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Ziel: Ziel der vorliegenden Untersuchung war die Bestätigung bereits bekannter sowie die Evaluierung neuer, bisher nicht beschriebener klinischer Bedingungen und präparatespezifischer Eigenschaften mit Einfluβ auf das posttransfusionelle Thrombozyteninkrement. Design: In einer retrospektiven Analyse wurde an 400 Einzelspender-Thrombozyten-transfusionen der Einfluβ von patienten- und präparatespezifischen Einfluβgröβen auf das posttransfusionelle Inkrement erfaβt. Die statistische Auswertung erfolgte mittels Varianzanalyse mit gemischten Effekten, wobei eine Irrtumswahrscheinlich-keit von 0,05 zugrunde gelegt wurde. Rahmen: Hämatologisch-onkologische Universitätsklinik. Patienten: 46 Patienten (24 männlich, 22 weiblich; Alter 17-80 Jahre). Intervention: Einzelspender-Thrombozytentransfusionen. Ergebnisse: Als klinische Parameter mit staíistisch gesichertem negativem Einfluβ auf das posttransfusionelle Inkrement konnten die bereits bekannten Effekte Spleno-megalie, Körpertemperatur und Knochenmarktransplantation verifiziert werden. Darüber hinaus lieβen sich weitere negative Einfluβgröβen wie Hepatomegalie oder die Applikation von Antibiotika nachweisen. Bei den präparatespezifischen Parametern zeigte sich, daβ sowohl die Leukozytenkontamination der Konzentrate als auch die prätransfusionelle Lagerungsdauer das posttransfusionelle Inkrement reduzieren. Schluβfolgerung: Während die klinischen Parameter wie Splenomegalie oder Körpertemperatur nur schwer beeinfluβbar sind, ist dies bei den präparatespezifischen Einfluβgröβen leichter möglich. Daher sollte bei der Auswahl der Konzentrate besonderer Wert auf eine geringe Leukozytenkontamination sowie eine kurze Lagerungsdauer gelegt werden.

Umfangreiche experimentelle Untersuchungen zur mit einer Bluttransfusion assoziierten Immunmodulation kristallisierten verschiedene immunologische Mechanismen heraus, wobei den Makrophagen eine zentrale Bedeutung zukommt. Um dieser Immunmodulation eine Bedeutung beim Verlauf einer Tumorerkrankung zuweisen zu können, ist es notwendig zu postulieren, daß das Immunsystem einerseits eine relevante Rolle bei der Kontrolle des Tumorwachstums ausüben kann und daß an-dererseits durch die Bluttransfusion eine über Monate andauernde Immunmodulation hervorgerufen werden kann. Für beide Thesen gibt es in der neueren Literatur Hinweise, so daß dies den Hintergrund zu bislang vorliegenden Ergebnissen klinischer Studien bildet, welche die Fremdbluttransfusion als einen Risikofaktor für die postoperative Infektion und das Tumorrezidiv darstellen. Der Problematik, daß bislang keine im Hinblick auf die Bluttransfusion kontrollierte Studie vorliegt, wurde mit einem Protokoll begegnet, bei dem die Eigenbluttransfusion kontrolliert der Fremdbluttransfusion bei der Operation des kolorektalen Karzinoms gegenübergestellt wurde. Die Ergebnisse bestätigen einen immunsuppressiven Effekt der Fremdbluttransfusion, klinisch durch eine signifikant höhere Infektkomplikations-rate dargestellt. Die bislang zu verzeichnende Tumorrezidivrate zeigt einen Trend zur verlängerten tumorfreien Überlebenszeit in der mit Eigenblut behandelten Gruppe, der sich in der Subgruppenanalyse als signifikant erweist. Dieses Ergebnis ist vorläufig und muß bis zur Bestätigung an einer größeren Fallzahl und durch längere Nachbeobachtungszeiten kritisch gewertet werden.

Background. Blood transfusions are associated with higher postoperative morbidity and tumor recurrence rates in colorectal cancer surgery. To reduce the need for transfusions in patients with tumor-induced anemia who are not suitable for autologous blood donation, it was tested whether perisurgical erythropoietin application would be able to stimulate hematopoiesis adequately. Methods. In a double-blind randomized study 150 IU/kg body weight erythropoietin was given subcutaneously every 2 days beginning 10 days before operation and continuing until postoperative day 2. Twenty patients were randomized into the erythropoietin group with three observed dropouts and 10 patients into the placebo group. Results. In the erythropoietin group two episodes of hypertension and one deep venous thrombosis were observed. Preoperative hemoglobin response in the erythropoietin group (p=0.069) was paralleled by a highly significant reticulocyte increase (p=0.0004). However, frequency of blood transfusion was not different between both study groups (erythropoietin, 1.82+/-0.80 units/patient; placebo, 1.80+/-0.97 units/patient). If iron availability was analyzed, a strong correlation between ferritin blood levels and transferrin iron saturation with hemoglobin response was observed in regression analysis (p<0.001). Conclusions. These results indicate that hematopoiesis in anemic patients with colorectal cancer can be stimulated by erythropoietin; however, clinical efficacy is to be expected only in selected patients with high iron availability, which calls for further studies combining erythropoietin and parenteral iron application.

Summary Eleven adults have been transplanted for various reasons between July 1979 and July 1982: 2 with aplastic anemia (AA), 1 with paroxysmal nocturnal hemoglobinuria (PNH), 8 with acute leukemia (AL). Four patients suffered from acute lymphocytic leukemia (ALL) and four from acute non-lymphocytic leukemia (ANLL). Two of them were transplanted in relapse, 1 in a partial remission, and 5 in complete remission. All patients were in their late stage of disease. The PNH-patient had an identical twin, 8 patients had an HLA- and MLC compatible sib, 1 an unrelated donor, and 1 was transplanted from his father. Four patients are alive, 2 more than 3 years: 1 with AA and 1 with ALL who was transplanted in relapse. Six patients died of infectious complications (4 of interstitial pneumonia, 1 of a candidasepsis, 1 of acute toxoplasmosis). Patients living more than 3 weeks had a take. Acute graft-versus-host (GvH) disease did not present a major problem. All patients received methotrexate for GvH-prophylaxis, in three instances the marrow was additionally pre-incubated with anti-T-cell globulin.

During storage of CPD-A1 preserved whole blood factors of the complement cascade become activated, as evidenced by a rapid increase in the concentrations of C3a-desArg and C4a-desArg. After 10 to 14 days of whole blood storage, the elevations of C3a and C4a levels were highly significant. This increase was paralleled by an increase in the concentration of the lysosomal proteinase elastase from polymorphonuclear (PMN) granulocytes. By contrast, the concentration of the C3 activator complex C4b2b remained unchanged even after 3 weeks of storage. The supplementation of the anticoagulant CPD-A1 with the polyvalent-proteinase-inhibitor aprotinin and the specific elastase- inhibitor eglin C failed to inhibit complement activation, whereas leukocyte depletion could partially abolish the increase of the concentration of C4a, but had no effect on C3a concentrations. These observations support the notion that cleavage of C4 during storage of whole blood is partially leukocyte dependent, whereas the activation of C3 is possibly caused by the activation of the alternate pathway of the complement system by contact of plasma with plastic surfaces.

Summary Functional characterization of subsets of T lymphocytes is essential for transplantation studies in dogs, as it is in other species. We studied the function of T cells separated by two mouse monoclonal antibodies recognizing complementary subsets — an antibody directed to canine T cells (MdT-P1) with an up-regulating function, and an antibody directed to human CD 8 (MT811) that cross-reacts with down-regulating canine T cells. Immunorosetting with sheep red blood cells and Percoll gradient allowed us to study depleted and enriched fractions. Their function was tested in mixed lymphocyte culture (MLC), cell-mediated cytotoxicity (CML), and coculture with B cells in a hemolytic plaque assay (PFC). In MLC, MdT-P1-positive cells showed a high proliferative response, and MT811-positive cells responded poorly to allogeneic cells. Vice versa, MT811-negative cells responded strongly, and MdT-P1-negative cells were poor responders but strong stimulators. Effector cells of CML were separated following 8 days of culture and prior to mixing with target cells. Enriched and depleted fractions with either antibody showed low cytotoxic activity as compared with unseparated cells. When added to unseparated effector cells MT811-positive cells suppressed cytotoxicity. B cells were obtained by resetting with staphylococcal protein A (SPA). Their immunoglobulin production was studied following 6 days of culture stimulated by pokeweed mitogen in a reverse hemolytic plaque assay. Again, MT811-positive cells added to the culture suppressed, and MT811-negative cells enhanced immunoglobulin production. In conclusion, immunorosetting with two monoclonal antibodies allowed us to distinguish subpopulations of canine T cells with up-regulating (helper/inducer) from those with down-regulating (suppressor) activity.

Indicator cells — tanned, surface stabilized sheep erythrocytes — were incubated for 1 h in supernatants of 3 h MLCs. Their electrophoretic mobility was measured by an analytical, carrier free electrophoresis system. The change in their mobility compared with an appropriate control was calculated in per cent and correlated with the conventional measured MLR-cpm. The correlation of the two quantities is statistically highly significant (p<0.01). Furthermore, the difference of the electrophoretic mobility values of the group of HLA-D-identical and the groups of HLA-D-haploidentical or -different donors is significant beyond the 1‰ level (p<0.0005). Our method enables, therefore recognition of a positive or negative MLC after only 4 h. Typing for HLA-D-determinants seems to be possible. This could be of great importance for histocompatibility testing and organ transplantation.

Storage of single-donor platelet concentrates is currently limited to 5 days. During this period, however, numerous morphologic and biochemical changes have been observed. These changes result in functional impairment of stored platelets. The present study describes increased binding of a monoclonal antibody against GMP 140 on the surface of stored single-donor platelets revealing an activation process. In contrast, binding of monoclonal antibodies directed against glycoprotein complex (GP) IIb-IIIa and ligand-induced binding site (LIBS1) is slightly diminished during storage. When platelets are stimulated with ADP GMP 140, GP IIb-IIIa, and LIBS1 are expressed to a higher extent than on the surface on non-stimulated platelets. The quantity exposed, however, depends upon the storage time. It is significantly reduced when platelets are stored for longer than 1-2 days. The present data indicate that storage of single-donor platelet concentrates affects fibrinogen binding, cell to cell cohesion, and release reaction. The results are in good agreement with conventional aggregation and in vitro bleeding time measurements.

Summary From March 1975 until May 1980 twelve patients with severe aplastic anemia were grafted with bone marrow from HLA-identical siblings by the Munich Cooperative Group for Bone Marrow Transplantation. Six patients are alive between 10 months and more than 5 years after grafting with normal blood values and marrow. One patient is treated as an out patient for chronic localized graft-versus-host disease (GvHD), five patients are well and without treatment. Six patients have died, one patient with a cerebral hemorrhage the day before transplantation, three patients following rejection of grafts 32, 40 and 55 days after grafting, one patient with severe GvHD 85 days after grafting and one patient, probably with interstitial pneumonia, following cerebral hemorrhage. Three of 6 patients who were conditioned with Cyclophosphamide (CY) only died following rejection of the graft. Two adults who were conditioned with CY and “total lymphoid irradiation” and three children, who were given unirradiated leukocyte concentrates from the marrow donor after grafting, did not reject their grafts. The results of the Munich-Cooperative Group for Bone Marrow Transplantation are comparable to those of large, specialized centers for bone marrow transplantation, they indicate possibilities of cure of severe aplastic anemia by marrow grafts from HLA-identical siblings. They confirm that better results are obtained with earlier transplantation in the course of the disease.

Summary Seventeen patients with relapsed, acute leukemia were grafted with bone marrow from HLA-identical siblings by the ‘Munich Cooperative Group for Bone Marrow Transplantation’ during the period from August 1975 to June 1980. The antileukemic and immunosuppressive conditioning treatment consisted of high doses of Bischlorethyl nitrosourea, Cytosine-Arabinoside and Cyclophosphamide, as well as, total body irradiation of about 9 Gy (midline body dose) from dual60Cobalt sources. Methotrexate was given to all patients for prophylaxis of graft-versus-host disease (GvHD). Nine patients received marrow that was treated with anti-T-cell globulin (ATCG) “in vitro”. — Crossreacting antibodies against hemopoietic stem cells were removed by absorption. Two of 5 evaluable patients given untreated marrow developed chronic GvHD, while patients given ATCG-treated marrow did not show unequivocal symptoms of GvHD. Six patients are in complete remission one to 33 months following bone marrow transplantation (b.m.t.) Five patients died with relapses of leukemia between 3 1/2 and 24 months following b.m.t., 3 patients died with interstitial pneumonia within 3 months of b.m.t. and 3 patients died with insufficient graft function within 4 weeks of b.m.t. Four of thirteen patients that were grafted more than 6 months ago are presently alive and in continuous complete remission at 11, 14, 29 and 33 months following b.m.t. Our results confirm that longterm remissions can be obtained with b.m.t. in patients with acute leukemia in advanced stage.

Zusammenfassung EMT-Indikator-Zellen wurden für 1 h im Überstand einer 3 h alten MLC inkubiert. Dann wurde ihre elektrophoretische Beweglichkeit mittels eines analytischen, trägerfreien Elektrophoresesystems gemessen. Die Änderung ihrer Beweglichkeit gegenüber einem Mediumstandard wurde in Prozent berechnet und mit den konventionell gemessenen MLR-cpm korreliert. Die Korrelation beider Größen ist entsprechend unseren Ergebnissen statistisch hoch signifikant. Es scheint möglich, mit dieser Methode bereits nach 4 h eine MLC als positiv oder negativ zu erkennen. Dieses Faktum könnte für die Histokompatibilitätstestung und die Transplantation von großer Bedeutung sein.

Allogeneic transplantation of human cancellous and cortical bone is a controversially discussed concept in trauma and orthopaedic surgery. Biological and immunological arguments support transplantation of autologous material whenever this is technically possible. On the other hand, synthetic alloplastic materials for bone substitution are available free of immunological and hygienic hazards. In this context the value of allogeneic bone grafts is discussed, especially considering the problem of AIDS. If autologous corticospongious bone is to be used its supply is limited. On the other hand, alloplastic synthetic artificial bone does not meet all the requirements demanded for substitution of large osseous defects up to now. The problems of geometric and mechanical stability of these alloplastic materials still remain. Therefore, no alternative to allografting of large, stable, corticospongious fragments exists in some cases. Bone transplantation is performed without vital indication in nearly every case. Thus an optimum of hygienic security has to be claimed for recipients of allogeneic bone. The “Munich model” for bone transplantation is presented and discussed.

Among gastric cancer patients, the Rhesus D-negative phenotype correlated with increased tumour recurrence [all patients, n = 83, P = 0.026; curatively (R0) resected patients, n = 51, P = 0.093] and reduced overall survival time (all patients, log-rank P = 0.0028; R0 patients, log-rank P = 0.0003) and was identified in multivariate analysis as the most important independent prognostic marker in the R0 patient group (relative risk 9.1, P = 0.0013).

Clinical and experimental studies indicate, that blood transfusion can modify the recipient's immune system. While beneficial to renal allograft survival, these immunomodulating effects may, however, prove detrimental to cancer patients. Recently, an adverse relationship between blood transfusion and cancer recurrence was reported in colon, lung, breast, kidney and gastric cancer. Moreover, a higher postoperative infection rate was observed when blood units were administered intraoperatively. We retrospectively reviewed the records of 174 patients with squamous cell carcinoma of the larynx undergoing curative resection between 1979 and 1985. One hundred and forty-one patients received blood transfusions, 33 did not. Recurrence rate (16.7%) was significantly related to clinical stage (p=0.008) and lymph node status (p=0.000); cumulative survival time depended significantly on clinical stage (p=0.003), lymph node status (p=0.004) and tumor size (p=0.017). In contrast, when corrected for these baseline prognostic factors, no significant correlation could be detected between blood transfusion and cancer relapse. Also, survival time did not depend on blood transfusions. No correlation could be found between postoperative infection rate and intraoperative application of blood transfusion (p=0.694). The present study does not support the hypothesis that blood transfusion adversely affects the prognosis of patients with laryngeal cancer. It indicates, that risk factors other than blood transfusion have a greater influence on recurrence and survival time.

Zusammenfassung Periphere Lymphozyten von 32 Patienten mit monoclonaler Gammopathie (16 IgG, 9 IgA and 7 IgM) und von 15 gesunden Spendern wurden hinsichtlich ihrer in vitro-Antwort auf folgende Stimuli untersucht: unspezifische Mitogene wiePhytohämagglutinin (PHA),PokeweedMitogen (PWM) andConcanavalinA (COnA), sowie die spezifischen Antigene Tuberkulin, Candida, Varidase, Tetanus Toxoid, Vaccinia-Antigen and Vaccinia-Kontroll-Antigen.

Summary 51Cr-platelet kinetic studies were performed in 77 patients with idiopathic thrombocytopenic purpura. The sequestration site was splenic for 63, splenic/hepatic for 7 or hepatic for remaining 7 patients. In 20 patients platelet survival was extremely shortened to 0–3 h, whereas only 26 patients had a survival time of more than 24 h. Those patients with low platelet counts also had a very short platelet survival time, whereas patients with higher platelet counts (>50×109/l) had longer platelet survival times. 51 patients (66%) were splenectomized following the kinetic studies. 25 patients who had a splenic sequestration site had normalized platelet counts and 6 patients had platelet counts between 80–149×109/l 12 months after splenectomy (i.e. in 92% of cases with splenic sequestration site a full or partial remission). Of the 11 patients with a hepatic or splenic/hepatic sequestration site, 2 patients had full remission, 1 partial remission, 3 patients had minimal improvement and 5 other patients were treatment failures in respect to the splenectomy.

Zusammenfassung Das Sckundärsyndrom, welches nach Transfusion von Knochenmark von C57BI Mäusen auf supraletal (900r) bestrahlte A/Jax Mäuse auftrat, wurde durch Kaninchen-anti-Mäuselymphozytenserum verzögert, in einigen Fällen unterdrückt.

Zusammenfassung Eine spontane Leukämie in C57Bl6-Mäusen wurde mit Antilymphozytenserum (ALS) allein und in Kombination mit Cyclophosphamid behandelt. Antilymphozytenserum allein bewirkte eine Erniedrigung der peripheren Leukozytenzahl und verlängerte die Überlebenszeit der Tiere. Durch die Kombination Antilymphozytenserum-Endoxan ließ sich die Überlebenszeit der Tiere auf das Doppelte verlängern. Die Ergebnisse der Versuche sprechen für eine Potenzierung der Wirkungen beider Mittel.

Zusammenfassung Im Serum einer gesunden Mutter wurden homologe, leukocytäre Isoantikörper festgestellt mit einem Agglutinationstiter von 1/32 – 1/256 und einem Immunadhärenztiter von 1/2. Absorptionsstudien ergaben ein Antikörpergemisch, das mit 97% unausgewählter Leukocytenspender reagierte und in der ? G-Fraktion lokalisiert war.

Summary The mixed leucocyte culture is an important test in the immunologic donor selection. It is particularly suited to find identical living donors within the family. The in vitro homograft reaction serves as a good indicator for the immunological reaction in vivo. The method and its advantages and disadvantages are described; its scope of application in the field of transplantation is discussed, including also the author's own observations.

Hintergrund:Der zunehmende Bedarf an Thrombozytenkonzentraten erfordert aus logistischen Gründen, die Thrombozytenspende frühzeitig durchzuführen und die Thrombozyten über mehrere Tage zu lagern. Ein besonderes Problem stellt allerdings die Qualitätskontrolle derartig gelagerter Thrombozyten dar. Die bisher zur Verfügung stehenden Methoden sind zeit- und personalaufwendig und erlauben zum Teil nur Aussagen über einzelne biochemische und physiologische Parameter. Die vorliegende Arbeit beschreibt eine neuartige Methode zur Qualitätskontrolle von Thrombozytenkonzentraten. Material und Methoden:Einzelspender-Thrombozytenkonzentrate wurden über 7 Tage gelagert; täglich wurden Aliquots entnommen und die In-vitro-Blutungszeit (Thrombostat 4000®) sowie die Thrombozytenaggregation (Aggregometer) bestimmt. Ergebnisse:Der In-vitro-Blutungstest ist schnell und einfach durchführbar, die Ergebnisse entsprechen denen der konventionellen Aggregometrie. Schlußfolgerung:Der In-vitro-Blutungstest erscheint als geeignete Methode, die Funktionsfähigkeit gelagerter Thrombozyten auch im Routinelabor zu überprüfen.

As reported earlier, factors of the complement cascade get activated in CPD-A1-stabilized whole blood. As early as after 10 days of storage under normal blood bank conditions the elevations of the concentrations of C3a-desArg and C4a-des-Arg were highly significant. By contrast, the concentration of the C3 activator complex C4b2b remained unchanged even after 3 weeks of storage. Leukocyte depletion partially inhibited the activation of C4 but had no effect on C3a concentrations. Therefore, cleavage of C4 during storage of whole blood seems to be partially leukocyte-dependent, whereas the activation of C3 is possibly due to the activation of the alternate pathway of the complement system by contact of blood to plastic surfaces. Even though the radioimmunologically measured C3a might be inactive as an anaphylatoxin, these observations are of clinical importance since the inactivated C3a-desArg still possesses biological activities such as activation of platelets which may lead to hypercoagulability and thrombosis.

Summary Two cases of immune haemolytic anaemia and acute renal failure induced by nomifensine are reported.

Ohne Zusammenfassung:

Since 1985, after excluding nonimmunological reasons for habitual abortion, we have been performing an MCL, a cross-match, and an HLA typing in these patients. Those with a low MLC response and a negative cross-match are considered to be immunological aborters. Only three out of 42 examined couples showed a low response and in 21 of 24 tested couples the cross-match was negative. Of 36 patients with a high response, 10 had successful pregnancies; seven of them shared more than two HLA antigens. An immunological treatment is indicated in patients with low response and a negative cross-match. High responders should have another pregnancy without stimulation. HLA typing is without any consequences.

Ohne Zusammenfassung: