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Biomedical subjects

J G Tew

Publications and source records attributed to J G Tew.

At least 91 records · Page 5Linked to original sources
PubMed1990

Serum antibodies to periodontal bacteria.

The purpose of this study was to determine how serum antibodies reactive with periodontitis-associated bacteria with relates to the diagnosis of periodontitis subjects. Study groups included localized juvenile periodontitis (LJP) subjects, severe periodontitis (SP) subjects, chronic adult periodontitis (AP) subjects, and age matched controls. Twenty-two bacterial strains, representing 18 different species most commonly found in early onset periodontitis were evaluated using serum from LJP, SP, and age matched controls. Serum IgG reactive with these organisms was determined using a radioimmunoassay (RIA). Serum antibody reactive with 13 bacterial strains differed significantly (P less than 0.01) between the three clinical groups. Discriminate analysis revealed that antibodies reactive with 5 bacterial strains of the 13 were able to identify the clinical group to which subjects belonged 79% of the time with control subjects being correctly identified 100% of the time, LJP subjects 78% of the time, and SP subjects 60% of the time. These strains included two strains of Actinobacillus actinomycetemcomitans (Y4 and N27), Fusobacterium nucleatum (E1D1), Eubacterium brachy, and Bacteroides gingivalis. The low classification rate of SP subjects suggested heterogeneity. The SP group could be divided into three subgroups using the serological data. One subgroup, with "super" severe attachment loss, generally lacked antibody reactive with these five organisms, another subgroup was serologically similar to LJP subjects, while the third subgroup had antibodies to additional organisms. This suggests that some SP subjects may represent a more advanced form of LJP. Comparison of antibody reactivity of AP subjects with age matched controls to 23 bacterial types revealed that mean serum antibody reactivity to only Bacteroides gingivalis was higher in AP subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinobacillus↗
PubMed1989

Germinal center B cells and mixed leukocyte reactions.

The present study was undertaken to determine if germinal center (GC) B cells are sufficiently activated to stimulate mixed leukocyte reactions (MLR). Percoll density fractionation and a panning technique with peanut agglutinin (PNA) were used to isolate GC B cells from the lymph nodes of immune mice. The GC B cells were treated with mitomycin C or irradiation and used to stimulate allogeneic or syngeneic splenic T cells in the MLR. Controls included high-density (HD) B cells prepared from spleens of the same mice and HD B cells activated with lipopolysaccharide (LPS) and dextran sulfate. GC B cells bound high amount sof PNA (i.e., PNAhi). Similarly, the LPS-dextran sulfate-activated B cells were PNAhi. Treatment with neuraminidase rendered the PNAlo HD B cells PNAhi. GC B cells and the LPS-dextran sulfate-activated HD B cells stimulated a potent MLR, while the untreated HD B cells did not. However, following neuraminidase treatment, the resulting PNAhi HD B cell population was able to induce an MLR. The PNA marker appeared to be an indicator of stimulatory activity, but incubating the cells with PNA to bind the cell surface ligand did not interfere with the MLR. GC B cells were also capable of stimulating a syngeneic MLR in most experiments although this was not consistently obtained. It appears that germinal centers represent a unique in vivo microenvironment that provides the necessary signals for B cells to become highly effective antigen-presenting cells.

Animals↗
PubMed1989

The alternative antigen pathway.

In this article, John Tew, Marie Kosco and Andras Szakal describe progress in deciphering a novel antigen pathway that represents an alternative to classical clearance by phagocytes. Upon booster immunization, immune complexes form rapidly and these complexes are handled not only by phagocytes but also by a series of non-phagocytic cells in a distinct pathway. Ultimately, immunogen in this alternative pathway is delivered to antigen-specific B cells in lymphoid nodules which process and present antigen to T cells. B cells obtain immunogen from follicular dendritic cells in the form of immune complex coated bodies or 'iccosomes'. Antigen-specific B cells appear to find these newly discovered iccosomes especially palatable.

Animals↗
PubMed1989

Immunodominant antigen of Actinobacillus actinomycetemcomitans Y4 in high-responder patients.

This study was undertaken to look for characteristics of the immunodominant antigen(s) of Actinobacillus actinomycetemcomitans Y4 that might help explain the high antibody titers in periodontitis patients. Radioimmunoassays (RIA) were performed on sera from 481 patients; sera from the 32 patients with the highest anti-Y4 titers (above 128,000 RIA U/ml) were further analyzed. Y4 antigen was boiled for 45 min or treated with papain, and antibody responses were analyzed by RIA and Western blotting (immunoblotting). In addition, carbohydrate was purified from Y4 and examined by Western blotting. The results indicated that the immunodominant antigen of Y4 in high responders was stable after papain treatment or boiling for 45 min. Papain or boiling eliminated protein bands but a large diffuse band persisted on Western blots. With increasing dilutions of sera, bands on Western blots corresponding to protein antigens disappeared, while the large diffuse band resembling that of carbohydrate persisted. Partially purified Y4 carbohydrate contained the large diffuse band. Double-immunodiffusion analysis indicated that rabbit serotype b-specific antiserum and patient sera recognized the same antigen. When the carbohydrate extract was passed over a lipid A-binding column to remove lipopolysaccharide, the smear corresponding to the immunodominant antigen was still present on Western blots. The immunodominant antigen of Y4 in high-responder individuals appears to be a carbohydrate and is possibly the capsular polysaccharide.

Actinobacillus↗
PubMed1989

Antibody-forming cell induction during an early phase of germinal centre development and its delay with ageing.

The present study was initiated to determine if an early phase of germinal centre (GC) development is associated with the generation of antibody-forming cells (AFC). Germinal centres in draining lymph nodes from immune mice were examined histochemically after secondary immunization for the presence of AFC at both the light and electron microscopic levels. Additionally, peanut agglutinin (PNA) high (Hi) GC B cells were isolated, placed in cell culture and specific antibody production was monitored at successive intervals. Electron microscopy showed that plasma cells in all stages of differentiation were present within GC at 3-5 days and to a lesser extent at 7 days following antigenic challenge. Furthermore, PNAHi GC B cells obtained between Days 3 and 5 spontaneously produced specific IgG when placed in culture. Germinal centre B cells isolated either before or after this period did not produce antibody without the addition of T-cell cytokines. Induction of AFC in GC occurred at the time when GC B cells acquire follicular dendritic cell (FDC)-derived, immune complex-coated bodies (iccosomes) and process and present this antigen to helper T cells. This suggested a causal relationship between iccosome release and AFC induction. Support for this was obtained by examination of AFC induction in aged mice where iccosome release has not been observed. Peanut agglutinin-positive GC B cells isolated from aged mice on Days 3-5 after antigen challenge failed to spontaneously produce specific antibody. Collectively, these data show that GC development 3-5 days after booster immunization results in AFC generation and suggests a role for FDC iccosomes in their induction.

Aging↗
PubMed1988

A novel in vivo follicular dendritic cell-dependent iccosome-mediated mechanism for delivery of antigen to antigen-processing cells.

Recent scanning electron microscopic studies on isolated follicular dendritic cells (FDC) showed that dendrites of certain FDC were "beaded," i.e., consisting of a series of interconnected immune complex coated bodies (termed "iccosomes," measuring 0.3 to 0.7 micron diameter). In vitro these iccosomes detach from one another with ease. The major objectives herein were to establish whether these structures can be detected in sections and whether iccosomes serve to disseminate antigen in vivo. Beginning at day 1, the time point used for isolating beaded FDC, the popliteal lymph nodes of immune C3H mice were studied with light and transmission electron microscopy for 2 wk (i.e., at days 1, 3, 5, 8, and 14) after hind footpad injection of the histochemically detectable antigen, horseradish peroxidase (HRP). Iccosomes (0.25 to 0.38 micron diameter), contoured by a peroxidase (PO)-positive coat of HRP-anti-HRP complexes, were first detected by transmission electron microscopy at day 1 adjacent to cell bodies of certain FDC. Within their limiting membrane they contained flocculent material that was PO positive. At day 3 by light microscopy, germinal centers were seen enlarged and the antigen-retaining reticulum, composed of antigen-bearing FDC, appeared diffuse. This coincided with the transmission electron microscopic visualization of a dispersed state of iccosomes among the follicular lymphocytes. At that time iccosomes were seen attached to the surface of lymphocytes via PO-positive immune complexes and were surrounded by microvillous processes of these cells. Germinal center lymphocytes and tingible body macrophages both responded to contact with iccosomes by endocytosis. Antigen-containing tingible body macrophage were most conspicuous by light microscopy at day 5, when transmission electron microscopy showed that the majority of germinal center lymphocytes contained endocytosed HRP in secondary lysosome-like granules associated with the Golgi apparatus. The number of dispersed iccosomes was markedly reduced by day 5. In controls injected with HSA, a PO-negative antigen, lymphocytes and tingible body macrophages were PO-negative. The presence of antigen in both cell types was confirmed through the use of a gold-conjugated antigen (goat IgG). Simultaneous immunoperoxidase labeling of the same tissues with anti-Ia showed the gold conjugate containing B cells to be Ia+. Antigen-positive B cells and tingible body macrophages were greatly reduced in numbers by day 14, suggesting the intracellular fragmentation of the antigen.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

In vivo obtained antigen presented by germinal center B cells to T cells in vitro.

Shortly after secondary immunization germinal center (GC) B cells obtain antigen from follicular dendritic cells (FDC) in the form of immune complexes. This antigen appears to be degraded by the GC B cells and may be processed for presentation to T cells. The present study was undertaken to determine whether GC B cells can process and present antigen obtained from FDC in vivo to appropriate T cells in vitro. GC B cells were isolated from immune mice with the use of Percoll density separation followed by a panning procedure which utilizes the ability of the plant lectin, peanut agglutinin (PNA), to selectively bind to GC B cells. The enriched GC B cells were approximately 80% highly positive for PNA, 97% positive for Ia and surface IgM, but less than 0.01% positive for Thy-1.2 or esterase. In some experiments, this population was further purified to near 100% highly PNA-positive cells with the use of fluoresceinated PNA and a fluorescence-activated cell sorter. Cell sorting analysis indicated that the antigen (125I-labeled ovalbumin (OVA)) was restricted to the highly PNA-positive cell fraction. The capacity of these highly PNA-positive B cells to present antigen was assessed by monitoring interleukin 2 (IL-2) production by the OVA-specific T cell hybridoma, 3DO-54.8. GC B cells obtained from mice 3 wk or more after secondary immunization did not elicit IL-2 production in the absence of added OVA. However, GC B cells isolated as early as 1 day and for over 1 wk after a challenge with OVA, were able to stimulate high levels of IL-2 production, in the absence of adding OVA to the cell cultures. This response was maximal on day 5 and corresponded precisely with the kinetics of the ultrastructural studies which document the uptake of antigen by GC B cells in vivo. The FDC-derived antigen was remarkably immunogenic when compared with exogenous antigen. These findings demonstrated that antigen obtained in vivo by GC B cells could be processed and presented to T cells. In vivo, GC B cells may induce the T cell help needed for the germinal center reaction, generate B memory cells, and help induce the high titers of antibody associated with the secondary antibody response.

Animals↗
PubMed1988

Thy-1 positive tingible body macrophages (TBM) in mouse lymph nodes.

This study was prompted by the observation that cells in mouse lymph nodes (LN) with cytological characteristics of tingible body macrophages (TBM) appeared to be Thy-1 positive. The objective of this study was to determine if these large cells were TBM and to conclusively demonstrate their reactivity for Thy-1. The cells were studied using monoclonal antibodies (MoAb) against Thy-1 and macrophage markers including F4/80 and Ia antigens at both light microscopic (LM) and electron microscopic (EM) levels. Immunocytochemical reactivity by TBM for Thy-1 antigen specific MoAb was demonstrated by LM in both in situ and in vitro LN preparations. Furthermore, ultrastructural examination of these germinal center cells in situ demonstrated that the Thy-1 reactivity visualized at the LM level was associated with ribosomes and endoplasmic reticulum as well as their plasma membrane. Similarly, these cells expressed intracytoplasmic and membrane reactivity for Ia antigens and also for the macrophage specific antigen F4/80. This indicates that the reactivity is due to active synthesis of the Thy-1 antigen and not attributable to reactivity of any phagocytosed Thy-1 positive cells. As defined by their germinal center location and morphological characteristics, these Thy-1 reactive macrophages were identified as TBM. Germinal center TBM thus represent a unique, vigorously phagocytic subset of mature macrophages which express both macrophage and thymocyte markers.

Animals↗
PubMed1988

Germinal center B cells present antigen obtained in vivo to T cells in vitro and stimulate mixed lymphocyte reactions.

In the present study we sought to test the ability of GC B cells to activate T cells by antigen presentation and mixed lymphocyte reactions. The first set of experiments demonstrated that GC B cells obtained three or more weeks after immunization could induce IL 2 production by T cells only by adding antigen to the cell cultures. In the next (table; see text) series, it was demonstrated that GC B cells isolated as early as 1 day and for over a week following an antigenic challenge with OVA (i.e. where they obtained antigen from the FDC-derived iccosomes in vivo), were able to stimulate high levels of IL 2 production in the absence of exogenous OVA. This response was maximal on Day 5 which corresponded precisely with the keinetics of the ultrastructural studies which document the uptake of antigen by GC B cells in vivo (see Szakal et. al, this volume). Furthermore, it was shown that the FDC-derived antigen was remarkably immunogenic when compared with exogenous antigen. The data from the MLRs demonstrated that GC B cells, which are low density cells, are potent stimulators of T cells compared to the resting, low density B cells. In addition, GC B cells even stimulated a significant syngeneic response. These results indicate that GC B cells can present antigen and interact with T cells efficiently. This is believed to be important in the germinal center reaction and in the induction and maintenance of the immune responses.

Animals↗
PubMed1988

A subpopulation of germinal center B cells differentiate directly into antibody forming cells upon secondary immunization.

In the present study we sought to test the hypothesis that GC B cells may be stimulated to differentiate into AFCs during the induction of the secondary antibody response. To test this we gave HRP immune mice a booster immunization and looked for AFCs in the GCs making anti-HRP specific antibody. Within 3 days cells in the GCs were making anti-HRP. Most of the cells were just starting to make immunoglobulin in the rough endoplasmic reticulum but some were nearly mature plasma cells like those in the medullary cords. Furthermore, we isolated GC B cells after a boost with OVA and tested their ability to differentiate directly into AFCs. Data obtained using an OVA specific RIA indicated that purified GC B cells would differentiate and produce specific antibody in the absence of added T cells or antigen. Examination of the cells after 5 days in culture indicated that many of them had differentiated into large plasma cells. These results obtained both in vivo and in vitro demonstrate that shortly after booster immunization GC B cells receive the appropriate antigen and T cell signals in vivo to differentiate and become AFCs.

Animals↗
PubMed1988

Antibody synthesis specific for nonoral antigens in inflamed gingiva.

In vitro experimentation indicates that periodontitis-associated bacteria contain potent polyclonal B-cell activators (PBA). We reasoned that if PBA were operative in vivo, plasma cells specific for nonoral antigens should be present in the inflamed gingival tissues, which are characterized by a plasma cell infiltrate. To test this, rabbits with experimental periodontitis were immunized in the hind legs with the histochemically detectable antigen horseradish peroxidase (HRP) or glucose oxidase (GO). At various times after secondary immunization, inflamed gingival tissue was removed, sectioned, and treated histochemically to reveal plasma cells that specifically bound HRP or GO. Remarkably, by 9 days after secondary immunization, hundreds of HRP- or GO-binding plasma cells were found in the inflamed gingival tissue of immunized rabbits. The presence of these plasma cells, observed 7 to 10 days after booster immunization, was further substantiated by the presence of large amounts of locally produced HRP- or GO-specific antibody in gingival crevicular fluid. By 1 month after secondary immunization, the number of antigen-binding plasma cells had decreased dramatically, but a small number of antigen-specific plasma cells were detected for as long as 9 months after secondary immunization. The large number of HRP- or GO-specific plasma cells observed 9 days after immunization led us to see whether recently stimulated cells were more susceptible to PBA. Peripheral blood lymphocytes (PBL) were obtained at different times after booster immunization and cultured in the presence or absence of a PBA from Fusobacterium nucleatum. At 7 days after immunization, PBL spontaneously differentiated into antibody-forming cells in culture, and this process was enhanced by PBA. In contrast, PBL taken months after immunization produced little antibody in culture, and enhancement by PBA was difficult to detect. Compared with resting B cells, the recently stimulated B cells clearly differentiated more readily into antibody-forming cells. In conclusion, antibody synthesis specific for nonoral antigens did occur in inflamed gingival tissue, and a number of mechanisms, including PBA, probably contributed to this phenomenon.

Animals↗
PubMed1987

Relationship of serum antibody to attachment level patterns in young adults with juvenile periodontitis or generalized severe periodontitis.

The goal of this study was to relate attachment loss patterns in early onset periodontitis subjects (juvenile periodontitis n = 47 and severe (generalized) periodontitis n = 52) with antibody reactivities to 25 bacterial strains which were suspected periodontal pathogens. The 25 antibody reactivities were screened by correlation analysis. Eleven strains were found to be significantly related to attachment loss. Using these 11 reactivities, stepwise multiple linear regression with plaque and age as covariates was used to further relate the reactivities within each subject group. Plaque was significantly related to the number of teeth with slight, moderate, or severe attachment loss. A significant inverse relationship was found between antibody reactivity with Haemophilus actinomycetemcomitans Y4 and the number of teeth having slight or moderate attachment loss. Similarly a significant inverse relationship between antibody reactivity with Bacteroides gingivalis and the number of teeth having moderate or severe attachment loss was found. The inverse relationship between the two antibody reactivities and attachment loss patterns were independent of the positive relationship of plaque. These relationships suggest that the failure to mount a substantial antibody response to these organisms leads to greater and more widespread periodontal disease in early onset periodontitis subjects.

Adolescent↗