneic spleen cells from CBA mice, although the presence of allogeneic spleen cells had a large amplifying effect on the response of control spleen cells. These experiments demonstrate a mechanism for the tolerization of bone marrow-derived cells by thymus-dependent antigens in the absence of the thymus.

Animals↗
PubMed1974

Role of complement in induction of antibody production in vivo. Effect of cobra factor and other C3-reactive agents on thymus-dependent and thymus-independent antibody responses.

In an in vivo study in mice, suppression by the C3-cleaving protein of cobra venom (CoF), and other C3-reactive agents (zymosan, aggregated IgG, anti-C3 antibodies, and type III pneumococcal polysaccharide) of the thymus-dependent antibody responses to sheep erythrocytes, ovalbumin, and human IgG was demonstrated. The thymus-independent antibody response to polyvinyl-pyrrolidone was however unaffected by CoF. These and other published observations suggest that there may be a requirement for functional C3 in induction of thymus-dependent but not thymus-independent antibody production. A model for the role of C3 in lymphocyte cooperation is proposed based on these data analyzed in the light of existing knowledge of this process. It is postulated that fixed C3 interacting with macrophage See PDF for Structure and B-cell C3 receptors might enhance or facilitate T-dependent presentation of antigen to B cells.

Animals↗
PubMed1975

Subpopulations of thymus dependent and thymus independent lymphocytes in human gut associated lymphoid tissues.

Human gut associated lymphoid tissues were studied in terms of the presence of subpopulations of thymus dependent and thymus independent lymphocytes. Both thymus dependent and thymus independent lymphocytes were present in substantial numbers, as judged from their ability to form spontaneous and complement mediated rosettes with sheep red blood cells and the presence of surface bound immunoglobulins. The relationship between this finding and the question of what is the lymphoid organ which may be the human equivalent of the avian bursa is discussed.

Animals↗
PubMed1978

Differential tolerance of thymus-independent and thymus-dependent antibody responses.

The effect of pre-immunization with soluble haptenated serum proteins or haptenated carbohydrates on the subsequent responses to the hapten in thymus-dependent or thymus-independent form was investigated. In certain cases both thymus-dependent and -independent responses were suppressed whilst in others a differential suppression of thymus-independent responsiveness was observed. These results are discussed in terms of B-cell subpopulations and mechanisms of tolerance induction.

Animals↗
PubMed1983

The expression of surface IgD on B cells responsive to thymus-independent and thymus-dependent antigens and its requirement for B-cell triggering.

Cells separated by the fluorescence activated cell sorter on the basis of their surface IgD (sIgD) phenotype have been examined for responsiveness to thymus-dependent and thymus-independent antigens. The ability of monoclonal anti-IgD alloantibodies to inhibit responses in vitro to the various classes of antigen has also been investigated. Evidence is presented indicating that both sIgD positive and sIgD negative cells can respond to all types of antigen tested. However, although the presence of sIgD was necessary for the response of sIgD positive cells to thymus-dependent antigens, the presence of this isotype was not obligatory for the response of the sIgD positive population to thymus independent antigens. The possible role of sIgD as the obligatory purveyor of a B-cell activation signal is discussed in the light of these findings.

Animals↗
PubMed1979

Myosin and actin containing cells in the human postnatal thymus. Ultrastructural and immunohistochemical findings in normal thymus and in myasthenia gravis.

Samples of normal human thymus of different ages (4-63 years old) were studied by immunofluorescence microscopy (using antibodies to smooth muscle myosin, to actin from the chicken gizzard, and antibodies to myosin from human striated muscle) as well as by routine electron microscopy. Thymus tissue from myasthenia gravis patients was also investigated for comparative reasons. Epithelial cells reacted with anti-smooth, but not with anti-striated muscle myosin, whereas myoid cells reacted with antibodies to striated, but not to smooth muscle myosin. Both epithelial and myoid cells displayed a strong immunoreactivity with antiactin. Corresponding to this immunoreactivity, both cell types contained bundles of thin, actin-like filaments. Myoid cells occurred in the rounded and elongated variety, and they were a normal constituent of all thymuses investigated in this study. Ultrastructurally, this non-innervated, striated muscle-like cell type possessed bundles of thin and thick filaments as well as Z lines in a rather disorganized arrangement, resembling striated muscle after denervation or various other pathologic conditions. There were no overt differences in the number and structure of myoid cells between healthy and myasthenic patients.

Acetylcholine↗
PubMed2002

CuZn-superoxide dismutase in human thymus: immunocytochemical localisation and secretion in thymus-derived epithelial and fibroblast cell lines.

CuZn-superoxide dismutase (SOD) plays a key role in mediating the cellular response to oxidative stress. Its expression in normal thymus was investigated by immunohistochemical techniques and CD3-, CD68- and cytokeratin-specific staining, in order to identify thymocytes, macrophages and epithelial components. Immunocytochemical studies showed an overall CuZn-SOD thymic distribution with a prevailing concentration within thymic medulla. The analysis of CuZn-SOD release by thymus-derived epithelial and fibroblast cell lines showed the ability of both cell lines to release the anti-oxidant enzyme, especially in the presence of stress conditions as represented by serum and nutrient deprivation. These data suggest that CuZn-SOD could be a relevant antioxidant paracrine molecule in human thymus.

Aged↗
PubMed1982

Autoradiographic study on the distribution of thymus-derived and thymus-independent lymphocytes in the spleen of Xenopus laevis.

Non-thymectomized and early-thymectomized Xenopus laevis were injected with 3H-thymidine, and labelled lymphocytes from thymus, spleen, and peripheral blood were transferred to histocompatible recipients to study their distribution by autoradiography. An extremely high proportion of labelled thymocytes was localized in the splenic red pulp of both non-thymectomized and thymectomized recipients after transfer. Labelled splenic lymphocytes were localized in a significantly higher density in the splenic white- than red pulp, particularly 24 hr after cell transfer. This preference of labelled lymphocytes in the white pulp was more evident when early-thymectomized toads were used as lymphocyte donors. These results strongly suggest that in Xenopus, thymus-independent lymphocytes preferentially localize in the splenic white pulp, and thymus-derived lymphocytes possibly in the red pulp.

Animals↗
PubMed1997

Kinetics and repertoire selection of T cells derived from the early waves of fetal thymus colonization after thymus grafting in allogeneic nude recipients.

The survival of T cells derived from the early waves of thymus colonization by haemopoietic cell precursors was investigated by grafting thymus from B6.Thy1.1 day 14 embryos (E14) (first wave) or E17 or newborn thymus (subsequent waves) into allogeneic athymic BALB/c (Thy1.2) nude recipients. The survival of donor-derived Thy1.1 cells was longer when they were derived from early thymocytes. Donor B6.Thy1.1 V beta 5 and V beta 11 T cells, although maturing in BALB/c host presenting Mls2a superantigens, were not deleted, in contrast to host Thy1.2 T cells differentiating from endogenous stem cells. These results show that the population of T cells derived from early precursors undergoes particular selection characteristics, which favour the inclusion of potentially autoreactive cells with prolonged survival, even in H-2 allogeneic conditions which normally do not allow the survival of peripheral T cells.

Animals↗
PubMed2001

In search of the origin of the thymus: the thymus and GALT may be evolutionarily related.

The thymus is the major primary immune tissue for the production of functional T lymphocytes in vertebrates. However, its evolutionary origin is unknown. It has recently been shown that the generation of local T cells also occurs in gut-associated lymphoid tissues (GALT). This suggests that the thymus and GALT have similar functions and that they might be evolutionarily related. We discuss the possibility that the thymus may have evolved from mucosa-associated lymphoid tissues (MALT) located in the gill region in early vertebrates. Various facts supporting this proposal are summarized.

Animals↗
PubMed1965

Role of the thymus in tolerance. I. Tolerance to bovine gamma globulin in thymectomized, irradiated rats grafted with thymus from tolerant donors.

Rats thymectomized and irradiated as adults were restored to immunologic reactivity by grafts of normal adult rat thymus and bone marrow. Reactivity of the delayed (cellular) type and formation of mercaptoethanol-sensitive (MES) and mercaptoethanol-resistant (MER) antibody returned within 3 weeks, while Arthus reactivity remained subnormal till 9 weeks after irradiation and grafting. When the thymus donor was tolerant to BGG, the recipient showed specific non-reactivity to this antigen 3 weeks and, to a much lesser extent, 6 weeks after grafting. This non-reactivity affected delayed responses and MER antibody. No effect was noted on Arthus reactivity and a slight effect on MES antibody. Controls showed that the non-reactivity was not due to transfer of free antigen at the time of grafting. It was concluded that different source organs are responsible for different immune functions and that specific immunologic tolerance may be induced within such an organ as the thymus.

Animals↗
PubMed1988

Thymus and autoimmunity. Transplantation of the thymus from cyclosporin A-treated mice causes organ-specific autoimmune disease in athymic nude mice.

Organ-specific autoimmune diseases such as gastritis, oophoritis, thyroiditis, or insulitis developed in athymic nu/nu mice after engraftment of the thymus from euthymic nu/+ mice treated with cyclosporin A (CsA), a potent immuno-suppressant. The development of autoimmune disease in the nu/nu mice was prevented by inoculation of thymocyte suspensions prepared from normal nu/+ mice, but not by thymocyte suspensions from CsA-treated nu/+ mice. Cotransplantation of normal nu/+ mouse thymus with CsA-treated thymus also suppressed the development of autoimmune disease. Inoculation of spleen cell suspensions prepared from normal adult nu/+ mice prevented autoimmune disease, but inoculation of those from newborn nu/+ mice did not. Thus, CsA appears to interfere selectively with the thymic production of certain suppressor T cells controlling self-reactive (autoimmune) T cells, allowing the latter to expand and cause autoimmune disease.

Animals↗
PubMed1998

Indefinite survival of skin allografts in adult thymectomized, antilymphocyte serum-treated mice given bone marrow and thymus grafts of donor origin: tolerance induction by donor bone marrow and thymus.

BACKGROUND: Perigraft immunosuppression with antilymphocyte serum (ALS) and postgraft infusion of donor bone marrow cells (BMC), which induces allograft unresponsiveness in a class I major histocompatibility complex (MHC)-disparate mouse combination, is much less effective in class I and II MHC-disparate strain combinations. Because the thymus plays a central role in the maturation, differentiation, and education of T lymphocytes, which are the principal mediator of allograft responses, we examined whether transplantation of donor-specific thymus combined with adult thymectomy of recipients enhances the tolerogenic effect of ALS and donor BMC infusion in a strongly histoincompatible class I and II MHC-disparate DBA/2-to-B6AF1 mouse strain combination. METHODS: Adult thymectomy was performed 4 weeks before grafting. B6AF1 mice received ALS (days -1 and 2), skin allografts (day 0), and BMC and/or thymus grafts (ThyTx) (day 7). Flow cytometric analysis was used to detect donor cells in tolerant mice. Limiting dilution assay was used to calculate the frequencies of precursor cytotoxic T lymphocytes against donor and third-party alloantigens. The presence of suppressor cells was determined by in vivo adoptive transfer assay and in vitro coculture mixed lymphocyte culture. RESULTS: When adult thymectomy and postgraft donor ThyTx were combined with ALS and donor BMC, DBA/2 allograft survival was prolonged with all grafts surviving for >122 days. Third-party (DBA/1) or recipient ThyTx was not effective. The tolerant mice accepted the second donor skin allografts but acutely rejected the third-party grafts. No significant chimerism was detected. Marked reduction of precursor cytotoxic T lymphocyte frequencies continued only against donor alloantigens after second donor and third-party skin grafting. No suppressor cell activity was detected. Immunopathological analysis revealed that the cells in the ThyTx of tolerant mice are of donor origin. CONCLUSION: Donor ThyTx combined with donor BMC infusion in adult thymectomized, ALS-treated mice induced clonal deletion of donor-reactive T cells.

Animals↗
PubMed2004

Role of the thymus in transplantation tolerance in miniature Swine: IV. The thymus is required during the induction phase, but not the maintenance phase, of renal allograft tolerance.

BACKGROUND: The authors' laboratory previously demonstrated that long-term tolerance to class I-disparate renal allografts in miniature swine can be induced by a short course of cyclosporine A (CsA), and that this stable tolerance is dependent on the presence of an intact thymus. In the present study, the authors have examined the requirement for a thymus during the pretransplant, induction, and maintenance phases of tolerance. METHODS: Twenty-two miniature swine underwent class I major histocompatibility complex-mismatched renal transplantation, with a 12-day course of CsA. Thymectomies were performed on days -21, 0, +8, +21, and greater than or equal to +42, in relation to the day of transplantation. Historical controls consisted of euthymic and sham-thymectomized recipients. RESULTS: Euthymic, sham-thymectomized, and day-greater than or equal to +42 thymectomized recipients demonstrated stable renal function and minimal anti-donor cytotoxic T-lymphocyte (CTL) responses. In contrast, day -21 and day 0 thymectomized recipients demonstrated allograft dysfunction, marked cellular infiltrates, with severe vasculitis and glomerular changes, and strong anti-donor CTL responses. Animals thymectomized on days +8 and +21 did not undergo severe rejection, but likewise did not demonstrate a stable clinical course. CONCLUSIONS: These data indicate that the requirement for thymic function in the induction of rapid and stable tolerance is greatest during the first 8 days and then diminishes over the next 2 weeks posttransplant. Failure of thymectomy to affect the course of tolerance after day +21 suggests that thymic function is not required for the maintenance of tolerance. Understanding the role of the thymus in establishing tolerance may permit the development of tolerance induction strategies, especially for pediatric transplant recipients.

Animals↗
PubMed1987

Thymus localization of monoclonal antibodies circumventing the blood-thymus barrier.

The binding in the rat thymus of mouse monoclonal antibodies which recognize determinants present on class II major histocompatibility complex antigens (HIS 19) and all T cells (HIS 17) was studied at several intervals after in vivo injection of a single dose (8 mg) of the monoclonal antibodies (MoAb). The MoAb, injected either intravenously or intraperitoneally, penetrated the thymus initially across the thymic capsule from the extravascular space. The extent to which the MoAb penetrated both cortex and medulla correlated with serum presence of the monoclonal antibody. These results suggest that the thymus cortex and medulla are permeable to MoAb present in the extravascular compartment. If this situation is a general phenomenon, which applies to circulating self non-major histocompatibility complex antigens under physiological conditions, the current dogma relating to self-tolerance needs to be reviewed.

Animals↗