bluuebunny/crossref_metadata_2025 huggingface dataset:
Guinea Pig Mast Cells: Comparative Study on Morphology, Fixation and Staining Properties
Authors: Ghanem N.S. , Assem E.S.K. , Leung K.B.P. , Pearce F.L.
Published: August 2009

We have studied the distribution of mast cells (MC) in different tissues of the guinea pig (GP), and certain aspects of their histochemical and cytochemical properties. In preparations fixed with Carnoy’s fluid, MC were extremely well-preserved and were detected in all tissue sections and cytocentrifuge smears obtained from enzymically dispersed cells after staining with alcian blue at pH 0.5–2.2. In formol-saline-fixed preparations, there was a substantial reduction in MC counts. Most of the tissue MC were located interstitially and in the perivascular connective tissue. A small proportion of MC was located in certain organ- or tissue-specific structures. When counted manually, MC comprised 0.43–3.1% of the cell population obtained by enzymic dispersion. This proportion was higher than that obtained by the automated Technicon H6000 basophil system, but the two counting systems confirmed organ differences in MC counts. MC in different GP tissues varied in size, morphology, histamine content and degree of susceptibility to formol-saline fixation. The granules of GP connective tissue MC differed from those of rat peritoneum in failing to take the fluorescent stain berberine sulphate which has greater affinity for the highly sulphated glycosaminoglycans such as heparin, and in having some morphological and histochemical similarity to ‘mucosal’ MC.
Evidence for in vitro Regulation of IgE Receptors on the Human Basophil Membrane by Their Removal and Reexpression: Effects of Ca2+, Mg2+, Some Metabolic Inhibitors and Fetal Calf Serum
Authors: Pruzansky Jacob J. , Patterson Roy
Published: August 2009

Human basophils free of receptor-bound IgE and suspended in RPMI-1640 + 0.2% EDTA or in a buffered salt solution lacking Ca2+ and Mg2+ bound optimal quantities of 125I IgE. In media containing Ca2+ and Mg2+ such as RPMI-1640 alone or a buffered physiologic salt solution binding was reduced significantly in the first 60 min and remained essentially unchanged in the next 120 min. Scatchard analysis showed the reduction to be due both to loss of number and affinity of receptors. The presence of both 2-deoxy-d-glucose and 2,4-dinitrophenol usually prevented the reduction of binding in RPMI-1640. Suspension of cells in RPMI-1640 supplemented with fetal calf serum (FCS) demonstrated reduction in binding at 60 min characteristic of divalent-cation-containing media but this was followed by a rise in binding to approximately control levels in the next 120 min. This pattern of binding occurred in about 70% of the experiments. In most of the remaining experiments there was no difference from control binding in that there was no early decrease and no late rise in binding. This variability was not leukocyte-donor-dependent. When cells were preloaded with 125I IgE and incubated in RPMI-1640 dissociation of IgE was more rapid than in media free of Ca2+ and Mg2+ The results suggest that membrane-bound IgE receptors on basophils may be shed by an energy and Ca2+ and Mg2+ requiring process and reexpressed dependent on a factor or factors present in FCS.
Hapten-Specific Antibodies in Allergic Contact Dermatitis in the Guinea Pig
Authors: Boerrigter G.H. , Bril H. , Scheper R.J.
Published: August 2009

A guinea pig model was used to investigate the potential role of hapten-specific antibodies in allergic contact dermatitis. Hapten-specific antibodies of both the IgG1, IgG2 and IgM (sub)classes could be readily detected by ELISA in sera obtained after single or repeated exposure to the allergens 2,4-dinitrochlorobenzene (DNCB) or 4-ethoxymethylene-2-phenyloxazolone. Experiments in which animals were strongly boostered with Freund’s complete adjuvant support previous reports that hapten-specific antibodies are able to transfer contact skin reactions with a delayed time course into naive recipients. In contrast, epicutaneous sensitization procedures never allowed transfer of such reactivity. Upon transfer of immune sera to previously sensitized animals, existing allergic contact dermatitis was unaffected, except for a distinct suppression of induration, but not erythema, in the DNCB system. In actively sensitized animals, antibody-related suppression of induration was never observed. Hapten-specific hyposensitivity, as induced by repeated epicutaneous exposure, was also not related to circulating antibody levels. Our findings do not support the view that hapten-specific antibodies should be reconsidered as playing a potentially important role in allergic contact dermatitis.
IgE-Isotype-Specific Suppressor Cells in the Mouse: Characterization Using Tetraparental Chimera Mice of High (DBA/2) and Low (SJL) IgE Responder Embryos
Authors: Okudaira Hirokazu , Terada Eiji , Yokoyama Minesuke , Miyamoto Terumasa
Published: August 2009

Tetraparental chimera mice were developed by aggregation of IgE high responder (DBA/2) and IgE low responder (SJL) embryos. Anti-dinitrophenyl (DNP) IgE antibody response in such mice (SJL↔DBA/2) upon challenge with DNP-keyhole-limpet hemocyanin (KLH) in alum was clearly suppressed, while anti-DNP IgG antibody response was not. High-titer anti-DNP IgE and IgG antibody response developed in Fl hybrid mice of SJL and DBA/2 (SDF1) mice. The experimental results suggest that high IgE antibody production is the dominant trait, and the IgE-specific suppressor gene in SJL mice is autosomal recessive. IgE-specific suppressor T cells in SJL mice actively suppressed IgE antibody formation by DBA/2 immunocompetent cells across the histocompatibility barrier. Hapten-specific B cells and carrier-specific T cells were prepared in SJL↔DBA/2 and SDF1 mice by immunization with DNP-KLH or ovalbumin (OA) in alum and transferred to irradiated SDF1 mice followed by challenge with DNP-OA. Hapten-specific B cells and carrier-specific helper T cells clearly developed in SDF1 mice. Recipient mice transferred with DNP-KLH-primed SDF1 spleen cells and OA-primed SDF1 spleen cells showed high-titer anti-DNP IgE and IgG antibody responses. OA-primed SJL↔DBA/2 spleen cells cotransferred with DNP-KLH-primed SDF1 spleen cells and OA-primed SDF1 spleen cells completely abolished secondary anti-DNP IgE antibody response. The data suggest that carrier-specific helper T cells for IgE and IgG antibody responses are distinct. The regulatory role of IgE-isotype-specific suppressor cells were considered to be the interference of cooperative cellular interaction between IgE B cells and carrier-specific, IgE-specific helper T cells.
Effect of Microwaves on the Activity of Murine Macrophages in vitro
Authors: Zafra C. , Peña J. , de la Fuente M.
Published: August 2009

The effect of nonionizing electromagnetic radiation in the microwave range was studied on the adherence, opsonization, phagocytosis, nitroblue tetrazolium reduction and antibody-dependent cell cytotoxicity of peritoneal macrophages from BALB/c mice. After 15 min of irradiation with microwaves modulated in pulses at a frequency of 2,450 MHz and a power density of 32 mW/cm2, an increase in all the parameters studied was observed. The use of microwaves as a possible tool for the stimulation of the immune system is discussed.
IgE-Dependent Killing of Brugia malayi Microfilariae by Human Platelets and Its Modulation by T Cell Products
Authors: Pancré V. , Cesbron J.Y. , Auriault C. , Joseph M. , Chandenier J. , Capron A.
Published: August 2009

Platelets isolated from patients infected with filariasis were cytotoxic for microfilariae in vitro. Moreover, platelets from normal donors acquired killing properties in the presence of serum from infected individuals. The humoral factor involved in this cytotoxic process was shown to be IgE. This IgE-dependent cytotoxicity of platelets was strongly inhibited by antigen-stimulated T lymphocyte supernatants from filarial patients.
Characteristics of the Induction Phase of Antithymocyte-Globulin-Induced Heart Allograft Tolerance in the Rat
Authors: Olausson Michael , Mjörnstedt Lars , Wramner Lars , Lindholm Leif , Söderström Tommy , Brynger Hans
Published: August 2009

Transplantation tolerance was induced in 50% of Wky rats treated with antithymocyte globulin (ATG) 2 days prior to transplantation of a PVG/c heart. The induction period immediately following ATG treatment and transplantation was characterized by instability, with 50% of the allografts being lost in rejection within 30 days after transplantation. Second syngeneic transplantation to ATG-treated recipients 5 days after the first transplantation facilitated tolerance induction and permanent acceptance of both transplants. Thirty days after ATG treatment and the primary transplantation, 50% of second syngeneic allografts were accepted. During the early period after transplantation a continuous antigen presence together with ATG was necessary. Graft removal and absence of the transplant for 3 days, close to ATG administration, resulted in rejection of a second PVG/c transplant. Furthermore, the induction phase could easily be manipulated by the addition of sensitized or tolerized spleen cells to the recipient, causing allograft rejection or tolerance, respectively. Sixty days after ATG treatment and transplantation, a longer period between graft removal and retransplantation was required to break tolerance. Injection of sensitized spleen cells at this time did not affect graft survival, indicating an increased stability of the unresponsive state 60 days after ATG treatment and transplantation.
Ttypanosoma cruzi: Fibronectin Promotes Uptake of Epimastigote Culture Forms by Human Neutrophils and Monocytes
Authors: Cornette J. , Capron A. , Ouaissi M.A.
Published: August 2009

Treatment of human neutrophils and monocytes with human plasma fibronectin (Fn) enhanced their association with Trypanosoma cruzi epimastigote culture forms, a stage of parasite which activates the alternative complement pathway, and this related to the concentration of Fn in the culture medium. By increasing the incubation time, the parasite interiorization by phagocytic cells was observed. An enhancing effect of this latter phenomenon was obtained in the presence of Fn, while the addition of anti-Fn antibodies exerted an inhibitory effect. Moreover, the velocity of phagocytosis of complement-coated epimastigotes in the presence of Fn appeared greater than that observed using non-coated epimastigotes or parasites preincubated in the presence of heat-inactivated C6-deficient rabbit serum. In addition, as a consequence of cell-Fn parasite interaction, cell activation was also seen. This has been demonstrated by a chemiluminescence assay. Using radiolabeled epimastigotes (3H-uridine), we demonstrated that a proportion of ingested parasites in the presence of Fn were killed. When neutrophils were used as effector cells, the cytotoxicity was greater than that observed with monocytes. This finding of increased trypanosome uptake by phagocytic cells in the presence of fibronectin suggests that this glycoprotein could act as a ligand or cofactor to mediate parasite-cell interaction.
Effects of Different Drugs on Passive Cutaneous Anaphylaxis Elicited in the Mouse Ear at 1.5 Hours
Authors: Inagaki Naoki , Nagai Hiroichi , Xu Qiang , Daikoku Michio , Nakatomi Ichiro , Koda Akihide
Published: August 2009

IgG1, antibody-mediated homologous passive cutaneous anaphylaxis at 1.5 h (1.5-hour PCA) was elicited in the ears of mice and the effects of different drugs were studied. The reaction, as measured by the amount of extravasated dye, was inhibited by antihistamines, antiserotonins, cyclic AMP-elevating agents, tranilast and ketotifen, but not by an SRS-A antagonist (FLP 55712), lipoxygenase inhibitors, cyclooxygenase inhibitors and disodium cromoglycate. These results suggest that the pharmacological profile of 1.5-hour PCA resembles that of 48-hour PCA mediated by IgE antibody in the mouse ear.
Mechanism Responsible for Alterations in Numbers of Autonomic Nerve Receptors in Experimental Asthma
Authors: Hayashi Kouichi , Taki Fumio , Sugiyama Satoru , Takagi Kenzo , Satake Tatsuo , Ozawa Takayuki
Published: August 2009

This study was designed to elucidate the mechanism responsible for alterations in the numbers of autonomic nerve receptors in experimental asthma. In the in vivo experiment, guinea pigs sensitized by exposure to aerosol of 2% ovalbumin for 7–8 min for 10 successive days were used as the experimental asthma group. The control group was exposed to saline. Beta-, alpha-1-adrenergic and muscarinic acetylcholine receptors in lung membranes were studied by direct binding techniques using l-3H-dihydroalprenolol, 3H-bunazosin and l-3H-quinuclidinyl benzilate, respectively. The experimental asthma group showed a 33% decrease in the number of beta-adrenergic receptors, a 37% increase in the number of alpha-1-adrenergic receptors and no change in the number of muscarinic acetylcholine receptors compared with the control group. The endogenous phospholipase activity was determined by high-performance liquid chromatography using tridecanoyl phosphatidylcholine as a substrate. The phospholipase activity in lung membranes in the experimental asthma group was elevated by 50% compared with that in the control group. Lung membrane phospholipid composition was analyzed by thin-layer chromatography with a flame ionization detector. In the experimental asthma group, the amounts of phosphatidylcholine and phosphatidylethanolamine decreased significantly compared with those in the control group. In the in vitro experiment after pretreatment of lung membranes with phospholipase A2, a decreased number of beta-adrenergic receptors, an increased number of alpha-1-adrenergic receptors and no change in the number of muscarinic acetylcholine receptors were observed. These results suggest that the activation of phospholipase in asthmatic subjects degrades the phospholipids in lung membranes, resulting in alterations in the numbers of autonomic nerve receptors.
Isolation and Characterization of a Relevant Aspergillus fumigatus Antigen with IgG- and IgE-Binding Activity
Authors: Kurup Viswanath P. , Ramasamy Muthiah , Greenberger Paul A. , Fink Jordan N.
Published: August 2009

An immunologically relevant antigen fraction was isolated from a cytoplasmic extract of Aspergillus fumigatus mycelium. The fraction was obtained by concanavalin-A affinity chromatography and hydrophobic interaction chromatography. Two protein bands were discernible in polyacrylamide gel electrophoresis while two precipitin peaks were detected in crossed immunoelectrophoresis. When tested against sera from patients with Aspergillus-induced diseases, the fraction showed both IgG and IgE antibody-binding activity. All of the sera studied from patients with allergic bronchopulmonary aspergillosis (ABPA) showed high IgG and IgE antibody titers, while sera from patients with aspergilloma and cystic fibrosis with ABPA demonstrated high IgG titers and only a moderate increase in IgE titers. None of the other groups showed any significant antibody titers against this antigen in their sera. Because of its binding to both IgG and IgE antibodies this fraction was found to be useful in the immunodiagnosis of Aspergillus-induced diseases.
Improved ELISA for Thyroid Microsomal Auto-Antibodies
Authors: Ohwovoriole A.E. , Wilkin T.J. , Scott-Morgan L. , Johnston P. , Mould W.
Published: August 2009

Thyroid microsomal antibodies (TMA) and thyroglobulin antibodies (TGA) are strongly associated with auto-immune thyroid disease. TMA and TGA have been mostly detected by means of either immunofluorescence (IF), tanned red cell haemagglutination (TRCH), or radio-immunoassay (RIA) until the recent development of the enzyme-linked immunosorbent assay (ELISA). The ELISA has not been as extensively used in TMA detection as in the assay for TGA. The RIA method, though more sensitive, is technically and materially very demanding while the TRCH and IF are simpler to perform but are less sensitive. The main problem with the ELISA for the detection of TMA appears to be the interference of TGA present in some test sera reacting with thyroglobulin present as a contaminant in the thyroid microsomal preparation. In this study, we compared the TMA results from an ELISA system designed to eliminate TGA interference with those of IF and TRCH. The ELISA system in which TGA interference was eliminated without concurrent inhibition of the test reaction that gives rise to false negatives was more sensitive than either IF or TRCH. A significant number of samples was falsely reported as negative by both TRCH and IF. The correlation of the degree of positivity between TRCH and ELISA was moderate and was higher than that between ELISA and IF, though both were highly significant. The ELISA technique for TMA detection described here is a more efficient, more sensitive and also more cost-effective system than either TRCH or IF, both of which should now be replaced by ELISA, provided steps are taken to avoid false positives and false negatives.
Natural Cytotoxicity in Adult Acute Leukemia
Authors: Sørskaar D. , Førre Ø. , Albrechtsen D. , Stavem P.
Published: August 2009

Natural cytotoxic activity was studied in 32 adult patients with acute non-lymphoid leukemia (ANLL) and 27 adult patients with acute lymphoblastic leukemia (ALL). Thirteen patients with active ANLL had a significantly reduced natural cytotoxicity in peripheral blood compared with that of healthy controls, 5.0 ± 3.4 versus 27.1 ± 11.5 (p < 0.0005). Patients with ANLL in remission had a normal natural cytotoxicity (p > 0.05). Patients with active ANLL had a significantly reduced number of Leu-7-positive cells (p < 0.001), while patients in remission had normal proportions of these cells (p > 0.05). Peripheral blood from patients with ALL showed reduced natural killer (NK) cell activity, both in active disease and in remission (p < 0.0005). The percentage of Leu-11b (CD 16)-positive cells was increased in patients in remission from ALL (p < 0.05). Bone marrow cells from patients with ALL in remission had a reduced natural cytotoxicity, 2.8 ± 4.0 versus 16.3 ± 9.0 in bone marrow controls (p = 0.01). In contrast, patients with ANLL in remission showed a normal bone marrow cytotoxicity (p > 0.05) while patients with active ANLL had a reduced NK cell activity (p < 0.03). The low NK activity observed in leukemic patients may be of importance for the pathogenesis of the diseases. Acute non-lymphoid leukemia (ANLL) is a malignant disease of the multipotential hemapoietic stem cell [1]. The stem cell is capable of differentiating into various cell lineages, i.e. erythroid, granulocytic, monocytic and megacariocytic cell lineages. ANLL may thus be divided into subgroups by using various cell markers and morphology [2, 3]. Acute lymphoblastic leukemia (ALL) is of lymphoid origin. Immunological approaches to the classification of ALL have demonstrated the heterogeneity of the disease. By means of morphology and various cell markers three major subtypes of ALL are recognized: pre-B-, B- and T-cell leukemia [4, 5]. Cells exerting natural cytotoxicity (most often denoted NK cells) can kill virus-infected cells [6] and some neoplastic target cells [7, 8]. The NK cells may thus constitute an important host-defence mechanism against virus infections, and against the development of certain malignancies including leukemia. We have thus previously demonstrated a reduced NK-cell activity in patients with smouldering leukemia [9], and in childhood ALL [10]. In the present study, patients with ANLL and ALL were studied with respect to NK-cell activity in peripheral blood and bone marrow. The active forms of ANLL and ALL had both decreased spontaneous cytotoxicity. In contrast, a difference was observed between ALL in remission and ANLL in remission. The former had a reduced spontaneous cytotoxicity, while the natural cytotoxicity in the latter was normal.
C5a-Induced Bronchoconstriction: Absence of a Role of Circulating White Blood Cells and Platelets
Authors: Regal Jean F.
Published: August 2009

C5a causes an intense bronchoconstriction when injected intravenously into the guinea pig. The present study was designed to determine if circulating white blood cells or platelets were important target cells for the induction of bronchoconstriction on intravenous injection of C5a. To accomplish this, we examined the effect of C5a on numbers of circulating cells as well as the effect of depletion of various circulating cell populations on C5a-induced bronchoconstriction. Intravenous injection of C5a caused a precipitous drop in circulating granulocytes which preceded the maximum bronchoconstrictor response. The levels of circulating platelets and mononuclear cells were unchanged by intravenous injection of C5a. Depletion of either white blood cells or platelets did not significantly alter either the maximum bronchoconstrictor response to C5a or the time course of the response. Thus, these studies suggest that C5a-induced bronchoconstriction in the guinea pig is not dependent on an interaction of C5a with circulating white blood cells or platelets.
A Factor Activating Complement via the Alternative Pathway in the Supernatants of B Cell Lines Transformed by Epstein-Barr Virus and in Sera Obtained from Patients with Systemic Lupus erythematosus
Authors: Hiramatsu Misako , Tsokos George C.
Published: August 2009

Serum factors activating the alternative pathway of the complement (APC) were detected in 5 of 14 patients with systemic lupus erythematosus (SLE). Epstein-Barr virus (EBV)-transformed B cell lines were subsequently established from these patients and 6 of these produced factors capable of activating the APC. Using a limiting dilution technique, we obtained a clone which was producing a factor activating the APC (AF); by affinity column fractionation and polyacrylamide gel electrophoresis, the AF was found to have heavy and light chains comparable to those of normal human IgG. Normal human serum exhibited C3 split products (demonstrated by immunoelectrophoresis) in the presence of AF and under conditions permitting activation of the APC. Sera devoid of factor B, but not of C2 and C4, failed to catabolize C3 in the presence of AF. The AF failed to stabilize erythrocyte-bound C3bBb or C4b2a convertases, indicating that it was not a nephritic-factorlike molecule. We conclude that IgG molecules present in the sera and produced by EBV-transformed B cell lines from patients with SLE are apparently responsible, at least partially, for complement consumption in these patients.
Heparin and Acute Inflammation in the Rat
Authors: Hanahoe T.H.P. , Jones D.R.
Published: August 2009

The subcutaneous injection of heparin into the dorsal cervical region of rats 30 min before the intrapedal administration of carrageenin significantly impaired the local inflammatory reaction from 2 h through to 6 h. This observation implies that the release of endogenous heparins from tissue mast cells may play a role in modulating the extent of the response to inflammatory insults.
Detection of Human Auto-Anti-Idiotypic Antibodies (Ab2)
Authors: Castracane John M. , Rocklin Ross E.
Published: August 2009

This study documents, by means of both solid and liquid phase assays, the presence of anti-idiotypic antibodies (Ab2) in the serum of one allergic individual undergoing ragweed immunotherapy. Serum from this individual was collected and F(ab′)2 fragments specific for ragweed antigen E (AgE) were prepared (Ab1). These AgE-specific F(ab′)2 Ab1, following absorption with normal immunoglobulin, were studied for their capacity to specifically bind autologous Ab2. The binding of Ab1 to Ab2 was demonstrated to be inhibitable by ragweed AgE but not by another allergen (oak) to which the patient reacted. These findings demonstrate the presence and binding specificities of Ab2 in the serum of a ragweed immunotherapy patient and may serve as a model to study the role of Ab2 in the regulation of Ab1 in allergic patients.
Phenotypic Analysis of Guinea Pig Langerhans Cells with Antibodies Directed against Leucocyte Surface Antigens
Authors: Baker D. , Healey D.G. , Verghese S. , Schäfer H. , Turk J.L.
Published: August 2009

The epidermis was stained with a panel of recently produced anti-guinea pig leucocyte antibodies. Guinea pig Langerhans cells were not detectable with antibodies directed against B lymphocytes (MSgp9), T lymphocytes (CT7 and MSgp7), T-helper/inducer (MSgp12) and putative T-suppressor/cytotoxic (CT6 and MSgp6) subsets. Langerhans cell expressed both major histocompatibility complex (MHC) class-I and II antigens and also an epitope (CT4) associated with lymphocyte migration, thus suggesting the migratory potential of this cell. Although the Langerhans cell did not express macrophage specific antigens, MSgp5, which detects lymphoid dendritic cells, was weakly expressed on the Langerhans cell. The Langerhans cell expressed a leucocyte-common antigen detected by H201. Double-labelling studies with anti-MHC class-II antibodies indicated that only 0.4 ± 0.3% of the pan leucocyte-positive epidermal cells were Ia-negative, indicating that it is unlikely that a guinea pig analogue of the murine Thy-1 + dendritic epidermal cell (Thy-1 +dEC) exists.
Antigenic Profile of ICRC Bacilli with Special Reference to Isolation of Immunogenic Subunit
Authors: Chirmule Narendra B. , Mulherkar Rita , Deo Madhav G.
Published: August 2009

A vaccine containing ICRC bacilli induces persistent immune conversion in lepromatous leprosy (LL) patients and lepromin-negative healthy subjects, in association with upgrading of the tissue responses in the former. With an idea to isolate the immunogenic ‘subunit(s)’, antigenicity of ICRC sonicate and its fractions were tested, with reference to both B- and T-cell responses. A very high molecular weight glycolipopro-tein, named PP-I with an apparent molecular weight of 1,000,000, has been isolated using gel permeation high performance liquid chromatography (HPLC). PP-I, which focussed as a single band at pH 5 in an LKB isoelectric focussing column, quantitatively interacted with 80% of the circulating antibodies in pooled LL sera, and also induced a late (3 weeks) Mitsuda-type skin response which shows excellent correlation with host immunity against Mycobacterium leprae. These observations suggest that PP-I is a complex bifunctional antigen containing epitopes for both B and T cells. The PP-I fraction of ICRC and a similar high molecular weight HPLC fraction of M. leprae produced a line of identity against rabbit anti-ICRC serum in Ouchterlony gel diffusion and gave comparable skin responses in healthy volunteers in leprosy endemic areas. The data indicate that the PP-I fractions from the two organisms are antigenically closely related. Preliminary studies in human volunteers showed that administration of PP-I of ICRC resulted in immune conversion in lepromin-negative healthy subjects. PP-I thus appears to be the appropriate immunogen that could be used in preparation of a ‘subunit’ anti-leprosy vaccine.
Immunosuppression in Malaria: Effect of Hemozoin Produced by Plasmodium berghei and Plasmodium falciparum
Authors: Morakote Nimit , Justus David E.
Published: August 2009

To a considerable degree, malaria-induced immunosuppression has been attributed to an inhibition of macrophage accessory cell function. In this study hemozoin, a plasmodium hemoglobin degradation product which readily accumulates in phagocytic cells and tissues during infection, was examined for its influence on immune responses. Hemozoin-laden liver and splenic macrophages from Plasmodium berghei-infected mice, displayed accessory cell dysfunction which was likely due to hemozoin loading by these phagocytic cells. This is indicated by the observation that hemozoin obtained from livers and spleens of infected mice as well as from Plasmodium falciparum cultures greatly inhibited splenic plaque-forming cell responses to sheep red blood cells. The results of the present study strongly suggest that the inhibition of macrophage accessory cell activity is due, at least in part, to the uptake and accumulation of hemozoin in their cytoplasms.
Allergenic Cross-Reactivity between Callistemon citrinis and Melaleuca quinquenervia Pollens
Authors: Stanaland Brett E. , Gennaro Robert N. , Bausher Michael G. , Klotz Sol.D. , White Roseann S. , Sweeney Michael J.
Published: August 2009

Aqueous extracts of both Callistemon citrinis (bottlebrush) and Melaleuca quinquenervia (melaleuca) were analyzed for allergenic cross-reactivity. Inhibition analysis using the radioallergosorbent test (RAST) was performed on the ammonium bicarbonate extracts of bottlebrush (NH4B) and melaleuca (NH4M) pollens. RAST inhibition analysis demonstrated that the extracts contained allergenically cross-reactive components. Sephadex G-100 column chromatography of NH4B and NH4M extracts resulted in at least 4 distinct peaks for each extract analyzed. These fractions were designated NH4B1-NH4B4 and NH4M1-NH4M4. A modified dot-blot assay for detection of allergenic components was utilized to show that the first elution peaks of bottlebrush and melaleuca, NH4B1 and NH4M1, respectively, contained allergenic components. These allergenic components, NH4B1 and NH4M1, had estimated molecular weights of 50,000 and 35,000 daltons, respectively.
Use of Antisera to the Isolated Alpha and Beta Subunits of C3 as Probes to Study Functional Sites Present on Particle-Bound C3b but Absent on Native Soluble Forms of C3
Authors: Whaley K. , Nilsson U.
Published: August 2009

The effect of antisera to the isolated α and β chains of C3 on certain C3b-dependent reactions has been studied. C5-mediated haemolysis of EAC 1423b was inhibited preferentially by antiserum to the α chain, whereas antiserum to the β chain inhibited the formation of C3bBb. The anti-β chain antiserum also stabilised C3bBbP, and rendered the enzyme relatively resistant to accelerated decay in the presence of factor H. These and previous findings that anti-α and anti-β IgG bind to restricted subsets of antigenic determinants on C3/C3b suggest that these antisera affect C3b function through the binding of antibodies to active binding sites exclusively exposed by bound C3b. The anti-α and anti-β antibody probes are currently being further developed to verify this interpretation.
Synergistic Enhancement of Mitogen Responses of Human Lymphocytes by Inhibitors of Cyclo-Oxygenase and Lipoxygenase
Authors: Blomgren Henric , Hammarström Sven , Wasserman Jerzy
Published: August 2009

Previously we reported that inhibitors of the enzyme cyclo-oxygenase, which reduce biosynthesis of prostaglandins, may enhance mitogenic responses of human blood lymphoid cells, whereas only marginal effects were observed with 5,8,11-eicosatriynoic acid (ETI), which inhibits 12-lipoxygenase and leukotriene biosynthesis. However, sharply enhanced mitogen responses were observed when lymphoid cells were cultured with both a cyclo-oxygenase inhibitor and ETI, suggesting a synergy between the two drugs. To test whether this is a more general phenomenon occurring between inhibitors of cyclo-oxygenase and lipoxygenase, we have now extended these studies to include the following lipoxygenase inhibitors: cirsiliol; esculetin; 5,8,11,14-eicosatetraynoic acid, and nordihydroguaiaretic acid. The results have shown that any of these drugs combined with a cyclo-oxygenase inhibitor may enhance mitogen responses more than one would expect by summing the effects of the inhibitors tested separately. We conclude that inhibitors of cyclo-oxygenase and lipoxygenase may synergistically enhance mitogen responses of lymphoid cells.
Complement Subcomponent C1q in Various Strains of Mice
Authors: Yonemasu K. , Sasaki T. , Hashimoto H. , Tsukao N.
Published: August 2009

The protein amount and the hemolytic activity of the initiating complement component (C1q) in the classical pathway, and the level of immunoglobulin G (IgG) in sera of various inbred mouse strains were measured in parallel with those in sera of closed colony mice. C1q levels were significantly high in male A/He and C57BL/10Sn mice, and conspicuously high in female C57BL/10Sn mice. In these strains and C57BL/6J mice of both sexes, IgG levels were also significantly high. IgG levels were strikingly high in female NZB mice and NZW mice of both sexes. On the other hand, both C1q and IgG levels were conspicuously low in BALB/c mice of both sexes and in male CBA mice, and surprisingly low in BALB/c-nu (nude) mice of both sexes. A significant correlation was observed between serum levels of C1q and IgG. The C1q specific activity, however, was equivalent in all serum samples with approximately 2 × 1013 effective molecules/mg. The subunit composition of C1q analyzed by polyacrylamide gel electrophoresis was well comparable within all strains of mice tested. These results may suggest that mouse serum C1q does not have any phenotypic polymorphism.
Mast Cell Degranulation as a Major Event in the Effector Phase of Delayed-Type Hypersensitivity Induced by Cloned Helper T Cells
Authors: Herrmann Peter , Schreier Max H. , Borel Jean-François , Feurer Camille
Published: August 2009

A kinetic study (0–72 h) was performed on the cellular composition of the exudate in delayed-type hypersensitivity (DTH) induced by the injection of cloned helper T cells into the footpad of C57BL/6J mice. In this model the reaction was maximal at 24 h, as assessed by local swelling. After an initial phase of inflammatory cell infiltration, we observed pronounced degranulation of mast cells, occurring at 15–19 h. Administration of antihistamines shortly before this time significantly inhibited the increase in footpad thickness at 24 h, suggesting mast cell mediators as a major cause of swelling in DTH. Furthermore, a new arrival of inflammatory cells, particularly eosinophils, was clearly correlated with mast cell degranulation. The latter, which might function to suppress local reactions, persisted as the only intact cell type after 48 h. Supplementary examination of peripheral blood smears showed successive peaks of neutrophils (at 3 h), monocytes (at 15 h) and finally eosinophils (after 20 h). These results suggest a local and systemic cascade in DTH which depends on mast cell degranulation for its full expression.
Endotoxin-Induced Auto-Immunity in Mice
Authors: Bloembergen Peter , van Dijk Hans , Hofhuis Frans M.A. , van den Heiligenberg Pauline , Willers Jan M.N.
Published: August 2009

Auto-antibody responses and circulating immune complex levels of mice with abnormal reactions to endotoxin were investigated after injection with the bacterial product. It was observed that C3H/HeJ mice displayed very high background plaque-forming cell responses towards bromelain-treated isologous erythrocytes which were slightly enhanced by endotoxin treatment. The same animals, however, did not bear auto-haemolysins in their serum, but became so upon endotoxin injection. A possible relationship between the high background reactivity of C3H/HeJ mice and the low toxicity of endotoxin in these animals is discussed. Neither untreated nor lipopolysaccharide-injected C3H/HeJ mice showed significant immune complex levels in their sera. This may be explained by their hyporesponsiveness, but by a low sensitivity to the toxic effects of endotoxin as well. C5-deficient and C5-sufficient mice showed similar auto-immune reactions, indicating that C5a, which is responsible for other effects of endotoxin, is not involved in endotoxin-induced auto-immunity.
Characterization of Basophil-Like Cells Derived from Nonhuman Primate Bone Marrow
Authors: Kirshenbaum Arnold S. , Bressler Robert B. , Friedman Marc M. , Metcalfe Dean D.
Published: August 2009

In order to determine whether mast cells or basophils could be derived from nonhuman primate bone marrow, cells from bone marrow aspirates were cultured in the presence of concanavalin A-stimulated nonhuman primate spleen cell supernatants (CAS). Culture conditions were identical to those used for culturing mucosal-like mast cells from mouse bone marrow. In this situation, basophil-like cells (BLC) could be identified in liquid cultures and averaged 14–19 μM in size, were round or oval in appearance, had lobulated nuclei, and contained less than 100 metachromatically staining granules per cell. By electron microscopy, granules had dense oval or semilunar cores with surrounding fibrous whorls. BLC were peroxidase positive, chloroacetate esterase negative, stained positively with acid toluidine blue, and contained 0.1–0.3 pg histamine per cell. BLC expressed IgE receptors and were Leu 5b and Leu 16 negative. IgE-sensitized BLC released histamine after stimulation with antihuman IgE or the calcium ionophore A23187. [35S]-labeled proteoglycans were degraded with chondroitinase ABC but not with heparinase, indicating the absence of heparin in BLC. Thus, culture conditions that include the use of CAS and lead to the growth of mast cells from rodent bone marrow result in the growth of BLC from nonhuman primate bone marrow. These observations suggest that fundamental differences exist in the type of histamine containing cells that arise from rodent and primate bone marrow when such bone marrow cells are cultured under identical conditions.
Potent Mitogenic Activity of Staphylococcal Enterotoxin A Requires Induction of Interleukin 2
Authors: Johnson Howard M. , Magazine Harold I.
Published: August 2009

Staphylococcal enterotoxin A (SEA), the most common cause of food poisoning, is capable of stimulating human T lymphocyte proliferation at concentrations as low as 10––13 to 10––16M. SEA also induces the lymphokines interleukin 2 (IL 2) and interferon gamma (IFNγ) at similarly low concentrations. HPL cultures were stimulated with SEA in the presence of antibodies to IL 2 to determine the possible role of this lymphokine in its potent mitogenic effects. Polyclonal and monoclonal antibodies to human IL 2 blocked SEA-induced mitogenesis. Treatment of cultures with higher concentrations of SEA overcame the anti-IL 2 blockage, corresponding to induction of higher concentrations of IL 2. Blockage of HPL mitogenesis by anti-IL 2 antibodies also resulted in inhibition of IFNγ production, which is dependent on IL 2. Neutralizing monoclonal antibody to IFNγ failed to block SEA-induced proliferation. The data indicate that the induction of IL 2, but not IFNγ, is a requirement for SEA induced lymphocyte proliferation. SEA food poisoning and IL 2 therapy for cancer result in similar toxic symptoms, characterized by malaise, fever, nausea or vomiting, and diarrhea. The similarity between SEA and IL 2 toxic effects, the fact that SEA is a potent inducer of lymphokines such as IL 2, and the fact that IL 2 induction is a prerequisite for the mitogenic effects of SEA raises the intriguing question of the role of lymphokines such as IL 2 in SEA-induced food poisoning.
Oxatomide Inhibits the Release of Chemical Mediators from Human Lung Tissues and from Granulocytes
Authors: Manabe Haruhiko , Ohmori Kenji , Tomioka Hisao , Yoshida Sho
Published: August 2009

The effects of oxatomide on the release of histamine and leukotriene C4 (LTC4) from human lung fragments and granulocytes were examined and the findings compared with the effects of the antiallergic drugs ketotifen, azelastine and tranilast. Oxatomide inhibited the release of both histamine and LTC4 from human lung fragments in cases of passive sensitization with human IgE myeloma serum upon anti-ε stimulation. IC50 values for the release of LTC4 from human lung fragments were as follows: oxatomide, 2.35 × 10––5M; azelastine, 27.2 × 10––5M; ketotifen, 52.1 × 10––5M; tranilast, 62.9 × 10––5M. Oxatomide also inhibited the release of both histamine and LTC4 from human mixed leukocytes stimulated by the calcium ionophore A23187. IC50 values for the release of LTC4 from human mixed leukocytes were as follows: oxatomide, 1.67 × 10––5M; azelastine, 3.65 × 10––5M; ketotifen, 12.2 × 10––5M; tranilast, 15.1 × 10––5M. The effects of oxatomide on the release of LTC4 from purified human neutrophils and eosinophils were also given attention. Oxatomide inhibited the release of LTC4 from eosinophils more effectively than from neutrophils and mixed leukocytes. As there is evidence that eosinophils play an important role in the development of late asthmatic responses and/or in the aggravation of asthma, oxatomide is expected to be an effective treatment for such conditions.
Evaluation of Specific Immunoglobulin E by Enzyme-Linked Immunosorbent Assay in Hydatid Disease
Authors: Wattal Chand , Mohan Chander , Agarwal Satish Chandera
Published: August 2009

The diagnostic significance of elevated hydatid-specific immunoglobulin E (IgE) as measured by enzyme-linked immunosorbent assay (ELISA) was evaluated against hydatid-specific IgG ELISA and Casoni’s intradermal test in surgically proven cases of hydatidosis. A specific IgE ELISA did not correlate with IgG ELISA or Casoni’s intradermal test, although its recorded sensitivity was 87%. The specific IgE ELISA was at the same time false-positive in 57.14% of the cases because of its cross-reactions with cases of ascariasis and taeniasis, unlike Casoni’s intradermal test and specific IgG ELISA. It is suggested that, in a helminth-infested population, the determination of specific IgE levels is not better than Casoni’s intradermal or specific IgG ELISA tests.
Demonstration of IgE-Sensitized Mast Cells in Human Heart and Kidney
Authors: Assem E.S.K. , Ghanem N.S.
Published: August 2009

We report the direct demonstration of IgE-bearing mast cells in human heart and kidney, as shown by immunohistochemical methods. This finding lends further support to the role of mast cells and IgE in the direct involvement of these two organs in immediate-type allergic reactions
Relationship between Neutrophil Infiltration and Tissue Eosinophilia in the Rat
Authors: Cook R.M. , Musgrove N.R.J. , Smith H.
Published: August 2009

Infection of rats with the parasite Mesocestoides corti increased the numbers of neutrophils, eosinophils and mononuclear cells in the blood. These peaked 11 days after infection and had declined to control levels by day 24. Increased numbers of eosinophils and mononuclear cells were also present in the peritoneal cavities of rats 24 days after infection. These gradually declined to reach control values by day 81. Intraperitoneal administration of glycogen to uninfected rats and to rats that had been infected for 24 and 81 days caused a transient increase in blood neutrophil numbers, maximal at 4 h. Although glycogen increased the numbers of neutrophils in the peritoneal cavities of uninfected animals and animals infected for 81 days, it did not increase the number of peritoneal neutrophils in rats that had been infected 24 days earlier. These results suggest that neutrophil infiltration can be impaired in animals undergoing an inflammatory response characterised by increased numbers of eosinophils.
Increased Superoxide Anion Production and Glutathione Peroxidase Activity in Peritoneal Macrophages from Autoimmune-Prone MRL/Mp-lpr/lpr Mice
Authors: Rokutan Kazuhito , Hosokawa Tomohide , Nakamura Keiya , Koyama Kunihiko , Aoike Akira , Kawai Keiichi
Published: August 2009

We studied the release of superoxide anion (O2––) in peritoneal macrophages from autoimmune-prone MRL/Mp-lpr/lpr (MRL/l) mice. Compared to resident peritoneal macrophages from control MRL/ Mp- +/+ (MRL/n) mice, macrophages from MRL/l mice exhibited an age-related increase of spontaneous and PMA-induced O2–– secretion in association with the development of the autoimmune process. Analysis of the kinetic parameters of NADPH oxidase in macrophages revealed that MRL/l macrophages were in a primed state, as shown by the decreased Km value of the oxidase for NADPH. Furthermore, we studied several key enzymes for their ability to scavenge the oxygen radicals in the macrophages. Among the enzymes studied, only glutathione peroxidase (GSH Px) activity was increased in peritoneal macrophages from MRL/l mice and this change was closely correlated with the increase in O2––; production. Thus, GSH Px activity in macrophages seems to play an important role in macrophage functions under increased oxidative stress.
Immunoglobulin E and Immunoglobulin G Subclass Distribution in vivo and Relationship to in vitro Generation of Interferon-Gamma and Neopterin in Patients with Severe Atopic Dermatitis
Authors: Reinhold U. , Pawelec G. , Wehrmann W. , Herold M. , Wernet P. , Kreysel H.W.
Published: August 2009

In vitro interferon-gamma (IFNγ) and neopterin generation by peripheral blood mononuclear cells (PBMC) from 15 patients with severe atopic dermatitis (AD) and 10 healthy controls was investigated. A significant proportion of patients had an impaired capacity to secrete IFNγ after phytohemagglutinin (PHA) stimulation in vitro and therefore IFNγ production was significantly lower compared to controls. Neopterin generation in vitro did not differ significantly from that of controls and no correlation between in vitro IFNγ and neopterin production could be observed in either group. Analysis of serum IgG subclass distribution showed that patients with AD had increased IgG4 serum concentrations while IgG1, IgG2 and IgG3 levels did not differ significantly from those of controls. Surface marker analysis revealed increased numbers of CD23+ lymphocytes in patients with AD which was positively correlated with the serum IgG4 and IgE concentration. Furthermore, a significant correlation was found between IFNγ generation in vitro and IgE and IgG4 concentration in vivo in AD. The data suggest that a possible dysregulation of IFNγ, interleukin-4 or other lymphokine production may be related to increased IgE and IgG4 production and seems to be an important factor in the pathogenesis of AD.
Behaviour of Guinea Pig T Cells Stimulated by Antigen, Allo-Antigen, and Mitogen
Authors: Healey D.G. , Agha Naveeda , Turk J.L.
Published: August 2009

The expression of major histocompatibility (MHC) antigens on guinea pig T cells was used as a functional marker for lymphocyte activation. Antigen-stimulated lymphocytes were recovered from guinea pigs responding to the contact sensitizer DNFB, and isolated T cells were then phenotyped using a new anti-guinea pig monoclonal antibody, MSgp7. The level of expression of MHC class II, as defined by the monoclonal antibody, MSgp8, was increased on T cells recovered 4 days after sensitization, as compared with unsensitized controls. The value of this experiment was extended by measuring MHC class II expression on T cells stimulated in vitro by the mitogen concanavalin A, where a clear increase in MSgp8 binding was also observed. Confirmation of the specificity of MSgp8 for guinea pig MHC class II antigens was achieved by studying the inhibitory capacity of this antibody on an MHC class II restricted mixed leucocyte reaction. The combination of antibodies MSgp7 and MSgp8 with flow cytometry could be applied to other guinea pig experimental models to quantitate the expression of MHC class II antigens on T cells to determine their putative value in disease manifestation.
Sensitization of Dairy Farmers to Bovine Antigens and Effects of Exposure on Specific IgG and IgE Titers
Authors: Virtanen T. , Vilhunen P. , Husman K. , Mäntyjärvi R.
Published: August 2009

Sensitization of dairy farmers to bovine-derived allergens and factors influencing this process were studied during the indoor cattle-feeding season. Eighteen typical family farms in eastern Finland were included in the study. Samples of airborne particulate material from stationary sites and the breathing zones of farmers were analyzed for bovine epithelial antigen (BEA) by the immunochemical enzyme-linked immunosorbent assay (ELISA) inhibition test. The levels of specific IgG and IgE antibodies against BEA and bovine urinary antigen (BUA) were measured from serum samples collected from farmers three times during the study. An association was found between anti-BUA IgG titers and BEA concentrations in the breathing zone. No major changes were observed between the antibody levels of sera collected at different times. In the samples from subjects with allergic rhinitis, however, a slight tendency toward descending anti-BUA IgE titers was found.