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Intravenous immunoglobulin suppresses IL-10 production by activated B cells in vitro

Open Journal of Immunology 2012年第4期2卷 149-160页

作者： Jun Tanaka Kumiko Hirano Yuzuru Sakamoto Akiko Sugahara-Tobinai Shota Endo Yumi Ito-Matsuoka Atsushi Nakano Masanori Inui lars nitschke Toshiyuki Takai Department of Experimental Immunology Institute of Development Aging and Cancer Tohoku University Sendai Japan Department of Genetics University of Erlangen Erlangen Germany Osaka Research Laboratory Benesis Corporation Osaka Japan

A therapeutic preparation of polyclonal human IgG, i.e., intravenous immunoglobulin (IVIg), has been employed to treat several inflammatory and autoimmune disorders. B cells are supposed to be a target of IVIg, but the molecular mechanism is elusive because of the lack of a suitable experimental system. To gain an insight into the beneficial effect of IVIg on B cells, we first established an experimental setting in which IVIg modulates a murine B cell function in vitro, and then aimed at identifying the mechanistic features at the molecular level. Here we show that IVIg down-regulates IL-10 production by CpG-activated B cells in vitro. The responsible component of IVIg was identified as the F(ab’)2 portion, whose polyclonality is mandatory for the suppressive effect. IVIg, bound to the surface of activated B cells, was found to be co-localized with intracellular SHP-1 on confocal laser microscopy, suggesting that B cell-surface immunoreceptor tyrosine-based inhibitory motif-harboring receptors that recruit SHP-1 are target molecules for IVIg in our experimental setting. Overall, we postulate a scenario in which IVIg attenuates B cells by suppressing IL-10 production, a B cell growth factor, and thus down-regulates the production of pathogenic antibodies.

关键词： F(ab’)2 Fragment Immune Inhibition ITIM Receptors Polyclonal IgG SHP-1

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Determination of Calcineurin Inhibitors in Dried Blood Spots from Kidney Transplant Recipients

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American Journal of Analytical Chemistry 2013年第10期4卷 27-35页

作者： lars Wilhelm Martin nitschke Markus Meier Reinhard Vonthein Jan Kramer Institute of Medical Biometry and Statistics University of Lübeck Lübeck Germany LADR GmbH Medizinisches Versorgungszentrum Dr. Kramer und Kollegen Geesthacht Germany LADR GmbH Medizinisches Versorgungszentrum Dr. Kramer und Kollegen Geesthacht Germany Medical Clinic I Transplant Center University Hospital of Schleswig-Holstein Lübeck Germany Medical Clinic I Transplant Center University Hospital of Schleswig-Holstein Lübeck Germany Medical Clinic I Transplant Center University Hospital of Schleswig-Holstein Lübeck Germany Nephrology Center Reinbek and Geesthacht Reinbek Germany

Background: Determination of cyclosporine A (CsA) and tacrolimus (Tac) in dried blood spots (DBS) could enable drug monitoring in transplanted patients without the necessity of having to take venous blood samples. Therefore, we have developed a method for quantitative determination of calcineurin inhibitors (CNI) by liquid-chromatography-tandem mass spectrometry (LCMS). Methods: In a study with 68 kidney transplant recipients (KTR, 34 CsA, 34 Tac), we tested the clinical application of LCMS monitoring in DBS in comparison to LCMS in whole blood. Results: The measuring range is proven for 27.33 to 1345 ng/ml for CsA and for 1.63 to 39.7 ng/ml for Tac. The requirements for clinical chemical analyses for precision and accuracy are complied with. Stability is documented for a period of 14 days. The study showed the following deviations from LCMS in whole blood for determination of CsA and Tac in DBS after introducing a correction factor by the haematocrit (Hct) value (CsA trough level: mean = 4.7%, ±1.96 standard deviation (SD) -52.1% to 61.4%, N = 96;CsA peak level: mean = 7.3%, ±1.96 SD -39.7% to 54.4%, N = 95;Tac trough level: mean = -0.5%, ±1.96 SD -76.4% to 75.3%, N = 88;Tac peak level: mean = 3.9%, ±1.96 SD -80.1% to 88.7%, N = 92). Conclusions: Our data show comparable results with the reference method by means of LCMS in whole blood. Therefore, DBS of KTR for determination of CNI levels could be transported on filter cards by mail to the respective laboratory resistant to breakage and the hazard of infection.

关键词： Cyclosporine A Tacrolimus LCMS DBS Immunochemistry Kidney Transplant

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