Streptomycin-induced anaphylactic reaction during in vitro fertilization (IVF)

Indications for in vitro fertilization (IVF) have been cautiously extended over the years. IVF is usually considered to be a technically complex method with only minimal side-effects. We report the case of a woman who developed an anaphylactic reaction during IVF immediately after the embryo transfer.     

Activated T cells induce expression of E-selectin in vitro and in an antigen-dependent manner in vivo

E-selectin is an endothelial adhesion molecule for polymorphonuclear cells, monocytes and skin-homing T cells. We have analyzed whether murine T cells are able to induce expression of E-selectin in vitro and in vivo. Using models of inflammation in which T cells play either a significant or only a minor role, we compared induction of E-selectin between normal mice and mice lacking functional T cells (athymic nude mice). In irritant contact dermatitis, a model without a major role for T cells, E-selectin was transiently expressed within the first 24 h in both normal and nude mice. In experimental leishmaniasis (where specific T cells play an important role), a high expression of E-selectin was maintained for 48 h in normal mice, whereas in nude mice expression was only transient. However, reconstitution of nude mice with 10⁸ T cells from draining lymph nodes (LN) of Leishmania-infected normal mice could restore sustained expression of E-selectin. Transfer of T lymphocytes from normal LN or from LN of mice sensitized to the contact allergen trinitrochlorbenzene (TNCB) did not have this effect. T cells from TNCB-sensitized mice, however, did induce sustained expression of E-selectin in nude mice when TNCB was applied locally; here, reconstitution with Leishmania-specific T cells had no effect. In vitro, T cells from infected or TNCB-sensitized normal mice increased expression of E-selectin on microvascular endothelial cells after 4 h of co-culture. T cells from untreated mice were less effective. Induction was dependent on direct cell-cell contact, but not on the action of interleukin-1α, interleukin-1β, tumor necrosis factor-α or interferon-γ. We conclude that sensitized T cells induce sustained expression of E-selectin in vivo in an antigen-dependent manner. This novel way of regulation could be relevant for cell-mediated immunity and chronic disease. The mechanisms are unknown, but, as in vitro, might require direct cell-cell contact.     

玄参提取物对大鼠血液流变性、凝固性和纤溶活性的影响

目的探讨玄参提取物对大鼠血液流变性、凝固性和纤溶活性的影响.方法观察玄参的石油醚、乙醇、水提取物灌胃12 d后,对大鼠血小板聚集率、血液黏度、血液凝固性、纤维蛋白溶解活性等方面的影响.结果玄参醚、醇、水提取物有显著抑制血小板聚集性、降低PAI-l的作用.结论玄参醚、醇、水提取物有抗血小板聚集、增强纤维蛋白溶解活性作用.     

Cowpox virus induces interleukin-10 both in vitro and in vivo

Cowpox virus infection induces interleukin-10 (IL-10) production from mouse bone marrow-derived dendritic cells (BMDCs) or cells of the mouse macrophage line (RAW264.7) at about 1800 pg/ml, whereas infections with vaccinia virus (strains WR or MVA) induced much less IL-10. Similarly, in vivo, IL-10 levels in bronchoalveolar lavage fluids of mice infected with cowpox virus were significantly higher than those after vaccinia virus infection. However, after intranasal cowpox virus infection, although dendritic and T-cell accumulations in the lungs of IL-10 deficient mice were greater than those in wild-type mice, weight-loss and viral burdens were not significantly different. IL-10 deficient mice were more susceptible than wild-type mice to re-infection with cowpox virus even though titers of neutralizing antibodies and virus-specific CD8 T cells were similar between IL-10 deficient and wild-type mice. Greater bronchopneumonia in IL-10 deficient mice than wild-type mice suggests that IL-10 contributes to the suppression of immunopathology in the lungs.     

Ribonuclease-gold ultrastructural localization of heparin in isolated human lung mast cells stimulated to undergo anaphylactic degranulation and recovery in vitro

BACKGROUND: Human mast cells are a rich and unique source of heparin, which is stored in cytoplasmic secretory granules and accounts for metachromasia, a staining property used to identify mast cells by light microscopy. OBJECTIVE: The sub-cellular locations of heparin in secretory human mast cells and human mast cells recovering from secretion are not known. Acquisition of this knowledge requires ultrastructural imaging of well-preserved cells with a visible probe which binds to heparin. We sought to develop this knowledge regarding human mast cell secretion by using a labelling method for heparin that depends on the well-known property of ribonuclease inhibition by heparin. METHODS: Human lung mast cells were isolated, partially purified, either stimulated or not stimulated to secrete with anti-IgE, and recovered 20 min or 6 h later for routine electron microscopy. Histamine secretion was also determined. A previously developed post-embedding, enzyme-affinity-gold electron microscopic technique to image ribonucleic acid (RNA) with ribonuclease-gold (R-G), which also binds to the enzyme inhibitor, heparin, was employed to determine the sub-cellular locations of heparin in non-secretory and secretory mast cells as well as in mast cells recovered from short-term cultures after secretion. Specificity controls for the novel use of this method and quantification of granule labelling in these controls were performed. RESULTS: Heparin was labelled by R-G in electron-dense granules within non-secretory human lung mast cells (HLMCs), in electron-dense granules that persisted in secretory HLMCs at the maximum histamine secretion time (20 min), and in electron-dense granules within recovering HLMCs. Specificity controls showed that gold alone did not label HLMCs and that absorption with heparin significantly reduced or abrogated HLMC granule staining with R-G, but that RNA absorption did not. Heparin stores were absent in newly formed, electron-lucent intracytoplasmic degranulation channels in secretory HLMCs. Electron-dense granule matrices in the process of extrusion to the cell exterior still retained heparin at the instant of cellular secretion. Non-granule heparin stores bound R-G in recovering HLMCs. These locations included resolving degranulation channels, as newly emergent granules partitioned and condensed within them, and electron-dense content-containing vesicles and progranules within synthetic mast cells. Ultimately, all known ultrastructural patterns of HLMC granules developed in recovering cells, and each of them contained heparin. CONCLUSION: Heparin was secreted from HLMCs which were stimulated by anti-IgE, and heparin was recovered by a combination of conservative and synthetic mechanisms in HLMCs after a secretory event.     

Coronary artery spasm and acute myocardial infarction in naproxen-associated anaphylactic reaction

We present the case of a 43-year-old man who suffered an acute myocardial infarction after oral administration of 250 mg of naproxen, prescribed as antiinflammatory-analgesic agent after tooth extraction. Both intradermal skin test and human basophil degranulation test were positive to naproxen. These findings suggest a naproxen-associated anaphylactic reaction with concomitant coronary artery spasm and posteroinferior infarction, a clinical event previously not reported with the use of this drug.     

Mast cells and the nodal sinus reaction in breast cancer

Mast cell counts in the sinuses of axillary nodes from breast cancer patients are shown to vary with the type of sinus reaction present. Few scattered mast cells were found in association with sinus histiocytosis. A mixed reaction, in which fewer histiocytes were present and more round cells, was accompanied by a significant increase in mast cell count, which in sinus catarrh showed a decrease. The possible role of sinus mast cells in relation to tumour spread is discussed.     

Effects of cromolyn on codeine-induced histamine release in vivo

Cromolyn has been shown to inhibit histamine release from mast cells induced by various stimuli in vitro. However, the local effects of cromolyn on codeine-induced wheal and flare skin reactions are not well understood. Intradermal injection of codeine induced prominent whealing in almost all humans. We studied the effect of local cromolyn injection on codeine-induced skin reactions, histamine release, and ultramicroscopic changes in mast cells in 10 volunteers. The finding in this study showed that injection of a 2% cromolyn solution before or together with the codeine injection does not affect the subsequent skin reactions, histamine release and ultramicroscopic changes of mast cells.     

Ligation of IgE receptors causes an anaphylactic response and neutrophil infiltration but does not induce eosinophilic inflammation in mice

Background: Eosinophils are selectively recruited into the tissues during chronic allergic inflammation. IgE is considered an initiator of the allergic reaction; however, the roles of IgE in allergic inflammation are not fully understood. Objective: We tested the hypothesis that antigen interaction with specific IgE antibody provokes eosinophilic inflammation. Methods: BALB/c mice were actively sensitized with ragweed extract and passively sensitized with anti-dinitrophenyl (anti-DNP) mouse IgE and challenged intraperitoneally by injecting either ragweed extract or DNP-ovalbumin (OVA). Immediate anaphylactic responses were examined by monitoring vascular permeability and by measuring histamine content in peritoneal lavage fluids. Late-phase allergic responses were examined by total cell counts and cell differentials. Results: Mice sensitized and challenged with ragweed showed immediate anaphylactic responses followed by temporal increases in neutrophils at 3 to 12 hours and sustained ncreases in eosinophils in their peritoneal cavities after 24 hours. Double-sensitized mice (ie, sensitized actively for ragweed and passively for DNP-OVA) challenged with ragweed showed immediate anaphylactic responses and peritoneal eosinophilia at 48 hours. Double-sensitized mice challenged with DNP-OVA showed comparable immediate anaphylactic responses but no peritoneal eosinophilia. Furthermore, at 8 hours, ragweed-challenged animals recruited both eosinophils and neutrophils, but DNP-OVA–challenged animals recruited only neutrophils. Finally, after active sensitization and challenge with ragweed, mast cell–deficient mice (WBB6F1-W/W^v) lacked the immediate response but showed comparable eosinophil accumulation as their litter mate controls (WBB6F1-+/+). Conclusion: Interaction of antigen with IgE antibody is insufficient to provoke eosinophilic inflammation in mice. (J Allergy Clin Immunol 2000;105:1202-10.)     

维生素E在体内外具有抗氧化作用

目的探讨维生素E在体内外的应激损伤。方法体外实验:人肝RBL细胞系根据H_2O_2损伤及损伤前后给予维生素E分为4组,对照组(C)、H_2O_2损伤组(Ec)、H_2O_2损伤前(Eb)及后(Ea)给维生素E组;体内实验:将20只清洁级雄性Wistar大鼠分为对照组和大及小剂量维生素E组,每天进行一次35和15 mg/kg溶液2 m L灌胃,连续3 d。体外实验应用MTT和TUNEL法检测细胞存活率和凋亡率,免疫印迹和免疫组化方法检测细胞中NF-κB、Hsp-70、Bcl-2、Bax及caspase-3的表达水平;体内实验应用生化法检测3和6 d后血浆内T-AOC、SOD、GSH和MDA的水平。结果 H_2O_2损伤组(Ec)细胞凋亡率增加(P0.01),细胞内Bax、Hsp-70、NF-κB及caspase-3显著升高(P0.01),而Bcl-2显著下降(P0.01),维生素E干预后能显著缓解上述变化(P0.01),H_2O_2损伤前干预效果优于后干预。灌胃后3 d血浆中T-AOC、SOD、GSH的水平增高,MDA降低(P0.01),6 d后其相关指标变化更加显著(P0.05)。结论维生素E可通过调节人肝RBL细胞相关蛋白表达水平和Wistar大鼠血浆中抗氧化酶体系而发挥抗氧化作用。     

Haptoglobin dampens endotoxin-induced inflammatory effects both in vitro and in vivo

We report that haptoglobin, an acute-phase protein produced by liver cells in response to interleukin-6 (IL-6), can modulate the inflammatory response induced by endotoxins. We provide evidence that haptoglobin has the ability to selectively antagonize lipopolysaccharide (LPS) effects in vitro by suppressing monocyte production of tumour necrosis factor-alpha, IL-10 and IL-12, while it fails to inhibit the production of IL-6, IL-8 and IL-1 receptor antagonist. In two animal models of LPS-induced bronchopulmonary hyperreactivity and endotoxic shock, haptoglobin knockout mice were more sensitive to LPS effects compared to their wild-type counterparts. The present data suggest that haptoglobin regulates monocyte activation following LPS stimulation. The increase in haptoglobin levels during an acute-phase reaction may generate a feedback effect which dampens the severity of cytokine release and protects against endotoxin-induced effects.     

Ocular anti-allergic compounds selectively inhibit human mast cell cytokines in vitro and conjunctival cell infiltration in vivo

BACKGROUND: Conjunctival mast cells (MCs) are important effector cells in seasonal allergic conjunctivitis, via histamine and cytokine secretion. Several new anti-allergic eye drops stabilize MCs and block histamine receptors, but their anti-inflammatory effects are unclear. OBJECTIVE: Anti-allergic drugs were compared for their anti-inflammatory effects in an in vitro model of human MC activation and in an experimental murine model of allergic conjunctivitis. METHODS: Human cord blood stem cell-derived (CBMC) and conjunctival biopsy-derived MCs were stimulated via FcepsilonRI, degranulation and histamine release were assayed at 1 h and cytokine secretion at 24 h using multiplex arrays. Mice sensitized to short ragweed pollen were given anti-allergics topically before allergen challenge, and conjunctival immuno-staining was performed at 24 h. RESULTS: After a 1 h stimulation, 80% of the CBMC had degranulated and secreted histamine (27.9+/-4.7 ng/10(6) cells; P<0.05). Pre-treatment by all drugs significantly reduced histamine and TNF-alpha, whereas IL-5, IL-8, IL-10 and TNF-beta profiles were differentially decreased. For conjunctival biopsy-derived cultures (n=11), FcepsilonR1 stimulation increased histamine, TNF-alpha, TNF-beta, IL-5 and IL-8 levels and the production of IL-5, IL-6 (P<0.05), histamine and IL-8 (P<0.01) was inhibited by epinastine. In vivo, epinastine and olopatadine pre-treatment significantly reduced the clinical scores and eosinophil numbers (n=6; P<0.05) while epinastine also reduced neutrophils (P<0.02). CONCLUSION: Differential effects on MC cytokine inhibition were observed, with epinastine inhibiting MC secretion of IL-5, IL-8, IL-10 and conjunctival neutrophil infiltration. The anti-allergic drugs have anti-histamine and mast-cell stabilizing properties but might differ in clinical improvement depending on the individual and the cytokines involved.     

The expression of murine eutaneous late phase reaction requires both IgE antibodies and CD4 T eells

Background Exposure of atopic patients to a specific allergen evokes an immediate response which is followed, in many cases, by a late phase reaction (LPR) some hours later. Here we have examined the immunological mechanisms required for the expression of cutaneous LPR in mice. Methods BALB/c mice were immunized by i.p. injection of ovalbumin (OVA) and alum actively or by i.v. injection of anti-OVA IgE monoclonal antibody (mAb) passively. After challenge by intradermal injection of OVA into ears, the changes in ear thickness, the number of eosinophils, and the levels of IL4 and IFN-γ protein at the site of antigen challenge were examined. Results Actively immunized mice developed a biphasic response at the site of OVA injection, while mice passively immunized with IgE anti-OVA mAb displayed a strong early response but no LPR. Cell transfer experiments using BALB/c nu/nu mice revealed that both OVA-specific IgE mAb and OVA-primed CD4 T cells were required to evoke LPR. Moreover, LPR was associated with increased levels of IL-4 production concomitant with reduced IFN-γ production and was abolished by pretreatment with anti-IL-4 neutralizing mAb. Conclusion It is suggested that murine cutaneous LPR against OVA is a type 2 inflammatory response in which both IgH antibodies and CD4 T cells play an obligatory role.     

Lysophosphatidic acid enhances antimycobacterial activity both in vitro and ex vivo

Lysophosphatidic acid (LPA) is a polar lipid metabolite which is involved in a wide range of biological processes, including cell proliferation and migration, wound healing, and increase of endothelial permeability. The present study reports evidences showing that LPA is able to enhance the antimicrobial activity of human macrophages and of bronchoalveolar lavage cells from tuberculosis patients leading to intracellular growth control of Mycobacterium tuberculosis. Such antimicrobial activity is mediated by the activation of phospholipase D which in turn induces acidification of M. tuberculosis containing phagosomes and is associated with the enhanced expression of Cathepsin D. These results suggest the possible protective role of this lysophospholipid in the activation of innate antimycobacterial response.