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PubMed1978

The immune response in NZB mice of different ages to thymus-dependent and thymus-independent phosphorylcholine antigens.

This study was designed to examine aberrations of immune responses in autoimmune NZB strain mice during ageing, at the level of individual B cell clones. The response to phosphorylcholine (PC) was chosen because murine responses to PC are restricted to a few B cell clones and can be characterized with idiotypic markers. Responses to both thymus-dependent (TD) and thymus-independently (TI) PC-containing antigens were measured in mice ranging from 1 to 62 weeks of age. We found that: (1) TD responses to phosphorylcholine keyhole limpet haemocyanin (PC-KLH) decreased markedly (about 17-fold) between 28 and 62 weeks of age. TI responses to the R36a strain pneumococcus decreased only slightly during the same period. (2) The PFC responses to both antigens became markedly prolonged in mice older than 24 weeks. (3) The NZB response to either antigen is essentially monoclonal, as measured by inhibition of PFC with specific anti-idiotype serum and PC hapten. No age-related alteration in avidity or idiotype expression was observed. Our results demonstrate that no aberrant PC-reactive B cell clones appear in old NZB, and lend support to the notion that the abnormalities observed are due to defective regulatory mechanisms.

Aging↗
PubMed1986

Rat thymus macrophages: an immunohistochemical study on fetal, neonatal and adult thymus.

The pre- and postnatal development of the macrophage population of rat thymus is investigated applying enzyme-histochemical and immunohistochemical techniques, both on tissue sections and cell suspensions. A set of three monoclonal antibodies (ED1, ED2 and ED3), each of which recognizes cells of the monocyte-macrophage lineage in the rat, enabled us to distinguish between macrophages in the various compartments of the thymus. The medulla is characterized by ED1-positive dendritic cells, the corticomedullary region comprises numerous monocyte-like ED1-positive macrophages and the cortex contains a particular subpopulation of branched ED2-positive macrophages. Both the medullary dendritic cells and the cortical branched cells show Ia-membrane staining. ED3-positive cells are only occasionally present. During fetal life ED1-positive monocyte-like macrophages and dendritic cells are present. Just after birth ED2-positive cortical macrophages start to develop. Their number increases strongly during the first week after birth. The role of the various subpopulations of thymic macrophages is discussed.

Animals↗
PubMed1985

Microenvironments in the normal thymus and the thymus in myasthenia gravis.

The disposition of epithelial cells and extracellular matrix, in the thymus of 8 cases of myasthenia gravis (MG) and in controls (over a wide age range) was studied. In the controls, the subcapsular epithelium was strongly Leu-7-positive in the fetus, negative in childhood, and positive again in adults. Another antibody, RFD4, also labeled the subcapsular epithelium in childhood and adults, but not fetal samples. The samples from MG cases showed the same staining pattern as adult control samples. The medullary epithelium was also RFD4+, and at all ages. The most striking changes in the advanced cases of MG were the unusual arrangement and hypertrophic appearance of medullary epithelial cell areas, separated by laminin-positive basement membranes from the alternating multiple bands of peripheral lymph-node-like areas. The latter had regions resembling the paracortex of lymph nodes as well as germinal centers (GCs). The T-cell zones contained heavy deposits of fibronectin. These T-cell zones were unique to the thymus in MG and were absent in the two normal thymic samples with isolated GCs. In MG the laminin-containing basement membrane, which separated the medullary epithelial and peripheral lymph-node-like areas, was fenestrated at circumscribed points closest to the GCs, thus apparently permitting communication among the medullary epithelium, the T-cell zones, the GCs and the associated antigen-presenting cells. Large numbers of interdigitating cells and some lymphocytes of cortical thymocyte phenotype were also found at these special sites, where opportunities for autosensitization may persist in MG.

Adult↗
PubMed1984

Hapten-specific B cell blockade of the immune response to a thymus-independent-1 antigen produced by concomitant administration of a thymus-independent-2 antigen.

CBA/N mice harbour an X-linked B cell defect which is transmitted by CBA/N female mice to their hybrid male progeny. These mice mount normal responses to thymus-dependent (TD) and some thymus-independent (TI-1) antigens, while the response to TI-2 antigens is absent. Hapten-specific plaque-forming cell (PFC) responses to TD antigens can be blockaded by concomitant exposure of these mice to TI-2 antigens bearing the same hapten. This paper investigates in defective mice the blockade of their response to TNP3-LPS (trinitrophenylated lipopolysaccharide, a TI-1 antigen), imposed by DNP59-Ficoll (dinitrophenylated Ficoll, a TI-2 antigen). The effectiveness of the blocking agent, DNP59-Ficoll, differed in various inbred mouse strains: CBA/N X C3H/HeN F1 male greater than CBA/N female greater than CBA/N X C3H/HeN F1 female. The role of T cells in the observed hapten-specific blockade phenomenon was investigated using athymic CBA/N nude mice and a B cell tolerogen. Our findings indicate that T cell participation is not essential for the blockade of CBA/N PFC responses and they suggest that direct blockade of TI- and TD-responsive B cell populations is likely to occur.

Animals↗
PubMed1980

In vitro and in vivo response of lymphoid cells from LHC hamsters to murine thymus-independent and thymus-dependent antigens.

Lymphoid cell populations (spleen, lymph node, peripheral blood, thymus, and bone marrow) from LHC inbred hamsters were studied in order to characterize further the immune response of this species. The direct PFC response to several thymic-dependent or thymic-independent antigens was evaluated. A specific direct PFC response occurred 4 days after immunization with SRBC, DNP-BSA, DNP-lys-Ficoll, TNP-LPS, TNP-BA, and SSS-III. Attempts to induce a polyclonal antibody response with LPS, TNP-LPS, SSS-III, and DNP-lys-Ficoll were unsuccessful. A weak polyclonal response was induced with TNP-BA. Spleen cells and PBL responded strongly in vitro to the T-cell mitogens Con A and PHA-P, but gave weak and inconsistent responses to the B-cell mitogens LPS and PI-PC. LHC hamster lymphoid cell populations bore sIg and receptors for C3 (EAC rosettes) in approximately the same ratio as various murine species. However, the profile of the number of cells bearing low-to-intermediate densities of sIg differed significantly from those of murine species when analysed with the FACS. There was a sharp reduction in the number of cells with low-to-intermediate densities of sIg. These data suggest that B cells in this strain and species lack the ability to translate signals which lead to polyclonal antibody synthesis or lack the appropriate populations of B cells that have membrane receptors for mitogens which are thought to induce such activity in murine systems and provide evidence for separate signals that induce thymus-independent and mitogenic responses. The importance of this model for studying mechanisms involved in B-cell activation is discussed.

Animals↗
PubMed1996

Ontogeny and immunohistochemical localization of thymus-dependent and thymus-independent RT6+ cells in the rat.

RT6 is a cell surface alloantigen that identifies a regulatory subset of peripheral T cells in the rat. Diabetes-prone BB rats are deficient in peripheral RT6+ T cells and develop spontaneous autoimmune insulin-dependent diabetes mellitus. Diabetes-resistant BB rats have normal numbers of RT6+ T cells, and insulin-dependent diabetes mellitus can be induced in these animals by in vivo depletion of peripheral RT6+ cells. Athymic rats are also severely deficient in peripheral RT6+ T cells. Although very different with respect to the peripheral RT6+ cell compartment, normal, athymic, and diabetes-prone BB rats all generate RT6+ intestinal epithelial lymphocytes (IELs). The goal of these studies was to analyze the ontogeny of RT6+ IELs and peripheral lymphoid cells by in situ immunohistochemistry. We observed the following. 1) RT6+ IELs appear before alpha(beta) T-cell-receptor- expressing IELs in diabetes-prone BB, diabetes-resistant BB, and athymic WAG rats. 2) In vivo depletion of peripheral RT6+ T cells in diabetes-resistant BB rats using a cytotoxic monoclonal antibody is not accompanied by depletion of RT6+ IELs. 3) A population of RT6+ T-cell-receptor-negative IELs is present in normal, euthymic diabetes-resistant BB rats, constitutes a larger percentage of the euthymic but lymphopenic diabetes-prone BB rat IEL population, and is the predominant IEL phenotype in athymic WAG rats. These results suggest that RT6+ cells are composed of both thymus-dependent and thymus-independent cell subsets that have different developmental characteristics and may differ in function.

ADP Ribose Transferases↗
PubMed1989

Primary in situ immune response in popliteal lymph nodes and spleen of mice after subcutaneous immunization with thymus-dependent or thymus-independent (type 1 and 2) antigens.

Mice were immunized subcutaneously with thymus-independent (TI)-type 1 antigen trinitrophenylated lipopolysaccharide (TNP-LPS), TI-type 2 antigen TNP-Ficoll or thymus-dependent (TD) antigen TNP-keyhole limpet haemocyanin (TNP-KLH) in order to study the primary in situ immune response in popliteal lymph nodes (PLN) and spleen. The spleen responded more rapidly in developing specific antibody-forming cells (AFC) than the lymph nodes did, in spite of the fact that antigens reach the spleen only after passing several lymph node stations. This difference between lymph nodes and spleen in developing AFC was particularly significant with respect to the responses to TI (both type 1 and type 2) antigens. No differences in the distribution of specific AFC in PLN and spleen were observed after immunization with TI and TD antigens. Results are discussed with respect to the relative contributions of lymph nodes and spleen to immune responses to antigens injected subcutaneously.

Animals↗
PubMed1986

Differences in the recruitment of virgin B cells into antibody responses to thymus-dependent and thymus-independent type-2 antigens.

The bone marrow of mammals generate large numbers of B cells throughout life. Most of these have a short life span. The subject of this report is to investigate the extent to which newly formed virgin B cells can be activated by thymus-dependent (TD) and thymus-independent type 2 (TI-2) antigens carrying the hapten 2,4-dinitrophenyl. The experimental approach used chimeras made between congenic rats of different kappa immunoglobulin light chain allotype. Host (kappa la) rats were depleted of peripheral B cells by whole body irradiation but had B lymphopoietic capacity conserved by shielding the hind limbs. Their peripheral B cell pool was reconstituted by transfer of kappa lb thoracic duct lymphocytes from donors immunized previously with the TD carrier. This provides test animals where newly produced virgin B cells only express kappa la but where initially most peripheral B cells are kappa lb. The TD antigen tested was able to activated both virgin and memory B cells in the period immediately following immunization. However, long-term antibody production was attributable to repeated activation of memory B cell clones without further virgin B cell recruitment. By contrast, antibody evoked by the TI-2 antigen initially was almost exclusively due to activation of donor peripheral B cells. However, over a period following TI-2 immunization there was a progressive increase in the amount of host antibody produced with corresponding decline of the donor component of the response so that the host response was dominant by six weeks. Control experiments were conducted to show that these effects could not be explained by allotype or isotype-directed suppression. The cellular basis of these differences was investigated further by studying the rate of repopulation of different B cell compartments in these chimeras by newly formed host and mature donor B cells. The results indicate that the onset of host antibody production to the TI-2 antigen closely correlated with the appearance of host B cells in the marginal zones of the spleen, whereas good TD host anti-2,4-dinitrophenyl responses antedated the appearance of host B cells in this compartment. These results are discussed in relation to other data implicating marginal zone B cells in responses to TI-2 antigens.

Animals↗
PubMed2004

The association between life events, social support, and antibody status following thymus-dependent and thymus-independent vaccinations in healthy young adults.

This study determined whether stressful life events and social support were related to antibody status following both thymus-dependent and thymus-independent vaccinations. Life events in the previous year and customary social support were measured in 57 healthy students at baseline. Antibody status was also assessed at baseline and at five weeks and five months following vaccination with the trivalent influenza vaccine and the meningococcal A+C polysaccharide vaccine. Taking into account baseline antibody titre, high life events scores prior to vaccination were associated with lower responses to the B/Shangdong influenza strain at both five weeks and five months and meningococcal C at five weeks. Life event scores were not associated with response to the other two influenza viral strains nor response to meningococcal A. Those with high social support scores had stronger 5-week and 5-month antibody responses to the A/Panama influenza strain, but not to any of the other strains. These associations could not be accounted for by demographic or health behaviour factors, and also emerged from analyses comparing those who exhibited a fourfold increase in antibody titre from baseline with those who did not. Life events and social support were related to antibody status following influenza vaccination in distinctive ways that may be partly determined by vaccine novelty and prior naturalistic exposure. Life events also predicted poor antibody response to meningococcal C polysaccharide vaccination after previous meningococcal C conjugate vaccination. Neither psychosocial factor was associated with response to primary meningococcal A polysaccharide vaccination.

Adult↗
PubMed1990

The in situ immune response in popliteal lymph nodes of mice after macrophage depletion. Differential effects of macrophages on thymus-dependent and thymus-independent immune responses.

Mice were subcutaneously (SC) injected in the left hind footpad with dichloromethylene diphosphonate (Cl2MDP)-containing liposomes to eliminate macrophages lining the subcapsular sinus (SCS) and those in the medulla of draining popliteal lymph nodes (PLN). In order to study the effect of depletion of these macrophages on the in situ immune response in the PLN, liposome-treated mice were SC injected in the same footpad with thymus-independent (TI) type 1 antigen trinitrophenylated lipopolysaccharide (TNP-LPS), TI-type 2 antigen TNP-Ficoll or thymus-dependent (TD) antigen TNP-keyhole limpet haemocyanin (TNP-KLH). No major differences were observed in antibody-serum titers of liposome-treated and control animals. After primary as well as secondary immunization with the TD-antigen TNP-KLH, an increase in the number of antibody-forming cells (AFC) was found and the peak of response was delayed in the PLN of liposome-treated animals. Such differences were not observed with the TI-antigens. These results indicate that macrophages lining the SCS and those in the medulla of the PLN are not essential for the induction of an immune response. The positive effect of macrophage-depletion on the number of AFC may be explained by competition for the antigen by macrophages and other antigen-presenting cells.

Animals↗
PubMed2003

Bacterial susceptibility to and chemical composition of essential oils from Thymus kotschyanus and Thymus persicus.

Susceptibility of Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Klebsiella pneumonia, and Pseudomonas aeroginosa to the essential oils extracted from two varieties of Thyme, i.e., Thymus kotschyanus Boiss. and Hohen. and Thymus persicus L. at preflowering and flowering stages were studied. The disk diffusion method was used to evaluate the zone of microbial growth inhibition at various concentrations of the oils. Minimal inhibitory concentration and minimal bactericidal concentration of the oils were determined and compared with each other. The oils from the above plants were found to be strongly bactericidal with that of T. kotschyanus being more effective. T. kotschyanusand T. persicus oils analyzed by gas chromatography (GC) and GC/mass spectrometry (MS) lead to identification of 33 and 26 components, respectively. The profile of the oil components from T. persicuswas similar to that of T. kotschyanus in almost all of the compounds but at different concentrations. The major components of T. kotschyanus oil before and at the flowering stages were carvacrol (35.06, 22.75%), thymol (26.60, 16.52%), gamma-terpinene (7.81, 0.34%), gamma-terpinene (4.34, 0%), borneol (2.29, 4.52%), myrcene (0.26, 12.65%), thymolquinone (0, 11.39%), nerol (0, 6.10%), and beta-caryophyllene (0, 5.54%), respectively, and those of T. persicus at the same stages were carvacrol (38.96, 27.07%), thymol (6.48, 11.86%), P-cymene (7.51, 10.16%), gamma-terpineol (0, 9.51%), nerol (15.66, 9.41%), gamma-terpinene (6.11, 6.51%), and thymol acetate (5.29, 5.30%), respectively. The contribution of oil components to its antibacterial property is discussed. High aromatic compound content of the phenol-rich oils seems to account for strong antibacterial activity.

Anti-Bacterial Agents↗
PubMed1972

Mouse thymus-independent and thymus-derived lymphoid cells. I. Immunofluorescent and functional studies.

The simultaneous use on mouse lymphoid suspensions of heterologous antisera directed against thymus-derived (T) cell mouse-specific lymphocyte antigen and brain-associated theta antigen (MSLA and BAtheta) or thymus-independent (B) cell mouse-specific bone marrow-derived lymphocyte antigen (MBLA) surface antigens allowed direct proof of the different specificity of these antisera by double immunofluorescence (IF) staining with selective visualization of fluorochromes. These antisera and antisera against mouse Ig and its different types of chains were then used with technique of either double IF staining or IF combined with radioautography, allowing the following conclusions: (a) Surface Ig (sIg) was found exclusively on B cells and never on T cells, but not all B cells had sIg. Cells containing detectable amounts of Ig were MBLA+, but had less sIg than other B cells or none at all. There was evidence for the existence of a significant number of MBLA+ lymphocytes, neither bearing nor containing detectable Ig. (b) micro-Chains were the most frequent but not the only heavy chains found on spleen cells; however, it could not be decided with the technique used, if a single cell can bear more than one type of heavy chain. No cell containing gamma-chains was found to bear surface micro-chains, although a very few cells containing both micro- and gamma-chains were observed. (c) The antigen-binding cells detected after immunization with bacteriophage T4, bovine serum albumin, Maia squinado hemocyanin, and sheep erythrocytes were analyzed for MSLA, MBLA or sIg using double IF, a combination of IF and radioautography, or inhibition of "rosette" formation. Practically all the antigen-binding cells detected were MSLA-, MBLA+, sIg+. (d) More B cells than T cells were found among short-lived lymphoid cells labeled by repeated in vivo injections of tritiated thymidine, but the results did not support a simplified concept equating T cells to long-lived and B cells to short-lived lymphocytes. (e) Cells dividing rapidly in the lymph nodes draining the sites of immunization with various antigens were predominantly T cells 2 days after immunization and in majority B cells a few days later. (f) Incubation of lymphoid cells at 37 degrees C with rabbit anti-mouse Ig or anti-kappa chains led to complete disappearance of sIg and to decrease of MBLA ("antigenic modulation"). In the same conditions, anti-MBLA gave partial modulation of MBLA and of sIg; MBLA, however, reappeared much faster than sIg. No modulation of T cell surface antigens by the appropriate antisera was observed. Cell treatment with Pronase could remove MBLA, sIg, MSLA, and BAtheta, which reappeared within a few hours. Neuraminidase treatment was without detectable effect on these antigens.

Animals↗
PubMed2004

Understanding thymus-independent antigen-induced reduction of thymus-dependent immune responses.

Deficiencies in immune responses against polysaccharides can have direct consequences for patients, and therefore, a better understanding of these immune reactions is crucial. We have studied the immune response against the polysaccharide dextran B512 (Dx). Administration of immunogenic doses of thymus-independent (TI) Dx reduces the immunoglobulin G1 response to later challenges with a thymus-dependent (TD) form of Dx. We investigated if this suppression is a general phenomenon caused not only by Dx but also by other TI antigens, and examined possible mechanisms contributing to this unresponsiveness. We show that clonal exhaustion is not involved in modulating subsequent responses, nor is signalling via FcgammaRIIB or other antibody mediated pathways. The reduced TD response is not an exclusive Dx phenomenon; it is also induced by TI antigen oxazolone (Ox). However, responses against the hapten dinitrophenyl (DNP) are not affected, indicating that the TI priming negative effect is not a general process. This may be explained by the restricted immune response to both Dx and Ox, in contrast to the unrestricted DNP response. Our conclusion from these experiments is that the underlying mechanism for the TI-induced reduction of latter TD responses is a property of the TI activation itself.

Animals↗
PubMed1995

Stimulation of the same B-cell population by thymus-independent dextran and by thymus-dependent oligosaccharide-carrier.

The immune response of adult BALB/c mice against the bacterial antigen dextran B1355S (Dex) is well characterized as thymus independent (TI type 2) and Igha linked. The antisera consist of mainly IgM/lambda antibodies directed against the alpha(1-->3) glucosidic linkage. This study describes the immune response against the alpha(1-->3) linkage in the thymus dependent form (TD), i.e. tetra- or heptasaccharides (N4 or N7) of glucose as hapten coupled to chicken serum albumin (CSA) as carrier. Whereas athymic BALB/c-nu/nu mice did not respond to the TD antigens N4-CSA and N7-CSA, euthymic BALB/c showed high anti-Dex antibody titres of IgM and, after 2 degrees immunization, a class switch to IgG (mainly IgG1) isotypes with lambda light chains. The hapten N4 inhibited Dex-binding of M104E or of antisera from Dex or N4-CSA or N7-CSA immunized mice at 1.7-10 x 10(-4)M. The idiotype composition of these antibodies resembled those after Dex immunization. We conclude that the same Dex-specific precursor B cells have been stimulated by either form of antigen. The ontogenic development of a Dex-specific response could not be accelerated by the aid of T cells, even of adult origin. It seems, therefore, that the maturation of antigen specific B cells is the limiting step in ontogeny.

Age Factors↗
PubMed1972

Thymus-dependent and thymus-independent lymphocyte separation: relation to exposed sialic acid on cell surface.

On preparative cell electrophoresis mouse lymph node lymphocytes separate into fast-moving (T, thymus-dependent) and slow-moving (B, thymus-independent) fractions. After treatment with neuraminidase all lymphocytes move as one very slow fraction, indicating that the difference in the mobility of the two kinds of cells is due to differences in the density of exposed sialic acid on their surfaces.

Animals↗
PubMed1981

Rise in inulin-sensitive B cells during ontogeny can be prematurely stimulated by thymus-dependent and thymus-independent antigens.

We have shown that the delayed acquisition of competence in expression of an anti-inulin (IN) response by neonatal BALB/c mice is preceded by a natural increase in the frequency of IN-sensitive B cells between 3 to 5 weeks of life. Up until 3 weeks of age, BALB/c mice resemble adult germfree mice in their low frequency of IN-sensitive B cells (1 to 2/10(6) B cells). Thereafter, the population of IN-reactive B cells rises about 10-fold to the young adult level, without deliberate immunization. This naturally expanded population of IN-reactive cells has an isotype profile resembling populations arising after intentional priming with other antigens in that it contained a large proportion of cells which generated clones expressing immunoglobulin G and immunoglobulin A isotypes, often without detectable immunoglobulin M. The late rise in frequency of IN-sensitive precursors could be induced prematurely by deliberate priming at 3 days of age with either the thymus-dependent antigen IN-hemocyanin or the thymus-independent bacterial levan. However, no circulating anti-IN could be detected following this early administration of antigen. Thus, the delayed expression of anti-IN does not reflect an absence of B cells of the appropriate specificity which can be stimulated to divide by antigen but rather other events occurring subsequent to perinatal generation of antigen-sensitive cells. Our observations support a role for the beta2 --> 1 fructosyl group as an environmental determinant which selectively expands out preexisting antigen-sensitive B cells at 3 to 5 weeks of age and these include clonotypes which express the predominant anti-IN idiotypes.

Animals↗