G. Ehninger

Polymerase chain reaction to evaluate antiviral therapy for cytomegalovirus disease

A sensitive and specific method to monitor suppression of cytomegalovirus (CMV) replication is essential in patients treated with ganciclovir after allogeneic bone-marrow transplantation. In this study, antiviral therapy of eighteen episodes of symptomatic CMV infection in 15 such patients were followed up clinically and by virus culture and polymerase chain reaction (PCR). Clinical improvement, culture, and PCR were assessed for their ability to predict the efficacy of ganciclovir therapy in each patient. In eleven successfully treated episodes of CMV disease (disappearance of symptoms and improvement in biochemical variables) clinical improvement was associated with an effective suppression of virus replication as shown by negative culture and PCR assays of blood and urine specimens obtained after antiviral therapy. 1 patient who did not improve clinically when receiving antiviral therapy remained both culture positive and PCR positive for CMV. 6 patients with early relapse of CMV disease or who died after an initial clinical improvement were PCR positive but culture negative after termination of therapy. Demonstration of CMV in blood and urine by PCR after stopping antiviral therapy (even when culture is negative) points to incomplete suppression of virus replication. The findings show that PCR is a better predictor of the efficacy of antiviral therapy than are culture or clinical assessment.

Decreased Essential Antioxidants and Increased Lipid Hydroperoxides Following High-Dose Radiochemotherapy

The blood from 19 patients having bone marrow transplantation was examined for the essential antioxidants alpha-tocopherol and beta-carotene as well as lipid hydroperoxides before, at and after bone marrow transplantation (BMT). Conditioning therapy, preceding BMT in order to achieve marrow ablation and immunosuppression, consists of high-dose chemotherapy which is mostly combined with total body irradiation (TBI). In order to see a possible difference between patients with and without additional TBI, we divided the patients up into two groups; patients receiving TBI (RT+) and patients without TBI (RT-). All patients required total parenteral nutrition beginning one week prior to BMT. After conditioning therapy plasma levels of absolute and lipid-standardized alpha-tocopherol and beta-carotene decreased in both groups, presumably as a result of an enhanced breakdown of these antioxidants. The loss of these lipid-soluble antioxidants has to be considered as a possible cause for early post-transplant toxicity. Lipid hydroperoxides increase significantly in the group of patients with additional TBI, whereas the other group, receiving no additional TBI, showed no significant change. We suggest high-dose supplementation of essential antioxidants for patients undergoing BMT.

Effect of etoposide (VP16-213) on lipid peroxidation and antioxidant status in a high-dose radiochemotherapy regimen.

SummaryA total of 13 patients receiving bone marrow transplants (BMT) for treatment of different haematological diseases were investigated. Conditioning therapy preceding BMT consisted of fractionated total-body irradiation (12 Gy) and high-dose chemotherapy with cyclophosphamide (2±60 mg/kg). Patients stratified to be at high risk for relapse (6/13) were additionally treated with etoposide (30 mg/kg). Plasma concentrations of absolute and lipid-standardized antioxidants (α-tocopherol and β-carotene) decreased following conditioning therapy, presumably as the result of an enhanced breakdown of these antioxidants. Etoposide treatment did not amplify the loss of essential anti-oxidants but significantly increased lipid hydroperoxide concentrations in serum. We suggest that the abnormal generation of lipid hydroperoxides is the result of free radical formation.

IL2‐ and IL4‐dependent proliferation of T‐cell clones derived early after allogeneic bone marrow transplantation: Studies of patients with chronic myelogenous leukaemia

Abstract:  In an attempt to explore T‐cell functions shortly after allogeneic bone marrow transplantation more fully, IL2‐ and IL4‐dependent proliferation was assessed on CD4+ TCRαβ+ T‐cell clones derived 4–6 weeks after transplantation. Both allogeneic pooled peripheral blood mononuclear cells and Epstein‐Barr virus‐transformed B‐cell lines (BCL) could function as accessory cells (AC) for PHA activation of T‐cell clones. Although minimal clonal proliferation was seen when the T‐cell activation signal was BCL + PHA + IL4, a majority of the clones could undergo IL4‐dependent proliferation after previous activation with AC + PHA + IL2. For certain clones, IL4 also showed an additive effect with IL2. Thus, IL4 was a growth factor for a majority of the investigated posttransplant T‐cell clones, and in vivo modulation of IL4‐dependent T‐cell functions may thus become a future therapeutic possibility to enhance graft‐versus‐leukaemia effects in bone marrow transplant recipients.

Secretion of IL-2, IL-3, IL-4, IL-6 and GM-CSF by CD4+ and CD8+ TCRαβ+ T-Cell Clones derived early after Allogeneic Bone Marrow Transplantation

Secretion of different cytokines may be an important T‐cell effector mechanism for bone marrow engraftment, graft versus host disease and graft versus leukaemia effects after allogeneic bone marrow transplantation (BMT). Cytokine secretion and autocrine proliferative capacity of T‐cell clones derived from leukaemia patients 3–6 weeks after allogeneic bone marrow transplantation were investigated. Only a minority of post‐transplant T‐cell clones (23/120; 19%) was capable of undergoing autocrine proliferation. By contrast, 21/65 (32%) normal control clones from the marrow donors derived under the same conditions were autocrine proliferative. All clones were interleukin‐2 (IL‐2) responsive. A majority (12/17; 71%) of autocrine proliferating post‐transplant clones secreted detectable IL‐2. Compared with control clones, CD4+ T‐cell clones derived early after BMT produced decreased levels of interleukin‐4 (IL‐4) and interleukin‐6 (IL‐6), whereas secretion of interleukin‐3 (IL‐3) and granulocyte/macrophage colony‐stimulating factor (GM‐CSF) showed no significant difference. The small number (n = 8) of post‐transplant CD8+ clones showed decreased production of IL‐3, IL‐4 and IL‐6 compared with control clones, but normal secretion of GM‐CSF. Neither CD4+ nor CD8+ T‐cell clones secreted interleukin‐7 (IL‐7).

A multicentre study on intensive induction and consolidation therapy in acute myelogenous leukaemia

E. Kurrle 1, G. Ehninger 2, M. Freund 3, G. HeiP, D. Hoelzer 4, H. Link 3, P.S. Mitrou 4, S. Ohl s, W. Queisser 6, G. Schlimok 7, and H. Wandt 5 Abteilung Innere Mediziu III, Medizinische Universitfitsklinik, D-7900 Ulm, Federal Republic of Germany 2 Abteilung Hfimatologie, Medizinische Universit~tsklinik, D-7400 Ttibingen, Federal Republic of Germany 3 Abteilung H~matologie-Onkologie, Medizinische Hochschule, D-3000 Hannover, Federal Republic of Germany 4 Abteilung H~tmatologie, Zentrum der Inneren Medizin, Universit~t Frankfurt, D-6000 Frankfurt/Main, Federal Republic of Germany 5 5. Medizinische Klinik, Klinikum NOrnberg, D-8500 Nt~rnberg, Federal Republic of Germany 60nkologisches Zentrum, Klinikum Mannheim, D-6800 Mannheim, Federal Republik of Germany 7 2. Medizinische Klinik, Zentralklinikum Augsburg, D-8900 Augsburg, Federal Republic of Germany

High-dose cytosine arabinoside and daunorubicin postremission therapy in adults with de novo acute myeloid leukemia : long-term follow-up of a prospective multicenter trial

A total of 149 consecutive de novo AML patients aged 50 years or less (median age = 37 years) were enrolled in this prospective multicenter trial initiated in May 1985. All patients received the same induction and early consolidation therapy with daunorubicin (DNR), cytosine arabinoside (Ara-C), and etoposide (DAV). High-dose Ara-C/DNR therapy included Ara-C at 3 g/m2, in 12 doses (HD-Ara-C/DNR I) and eight doses (HD-Ara-C/DNR II), followed by DNR 30 mg/m2 for 3 days. A complete remission (CR) was achieved in 104 (70%) patients; 61 complete responders received at least one cycle with HD-Ara-C/DNR. If those patients who were transplanted in first CR (n=26), were not considered, the median relapsefree-survival (MRFS) of the remaining 78 patients was 15 months, with a probability of relapse-free survival (RFS) at 116 months of 30% (95% CI, 20–40%) after a median follow-up of 95 months. The MRFS of the HD-Ara-C/DNR consolidated patients was 25 months, with a probability of RFS at 116 months of 37% (95% CI, 24–50%). If all patients who were transplanted (n=44) were not considered, the median survival time (MST) was 18 months with a probability of being alive at 118 months of 24% (95% CI, 16–33%). MST of the HD-Ara-C/DNR consolidated patients was 58 months with a survival probability of 46% (95% CI, 31–60%) at 118 months. Prognostic factor analysis did not reveal any significant influence of age, sex, FAB subtype, white blood cell count, hemoglobin level, thrombocyte count, LDH, or response to the first induction course on RFS of the HD-Ara-C/DNR consolidated patients. In summary, HD-Ara-C/DNR consolidation can improve the long-term outcome of a subgroup of de novo AML patients. Further improvement of the outcome seems to depend on the identification of patients with an inferior outcome under that strategy who might benefit from alternative treatment strategies.

Correlation between low CSA plasma concentration and severity of acute GvHD in bone marrow transplantation

SummaryBetween 1982 and 1986 51 patients were treated with ciclosporin a (CSA) to prevent graft versus host disease (GvHD) after bone marrow transplantation (BMT). Major side effects of the drug were tremor, hypertension, hepatotoxicity and nephrotoxicity. Acute GvHD 0° to II° occurred in 80% of our patients, and GvHD III° and IV° in 20% despite the use of CSA. Two to four days before the onset of GvHD, CSA serum levels were significantly lower on the average in patients who developed GvHD III° and IV° compared to the others. Our data indicate that plasma CSA concentrations higher than 250 ng/ml should be achieved to reduce the severity of GvHD after BMT.

Synthesis and structural elucidation of biliary excreted thioether derivatives of mitoxantrone

1. Hplc-MS coupling has been used for the identification of thioether derivatives of the anticancer agent mitoxantrone in the bile of pig. 2. Three biologically relevant new thioether derivatives of mitoxantrone have been synthesized by a horseradish peroxidase-catalysed reaction. 3. The thioether derivatives have been characterized by means of ion-spray tandem mass spectrometry and nmr spectrometry including two-dimensional techniques. 4. The carbon-sulphur bond formation takes place at the hydroquinone moiety of the anthraquinone skeleton pointing to the importance of a tautomeric equilibrium between different species of the oxidized drug. 5. The occurrence of the synthesized compounds in biological systems suggests a metabolic pathway that may be relevant for the cytotoxicity of mitoxantrone (oxidative activation).

Bone marrow transplantation for acute leukemia in second or subsequent remission.

SummaryTwenty-one patients with acute leukemia in second to fifth remission were treated with bone marrow transplantation: 19 patients with transplants from HLA-matched siblings and two with transplants from identical twins. Twelve patients survived from 15 to 1,625 days after transplantation: six of 11 in the ALL group and six of 10 in the AML group. Recurrence of leukemia after marrow transplantation occurred in five patients. The cause of death in five patients was infection, in two patients combined with graft-versushost disease. Long-term disease-free survival can probably be achieved in 30%–35% of all patients with acute leukemia who receive a marrow transplant in second or subsequent remission.