Human Leukocyte Antigen Compatibility and De Novo Donor-Specific Antibodies in Long-term Renal Transplant Patients With Stable Graft Function.

PURPOSE: De novo donor-specific antibodies (DSA) are associated with antibody-mediated rejection leading to late renal transplant failure. The aim of this study was to evaluate whether HLA compatibility is associated with sensitization along with other risk factors. METHODS: Eighty-nine stable renal transplant recipients (47 men) were studied. Patients were classified into 2 groups according to HLA compatibility between donor and recipient, group A (1-4/8 matches) and group B (5-8/8 matches). Cold ischemia time (CIT) and delayed graft function (DGF) were recorded along with time with a functional graft. Anti-HLA antibodies were detected using a Luminex single-antigen bead assay and were further classified into DSA and non-DSA. RESULTS: HLA group A consisted of 49 (56%) transplant recipients while 38 (44%) were classified to group B, with functional grafts for 10.9 ± 6.7 and 14.8 ± 8.5 years, respectively (P = .019). Group A patients had more anti-HLA antibodies than group Β (P = .001) and this correlation was retained for DSA patients. De novo anti-HLA were detected in 40 patients; DSA were detected in 19 (21.8%). DSA (+) patients had recorded with functional renal grafts for 11 ± 5 years, compared to 14.4 ± 8.6 years (P = .048) for anti-HLA negative patients. Increased CIT and DGF were associated with anti-HLA antibodies detection but no with DSA. CONCLUSION: HLA compatibility is probably correlated with DSA in a context of a more general anti-HLA sensitization, and both have a negative effect on long-term renal graft outcome.

Purification and properties of bovine nasal hyaline cartilage collagenase.

1. Collagenase from bovine nasal hyaline cartilage was extracted with 1 and 3 M NaCl in Tris-CaCl2 buffer. 2. Two peaks of collagenase activity were revealed on DE52 ion exchange column, collagenase 1 and collagenase 2. 3. The apparent mol. wt of collagenase 1 and 2 as determined by SDS-PAGE were 68 and 43 kDa, respectively. 4. Both enzymes degrade native collagen type II into two characteristic products, TCA(3/4) and TCB(1/4), at 25 degrees C and pH 7.6. 5. Trypsin and aminophenylmercuric acetate were capable of increasing the collagenase 1 activity. 6. The two enzymes can be characterized as metalloproteinases since they were inhibited by EGTA and 1,10-phenanthroline. The use of proteinase inhibitors (N-ethylmaleimide, iodoacetic acid, phenylmethylsulphonyl fluoride, soybean trypsin inhibitor, pepstatin, dithiothreitol) showed that the enzymes do not contain serine, cysteine or aspartic acid in their active sites.

Regulation of cation transport pathways and glycolytic enzyme activity by alterations in red cell volume.

In the presence of NH4Cl and hypotonic solutions, Rana balcanica red cells respond by increasing their volume. The stimulation of cellular volume by hypotonicity is more rapid than that of NH4Cl, while the maximum value is less than that observed in the presence of NH4Cl. Depending on the cause of swelling, (nct uptake of NH4Cl or decrease in external osmolality) cells show specific responses. The NH4Cl treatment causes a significant increase in intracellular Na+, from 5.14 +/- 0.78 to 29.84 +/- 0.47 mmoles l-1 cell, while hypotonicity leads to a significant decrease of this cation, to 3.85 +/- 0.25 mmoles l-1 cell in relation to the control, after 30 min of incubation of Rana balcanica erythrocytes. In addition, amiloiride significantly reverses the NH4Cl effect with respect to intracellular Na+. Both treatments cause a significant K+ loss in comparison with controls. Two glycolytic enzymes glyceraldehyde phosphate dehydrogenase (GAPDH) and pyruvate kinase (PK) of Rana balcanica haemolysate were found to respond to the NH4Cl effect by significantly decreasing their activity.

Metabolic effects and cellular volume responses induced by noradrenaline in nucleated erythrocytes.

The mechanism of adrenergic swelling, cAMP accumulation and associated Na+ and K+ concentration changes was investigated in amphibian Rana ridibunda erythrocytes. The addition of noradrenaline to an isotonic suspension of red cells of frog Rana ridibunda in the presence of the phosphodiesterase inhibitor isobutyryl-methylxanthine (IBMX), induced a significant increase of the cell volume. Forskolin treatment showed analogous results. The removal of the Na+ from the incubation medium, inhibited the volume changes caused by either noradrenaline or forskolin. Under the same conditions a more than two-fold increase of lactate formation, 260% increase of glucose consumption and accumulation of cAMP were found. These effects are specific and rapid. The peak of lactate production at 7.5 min was followed by a slow further decrease, whereas cAMP reached a plateau after 15 min. The increased glycolytic rate is probably the consequence of an activation of phosphofructokinase by cAMP. When the red cells were incubated in the presence of either noradrenaline or the cAMP analog, dibutyryl-cAMP the intracellular concentration of Na+ was significantly increased by the first 7.5 min of incubation compared with the initial values. Both the adrenergic activation and dibutyryl-cAMP treatment induced an intracellular decrease in the K+ content by 15%. In the presence of amiloride the Na+ and the K+ content of erythrocytes remained unaltered. Cellular swelling may be a prerequisitive for activation of Na+ and K+ movements. These findings suggest a regulatory role of cAMP in the energy metabolism of Rana ridibunda erythrocytes. In addition, the adrenergic responses were rapid and specific to alpha 1 and beta-antagonists.