bluuebunny/crossref_metadata_2025 huggingface dataset:
Alterations in Function and Subpopulations of Peripheral Blood Mononuclear Leukocytes in Children with Portal Hypertension Leukocyte Subtypes and Function in Portal Hypertension
Authors: Ferrante Antonio , Davidson Geoffrey P. , Beard Lorraine J. , Goh David H.B.
Published: August 2009

Children with advanced portal hypertension usually have splenomegaly (often associated with hypersplenism) which may lead to a decreased life span of circulating blood leukocytes. In a group of 6 children (age range 4–15 years; mean age 8.5 years) with portal hypertension, we examined peripheral blood mononuclear leukocytes for possible changes in their proportions, numbers and function. Compared to normal subjects, children with portal hypertension had a marked reduction in the percentages and absolute numbers of T lymphocytes (CD3+), T ‘helper/inducer’ (CD4+) and T ‘suppressor/cytotoxic’ (CD8+) cells. However, the percentages of B lymphocytes (FMC1+), natural killer cells (Leu 11+) and monocytes (morphological) were increased, and the absolute numbers of B cells were significantly increased in these patients. Lymphocyte responsiveness to phytohemagglutinin, pokeweed mitogen and concanavalin A were depressed in the patients, while the natural killer cell cytotoxicity was slightly increased. Thus, children with portal hypertension show changes in the relative proportions and absolute numbers of peripheral blood mononuclear blood leukocyte subpopulations as well as changes in mononuclear leukocyte function.
Numbers and Heterogeneity of Mast Cells in the Male Genital Tract of the Rat
Authors: Fritz Franz Joachim , Pabst Reinhard
Published: August 2009

Normal adult rats were used to quantitate and characterize mast cells in the male genital tract. The tissues were either fixed in a fixative containing formalin (Schaffer solution) or with basic lead acetate (BLA) to identify ‘connective-tissue mast cells’ and ‘mucosal mast cells’, respectively. In the epididymis and seminal vesicle small numbers of mast cells were identified without any obvious heterogeneity. In the prostate, however, a mean of 45.1 ± 9.3 and 23.0 ± 4.0 mast cells/mm2 was found after BLA and Schaffer fixation, respectively. This difference might be of functional and clinical significance.
Cytochemical Evidence of Lack of Basic Protein in Mucosal Mast Cells of the Small Intestine in the Rat
Authors: Lunardi L.O. , Jamur M.C. , Vugman I. , Gonçalves R.P.
Published: August 2009

The ammoniacal silver method, which identifies basic proteins, gives a positive reaction in cytoplasmic granules of rat peritoneal mast cells. However, in cytoplasmic granules of mucosal mast cells in the small intestine of the rat, this reaction is negative.
Suppression of IgE Antibody Response by Irradiated Reticulum Cell Sarcoma Cells in SJL Mice
Authors: Watanabe Naohiro , Ovary Zoltan
Published: August 2009

A low number of highly irradiated spleen or lymph node cells from reticulum cell sarcoma (RCS)- bearing SJL mice brings about suppression of IgE antibody production in adequately immunized and sublethally irradiated recipients, probably by known proliferative action of RCS cells on the few Ly-1 T cells which escaped from the irradiation.
Circulating Immune Complexes and C1 Activation in Patients with Rapidly Progressive Glomerulonephritis, before and after Treatment with Immunosuppression and Plasma Exchange
Authors: Sjöholm A.G. , Brun C. , Larsen S. , Thysell H.
Published: August 2009

Well-known assays for detection of circulating immune complexes (CIC) (C1q binding assay, C1q deviation test, solid-phase C1q binding assay, solid-phase conglutinin binding assay, and the platelet aggregation test) were used in 30 patients with different kinds of rapidly progressive glomerulonephritis. In 89% of the patients evidence of CIC was found in at least one of the assays. 81% of the patients were positive in the C1q binding assay and/or the conglutinin binding assay with addition of complement. CIC were more often detected by several assays, and with higher values in patients with systemic disease than in patients with renal involvement alone. CIC were found in 2 patients with Goodpasture’s syndrome. 15 patients were reexamined after treatment with plasma exchange, when the disease was clinically inactive. The patients were still on immunosuppressive treatment. In most of these, CIC were detectable, but with lower values than at the start of treatment. Before treatment, high levels of Cī-Cīs-Cī inactivator complexes suggested increased C1 activation in 93% of the patients. C-reactive protein was raised, and the concentrations of C1q, C1s, C4, C3 and factor B were normal or high in most of the patients. Pronounced hypocomplementemia was found only in 2 patients with systemic lupus erythematosus (SLE). After treatment, the levels of C-reactive protein, C1q, C1s, C3 and factor B had decreased in the non-SLE patients, while the average levels of C4 and Cīr-Cīs-Cī inactivator complexes were essentially unchanged.
Inhibition of Cytotoxic T Lymphocytes by a Schistosome-Derived Inhibitory Factor Is Independent of an Inhibition of the Production of Interleukin 2
Authors: Mazingue C. , Dessaint J.P. , Schmitt-Verhulst A.M. , Cerottini J.C. , Capron A.
Published: August 2009

Schistosome-derived inhibitory factor (SDIF) was shown to inhibit lymphocyte proliferation. While SDIF is not toxic to lymphoid cells, the generation of cytotoxic effector cells was inhibited by SDIF in a mixed lymphocyte culture. This inhibitory effect was not attributable to the induction of suppressor cells, as SDIF also inhibited the development of nonspecific suppressor cell activity in 7-day cultures of unstimulated spleen cells. The interleukine 2-dependent proliferation of cytotoxic T lymphocytes of or of blast cells was markedly reduced by the addition of SDIF. In contrast, the production of interleukine 2 itself was not impaired by SDIF. These results support the hypothesis of an inhibition by SDIF of a particular step of the mitotic cycle, probably posterior to the G₀-G1 transition. An inhibitory activity of SDIF on the expression of the cytolytic activity of cytotoxic T lymphocytes was also observed.
Immediate Hypersensitivity Reaction to β-Lactoglobulin in the Epithelium Lining the Colon of Guinea Pigs Fed Cows’ Milk
Authors: Cuthbert A.W. , McLaughlan P. , Coombs R.R.A.
Published: August 2009

Isolated epithelia from the colons of guinea pigs fed cows’ milk have been studied in vitro under short circuit current conditions. When challenged with β-lactoglobulin (βLG) from the basolateral side, a large, transient inward flowing current was recorded. Pharmacological and ion flux measurements indicated this was due to inappropriate chloride secretion. There was no effect of βLG on normal epithelia. The effects of challenge with βLG were abolished by pretreatment with indomethacin, a prostaglandin synthetase inhibitor. By exposing epithelia from water-drinking animals in vitro to serum from milk-drinking guinea pigs sensitivity to βLG could be conferred.
Sensitivity of Passive Cutaneous Anaphylaxis in Rats
Authors: Watanabe Naohiro , Kobayashi Akio
Published: August 2009

Passive cutaneous anaphylaxis (PCA) sensitivity and IgE antibody production were examined in rats from four different outbred colonies: Wistar A, Wistar B, Sprague-Dawley, and Donryu. PCA sensitivity was the highest in Wistar B and the lowest in Wistar A rats using anti-Ascaris, antibovine serum albumin, and anti-Clonorchissinensis IgE antibodies. The PCA sensitivity related inversely to the reverse PCA reaction with rabbit antirat IgE antibody, suggesting that preexisting IgE molecules on the mast cells in the skin interfered with the PCA reaction. However, the skin sensitivity to compound 48/80 was comparable in rats from the four colonies indicating the same sensitivity to nonimmunological stimulation. Ability of rats to produce IgE antibody was tested by immunizing them with dinitrophenol-coupled Ascaris extract and Bordetella pertussis adjuvant. Essentially no difference in antidinitrophenol-coupled Ascaris extract IgE antibody production was observed among animals from the different colonies.
Sensitizing Ability of Derivatives of Picryl Chloride after Exposure of Mice on the Skin and in the Lung
Authors: Enander I. , Ahlstedt S. , Nygren H. , Björkstén B.
Published: August 2009

In the present study the immunizing efficacy of antigen or hapten administered by the respiratory route and the resulting inflammatory reactions in the lungs after challenge were studied. We compared the induction of delayed hypersensitivity (DH) and the IgG, IgM, IgA, and IgE antibody formation after immunization with three different trinitrobenzene derivatives by the epicutaneous, the intradermal, and the intraperitoneal routes with the intranasal route. Mice sensitized with picryl chloride (PiCl) epicutaneously or intradermally, or with picryl sulfonic acid (PSA) epicutaneously showed strong DH responses. Moderate DH responses were obtained after sensitization with PSA intradermally and with trinitrophenylated human serum albumin (TNP-HSA) intraperitoneally. Intranasal administration of PSA and TNP-HSA also resulted in moderate DH responses. Strong IgG responses were seen in mice sensitized with PiCl and PSA epicutaneously, intradermally, and also intranasally. Administration of TNP-HSA epicutaneously, intradermally, and intraperitoneally also gave high IgG responses, while intranasal administration only induced moderate ones. Considerable IgM responses were seen only in mice sensitized epicutaneously with PSA. Antibody responses of the IgA isotype were only seen after epicutaneous sensitization with PiCl or PSA and intradermally with PiCl. Histological examination of lung tissue from mice sensitized with PiCl epicutaneously or with PSA intranasally and challenged with PSA intranasally revealed some inflammatory cells around bronchioli, capillaries and in the interstitial tissue. Differentiation of mucous cells in the epithelium covering inflammatory cells was also seen, and a slight increase in mast cell numbers around vessels. However, after sensitization with PiCl epicutaneously, TNP-HSA intranasally, and challenge with 2% TNP-HSA intranasally, a weaker inflammation was seen. In conclusion, intranasal administration of PSA or epicutaneous administration of PSA and PiCl gave strong DH and IgG responses. These sensitization regimes also resulted in pronounced inflammatory reactions in the lungs.
Immune Responses in Rats Sensitized with Aerosolized Antigen
Authors: Ahlstedt S. , Smedegård G. , Nygren H. , Björkstén B.
Published: August 2009

Rats of BN × Wi/Fu strain were immunized by aerosol exposure to 0.001–1% of ovalbumin (OA) without using any adjuvant for 2- or 1-week periods with 4-week intervals. The immunization, particularly with the highest dose, induced antibody formation of IgE, measured with RAST, and of IgG and IgA, measured with ELISA. Regional lymph node cells from immunized animals showed spontaneous proliferation which was suppressed by syngeneic spleen cells. The cell mixture could be stimulated by addition of specific antigen or mitogen, while cultures of peripheral lymph node cells showed a less consistent increase in lymphoblast formation. Histological examination of the lungs of immunized animals revealed a rapid onset of inflammation with accumulations of mononuclear cells and also increased numbers of mast cells. There was some dose-response relationship between antigen concentration and cell accumulations. The immunization also seemed to induce differentiation of epithelial cells to mucous cells. Intravenous provocation of animals immunized with 0.01–1% OA aerosol resulted in clinical bronchial reactivity assessed as increased transpulmonary pressure. This airway response was attenuated by pretreatment with indomethacin. Animals immunized with 1% OA did, however, not respond to challenge. These animals showed a decreased response to serotonin compared with nonimmunized controls. In summary, the described animal model seems to provide a tool for studies of the asthmatic reaction.
Grass Conjuvac®
Authors: Taylor W.A. , Francis Daphne H.
Published: August 2009

Sera of rabbits injected with Conjuvac® developed high titres of pollen-specific haemagglutinating antibody. Similarly, sera of guinea pigs treated with Conjuvac developed high titres of pollen-specific IgG1 antibody, as assessed using guinea pig passive cutaneous anaphylaxis (PCA). In both species, the immunogenicity of pollen extract was increased by conjugation to alginate to an extent greater than or equal to that resulting from adsorption to aluminium hydroxide. In contrast, in rats, Conjuvac alone failed to induce pollen-specific IgE antibody, and it suppressed the IgE response induced by pollen extract adsorbed to aluminium hydroxide.
Local and Systemic Immune Responses to Intestinally Presented Antigen
Authors: Nicklin S. , Miller K.
Published: August 2009

We have investigated the ability of the gut-associated lymphoid tissues (GALT) to react against standard antigens presented via different routes of administration. When antigens were presented directly to GALT, either by an injection into the Peyer’s patches or via a mechanically damaged lamina propria, a rapid and vigorous immune response ensued. This was manifested systemically as an antigen-specific humoral antibody response in the serum and locally as an antigen-specific immunoglobulin A response in the bile. These results indicate that GALT is capable of reacting to foreign materials penetrating the gut wall. Additional experiments demonstrated that the extended feeding of degraded iota carrageenan, a material known to produce non-specific mucosal inflammation, caused GALT to react to antigenic molecules present within the gastrointestinal tract and enhanced both serum and biliary antibody responses against specific gut commensal microorganisms. The immunological significance of these observations is discussed.
Macrophages in Efferent Lymph of Sheep and Their Role in Lectin-Induced Lymphocyte Blastogenesis
Authors: De Martini J.C. , Fiscus S.A. , Pearson L.D.
Published: August 2009

Very low numbers (3.9 ± 3.0/105 cells) of macrophages (cells with intense cytoplasmic staining for nonspecific esterase) were found in seven popliteal lymph samples taken at surgery from 6 sheep. 67% of these cells were phagocytic for latex beads. Dose response and limiting dilution experiments indicated that efferent lymph lymphocytes responded to concanavalin A and pokeweed mitogen when at least 10 macrophages/105 cells were present, but macrophages appeared not to be necessary for phytohemagglutinin responses. Further depletion of macrophages from efferent lymph lymphocytes by nylon wool columns almost abolished concanavalin A and pokeweed mitogen responses and reduced phytohemagglutinin responses by 67%. The responses to all mitogens were restored to near normal efferent lymph lymphocyte levels by addition of 1% adherent cells and were elevated to near normal blood cell levels by 5% adherent cells. These results indicate that macrophages are present in efferent lymph and that, despite their low numbers, they may be of functional significance in regulation of immune responses.
Presence of Basophils in PHA-Reacted Skin Sites of Mice
Authors: Hurtado Inés , Urbina Cecilia
Published: August 2009

Subcutaneous injections of the lymphocyte mitogen PHA in mice induced a local reaction which was delayed in time course and characterized histologically by dermal infiltration with blood granulocytes. Basophils were identified in the skin sections by electron microscopy as their granules differ from those of other blood granulocytes and tissue mast cells.
IgA Isotype Restriction in the Mucosal but Not in the Extramucosal Immune Response after Oral Immunizations with Cholera Toxin or Cholera B Subunit
Authors: Lycke Nils , Lindholm Leif , Holmgren Jan
Published: August 2009

Intestinal mucosal as well as extramucosal antibody responses were studied in mice after peroral immunizations with cholera toxin or cholera B subunit. The immunizations with cholera toxin gave rise to a marked response with antitoxin-secreting cells (PFC) in Peyer’s patches (PP), mesenteric lymph nodes (MLN) and spleen showing isotype distribution of IgG > IgA > IgM and with PFC kinetics in MLN and spleen that suggested migration of cells from PP after peroral administration rather than cells stimulated in situ by adsorbed antigen. Highest numbers of PFC were obtained after 2 immunizations, and further administrations resulted in a decrease in the PFC response in MLN and spleen, while the PP responsiveness was relatively unchanged, and interestingly, protective immunity and IgA-dominated antitoxin titers in intestinal washings increased markedly by the additional boosters. Animals immunized with cholera B subunit, which lacks the adenylate cyclase-stimulating capacity of cholera toxin, showed similar PFC responses in extramucosal organs as those receiving cholera toxin but were poorly protected and had correspondingly lower IgA antitoxin titers in intestinal washings. These results suggest that the mucosal IgA antitoxin predominance is mainly due to regulatory mechanisms operating on the end-stage differentiation of the committed B cells in lamina propria and that this differentiation, as judged from the different results with cholera toxin and its B subunit, might be influenced by cyclic AMP.
Search for Thymocyte Growth-Stimulating Activity in Extracts of Different Tissues and among Established Growth Factors and Thymic Maturation Hormones
Authors: Söder O. , Ernström U.
Published: August 2009

A thymocyte specific growth factor has previously been prepared from calf thymus. Similar activity is now shown to be present in thymus and spleen of guinea pigs, but not in lymph nodes, liver and kidney. Purified thymic maturation hormones, interleukins and several growth factors for other cell types than lymphocytes were tested and found not to stimulate DNA synthesis of thymocytes. A slight activity was found in thymosin fraction 5, known to contain at least 35 polypeptides. Our factor lacked comitogenic effect when tested in combination with concanavalin A (ConA) and phytohemagglutinin, did not promote continuous growth of ConA-induced lymphoblasts and did not affect the cyclic AMP level of thymocytes. The data indicate that our factor is functionally distinct from the purified hormones and defined factors tested.
Immune Response to Trichomonas vaginalis
Authors: Yano A. , Yui K. , Aosai F. , Kojima S. , Kawana T. , Ovary Z.
Published: August 2009

The immune response of patients with trichomoniasis vaginitis to Trichomonas vaginalis was examined by using passive hemagglutination assay and skin reactions. Trichomonas-specifîc immune responses could hardly be detected in the patients by these assay methods. In order to develop a simple and reliable assay for demonstration of T. vaginalis-specific immune responses in the patients, proliferation responses of peripheral mononuclear cells from the patients were examined. Lymphocytes obtained from the patients were shown to incorporate 3H-methyl-thymidine when stimulated with T. vaginalis antigen in vitro. Furthermore, immunochemical analysis of T. vaginalis antigen has been done by Sephadex chromatography and sodium dodecyl sulfate gel electrophoresis. The data revealed that the murine and human IgM antibodies specifec to T. vaginalis recognize various antigens with a wide range of molecular weights (between 13,000 and 100,000 daltons), while human and murine IgG antibodies recognize molecules with a much narrower range of molecular weights (50,000–100,000 daltons).
The Optimum Conditions for Antibody Production in vitro by Peyer’s Patch Lymphocytes and a Comparison between Total Immunoglobulins and Antigen (TNP)-Specific Antibody Synthesis following Mitogen Stimulation
Authors: Layton G.T. , Smithyman A.M.
Published: August 2009

Using optimised conditions in terms of cell concentration (2 × 106/ml), duration of culture (7 days) and pH (7.0–7.1) and the highly sensitive enzyme-linked immunosorbant assay for antibody quantitation, we assessed the effects of lipopolysaccharide (LPS) and concanavalin A (Con A) on the levels of both total immunoglobulins and TNP-specific antibodies and compared the proportion of IgA anti-TNP to total IgA synthesised following the in vivo priming of mice, 5 days prior to culture, with trinitrophenylated sheep red blood cells (TNP-SRBC). LPS stimulation resulted in an increase in total IgA (ninefold), IgM (sixfold) and IgG (eightfold) synthesis over unstimulated cultures and similar increases were seen in IgA anti-TNP (eightfold), IgM anti-TNP (fivefold) and IgG anti-TNP (twelvefold). Con A stimulation resulted in a slight increase in total IgA (threefold), IgG (twofold) but a decrease in IgM (twofold) but did not appear to affect TNP-specific antibody levels. In unstimulated cultures, IgA anti-TNP comprised 7.8% of the total IgA synthesised. In LPS and Con A stimulated cultures the proportion was 6.6% and 3.2%, respectively. Inclusion of 10% fetal calf serum in the culture medium resulted in a substantial increase in the synthesis of IgG, IgM and IgA in unstimulated cultures but increased IgA levels only in LPS stimulated cultures. Our results emphasise the importance of optimising culture conditions when studying in vitro antibody synthesis and demonstrate that Peyer’s patches respond quite substantially to parenterally administered antigen and that these responses can subsequently be monitored in vitro.
Susceptibility of Two Colonies of Wistar Rats to Inflammation, with Particular Reference to Delayed Hypersensitivity
Authors: de Brito F.B. , Hanahoe T.H.P.
Published: August 2009

Two colonies of Wistar rats were tested for their abilities to produce delayed hypersensitivity reactions and other forms of inflammation. Tuck rats, which respond to dextran with an anaphylactoid reaction, produced delayed reactions to tuberculin protein and to ovalbumin in Freund’s incomplete adjuvant. On the other hand, NELP rats which do not respond to dextran also produced delayed reactions to tuberculin protein, but only to ovalbumin when this was contained in Freund’s complete adjuvant. Rats of both colonies responded to cotton pellet-induced inflammation, but the adult NELP rats showed resistance both to adjuvant-induced and to collagen-induced arthritis, as well as to the production of experimental allergic encephalomyelitis. NELP rats also showed a much greater reticulo-endothelial system phagocytic activity although antibody titres to sheep red blood cells and the mitogenic activity of concanavalin A and of lipopolysaccharide on spleen cells were similar in the two colonies of rats.
Rheumatoid-Like Synovitis in Rabbits by Intra-Articular Administration of α2-Macroglobulin · Trypsin Complexes and Alkylamine-Treated α2-Macroglobulini
Authors: Borth W. , Susani M.
Published: August 2009

The present animal experiments revealed that repeated intra-articular administration of Α2-macroglobulin trypsin complexes caused a rheumatoid-like synovitis in not preimmunized rabbits. In early stages of synovitis, acute to subacute pathomorphological alterations were observed. In later stages, the chronic infiltration was accomplished by early onset of fibroplasia in some experimental joints. Similar lesions were caused by administration of methylamine-treated Α2-macroglobulin, an inactive Α2-macroglobulin (Α2-M), very like that produced in the trapping of a proteinase. In contrast, administration of native plasma Α2-M was ineffective, whereas active trypsin produced moderate inflammation. It is felt that the present experimental model resembles immunopathological processes in human arthritides, where the occurrence of Α2-M·proteinase complexes was verified.
Comparison of Iodinated Antigens of Metastatic and Non-Metastatic Mammary Adenocarcinoma Cells
Authors: Abbas Mohammed K. , Yoo Tai-June
Published: August 2009

The surface and detergent-soluble proteins of metastatic (TMT-081) and nonmetastatic (MT-100) mammary adenocarcinoma cells were labeled using [125I]-lactoperoxidase. The iodinated proteins were analyzed by SDS-gel electrophoresis and autoradiography. The results revealed seven iodinated surface polypeptides in metastatic cells with molecular weights of 150,000, 130,000, 110,000, 78,000, 67,000, 40,000 and 22,000. The nonmetastatic cell surface revealed eight iodinated polypeptides with molecular weights of 175,000, 150,000, 130,000, 120,000, 110,000, 67,000, 40,000, and 24,000. The small molecular weight polypeptides, 22,000–24,000, of both metastatic and nonmetastatic cells were the most intensely labeled and appeared as a major surface component. lodination of the detergent-soluble proteins in tumor extracts results in the labeling of five polypeptides (175,000, 67,000, 44,000, 29,000, 22,000 MW) of metastatic tumor and six polypeptides (175,000, 78,000, 67,000, 40,000, 35,000, 22,000 MW) of nonmetastatic tumor. Immunoprecipitation analyses indicate that the iodinated tumor polypeptides are antigenic to a xenogenic host. The 175,000 MW surface polypeptide of nonmetastatic and the 40,000 MW polypeptide of metastatic cells were not detected in the immunoprecipitates. The results of immunoprecipitation suggest that the small molecular weight (22,000–24,000) polypeptides are the predominant antigens of both tumor cells.
Effect of a Purified Excreted Factor from Leishmania brasiliensis on Macrophage Activity
Authors: Rodriguez Noris , Hernández Angel G. , Merino Fernando
Published: August 2009

The effect of a purified glycoprotein excreted to the culture medium by Leishmania brasiliensis on the endocytic and degradative capacity of macrophages was studied. It was demonstrated that 125I-albumin incorporation as well as its intracellular fate by mouse peritoneal or tumor J774-G8 macrophages was greatly reduced when they were treated with excreted factor (EF). EF appears to alter also membrane properties in addition to its intracellular effect. The results are discussed in relation to the Leishmania survival mechanism inside the host macrophages and its pathogenicity.
Histamine Releasers and Rat Mast Cells
Authors: West G.B.
Published: August 2009

The release of histamine from rat tissues rich in mast cells, induced by antigen, concanavalin A, dextran, compound 48/80, A 23187, phosphatidic acid and chlortetracycline, has been compared with that from isolated peritoneal mast cells of rats. Whereas most of the agents were more active than dextran in in vitro experiments, the reverse was found when they were injected intradermally into the skin, or subcutaneously into the paws. In fact, A 23187 and chlortetracycline (both calcium ionophores), as well as phosphatidic acid (another non-cytotoxic releaser), failed to release significant amounts of histamine when injected into the animal. Thus, the experimental conditions in which comparisons of the activities of histamine releasers are made should always be stated.
The Reactivities of Bauhinia purpurea and Lens culinaris Lectins to Mouse B Lymphocytes and Their Subsets
Authors: Hishinuma Atushi , Imai Yasuyuki , Nakano Toshiaki , Osawa Toshiaki
Published: August 2009

The use of lectins for the separation of murine spleen lymphocyte subsets was examined. Bauhinia purpurea agglutinin was found to be effective for the enrichment of B cells by differential agglutination. When the differential agglutination technique using lectins was applied to the separation of B cell subsets, Lens culinaris agglutinin was found to be able to fractionate B cells into high responding and low responding subsets for dextran sulfate. A different expression of cell surface IgD was also observed between the cell populations showing different agglutinability with Lens culinaris agglutinin.
Heterogeneity of Human Peripheral Blood Eosinophils: Variability in Cell Density and Cytotoxic Ability in Relation to the Level and the Origin of Hypereosinophilia
Authors: Prin Lionel , Capron Monique , Tonnel André-Bernard , Bletry Olivier , Capron André
Published: August 2009

Considerable variations in the distribution of eosinophil populations were revealed by density gradient centrifugation of peripheral blood leukocytes from 14 normal subjects and 31 patients with a transient or persistent hypereosinophilia. Such a purification procedure enabled us to collect in healthy controls, eosinophils (77.1 ± 15.9% SD purity) with an appreciable cell recovery (21.7 ± 14.0% SD) in the densest gradients (‘normodense’ cells). A high degree of purity was obtained in the same gradients, with leukocytes from patients with hypereosinophilia (84.4 ± 11.9% SD pure eosinophils) but the cell recovery was significantly decreased (4.1 ± 3.4% SD; p < 0.001). A study of the various fractions found to contain eosinophils showed cells with an altered cellular density (‘hypodense’ cells) especially in marked hypereosinophilia. Furthermore, studies on leukocytes from patients with a very high hypereosinophilia (differential cell count > 70%) led to the recovery of almost pure fractions of eosinophils in the low density gradients. Functional studies were performed on each distinct cell fraction collected. They included investigations of their cytotoxic ability in a heterologous antibody-dependent cell-mediated cytotoxicity assay and their biochemical activity by using a previously described technique evaluating the membrane hexose transport. Variability in the functional potentialities was observed in relation to the cellular density: higher cytotoxic eosinophil abilities were noted in the case of the ‘hypodense’ cell population. According to biochemical criteria, these latter cells appear to be more activated and capable of responding to stimulation.
C5a-Induced Histamine Release
Authors: Regal J.F. , Hardy T.M. , Casey F.B. , Chakrin L.W.
Published: August 2009

C5a has long been known to cause histamine release from isolated perfused guinea pig lung, isolated human basophils, rat and guinea pig mast cells, and guinea pig skin. The purpose of the present study was to compare the ability of guinea pig C5a to cause histamine release from lung fragments of dog, rabbit and rat with that released from guinea pig lung fragments. In addition, C5a-induced histamine release from the skin of the guinea pig, rabbit, and rat was investigated by examining antihistamine inhibition of C5a-induced vascular permeability changes. Guinea pig C5a caused a concentration-related release of histamine from guinea pig lung fragments with only minimal amounts of histamine being released from dog, rat, and rabbit lung fragments over the same concentration range. Guinea pig C5a also caused increased vascular permeability in guinea pig skin which was markedly inhibited by H1 antagonists. In rabbit skin, guinea pig C5a increased vascular permeability in the presence of PGE2 but the vascular leakage was unaffected by antihistamines. Rat skin was virtually unresponsive to C5a with or without PGE2. This data emphasize the differences between reactivity of species to C5a and the danger of extrapolating results obtained in one animal species to models employed with another species.
Accessory Cell Function of Human Vascular Endothelial Cells in Pokeweed-Mitogen-Stimulated Immunoglobulin Production by Peripheral Blood Lymphocytes
Authors: Hashimoto Yoshimi , Nakano Keiichiro , Yoshinoya Sadayoshi , Tanimoto Kiyoaki , Miyamoto Terumasa
Published: August 2009

The possibility that human vascular endothelial cells (EC) can function as accessory cells for pokeweed mitogen (PWM)-induced immunoglobulin synthesis by peripheral blood lymphocytes was examined. Adherent cells (AC) were vigorously depleted from peripheral blood mononuclear cells (PBMC) and PBMC were cocultured with or without various numbers of EC or AC in the presence of PWM. EC which had been treated with interferon-γ (IFN-γ) expressed HLA class II antigens on the surface and promoted both IgM and IgG production by lymphocytes as effectively as monocytes did. Nontreated EC also enhanced immunoglobulin synthesis, but less effectively. Pretreatment of EC with monoclonal anti-HLA class II antigen antibody suppressed the accessory cell function of EC. These results indicate that EC have the ability to function as sufficient accessory cells in the mitogen-induced in vitro immunoglobulin synthesis, and that this ability was closely related to the expression of HLA class II antigen. Under the circumstance of an inflammatory response where IFN-γ would exist, the accessory cell function of EC would be enhanced and would modify the immune response of immunocompetent cells.
Eosinophilia in Mouse Regulated by Multiple Factors following Repeated Stimulations with Ascaris Eggs into the Peritoneal Cavity
Authors: Kano Shiro , Kasuya Shiro , Ohtomo Hiroshi
Published: August 2009

Repeated injections of viable eggs of Ascaris suum into the peritoneal cavity of BALB/c mice induced marked peritoneal eosinophilia (9.3 × 106 cells/mouse, 52.8%) without infection. Extracted antigen (Asc) and eggs that had been killed also were able to induce marked eosinophilia; however, their levels were significantly lower (p < 0.001). This experimental system was able to demonstrate how various multiple factors cumulatively affected the degree of peritoneal eosinophila. These factors included: mechanical stimulation, caused by the repeated peritoneal lavages (4.4 × 105 cells/mouse), against a background level of peritoneal eosinophils of 4.5 × 104 cells/mouse; T-cell-independent stimulation, observed when the viable eggs were injected repeatedly into athymic nude mice (nu/nu; 1.6 × 106 cells/mouse), while on the other hand, neither eggs that had been killed nor Asc displayed any T-cell-independent augmentation of eosinophilia; and, T-cell-dependent augmentation of peritoneal eosinophilia, observed in heterozygous athymic mice (nu/+), in thymocyte-reconstituted nu/nu, and in normal BALB/c mice. Egg culture supernatant could not act on bone marrow cells to support eosinophil colony formation. In conclusion, only viable eggs were considered prime inducers, consisting of T-independent and T-dependent parts exerting cumulative effects.
Analysis of Differing Patterns of Cross-Reactivity of Honeybee and Yellow Jacket Venom-Specific IgE: Use of Purified Venom Fractions
Authors: Wypych John I. , Abeyounis John , Reisman Robert E.
Published: August 2009

Prior studies of sera from insect sting-allergic patients have analyzed the relationship of coexisting honeybee venom- and yellow jacket venom-specific IgE. Radioallergosorbent (RAST)-inhibition tests with these venoms revealed four different patterns of activity. In this present study, purified fractions prepared from these venoms were used to analyze these varying patterns. The hyaluronidases of yellow jacket venom and honeybee venom showed extensive cross-reaction. The phospholipases from these venoms showed minimal cross-reactivity; antigen 5 was restricted to yellow jacket venom. There was a high molecular weight component in yellow jacket venom with immunologic properties similar to honeybee venom acid phosphatase. Sera from individual patients showed quantitative and qualitative differences in the reactions to the major components of both venoms. The differences in the RAST-inhibition patterns in patients with elevated levels of both honeybee venom- and yellow jacket venom-specific IgE are accounted for by these differences as well as by differences in the cross-reactivity between the individual components.
Lymphocyte Analysis in a Patient with X-Linked Agammaglobulinemia and Isolated Growth Hormone Deficiency after Development of Echovirus Dermatomyositis and Meningoencephalitis
Authors: Wagner D.K. , Marti G.E. , Jaffe E.S. , Straus S.E. , Nelson D.L. , Fleisher T.A.
Published: August 2009

A previously described patient with X-linked agammaglobulinemia and growth hormone deficiency developed an echovirus-associated meningoencephalitis and dermatomyositis-like syndrome while being treated with intramuscular gamma globulin and human growth hormone. Initiation of high-dose intravenous gamma globulin resulted in resolution of the clinical symptoms and the patient has remained asymptomatic over the past 55 months. Lymphocyte phenotype analysis at the time of presentation with echovirus infection revealed an increase in CD2+, CD16+, HNK-1+ lymphocytes, a decrease in CD4+ T cells as well as absence of B cells. This elevation in the LGL/NK phenotype resolved with clinical improvement. In addition, there was evidence of lymphocyte activation following the development of echovirus infection (increase in HLA-DR expression and elevated serum IL-2 receptor levels) which resolved with clinical improvement. A muscle biopsy obtained during the period of the dermatomyositis-like syndrome demonstrated a CD8+ lymphocytic infiltrate very similar to the observations in classical dermatomyositis. Taken together, these findings suggest that growth hormone therapy in this patient failed to alter the humoral immunodeficiency. In addition, serum IL-2 receptor levels and lymphocyte phenotyping may be useful adjuncts for monitoring echovirus disease in immunodeficient patients.
Effect of Magnesium Deficiency on Interleukin Production by Fisher Rats: Effect of Interleukins on Reduced in vitro Lymphocyte Responses to Concanavalin A and Lipopolysaccharide
Authors: Yiangou M. , Hadjipetrou-Kourounakis L.
Published: August 2009

The ability of rats fed a magnesium-deficient diet to produce interleukins (ILs) and the effect of ILs on in vitro lymphocyte mitogenesis have been studied in rats. Lack of magnesium resulted in a lower number of plastic-adherent spleen cells and in a reduction of IL-1 production. IL-2 production was not significantly affected, indicating differential sensitivity of T cells to magnesium deficiency. The diminished mitogenic response of splenocytes to concanavalin A (Con-A) was restored by the addition of IL-1 supernatant, while the addition of IL-2 supernatant and recombinant IL-2 resulted in significantly greater enhancement of proliferation in response to Con-A, compared with that of control spleen cells. The fact that recombinant IL-2 restored the T-cell responses to Con-A indicates that the active factor in the IL-2 supernatant is IL-2.
In vitro Reactivities of Blood Lymphocytes from Symptomatic and Asymptomatic Pigeon Breeders to Antigen and Mitogens
Authors: Bouic Patrick J. , Nel Noel C. , de Beer Pieter M. , Joubert James R.
Published: August 2009

The proliferation of blood lymphocytes in response to pigeon gammaglobulin in the presence or absence of IL2 was measured in 9 symptomatic and 12 asymptomatic pigeon breeders and 24 controls. The symptomatic breeders exhibited spontaneous lymphocyte proliferation (p < 0.001) when compared with controls whilst lymphocytes from asymptomatic breeders required the simultaneous addition of IL2 in order to proliferate (p < 0.001). IL2 alone did not induce cellular proliferation. Both patient groups had abnormal T-cell functions and response to lectins and, in the symptomatic group, there existed an inverse correlation (r = ––0.70, p < 0.001) between specific antibody titres to pigeon gammaglobulin and the Con A response. We conclude that both patient groups have circulating, presensitized antigen-specific lymphocytes but these require different stimulatory signals at the cellular level.
Histamine-Releasing Activity of Endogenous Peptides on Mast Cells Derived from Different Sites and Species
Authors: Arock M. , Devillier P. , Luffau G. , Guillosson J.J. , Renoux M.
Published: August 2009

We have examined the cytological and functional characteristics of mast cells grown in tissue culture from the bone marrow of mice and rats and compared them with mast cells isolated from the peritoneal cavity of these animals. In both species, bone marrow-derived mast cells (considered to be a model of mucosal mast cells) have fewer cytoplasmic granules and lower histamine content than peritoneal mast cells. Sprague-Dawley rat peritoneal mast cells were responsive to various endogenous peptides and to compound 48/80. However, peritoneal mast cells isolated from BDF1 mice (a strain widely used to obtain bone marrow-derived mast cells) were not responsive to the same secretagogues. Rat and mouse bone marrow-derived mast cells obtained from Sprague-Dawley rats and BDFI1 mice were also hyporesponsive to calcium ionophore as compared to peritoneal mast cells and unresponsive to compound 48/80 and peptides. Despite the similarity of the functional characteristics of mouse and rat bone marrow-derived mast cells, mouse bone marrow-derived mast cells could not be used as a model of responsiveness to peptides for rat mucosal mast cells because of the differences in responsiveness between the peritoneal mast cells in the two species. Obtention of homogeneous rat bone marrow-derived mast cells may provide a useful tool to study the functional heterogeneity in an intraspecies system since most of our knowledge on mast cell physiology and pharmacology is derived from studies on rat peritoneal mast cells.
Immunoglobulin Synthesis and Gene Rearrangement in Ataxia-Telangiectasia B-Lymphoblastoid Cell Lines
Authors: Kumar Sharad , Seymour G.J. , Lavin Martin F.
Published: August 2009

We have analysed the surface immunoglobulins, cell supernatant immunoglobulins and rearrangement of genes specifying these markers in Epstein-Barr virus-transformed lymphoblastoid cell lines, produced from normal individuals and patients with the genetic disorder ataxia-telangiectasia (A-T). Surface IgG and IgM were detected in both normals and A-T patients, while IgA was only present in the controls at a low level. Cell supernatant IgG or IgM was present in controls and some A-T patients, but the majority of A-T patients had both present. No IgA was detected in supernatants from either cell type. All of the cell lines studied showed rearrangement of immunoglobulin heavy chain genes and expression of mRNA. The results described here demonstrate that reduced IgA is not confined to A-T cells and they do not reflect the low levels of serum IgA in A-T patients.
Reactivity of Autoantibodies in Systemic Lupus etythematosus with Synthetic Core Histone Peptides
Authors: Muller Sylviane , Bonnier Dominique , Thiry Marc , Van Regenmortel Marc H.V.
Published: August 2009

The specificity of autoantibodies present in the serum of 151 patients with systemic lupus erythematosus (SLE) was investigated by ELISA using as antigen individual histones as well as 17 different core histone synthetic peptides. Many of the sera reacted with four terminal peptides (residues 1–21 and 130–135 of H3, 1–29 of H4 and 1–25 of H2B) while fewer reacted with internal peptides (residues 65–85 of H2A and 40–55 of H3). Of the 151 SLE sera, 88% reacted with one or more of the six core histone peptides whereas only 57% reacted with one or more of the complete core histone molecules. Antibodies to mononucleosomes from chicken erythrocytes were also prepared in rabbits. The rabbit antisera were tested by ELISA using as antigen chromatin subunits, native and denatured DNA, individual histones and 23 natural and synthetic peptides of histones. The antinucleosome antibodies were found to recognize the same peptide fragments as those recognized by the SLE sera.
Does Vaccination of Splenectomized Patients against Pneumococci Induce Dextran-Reactive Antibodies?
Authors: Ljungström K.-G. , Hedin H. , Richter W. , Flordal P.A.
Published: August 2009

Polyvalent pneumococcal vaccine containing polysaccharides from serotypes known to be cross-reactive with dextran was used for vaccination of 72 patients either shortly before or on the average 6.25 years after splenectomy. The titer of dextran-reactive antibodies was determined by passive hemagglutination before as well as 6 weeks and 6 months after vaccination. Already prior to vaccination the patients had moderately elevated titers of these antibodies and the titer was somewhat reduced during the study period. Small inter-group differences were observed but the titers of patients who had received cytostatic and/or radiation therapy did not appear to react differently to vaccination. It is concluded that this type of vaccine does not induce dextran-reactive antibodies when given to asplenic individuals.
Chemokinetic Effects of Exogenous and Endogenous Tumor Necrosis Factor Alpha on Human Blood Monocytes
Authors: Mushtaha Akram A. , Schmalstieg Frank C. , Hughes, Jr. Thomas K. , Rudloff Elizabeth , Goldman Armond S.
Published: August 2009

The possible chemokinetic effect of human tumor necrosis factor α (TNF-α) upon human monocytes was investigated by using a type 1 collagen gel system. Human recombinant TNF-α was found to be chemokinetic, and concentrations of that cytokine as low as 0.7 pg/μl enhanced the movement of those leukocytes. Furthermore, we explored whether endogenous TNF-α stimulates monocyte movement by incubating blood monocytes with a TNF-α-stimulating agent, human recombinant interleukin 1, with or without antibodies to human TNF-α. Human recombinant interleukin 1 enhanced the motility of blood monocytes, and that effect was blocked by rabbit polyclonal antibodies to human TNF-α. Furthermore, the movement of blood monocytes not exposed to exogenous cytokines was decreased after incubation with anti-TNF-α. Thus, endogenous as well as exogenous TNF-α is a chemokinetic agent for human monocytes.
Immunological Alterations Inducible by Mercury Compounds
Authors: Pietsch Peter , Vohr Hans-Werner , Degitz Klaus , Gleichmann Ernst
Published: August 2009

We determined the levels of total IgE, IgG and IgM in the sera of HgCl2 and gold sodium thiomalate (GST) treated A.SW, C57Bl/6, and DBA/2 mice. In HgCl2 treated A.SW mice, IgE and IgG levels increased up to 30 times above normal. In C57Bl/6 mice HgCl2 induced a slight IgE increase, but no change in IgG. Strain DBA/2, by contrast, did not show significant increases in either IgE or IgG. A.SW mice also proved susceptible to the immunostimulatory effects of GST in that they responded by significant increases in IgE, IgG, and IgM. C57Bl/6 mice responded to GST by an increase in IgM alone, and DBA/2 mice again, were resistant. The same strain-specific responses have been reported with respect to the autoimmunizing effects inducible by HgCl2 and GST.
Monoclonal Antibodies against Birch Pollen Allergens: Characterization by Immunoblotting and Use for Single-Step Affinity Purification of the Major Allergen Bet v I
Authors: Jarolim Erika , Tejkl Margarethe , Rohac Madeleine , Schlerka Gabriele , Scheiner Otto , Kraft Dietrich , Breitenbach Michael , Rumpold Helmut
Published: August 2009

Two monoclonal antibodies against birch pollen proteins were produced by immunizing BALB/c mice with birch pollen extract. In immunoblotting experiments, antibody BIP 1 reacted with a 17-kilodalton (kD) protein considered to represent the major birch pollen allergen Bet v I. A second monoclonal antibody, BIP 3, reacted with 3 different birch pollen proteins of molecular weights 32, 36 and 68 kD of which the 36- and 68-kD proteins corresponded to minor allergens of birch pollen. Two-dimensional electrophoresis/immunoblotting experiments revealed that BIP 1 reacted with all Bet v I isoallergens, also identified by human IgE antibodies. Using BIP 1 coupled to Sepharose 4B as reverse immunosorbent, Bet v I was obtained in a single-step procedure and characterized as single band by SDS-PAGE.
Allergenic Variability of Different Strains of Candida albicans
Authors: Savolainen J. , Viander M. , Einarsson R. , Koivikko A.
Published: August 2009

The allergenic variation of six Candida albicans strains was studied by immunoblotting with pooled sera from 20 C. albicans-RAST-positive subjects. The presence of a 46-kilodalton (kD) protein, the major allergen, and a 29-kD protein, an intermediate allergen, was shown in crude extracts from disrupted cells (six strains). There was a significant variation in the content of the major C. albicans allergen (46 kD) between the different strains. Surface extracts of all six C. albicans strains contained mannan. Immunoblotting of cell wall extracts of intact C. albicans (agar plate) of different age revealed the strongest IgE binding in cultures incubated for 36–50 h. Due to the varying proportions of allergens demonstrated in the different strains of C. albicans, the starting source material for extract production should be a pool of individual strains, which all should be analysed prior to pooling. Special attention should be paid to the selection of C. albicans strains, and a parameter such as the breakability of the strain should be considered.