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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 16 (1982), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: By analysis of thymidine uptake during the first 4 h of incubation and by examining the responsiveness of unfractionated mouse spleen cells upon mitogenic or allogeneic stimulation, some serum factors of major importance for in vitro cultivation of lymphocytes have been examined. Albumin and L-alanine are essential for the maximal preservation of the in vivo-initiated lymphocyte activity during the first hours of incubation in vitro. Transferrin plays a major role in the lymphocyte proliferation, induced by mitogens in vitro, and, finally, zinc and selenium exert a clear enhancing effect on the response to an allogeneic stimulation. The AATSZ medium (RPMI 1640 enriched with t-alanine, albumin, transferrin, zinc chloride, and sodium selenite) enables a proliferation of the same magnitude as or higher than PCS medium. The kinetics are the same, and the cell viability is comparable, but standardization is much simpler with AATSZ. This is primarily because FCS binds some mitogens and contains inhibitors. Consequently, the standardization of such a culture system is dependent on variations from serum batch to serum batch. On the other hand, the current composition of the AATSZ medium promotes the sticking capacity of T lymphocytes and does not support growth of all lymphoid cell lines. Consequently, this defined medium, although not yet suitable as optimal medium for all lymphocyte functions, can advantageously be used for short-term studies of most murine lymphocyte functional and cooperation studies in vitro.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 14 (1981), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Serum exerts several effects in lymphocyte cultures, one of them being manifested very early. The presence of fetal calf serum (FCS) results in a markedly higher thymidine uptake within a few minutes, as compared with serum-free cultures. On the other hand, the uptake of uridine and other purine bases seem to be little influenced by serum. Experiments comparing the uptake of thymidine into the cytoplasm or into trichloroacetic acid-precipitable material suggested that the intracellular thiymidine pool increases in size when FCS is added. Using a Lineweaver Burt plot for thymidine and uridine uptake over a 4-h period, no changes in uridine uptake were observed in the presence of FCS; on the contrary, serum induced an increased Vmax for thymidine, whereas Km remained constant. Cytofluorographic quantitation of G0 and G1 cells indicated that cells disappear more rapidly from the G1 phase in the presence of FCS. The addition of hydroxyurea to the cultures prevented this disappearance. The results taken together strongly suggest that serum contains a factor promoting the shift of G1 cells into the S phase. When a 4-h culture period was used, all sera from differed species tested at low concentrations appeared to contain this activity.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 35 (1992), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Human B lymphocytes express components of the superoxide generating system of phagocytes, NADPH oxidase. We studied regulation of this ‘B-cell oxidase’ during in vitro blast transformation, using Lucigenin-amplified chemiluminescence (CL) to detect superoxide release. While freshly isolated tonsil B lymphocytes showed no CL responses, culture with phorbol myristate acetate (PMA) and ionomycin induced susceptibility to CL triggering by anti-IgM and anti-HLA-DR. Maximal effects were observed after 3 days of culture with 0.4 ng/ml PMA+1μg/ml ionomycin. Cells from such B lymphoblast cultures showed no CL responses to opsonized zymosan. In contrast, peripheral blood mononuclear cells, where monocytes are the predominant oxidant source, showed CL responses to opsonized zymosan but not to anti-IgM and anti-HLA-DR, either before or afterculture with PMA+ionomycin.Culture of B cells with the surface immunoglobulin cross-linking agent staphylococcus aureus Cowan 1 also led to emergence of a CL response to anti-IgM, which was enhanced by interferon-γ. Interestingly, markedly fewer B blasts than freshly isolated B lymphocytes expressed cytochrome b-558 surface antigen. Thus, the B-cell oxidase is up-regulated during blast transformation and can be triggered via surface IgM and HLA-DR; however, this appears to be restricted to a subset of B lymphoblasts.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Scandinavian journal of immunology 32 (1990), S. 0 
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Normal human B lymphocytes and Epstein-Barr virus-transformed B-cell lines can produce reactive oxygen species such as superoxide if treated with phorbol myristate acetate (PMA) or with the surface immunoglobulin cross-linking agents protein A and anti-immunoglobulin. Here, we investigated under which conditions specific antigen, the natural ligand of surface immunoglobulin, can stimulate an oxidative burst in monoclonal Epstein Barr virus-transformed B-cell lines producing antibodies of known specificities. After a short lag time of 1-2 min, exposure to the specific antigen stimulated a prolonged oxidative burst (tmax, 30-90 min). as measured by Lucigenin-enhanced, superoxide dismutase-inhibitable chemiluminescence, in the corresponding line only. The effect was induced in each line if the specific antigen was immobilized to a solid support. Except in one line in which antigen also stimulated an oxidative burst if presented at relatively high density on a soluble carrier, soluble antigen did not induce B-cell oxidase activation. This suggests that normal, non-transformed B lymphocytes also require interaction with relatively dense deposits of specific antigen for activation of their oxidase.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 20 (1990), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: A simple method is described which enables measurement of anti-IgE antibodies in free form as well as in immune complexes of IgE and anti-IgE. Anti-IgE antibodies were purified from serum of one selected blood donor with highly elevated levels of such auto-antibodies. These purified anti-IgE auto-antibodies inhibited the measurement of myeloma IgE. Purification also revealed that 98% of the subject's serum IgE was masked by anti-IgE auto-antibodies. Our data suggest that IgE determinations in sera containing anti-IgE antibodies might be underestimated.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Hypersensitivity reactions to non-steroidal anti-inflammatory drugs (NSAIDs), manifested by cutaneous symptoms and/or airway manifestations represent 20–25% of all hypersensitivity reactions to drugs. Today, it is still claimed that no in vitro diagnostic tests exist for that condition and that the only way to confirm the diagnosis is a provocation challenge.Objective The objective of this study was to assess whether NSAIDs may provoke blood basophil activation in vitro in such patients, as detected by a flowcytometric technique.Methods Sixty NSAID hypersensitive patients (38 with cutaneous, 20 with airway and two with cutaneous and airway symptoms) and 30 control patients (15 asthmatics) were selected. Their hypersensitivity was confirmed by documented history indicating at least two clinical episodes to two or more different NSAIDs or by positive oral provocation challenge. Isolated buffy coat leukcocytes were stimulated in vitro with aspirin, paracetamol, metamizol, diclofenac, and naproxen. The percentage of activated basophils was evaluated by an anti-CD63.Results Aspirin showed a sensitivity of 43.3%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value of 99.4%. For the other NSAIDs, the sensitivity and specificity values were: for paracetamol 11.7% and 100%, for metamizol 15% and 100%, for diclofenac 43.3% and 93.3% and for naproxen 54.8% and 74.1%. When considering the first four NSAIDs, the global sensitivity raised to 63.3% and specificity to 93.3%. If the number of tests is to be limited for practical reasons, the combination of acetylsalicylic acid and diclofenac at two concentrations yields a sensitivity of 58.3% and a specificity of 93.3%.Conclusions Flowcytometric determinations of basophil activation following stimulation with NSAIDs show a high sensitivity (60–70%) with specificity above 90%. So this test may help avoiding some cumbersome and dangerous provocation challenges.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 20 (1990), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Although antihistamines were usually considered as ineffective in asthma, the appearance of new more potent anti-H1-antagonists with no or less side-effects leads to a reassessment of their use. Although mediators and mechanisms other than histamine are involved in natural or provoked reactions to allergens in the skin, the nose or the lungs, the new anti-H1-antagonists have been shown to be effective. For some of these drugs, the effect may go beyond a mere H1-receptor blockade and involve impairment of mediator release from mast cells and/or an inhibiting effect on other cell types (e.g. eosinophils) implicated in the late-phase reactions. Quantitative kinetics evaluation of late-phase reactions in the skin and of their inhibition by anti-allergic drugs will be helped considerably by the development of telethermography, a new skin evaluation technique.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 12 (1982), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Skin tests with histamine, the histamine-liberator, codeine, and various allergens as well as blood basophil degranulation by anti-IgE or anti-IgG4 and total serum IgE levels have been studied in two female populations of different ages (average 23-1 and 73-9 years). All thirty-one patients selected for this study were clinically non-allergic.We observed a trend towards reduced skin reactions to histamine and codeine in the higher age-group; on the other hand, we could not find any decrease of basophil degranulation as a sign of basophil impairment with age. Likewise no difference in total serum IgE levels have been noticed.No correlation between skin tests and basophil degranulation was observed; yet patients with isolated, positive skin tests to house dust and/or Candida albicans showed a statistically significant reduced blood basophil degranulation by anti-IgE or anti-IgG4.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Clinical & experimental allergy 11 (1981), S. 0 
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Quantification of human basophil degranulation induced by anti-IgE, anti-IgG4, and by ionophore was performed using a flow-cytometric system. It was shown that these antibodies and ionophore can degranulate basophils in a dose-dependent manner, and that there is a wide variation in the response of basophils obtained from different individuals to these stimuli. A significant correlation was observed between the degree of degranulation induced by anti-IgE and anti-IgG4, while this was not the case for anti-IgE and ionophore. It was also shown that IgG4 myeloma protein can passively sensitize basophils. In general, degranulating efficacy was in the order of ionophore 〉 anti-IgE 〉anti-IgG4, both in allergic and non-allergic individuals.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Type of Medium: Electronic Resource
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