Das Buch, geschrieben von Mitgliedern der Arbeitsgemeinschaft "Klinische Geweberegeneration" der Deutschen Gesellschaft für Orthopädie und Unfallchirurgie, vermittelt praxisnahes und aktuelles Wissen ...rund um Gelenkknorpelschäden. Schwerpunkte sind Diagnostik und moderne Therapieverfahren wie Knorpelzelltransplantation oder osteochondraler Transfer, die durch Grundlagen zur Biologie und Anatomie des Knorpels sowie Fragen der Dokumentation und Abrechnung abgerundet werden. Das Buch richtet sich an operativ tätige Orthopäden und Unfallchirurgen, aber auch Allgemeinchirurgen, Sportmediziner und Arthroskopeure.
The modification of low-density lipoprotein (LDL) by normal, myeloperoxidase (MPO)-deficient and NADPH oxidase-deficient granulocytes was investigated using the monoclonal antibody (mAb) OB/04, which ...was originally generated against copper-oxidized LDL. Incubation of LDL with normal granulocytes increased the reactivity of LDL with mAb OB/04. These effects were even more pronounced using MPO-deficient granulocytes. Inhibitors of oxidative reactions (the NADPH oxidase inhibitor diphenyleneiodonium chloride DPI, catalase, superoxide dismutase SOD) did not significantly reduce LDL oxidation by normal granulocytes. Furthermore, granulocytes of a patient with NADPH oxidase deficiency were almost equally effective as normal granulocytes, indicating that oxidative burst-derived reactive oxygen species are of only minor importance in the generation of mAb OB/04-detectable new epitopes on LDL in vitro. In contrast, incubation of LDL with iron and copper prior to and during incubation with normal granulocytes markedly enhanced the generation of OB/04-detectable epitopes. It is supposed that, besides superoxide (in normal and MPO-deficient granulocytes) or instead of superoxide (in NADPH oxidase-deficient granulocytes), lytic enzymes released by activated granulocytes may enhance the availability of transition metals for oxidation of LDL. Our results support the concept that transition-metal-dependent pathways of LDL oxidation in combination with degranulation products of granulocytes are important.
To quantify left ventricular function derived from retrospectively ECG-gated multislice spiral CT (MSCT) data sets in comparison to MRI.
In 16 patients (14 males, 2 females, mean age 56.8 +/- 11.5 ...years), retrospectively ECG-gated MSCT angiography of the coronary arteries and breath-hold steady state free precession cine MRI were performed. From MSCT data-sets, 20 axial image series were reconstructed every 5 % of the RR interval. Multiplanar images were reformatted in the short axis orientation from axial images. End-systolic and end-diastolic images were selected. From these images end-systolic volume (ESV), end-diastolic volume (EDV) and stroke volume (SV) as well as the ejection fraction (EF) and myocardial mass (MM) were determined using the Simpson's method and compared with MRI. Furthermore, image quality was assessed for both imaging modalities using a four point grading scale.
All parameters were found to have an excellent correlation between MSCT and MRI data (Pearson's correlation coefficient 0.95 - 0.99), without clinically relevant differences between both modalities. On average, the difference between both methods was 0.5 ml for ESV, 0.8 ml for EDV, 1.3 ml for SV, 0.9 % for EF and 2.3 g for MM. Image quality was slightly better for MRI (1.5 +/- 0.65) than for MSCT (1.64 +/- 0.74).
Retrospectively ECG-gated MSCT angiography can not only visualize the coronary arteries but also enables precise quantification of the left ventricular function from the same MSCT data set.
Complete experimental data sets of HLA-ligand motifs and T-cell recognition patterns can be derived from combinatorial peptide libraries. These data provide the exact molecular basis for a fast ...development of synthetic vaccines, T-cell superagonists and non-peptide antagonists. Patient-specific peptides, peptidomimetics and vaccines of highest reactivity can be derived directly from the data sets via our prediction programme EPIPREDICT. The resulting lead structures may be developed into valuable diagnostics and therapeutic tools for the treatment of viral infections, autoimmune diseases and tumors. As one example, antibody and T cell recognition in the intestinal auto-immune disease, coeliac disease was investigated in more detail concerning the deamidation of γ-gliadin peptides by tissue transglutaminase 9tTG) leading to autoreactive peptides specific for HLA-DQA1*0501, DQB1*0201.
Superantigens interact with and activate a sizeable fraction of T cells characterized by expression of specific V beta gene segments of their antigen receptor. The massive activation of T cells in an ...organism is considered responsible for clinical symptoms associated with superantigen-producing bacteria. Here we studied the in vitro activation of human T cells by the superantigen Staphylococcus Enterotoxin B on a cell by cell basis. Superantigen-reactive T cells were stained with a V beta 12-specific monoclonal antibody and analyzed in a cytofluorograph. Blast formation of SEB-reactive T cells occurs within 12 h and reaches a plateau after 24 h. Double-staining of V beta 12+ T cells with antibodies against different T cell activation or adhesion surface molecules revealed a time-dependent differential upregulation for CD2, CD11 = LFA-1, CD25, CD28, CD69, and HLA-DR. The expression of CD3, CD4 and CD5 was not influenced by the superantigen. The rapid phenotypic changes of superantigen reactive T cells in terms of marker expression and cell size could provide early tools in diagnosing diseases caused by superantigens.
TSST-1 is a
Staphylocoaus aureus-derived superantigen which has been implicated in the pathogenesis of toxic shock syndrome. In mice, superantigen-induced prolifetation is followed by deletion or ...anergy of reactive T cells. So far, superantigen-induced T-cell anergy has not been observed in humans. We therefore examined PBMCs derived from a 1 5-year-old patient suffering from severe toxic shock syndrome. Markedly elevated levels of circulating TSST-1-reactive T cells were found by cytofluorometric analysis. Upon in vitro re-stimulation with TSST-1, hyporesponsiveness of TSST-1-responsive Vβ2
+ T cells was detected, thus confirming results obtained in the murine system.
Superantigen-activated T cells can be targeted by monoclonal antibodies (mAb) to lyse MHC class II negative tumour cells. In this study we determined the susceptibility of the T-lymphoblastoid ...leukaemic cell line CCRF-CEM and its multidrug resistant sublines CCRF VCR100, CCRF VCR1000 and CCRF ADR5000 to lysis by monoclonal antibody-targeted and superantigen-activated T cells (superantigen-dependent cellular cytotoxicity, SDCC). A recombinant fusion protein of protein A and the superantigen Staphylococcus enterotoxin A (SEA) was used together with the mAbs anti-CD7, anti-CD38, anti-CD45RA and 4E3 (anti-P-glycoprotein) to correlate susceptibility to SDCC with expression of the MDR1-gene product. Our results demonstrated SDCC to be independent of MDR1-gene expression. This was further confirmed by blocking the function of Pgp in the leukaemic cell lines with a cyclosporine A derivative, which had no influence on SDCC. As expected, expression of the respective cell surface antigens on target cells had a strong impact on SDCC, although other factors seem to influence efficiency of SDCC as well.
Superantigens such as the staphylococcal enterotoxin A (SEA) are among the most potent T cell activators known. They bind to major histocompatibility complex (MHC) class II molecules and interact ...with T cells depending on their T cell receptor (TCR) Vβ expression. Superantigens also induce a variety of cytokines and trigger a direct cytotoxic effect against MHC-class-II-positive target cells. In order to extend superantigen-dependent cell-mediated cytotoxicity (SDCC) to MHC-class-II-negative neuroblastoma cells, SEA was linked to the anti-ganglioside GD
2
human/mouse chimeric monoclonal antibody (mAb) ch14.18. Ganglioside GD
2
is expressed on most tumours of neuroectodermal origin but is expressed to a lesser extent on normal tissues. The linkage of ch 14.18 to SEA was achieved either with a protein-A-SEA fusion protein or by chemical coupling. Both constructs induced T-cell-mediated cytotoxicity towards GD
2
-positive neuroblastoma cells in an effector-to-target(E∶T)-ratio-and dose-dependent manner in vitro. To reduce the MHC class II affinity of SEA, a point mutation was introduced in the SEA gene (SEAm9) that resulted in 1000-fold less T cell killing of MHC-class-II-expressing cells as compared to native SEA. However, a protein-A-SEAm9 fusion protein mediated cytotoxicity similar to that of protein-A-SEA on ch14.18-coated, MHC-class-II-negative neuroblastoma cells. Taken together, these findings suggest that superantigen-dependent and monoclonal-antibody-targeted lysis may be a potent novel approach for neuroblastoma therapy.
CTLs bearing certain T-cell receptor V beta-regions are directed by the bacterial superantigen Staphylococcus enterotoxin A (SEA) to lyse MHC class II-positive cells. In order to extend ...superantigen-dependent cytotoxicity to MHC class II-negative carcinoma cells, covalent conjugates of superantigen and mAbs against surface markers of these cells have been used. We now describe a novel strategy which allows rapid selection of mAb suitable for superantigen targeting against MHC class II-negative tumor cells. A recombinant fusion protein of protein A and SEA binding to the mAbs CD7 or CD38 was able to mediate T cell-dependent lysis of MHC class II-negative Molt-4 and CCRF-CEM acute lymphatic leukemia cell lines. Lysis was dose dependent and correlated with E:T cell ratio. In contrast, SEA alone did not induce any significant lysis. In order to decrease the MHC class II affinity of the protein A-SEA complex, a point mutation was introduced into SEA (protein A-SEA mu9). The mutated fusion protein had similar potency as protein A-SEA against Molt-4 cells but was 100-fold less active against MHC class II-positive cells. Considering the efficiency and specificity of the mutated SEA protein interacting with mAb in targeting T lymphocytes against MHC class II-negative leukemia cells while only marginally affecting normal MHC class II-positive cells, we suggest the development of SEA-mAb fusion proteins as a potential adjuvant therapy of leukemias.
PDF
