Search path mapping: a versatile approach for visualizing problem-solving behavior.
Explore the source record for details and available documents.
California↗
Biomedical subjects
R H Stevens
Publications and source records attributed to R H Stevens.
Interleukin 6 is essential for antibody secretion by human in vivo antigen-induced lymphoblastoid B cells.
In vivo immunization of normal subjects with a variety of antigens generates circulating lymphoblastoid (LB) B cells, which in vitro spontaneously secrete significant levels of specific antibody. Since activation and initial differentiation of these cells occurs in vivo, they provide a useful model for the study of the later stages of B cell maturation. In the present study, we investigated the requirement of interleukin 6 (IL-6) for the "spontaneous" in vitro production of IgG-Tet by LB B cells. Addition of IL-6 to cultures of LB B cells in medium supplemented with 10% fetal calf serum failed to increase the levels of IgG-Tet produced in vitro. However, addition of anti-IL-6 antibodies decreased IgG-Tet production as much as 70%, and this inhibition could be reversed by the addition of IL-6. LB B cells cultured in serum-free medium in order to restrict endogenous IL-6 production secreted only low levels of antibody, unless exogenous IL-6 was added. Addition of 2.5 units/ml of IL-6 to serum-free cultures induced an increase in IgG-Tet secretion nearly comparable to that seen in cultures supplied with serum. The magnitude of the increase in IgG-Tet secretion in response to exogenous IL-6 was inversely related to the number of cells in culture, which was due in part to increased endogenous IL-6 production in cultures with higher cell concentrations. Experiments including hydroxyurea in serum-free cultures indicated that IL-6-dependent enhancement of LB B cells' IgG-Tet secretion was not primarily mediated by cell growth. These observations suggest that in vivo generated LB B cells are not totally committed to antibody secretion, and that IL-6 is essential for in vivo antigen-induced LB B cells to reach the antibody-secreting stage.
Antibodies↗
Soluble CD23 containing B cell supernatants induce IgE from peripheral blood B-lymphocytes and costimulate with interleukin-4 in induction of IgE.
The role of soluble fragments of CD23 and their relationship to interleukin-4 (IL-4) in the in vitro production of IgE by normal human peripheral blood mononuclear cells was examined. Most donors' cells were induced to produce IgE in vitro by IL-4 during a 9- to 21-day culture. This stimulation was not observed in the absence of T cells. Inability of IL-4 to induce IgE in nonresponding cultures was associated with a failure to express CD23 on Lev-19+ natural killer cells; CD23 expression on B cells and monocytes was equivalent in responding and nonresponding subjects. Concentrated supernatants from Epstein-Barr virus-transformed B cell lines containing soluble fragments (sCD23) of the low-affinity Fc epsilon R (Fc epsilon R-II, CD23) induced IgE from all donors' cells in the absence of T cells. The sCD23 containing supernatants were demonstrated to be devoid of IL-4, and their effect could not be abrogated by anti-IL-4. IgE induction by both IL-4 and sCD23-containing supernatant were blocked by anti-CD23 monoclonal antibody. Affinity absorption of sCD23 removed the IgE-inducing activity. The cells most responsive to the sCD23 material were small, resting B cells rather than large in vivo activated cells. IL-4 synergized with sCD23-containing supernatant in the T cell-depleted cultures, and limiting dilution analyses demonstrated that IL-4 caused a more than tenfold increase in the precursor frequency of cells capable of responding to sCD23-containing supernatant with IgE production. These data are consistent with the hypothesis that IL-4 has multiple effects in the ultimate induction of human IgE including (1) commitment of B cells to IgE and (2) the generation of natural killer cell sCD23 fragments that subsequently drive IgE-committed cells to IgE synthesis.
Adult↗
AIDS Kaposi sarcoma-derived cells produce and respond to interleukin 6.
Cell lines derived from Kaposi sarcoma lesions of patients with AIDS (AIDS-KS cells) produce several cytokines, including an endothelial cell growth factor, interleukin 1 beta, and basic fibroblast growth factor. Since exposure to human immunodeficiency virus increases interleukin 6 (IL-6) production in monocytes and endothelial cells produce IL-6, we examined IL-6 expression and response in AIDS-KS cell lines and IL-6 expression in AIDS Kaposi sarcoma tissue. The AIDS-KS cell lines (N521J and EKS3) secreted large amounts of immunoreactive and biologically active IL-6. We found both IL-6 and IL-6 receptor (IL-6-R) RNA by slot blot hybridization analysis of AIDS-KS cells. The IL-6-R was functional, as [3H]thymidine incorporation by AIDS-KS cells increased significantly after exposure to human recombinant IL-6 (hrIL-6) at greater than 10 units/ml. When AIDS-KS cells (EKS3) were exposed to IL-6 antisense oligonucleotide, cellular proliferation decreased by nearly two-thirds, with a corresponding decrease in the production of IL-6. The decrease from IL-6 antisense in AIDS-KS cell proliferation was reversed by the addition of hrIL-6. We confirmed that AIDS-KS cells produced IL-6 in vivo by preparing RNA and tissue sections from involved and uninvolved skin from a patient with AIDS Kaposi sarcoma. We detected immunoreactive IL-6 in the involved tumor areas and to a lesser extent in the surrounding normal epidermis. Slot blot hybridization showed a great excess of IL-6 and IL-6-R RNA in involved skin compared to uninvolved skin. These results show that both IL-6 and IL-6-R are produced by AIDS-KS cells and that IL-6 is required for optimal AIDS-KS cell proliferation, and they suggest that IL-6 is an autocrine growth factor for AIDS-KS cells.
Acquired Immunodeficiency Syndrome↗
Human IgG anti-F(ab')2 antibodies possess rheumatoid factor activity.
Individual preparations of affinity purified anti-F(ab')2 antibodies and anti-Fc antibodies isolated from the sera of patients with rheumatoid arthritis (RA), were examined for reactivity with the Fab and Fc fragments of human IgG. Western blot assays demonstrated specific interaction of affinity-purified anti-Fab antibodies with both Fab and Fc molecules. Approximately one-half of the anti-Fab antibody preparations studied contained IgG antibodies reactive with Fab and Fc fragments in ELISA, suggesting the existence of naturally occurring epibody-like autoantibodies in these patients. Thirteen of 14 affinity-purified anti-Fc antibody preparations contained IgG cross-reactive with Fab molecules in ELISA. Double-adsorption assays on affinity columns demonstrated that a minimum of 14%, and possibly as much as 50%, of the IgG anti-Fab antibodies reacted with the Fc of IgG. Conversely, a minimum of 12%, and possibly as much as 70%, of the IgG anti-Fc antibodies reacted with IgG Fab molecules. Anti-Fab antibodies isolated from non-RA individuals also exhibited anti-Fc reactivity in ELISA, demonstrating the presence of these dual-reactive antibodies in other autoimmune and normal individuals. These studies establish the presence of naturally occurring IgG autoantibodies reactive with both the Fab and Fc fragments of human IgG. Their existence emphasizes the potential of anti-immunoglobulin antibodies to recognize a multiplicity of antigens, possibly including other members of the immunoglobulin supergene family.
Antibody Specificity↗
Structural proteins of the Actinobacillus actinomycetemcomitans bacteriophage phi Aa.
øAa is an A1 morphotype bacteriophage which infects certain strains of Actinobacillus actinomycetemcomitans. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of dissociated, purified phi Aa particles revealed 7 major structural proteins (P1-P7) ranging in size from 17.5 to 52.7 kilodaltons (Kd). Treatment of the intact phage particles with 67% dimethyl sulfoxide (DMSO) resulted in the separation of the virion head and tail subunits. Purification of the head subunits was accomplished by sucrose density gradient centrifugation of the DMSO-treated phage particles. The purified head subunits were composed of a single protein having an electrophoretic mobility which corresponded to a 39.5 Kd protein (P3) of the intact virus. Raising the pH of a purified phi Aa suspension to 12.7 disrupted the head subunits, as well as the tail tube and tail fibers, releasing intact contractile tail sheaths. The tail sheaths were collected by centrifugation. The purified tail sheaths were analyzed by SDS-PAGE and were found to be composed of two proteins (P1 and P2) having molecular weights of 52.7 and 41.2 Kd respectively. The location of each of the 4 remaining major structural proteins in the phi Aa virion remains to be determined.
Actinobacillus↗
Administering a microcomputer-based problem-solving examination.
Administering a microcomputer-based problem-solving examination in the basic sciences to medical students poses a series of practical concerns not associated with traditional types of testing. This article reviews the issues of planning, student preparation, exam scheduling, student verification, and software security. Also discussed are the effects on student scores of exam scheduling and of students' familiarity with computers, student evaluations of the exam, and security and technical difficulties.
Allergy and Immunology↗
Animal model of conjunctival primary acquired melanosis.
A condition clinically identical to human conjunctival primary acquired melanosis (PAM) was induced in 16 of 20 Dutch (pigmented) rabbits after weekly topical 60-microliters applications of a 1% solution of 7,12-dimethylbenz[a]anthracene (DMBA) in acetone. Pigment stippling appeared in the conjunctiva as early as 5 weeks after the initial carcinogen application. Confluent patches of flat pigmentation appeared over the palpebral conjunctiva 18 weeks after the onset of treatment and showed progressive lateral enlargement and darkening. Histologically, a spectrum of changes from increased melanin production and melanocytic hyperplasia without atypia (resembling the human condition of PAM without atypia) through atypical melanocytic hyperplasia (resembling human PAM with atypia) was identified. The development of this model permits further investigations to explore and explain the clinically observed phenomenon of waxing and waning of PAM and its promotion to conjunctival malignant melanoma.
9,10-Dimethyl-1,2-benzanthracene↗
Evaluating preclinical medical students by using computer-based problem-solving examinations.
A study was designed to determine the feasibility of creating and administering computer-based problem-solving examinations for evaluating second-year medical students in immunology and to determine how students would perform on these tests relative to their performances on concurrently administered objective and essay examinations. The results indicate that problem-solving questions of uniform difficulty can be created and that correct solution of the problems correlates with the early selection of the most relevant data. The mean scores on the three types of tests were not significantly different. The scores on the essay and objective examinations were most highly correlated, those on the computer-based and objective examinations were minimally correlated, and those on the computer-based and essay examinations were not correlated.
Allergy and Immunology↗
Detection and significance of anti-Fab autoantibody.
Explore the source record for details and available documents.
Antibodies, Anti-Idiotypic↗
Serum IgG4 anti-Fab antibodies in rheumatoid arthritis are constitutively expressed.
IgG4 comprises a significant proportion of the total anti-Fab antibody (aFABA) response in many but not all patients with rheumatoid arthritis (RA). Analyses of the dynamics of IgG aFABA subclass expression in 11 RA patients for periods of up to 11 months demonstrated that IgG4 aFABA was restricted to 6 of the 11 RA patients' sera initially studied and comprised approximately 25% (or more) of the total IgG aFABA response. Quantities of IgG4 aFABA in subsequent, serially obtained serum samples from these patients remained stable throughout the study period, whereas the remaining RA patients whose initial sera possessed small quantities of serum IgG4 aFABA failed to generate any augmented IgG4 aFABA response during the study. Elevated expression of IgG4 aFABA did not appear to be a consequence of a generalized polyclonal gammopathy or a generalized increase in autoantibody expression, though patients with higher total IgG4 serum levels expressed significantly greater quantities of IgG4 aFABA. These results indicate that the differential expression of IgG4 aFABA among RA patients reflects constitutive production within a subset of RA patients in whom IgG4 appears to comprise a significant proportion of the total IgG aFABA response.
Antibodies, Anti-Idiotypic↗
Recognition by anti-Fab antibodies in rheumatoid arthritis of structure(s) widely distributed on human Fab molecules.
Anti-Fab antibodies (aFABA) of restricted clonality and acidic spectrotypes were isolated from the sera of patients with rheumatoid arthritis (RA). These aFABA reacted with multiple populations of pooled human Fab molecules, which had been charge separated by chromatofocusing techniques (CF), indicating that the structures recognized by these aFABA were present on a polyclonal population of Fab molecules. The structures were also widely distributed among the Fab repertoires of normal individuals, as well as individual autologous and heterologous RA patients. Thus, the aFABA did not appear to recognize highly restricted epitope(s), i.e. a private idiotope, limited in its expression to RA individuals. The determinants of the Fab molecules recognized by affinity purified aFABA could be defined by linear and/or conformational structures, depending upon the individual from which the aFABA were isolated. Additionally, some of the affinity purified aFABA also reacted with Fc fragments, suggesting the presence of epibody-like autoantibodies in this population. Lastly, size analysis of the circulating IgG4 aFABA complexes indicated that these autoantibodies were not complexed with intact IgG, but rather with a molecule of 40-60 kDa, further suggesting the potential for these autoantibodies to react with multiple antigens.
Arthritis, Rheumatoid↗
The comparative cytotoxicity of periodontal bacteria.
The direct cytotoxicity of sonic extracts (SE) from nine periodontal bacteria for human gingival fibroblasts (HGF) was compared. Equivalent dosages (in terms of protein concentration) of SE were used to challenge HGF cultures. The cytotoxic potential of each SE was assessed by its ability to (1) inhibit HGF proliferation, as measured by direct cell counts; (2) inhibit 3H-thymidine incorporation in HGF cultures; or (3) cause morphological alterations of the cells in challenged cultures. The highest concentration (500 micrograms SE protein/ml) of any of the SEs used to challenge the cells was found to be markedly inhibitory to the HGFs by all three of the criteria of cytotoxicity. At the lowest dosage tested (50 micrograms SE protein/ml); only SE from Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Fusobacterium nucleatum caused a significant effect (greater than 90% inhibition or overt morphological abnormalities) in the HGFs as determined by any of the criteria employed. SE from Capnocytophaga sputigena, Eikenella corrodens, or Wolinella recta also inhibited cell proliferation and thymidine incorporation at this dosage; however, the degree of inhibition (5-50%) was consistently, clearly less than that of the first group of three organisms named above. The SE of the three other organisms tested (Actinomyces odontolyticus, Bacteroides intermedius, and Streptococcus sanguis) had little or no effect (0-10% inhibition) at this concentration. The data suggest that the outcome of the interaction between bacterial components and normal resident cells of the periodontium is, at least in part, a function of the bacterial species.
Bacterial Physiological Phenomena↗
Lymphocyte proliferative responses to mitogens in rats having an ancestry of a perinatal iodine-131 insult.
The possible existence of a genealogical memory consisting of altered lymphocyte proliferative responses to a perinatal iodine-131 insult has been investigated in two generations of inbred Fischer F344 rat offspring. The studies which involved exposure to the radioiodine during late pregnancy with concentrations ranging from 1.85 MBq (50 microCi) to 7.4 MBq (200 microCi) revealed that only the peripheral blood T lymphocytes of the first generation male animals were significantly affected. These animals were found to possess T lymphocytes which exhibited increased proliferative responses expressed toward the mitogens concanavalin A and phytohemagglutin; however, no significant changes were noticeable in their B cell population following exposure to lipopolysaccharide. Neither the first generation females nor the male and female offspring of the second generation developed through sibling interbreeding seemed to be affected, this was unlike the cellular, humoral, and natural immunity which had previously been observed to be changed in both the second and third generation animals. These observations suggest that the effects of the radiation insult upon immunocompetency as measured by lymphocyte proliferation do not appear to be inherited.
Animals↗
A bacteriocin of Actinobacillus actinomycetemcomitans.
An inhibitory factor from Actinobacillus actinomycetemcomitans Y4 was isolated, and its properties indicated that it was a bacteriocin (actinobacillicin). The bacteriocin was active against Streptococcus sanguis strains, Streptococcus uberis (FDC1), and Actinomyces viscosus T14 as well as other strains of A. actinomycetemcomitans, but not against other crevicular bacteria, including other streptococci and actinomycetes. The activity of this bacteriocin was inhibited by pronase, trypsin, and heat (45 min at 56 degrees C) but not by DNase, RNase, phospholipase, exposure to UV light, or low pH (1.0 to 6.5). Although actinobacillicin markedly inhibited glycolysis in S. sanguis, the primary mechanism of its bactericidal action appears to be alterations in cell permeability, with the resultant leakage of RNA, DNA, and other essential intracellular macromolecules. These findings provide an ecologic explanation for the reciprocal growth relationship between A. actinomycetemcomitans and S. sanguis/Actinomyces viscosus observed in localized juvenile periodontitis.
Actinobacillus↗
Purification and biochemical properties of a bacteriocin from Actinobacillus actinomycetemcomitans.
Extracts of certain strains of Actinobacillus actinomycetemcomitans are inhibitory to strains of Streptococcus sanguis such as S. sanguis ATCC 10556. The isolation of a protein from an A. actinomycetemcomitans sonic extract which copurified with the inhibitory activity was accomplished by preparative isoelectric focusing, Sephadex G-100 gel filtration chromatography, and preparative polyacrylamide gel electrophoresis (PAGE). The resulting isolated protein, which focused at a pH of 6.1 to 6.3, appeared as a single band in anionic nondissociating PAGE analysis. This protein could be dissociated into two subunits with molecular weights of 50,000 and 70,000, which were resolvable by PAGE analysis. A 1,758-fold increase in specific activity was seen in the purified inhibitory protein compared with the crude sonic extract starting material. The properties of the inhibitory activity in the A. actinomycetemcomitans extract are characteristic of a bacteriocin. Accordingly, we propose the name actinobacillicin for the inhibitory protein.
Actinobacillus↗
Defective memory B cell formation in patients with inflammatory bowel disease following tetanus toxoid booster immunization.
Inflammatory bowel disease patients with mild to inactive disease were studied for their ability to generate pokeweed mitogen responsive anti-tetanus toxoid IgG producing B cells in the peripheral blood 21 days following in vivo tetanus toxoid booster immunization. Compared to normals and disease controls, patients with inflammatory bowel disease had significantly impaired in vitro production of anti-tetanus toxoid antibody during an 8 day pokeweed mitogen stimulated culture period. There was poor correlation between the ratio of helper to suppressor T cells in the peripheral blood and pokeweed mitogen stimulated antibody production. Likewise, there was little correlation between in vitro antibody production and peripheral blood natural killer cell cytotoxic activity. Culture of the patients B cells with normal T cells failed to improve antibody production in vitro. These results indicate that many inflammatory bowel disease patients have an impaired humoral immune response to tetanus toxoid booster immunization. This impaired immune response may be due to an inability to generate B cell precursors of anti-tetanus toxoid IgG producing B cells rather than to abnormal circulating helper or suppressor T cell activity or natural killer cell regulatory activity.
Adult↗
Identification of environmental carcinogens utilizing T-cell mediated immunity.
There now exists a very large body of information that directly links the exposure to certain environmental factors with the ultimate development of specific types of human cancer. For this reason, a number of tests possessing varying degrees of biological complexity have been devised with the intent to first identify, and then ultimately reduce the risk of exposure to tumor causing agents. The presently employed chronic whole animal tests generally accepted as measures of human carcinogenicity have certain limitations in that they are lengthy to perform, very expensive, and require complicated pathological examinations of the various tissues. Consequently, there exists a need for short-term whole animal bioassays that can serve to complement such lengthy chronic studies; we are proposing that one such a test can be developed through utilizing procedures that have been designed for evaluating T-cell immunological responses.
1,2-Dimethylhydrazine↗