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- Minang, Jacob T., et al.
(författare)
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ELIspot displays a better detection over ELISA of T helper (Th) 2-type cytokine-production by ex vivo-stimulated antigen-specific T cells from human peripheral blood
- 2008
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Ingår i: Immunological Investigations. - : Informa UK Limited. - 0882-0139 .- 1532-4311. ; 37:4, s. 279-291
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Tidskriftsartikel (refereegranskat)abstract
- Detection of cytokines secreted by ex vivo antigen-stimulated peripheral blood mononuclear cells (PBMC) by ELISA is hampered by low frequencies of specific T cells and cellular receptor consumption. We investigated if ELISpot, measuring cytokine production at the single cell level, facilitated a better detection of the Th2 cytokines IL-4 and IL-5. PBMC from nickel-allergic (n = 31) and non-allergic subjects (n = 10) were stimulated with nickel or tetanus toxoid (TT) and cytokine production assessed by ELISpot and ELISA. By IL-4 ELISpot, 74% of the allergic and 0% control subjects responded to nickel and 56% of all subjects to TT. ELISA detected IL-4 after nickel stimulation only in 13% of the allergic subjects. Also detection of TT-induced IL-5 was improved by ELISpot with 54% subjects responding versus 24% in ELISA. In contrast, detection of nickel-induced IL-5 was more comparable between methods, most likely due to the 7-fold higher IL-5 production per cell in response to nickel versus IT. The low IL-5 response to TT was associated with a higher induction of the down-regulatory cytokine IL-10 by TT as compared to nickel (p < 0.001). Overall, ELISpot displayed a better detection of IL-4 as well as low intensity IL-5 responses thus emphasizing the importance of selecting suitable methods for the measurement of cytokine production ex vivo.
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- Nezlin, Roald, et al.
(författare)
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Presence of IgG-CD4 complexes in the circulation
- 2008
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Ingår i: Immunological Investigations. - : Informa UK Limited. - 0882-0139 .- 1532-4311. ; 37:2, s. 153-162
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Tidskriftsartikel (refereegranskat)abstract
- Many proteins of various origins are able to form complexes with Immunoglobulin G using reaction sites formed by residues of constant domains. Studies of IgG complexes of non-immune nature are important as biologically active proteins could escape from the circulation forming complexes with IgG as well as with other serum proteins whose concentration in serum is high. A quantitative characterization of circulating complexes in various diseases could give valuable information on the development of pathological processes. Immunoglobulins G are widely used for the treatment of a number of diseases and it is essential to understand what proteins are present in the preparations beside IgG. In the present study CD4 T-cell membrane glycoprotein was found in complexes with human IgG molecules isolated from donor sera as well as from sera of SLE patients via a sensitive quantitative dot-blot assay. According to immunoblotting experiments the CD4 part of the complexes had a molecular mass of about 50 kDa and is composed from all four extracellular domains. The CD4 content of the complexes varied among the studied human sera. There was no difference in IgG-CD4 complex concentration between the SLE patients and healthy controls. The data support the assumption that IgG molecules are able to act as scavengers and eliminate various proteins from the circulation including soluble CD4 protein.
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- Xie, Baiyi, et al.
(författare)
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Combined Costimulation Blockade Inhibits Accelerated Rejection Mediated by Alloantigen-primed Memory T Cells in Mice
- 2009
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Ingår i: Immunological Investigations. - : Informa UK Limited. - 0882-0139 .- 1532-4311. ; 38:7, s. 639-651
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Tidskriftsartikel (refereegranskat)abstract
- Donor-reactive memory T cells threaten the survival of transplanted organs via multiple pathways. This study was undertaken to induce tolerance of cardiac allografts in mice, in which alloreactive memory T cells were adoptively transferred, by combined costimulatory blockade of both effector and memory T cells. We found that the median survival time (MST) of the grafts was 5.17 days in the untreated group, 10.33 days in the CTLA4Ig- and antiCD40L- treated (2-combined) group, and more than 100 days in the CTLA4Ig-, anti-CD40L-, anti-LFA-1-, and anti-OX40L-treated (4-combined) group. Histological analysis revealed that the mean rejection level was Grade 4 in the untreated group, Grade 3 in the 2-combined treatment group, and Grade 0 in the 4-combined treatment group. CD44(high) T cells were detected only in the untreated group. The in vitro proliferation of lymphocytes of both untreated and 2-combined group was higher than that of the 4-combined treatment group (p < 0.01). Compared with the untreated group, the expression levels of IL-2, IFN-gamma, and Foxp3 were lower in the 2-combined treatment group; the expression levels of these genes were the lowest in the 4-combined treatment group. IL-10 expression was significantly higher in the 4-combined treatment group than in the other groups. These results demonstrate the inhibition efficacy of combined costimulation blockade in accelerated-rejection models and the possible mechanisms underlying the suppression of cellular immunity in mice receiving grafts as well as in inducing the activation of IL-10-producing Tr1 cells in grafts.
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