J. van Kuik

A 500-MHz 1H-NMR study on the N-linked carbohydrate chain of bromelain. 1H-NMR Structural-reporter-groups of fucose α(1-3)-linked to asparagine-bound N-acetylglucosamine

The 500-MHz1H-NMR characteristics of theN-linked carbohydrate chain Manα1-6[Xylβ1-2]Manβ1-4GlcNAcβ1-4[Fucα1-3]GlcNAcβ1-NAsn of the proteolytic enzyme bromelain (EC 3.4.22.4) from pineapple stem were determined for the oligosaccharide-alditol and the glycopeptide, obtained by hydrazinolysis and Pronase digestion, respectively. The1H-NMR structural-reporter-groups of the α(1–3)-linked fucose residue form unique sets of data for the alditol as well as for the glycopeptide.The 500-MHz1H-NMR characteristics of theN-linked carbohydrate chain Manα1-6[Xylβ1-2]Manβ1-4GlcNAcβ1-4[Fucα1-3]GlcNAcβ1-NAsn of the proteolytic enzyme bromelain (EC 3.4.22.4) from pineapple stem were determined for the oligosaccharide-alditol and the glycopeptide, obtained by hydrazinolysis and Pronase digestion, respectively. The1H-NMR structural-reporter-groups of the α(1–3)-linked fucose residue form unique sets of data for the alditol as well as for the glycopeptide.

Primary structure of the acidic carbohydrate chain of hemocyanin from Panulirus interruptus

The N‐glycosidic carbohydrate chains of hemocyanin from the spiny lobster Panulirus interruptus were liberated by hydrazinolysis of a pronase digest and subsequently reduced. Separation of the mixture of oligosaccharide‐alditols by high‐voltage paper electrophoresis resulted in a neutral (90%) and an acidic (10%) fraction. 500‐MHz 1H‐NMR spectroscopy of the acidic fraction revealed a single component with the following novel structure:

T cells in Cardiac Allograft Vasculopathy Are Skewed to Memory Th-1 Cells in the Presence of a Distinct Th-2 Population

Cardiac allograft vasculopathy (CAV) in heart transplantation (HTx) patients remains the major complication for long‐term survival, due to concentric neointima hyperplasia induced by infiltrating mononuclear cells (MNC). Previously, we showed that activated memory T‐helper‐1 (Th‐1) cells are the major component of infiltrating MNC in coronary arteries with CAV. In this study, a more detailed characterization of the MNC in human coronary arteries with CAV (n = 5) was performed and compared to coronary arteries without CAV (n = 5), by investigating MNC markers (CD1a, DRC‐1, CD3, CD20, CD27, CD28, CD56, CD68, CD69, FOXP3 and HLA‐DR), cytokines (IL‐1A, 2, 4, 10, 12B, IFN‐γ, and TGF‐β1), and chemokine receptors (CCR3, CCR4, CCR5, CCR7, CCR8, CXCR3 and CX3CR1) by immunohistochemical double‐labeling and quantitative PCR on mRNA isolated from laser microdissected layers of coronary arteries. T cells in the neointima and adventitia of CAV were skewed toward an activated memory Th‐1 phenotype, but in the presence of a distinct Th‐2 population. FOXP3 positive T cells were not detected and production of most cytokines was low or absent, except for IFN‐γ, and TGF‐β. This typical composition of T‐helper cells and especially production of IFN‐γ and TGF‐β may play an important role in the proliferative CAV reaction.

Donor-Specific Antibodies Are Produced Locally in Ectopic Lymphoid Structures in Cardiac Allografts

Cardiac allograft vasculopathy (CAV) is a transplant pathology, limiting graft survival after heart transplantation. CAV arteries are surrounded by ectopic lymphoid structures (ELS) containing B cells and plasma cells. The aim of this study was to characterize the antigenic targets of antibodies produced in ELS. Coronary arteries and surrounding epicardial tissue from 56 transplant recipients were collected during autopsy. Immunofluorescence was used to identify antibody‐producing plasma cells. Immunoglobulin levels in tissue lysates were measured by enzyme‐linked immunosorbent assay and analyzed for donor‐specific HLA antibodies by Luminex assay. Cytokine and receptor expression levels were quantified using quantitative polymerase chain reaction. Plasma cells in ELS were polyclonal and produced IgG and/or IgM antibodies. In epicardial tissue, IgG (p < 0.05) and IgM levels were higher in transplant patients with larger ELS than smaller ELS. In 4 of 21 (19%) patients with ELS, donor‐specific HLA type II antibodies were detected locally. Cytokine and receptor expression (CXCR3, interferon γ and TGF‐β) was higher in large ELS in the epicardial tissue than in other vessel wall layers, suggesting active recruitment and proliferation of T and B lymphocytes. ELS exhibited active plasma cells producing locally manufactured antibodies that, in some cases, were directed against the donor HLA, potentially mediating rejection with major consequences for the graft.

IL-4 promotor gene polymorphism in heart transplantation

test. Results: Overall survival was borderline significantly different between all groups (OKT3: 52.4%, F-ATG: 44.3 %, THG: 67.5% and BT563: 51.8%; p 0.095). Causes of death were different between the four groups. OKT3 treated patients had the highest rate of death from graftsclerosis (40%) and BT 563 the highest rate in death from acute rejection (23%). In the THG group there was the lowest incidence of acute rejections (57.3%, 29.5 %, 18% and 50%; p 0.002). There was no difference in overall freedom from severe infections (90.5%, 81.8 %, 92.4% and 88.9%; p 0.232) and CMV-disease (80.9%, 79.6 %, 83.6% and 77.8%; p 0.822). The incidence of cancer was similar in all groups (14.3 %, 22.8 %, 11.7% and 11.1%; p 0.146), whereas there was a clear trend of lower incidence of graftsclerosis in THG treated patients (52.2 %, 38.0 %, 25.7% and 41.2%; p 0.053). Conclusions: Different Antibody induction protocols seem to be dissimilar in their immunmodulating function. Although all three studies were powered to detect differences in early rejection, long-term data show different incidence of development of graftsclerosis and on survival. Long-term adverse events were comparable and tolerable. These data clearly show that early immunmodulating influence of induction antibodies have longrange effects on the immune system.