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  • 1
    Keywords: ENDOTHELIAL-CELLS ; CD8(+) T-CELLS ; DENDRITIC CELLS ; TOLERANCE ; IMMUNE-RESPONSE ; MOUSE MODEL ; INFECTIONS ; EXOGENOUS ANTIGEN ; HEPATITIS-B-VIRUS ; LIVER-DAMAGE
    Abstract: Viruses can escape cytotoxic T cell (CTL) immunity by avoiding presentation of viral components via endogenous MHC class I antigen presentation in infected cells. Cross-priming of viral antigens circumvents such immune escape by allowing noninfected dendritic cells to activate virus-specific CTLs, but they remain ineffective against infected cells in which immune escape is functional. Here, we show that cross-presentation of antigen released from adenovirus-infected hepatocytes by liver sinusoidal endothelial cells stimulated cross-primed effector CTLs to release tumor necrosis factor (TNF), which killed virus-infected hepatocytes through caspase activation. TNF receptor signaling specifically eliminated infected hepatocytes that showed impaired anti-apoptotic defense. Thus, CTL immune surveillance against infection relies on two similarly important but distinct effector functions that are both MHC restricted, requiring either direct antigen recognition on target cells and canonical CTL effector function or cross-presentation and a noncanonical effector function mediated by TNF.
    Type of Publication: Journal article published
    PubMed ID: 22939982
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  • 2
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background The aim of this study was to investigate in vitro lymphocyte proliferation in the diagnosis of allergy to phenoxymethylpenicillin (PcV), comparing chemically reactive PcV, added to cell cultures in unconjugated form, to a PcV-PLL (poly-L-lysine) conjugate as antigens. Side-chain specificity of lymphoproliferative responses was investigated with reactive benzylf)enicillin (PcG) and bacampicillin. Methods Seventeen patients with a history of hypersensitivity reactions in connection with PcV treatment were studied by means of the lymphocyte transformation test (LTT), the radioallergosorbent test (RAST). skin tests (prick and intracutaneous), and oral challenge with PcV. LTT was also performed in 20 control subjects exposed to PcV therapeutically, and in eight subjects with occupational exposure to this penicillin. Results Nine patients had a positive in vivo test to PcV (five by oral challenge, three by intracutaneous test, and one by both tests), and six were challenge-negative. When reactive PcV was used as antigen in LTT, positive LTT responses were observed in five of the nine patients with a positive in vivo test, and two of them were also side-chain specific. Positive LTT responses with reactive PcV also correlated with a positive RAST infiveof seven subjects. None of the six patients with a negative challenge test, and only one of the 28 controls showed a positive LTT result with reactive PcV. Thus, the specificity of LTT with reactive PcV was 96%. In contrast, when PLL-conjugated PcV served as antigen, four challenge-negative subjects and 11 controls were LTT-positive. Conclusions The results of this study indicate that LTT with chemically reactive PcV could be useful as an in vitro complement in the diagnosis of PcV allergy and as a tool to reveal the side-chain specificity of peripheral blood lymphocytes. A positive LTT to PLL-conjugated PcV may be an indicator of immunization, but not necessarily allergy, to the penicilloyl structure.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Antigen-specific cell lines or clones are often used as models of drug-specific allergy. However, cloning procedures are time consuming, and the repeated antigen stimulation cycles as well as the addition of various growth enhancers may affect the in vivo relevance of these systems.〈section xml:id="abs1-2"〉〈title type="main"〉ObjectiveUsing bacampicillin-allergic subjects, we wanted to investigate the applicability of primary recall in vitro lymphocyte responses to characterize type I and type IV allergy. The sensitivity and specificity of LTT (Lymphocyte transformation test), when used as an in vitro diagnostic tool, were also assessed.〈section xml:id="abs1-3"〉〈title type="main"〉MethodsA total of 39 patients with symptoms of type I (rhinitis) or type IV (allergic contact dermatitis, ACD) allergy following occupational exposure to bacampicillin, were included. Ten individuals without penicillin allergy or occupational exposure to bacampicillin served as controls. All subjects were LTT tested. Four patients with rhinitis and two patients with ACD were available for studying the immunophenotype and the TCR-Vβ repertoire of bacampicillin induced lymphoblasts as well as the cytokine profiles and expression of the activation markers CD23 and CD134 in primary PBMC cultures.〈section xml:id="abs1-4"〉〈title type="main"〉ResultsLTT was positive in 87% and at least one of the skin tests was positive in 85% of the patients with allergic symptoms. 69% of the patients with type I allergies were patch test-positive. Results from LTT and skin test correlated in 87% of the cases. The combined sensitivity of LTT and skin tests was 92%. The specificity of LTT was 90% in healthy controls. Bacampicillin induced lymphoblasts were mainly CD4 + in both ACD and rhinitis patients. The TCR-Vβ profiles of the predominant CD4 + lymphoblasts were heterogeneous with individual skewing towards Vβ2, Vβ3, Vβ5.1 and/or Vβ14. An increased expression of IFNγ was detected in bacampicillin treated PBMC cultures from the ACD but not from rhinitis patients. IL-5 was detected in bacampicillin exposed PBMC cultures from all patients but not from healthy controls. This Th2 environment could also be verified by CD23 and CD134 expression.〈section xml:id="abs1-5"〉〈title type="main"〉ConclusionLTT and skin tests are equally sensitive in identifying bacampicillin allergic subjects. When the two tests are combined, the sensitivity increases. The patch test is useful not only for detection of type IV but also for the identification of type I allergies. When using primary PBMC cultures, IFNγ is the most suitable cytokine to discriminate between type I and type IV allergy. IL-5 can possibly be used as a general marker for bacampicillin induced allergy. Thus, primary cell cultures may be considered as an alternative to T-cell lines or clones for the study of drug induced allergy.
    Type of Medium: Electronic Resource
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