SummaryThe functional differentiation of immune cells at early age plays a central role in immune physiology, e.g. for the sufficient eradication of pathogens. However, imbalances in effector cell responses may also have an impact in the pathophysiology of childhood diseases such as atopy and autoimmune disorders. As information on immune cell responses in infancy and early childhood is scarce, we conducted an observational, cross-sectional study in healthy newborns ( n = = = = 18), infants and young children ( n = = = = 54) aged 1-96 months and adult controls ( n = = = = 19) to assess cytokine mRNA and protein expression upon phorbol 12-myristate 13-actate/ionomycin stimulation and LPSinduced IL-12 expression in monocytes. The intracellular expression of interferon (IFN)-γ γ γ γ , tumour necrosis factor (TNF)-α α α α ( R = = = = 0·748, P < < < < 0·0001; R = = = = 0·784, P < < < < 0·0001, respectively) and interleukin (IL)-2 protein expression ( R = = = = 0·384, P = = = = 0·008) was demonstrated to increase progressively with age. While a correlation between IL-4 protein expression and age was noted ( R = = = = 0·342, P = = = = 0·007), the levels of IL-5 and IL-10 protein expression tended to be regulated on an individual basis during infancy and early childhood. An age correlation was also observed for intracellular IL-12 expression ( R = = = = 0·331, P = = = = 0·009) in monocytes. These findings are valuable for further assessment of normal variations and maturation processes in immune cell responses and for the clinical-therapeutic monitoring of immunological status in various childhood diseases.

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The Flow-GIFT method permits rapid detection of granulocyte antibodies requiring fewer donor test cells. This method is ideal for automation and will potentially open the way for screening of granulocyte antibodies in a large donor population.

Recently, we demonstrated an association of the IL-6 promoter polymorphism at position -174 (G→C) with kidney allograft survival whereby carriers of the −174GG genotype were identified as having superior graft survival. As two additional polymorphisms were discovered in the neighborhood at positions −572 (G→C) and −597 (G→A), respectively, and as functional studies revealed a cooperative impact of all three on the IL-6 gene transcription, we investigated whether there is a combined effect on kidney transplant outcome. We determined IL-6 promoter haplotypes −597 (G→C)/−572 (G→A)/−174 (G→C) ( −597/−572/−174 haplotype) using a PCR system with sequence-specific primers in 158 patients after primary cadaveric kidney transplantation. We here show that the −597 and −174 polymorphism are in tight-linkage disequilibrium and that homozygous carriers of the GGG −597/−572/−174 haplotype (GGG/GGG genotype) have superior 3-year graft survival rates compared with the 8.0-fold increased risk of premature graft loss in all other patients. Interestingly, patients carrying the GGG/GCG genotype had the lowest allograft survival rate. Thus determination of the combined −597/−572/−174 genotype allows for further differentiation of −174GG patients into subgroups and consequently for a more accurate identification of patients at risk. Our results indicate that the three polymorphisms act in a cooperative fashion and we provide evidence for an exceptional clinical impact of the IL-6 −597/−572/−174 genotype on the success of kidney transplantation.

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The aim of this study was to investigate whether moderate or exhaustive endurance exercise influences cytokine levels in whole-blood culture supernatants after stimulation. Therefore, eight healthy subjects were first exposed to moderate exercise on a cycle ergometer for 30 min at 70% of their 4-mmol/l lactic acid (anaerobic) threshold, and 1 week later to exhaustion (for 90 min) at their anaerobic threshold. Blood samples were taken before, 30 min after and 24 h after each exercise bout. The following lymphocyte subpopulations were determined: CD14-positive(+)/CD45+, CD4+, CD8+, and CD16+. Cytokine levels in the supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Production of interleukin (IL)-1beta, IL-6 and tumour necrosis factor (TNF)-alpha were induced with lipopolysaccharides (LPS), and that of IL-2 and interferon (IFN)-gamma with staphylococcal enterotoxin B (SEB) and phytohaemagglutinin (PHA). Cortisol levels were also determined by ELISA. The lymphocyte subset distribution was observed to be unchanged after moderate exercise. Thirty minutes after exhaustive exercise, the CD16+ count was found to be significantly lower, whereas 24 h later the CD4+ count was significantly higher than pre-exercise counts. Moderate exercise influenced the IFN-gamma production (PHA-stimulated), which increased significantly from 974 (391) pg/ml before exercise to 1450 (498) pg/ml 24 h later. Thirty minutes after exhaustive exercise the IFN-gamma level in the supernatants (SEB-stimulated) was significantly decreased (from 14470 (11840) pg/ml before exercise to 6000 (4950) pg/ml after exercise). The IL-1beta and TNF-alpha production per monocyte was also significantly reduced.

Summary Background and Methods: As a source of hematopoietic stem cells, cord blood (CB) is an alternative to bone marrow or peripheral blood stem cells (PBSC). The Mannheim CordBlood Bank has currently stored about 1,750 allogeneic CB units. Here we report our experiences and discuss future perspectives of CB banking. We analyzed CB units for nucleated cell (NC), mononucleated cell (MNC) and CD34+ cell count, volume, colony-forming units (CFU-GM) as well as ethnic background of the donor. Transplanted CB units were analyzed for patient and transplant characteristics and compared to stored CB units. Results: Only 25% of all collected CB units met storage criteria. Main reasons for exclusion were: i) insufficient volume (57.7%), ii) delayed arrival at the processing site (19.2%) and iii) little cell count (7.2%). Up to now 36 CB units have been released for transplantation mainly to children (62%). Transplant indications were hematological diseases, immune deficiencies and metabolic diseases. Transplanted CB units showed significantly higher cell counts compared to stored units (NC: 12.5 vs. 7.2 × 10 8 , MNC: 4.7 vs. 2.9 × 10 8 , CD34+ cells: 3.3 vs. 1.8 × 10 6 , mean; p < 0.001 each) and were found more often in ethnic minority groups (36 vs. 20%; p = 0.04). Conclusions: Even though cell count and volume are key parameters for the eligibility of CB units, our data indicate that the ethnic background of the donor also plays a major role. Collection and processing of CB should be optimized in order to gain maximum volume and cell counts.Schlüsselwörter Nabelschnurblut · Nabelschnurblut-Aufarbeitung · Stammzelltransplantation · Hämatopoetische Stammzellen Zusammenfassung Hintergrund und Methoden: Nabelschnurblut (CB) ist eine Alternative zur Gewinnung von hämatopoetischen Stammzellen aus Knochenmark oder peripheren Stammzellen. Die Mannheimer Nabelschnurblutbank hat bisher 1750 allogene Nabelschnurblutpräparate eingelagert. Wir berichten über unsere Erfahrungen und diskutieren Zukunftsperspektiven der Nabelschnurbluteinlagerung. Folgende CB-Parameter wurden untersucht: Volumen, nukleäre (NC), mononukleäre (MNC) und CD34+ Zellen, Kolonie bildende Einheiten (CFU-GM) sowie ethnische Herkunft des Spenders. Bei den transplantierten Präparaten wurden die Patienten-und Transplantatdaten ausgewertet und die Präparateeigenschaften mit eingelagerten CB verglichen. Ergebnisse: Nur 25% aller gesammelten Nabelschnurblutpräparate erfüllten die Einlagerungskriterien. Die Hauptursachen hierfür waren 1) zu niedriges Vollblutvolumen (57,7%), 2) zu spätes Eintreffen der Prä-parate im Verarbeitungszentrum (19,2%) sowie 3) zu niedrige Zellzahl (7,2%). Bisher wurden 36 Mannheimer Nabelschnurblutpräparate, vorwiegend an Kinder (62%) abgegeben. Die Transplantationsindikationen waren primär hämatologische Erkrankungen, Erkrankungen des Immunsystems und metabolische Erkrankungen. Die Zellzahlen der transplantierten Präparate waren signifikant höher, als die der eingelagerten (NC: 12,5 vs. 7,2 × 10 8 , MNC: 4,7 vs. 2,9 × 10 8 , CD34+ Zellen:...