Outcome of elderly patients with classical Hodgkin's lymphoma.

INTRODUCTION: The optimal management of elderly patients (pts) with Hodgkin's lymphoma is not yet defined. The aims of the present study were: 1) to evaluate clinical and laboratory characteristics of elderly pts; 2) to indentify risk factors for unfavorable outcome. PATIENTS AND METHODS: The outcome of 182 pts ≥ 60 years (y) was retrospectively analyzed (median age, 67y). Mixed cellularity histology was diagnosed in 49.5 %, advanced stage of disease was in 68.7 % pts, CIRS > 3 in 35.7 %, ECOG PS ≥ 2 in 22.9 % (60-69y) of pts. Chemotherapy (CMT) alone was used in 69.2 % and combination of CMT and radiotherapy in 26.9 % of pts. Anthracycline-based CMT received 83.5 % of pts. The median follow-up was 4.5y. RESULTS: The overall response/complete remission rate was 85.6/70.7 %. The median progression free survival (PFS) and overall survival (OS) were 10y and 11.3y, respectively. Estimated 5-y PFS and 5-y OS were 65.7 % (in contrast to 98.2 % in pts < 60y; p < 0.001) and 70.5 % (99.4 % in pts < 60y; p < 0.001). Overall 70 (38.5 %) elderly pts died. The independent risk factors for a shorter OS included CIRS > 3, lymphopenia < 8 % and anthracycline-free CMT, for a shorter PFS anthracycline-free CMT and lymphopenia < 8 %. CONCLUSION: CIRS > 3, lymphopenia < 8 % and anthracycline-free chemotherapy appear to be significant for unfavorable outcome.

[Long-term results of the UHKT-911 study of adult patients under 65 years of age with de novo acute myeloid leukemias without favorable karyotypes]. / Dlouhodobé výsledky lécby dospelých nemocných do 65 let s de novo akutními myeloidními leukémiemi bez príznivých karyotypu ve studii UHKT-911.

Between February 1991 and April 1994 induction chemotherapy of 32 adult consecutive patients under 65 years with de novo acute myeloid leukemias (AML) was started in the study UHKT-911. They were 19 women and 13 men, aged 18-63 (median 44) years. Their AML were classified according to the FAB classification: 3 M0, 3 M1, 9 M2, 14 M4, 3 M5. Induction chemotherapy consisted of 1-2 cycles with 3-4 doses of daunorubicin (DNR) 45 mg/m2/d i.v. and 14 doses of cytosine arabinoside (Ara-C) 200 mg/m2 per 3-h infusion every 12 hours. After the treatment patients, not being in complete remission, got the HD cycle with 10 high-doses of Ara-C 2000 mg/m2 per 3-h infusion every 12 hours i.v. and DNR 45 mg/m2/d i.v. on days 4 and 5, then the EMi cycle composed of etoposide 100 mg/m2/d i.v. for 5 days and mitozantrone 10-12 mg/m2/d i.v. on days 1, 3 and 5. Complete remission (CR) was achieved in 25 of 32 (78%) patients after 1-3 cycles. Five patients died between days 5 and 24 of treatment of infections, two patients were resistant to 4 cycles of induction therapy and survived 8.4 and 13.5 months. Three patients chose allogeneic bone marrow transplantation in their 1st CR from their relatives. Two of them have been living in CR for 115 a 110 months since diagnosis, the third died of sepsis on the day 52 after transplantation. Two patients in CR died of infections after their 2nd. consolidation cycle. Twenty patients in CR completed 2-4 consolidation cycles (1-3 HD, 1 EMi). Median of their CR duration was 17.8 (2-117) months. Relapse appeared in 12 cases after 4.4-34.8 (median 12.5) months, 8 patients (6 women and 2 men, aged 29-63 years) have remained longer than 5 years in their 1st. CR. Cytogenetic examination of their bone marrow showed a normal karyotype in 4 cases, 1x 46,XX,del(1)(p32p34), 1x 46,XX,16p+, 1x 47,XX,+mar, 1x 46,XX,del(5)(q22q33). After 62 months in CR a pancytopenia with dysplastic bone marrow changes developed in one of them, probably a secondary myelodysplastic syndrome, lasting for further 33 months. Event-free survival at 5 years was 27.5% (8/29 patients), significantly better (p = 0.046) against 7.5% (3/40) patients treated without HD cycles in the years 1982-1987. The same difference was observed in 7.5-year overall survival (p = 0.036) between the two studies, when 3 of 6 patients 60-64 years old remain in their 1. CR.

Prediction and reversion of post-transplant relapse in patients with chronic myeloid leukemia using mixed chimerism and residual disease detection and adoptive immunotherapy.

In the prospective study, we examined hematopoietic mixed chimerism (using polymerase chain reaction (PCR) of variable number of tandem repeat-VNTR sequences) and minimal residual disease (MRD) status (using qualitative and in the case of positivity quantitative reverse transcriptase polymerase chain reaction (RT-PCR) for the BCR/ABL fusion mRNA) in serial peripheral blood samples taken from 25 patients after bone marrow transplantation (BMT) for chronic myeloid leukemia (CML). Increasing mixed chimerism in correlation with increasing signal of MRD was detected in 10 patients. In two patients mixed chimera status and BCR/ABL rearrangement led to hematologic relapse, in five patients molecular relapse was followed by reappearance of Ph chromosome and three patients developed molecular relapse only. Adoptive immunotherapy-donor lymphocyte infusion (DLI), interferon (INF) and discontinuation of post-transplant immunosupression-separately or in different combinations was used in nine patients with molecular, cytogenetic or hematologic relapse of CML. The results demonstrate that significant response at the molecular level can be achieved for a majority of CML patients and that using of all forms of adoptive immunotherapy controlled by MC and MRD is more efficient in patients treated in early molecular relapse-with minimal disease burdens.

Simple competitive two-step RT-PCR assay to monitor minimal residual disease in CML patients after bone marrow transplantation.

Quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) assessing the amount of transcripts of the BCR/ABL gene, the molecular marker of chronic myeloid leukemia (CML), is the only method sensitive enough for monitoring of minimal residual disease (MRD) in CML patients after bone marrow transplantation (BMT). In this study we present a simple modification of competitive Q-RT-PCR using natural competitors from cell lines K562 and BV173. The competitors were used in the form of unpurified RNA in cell lysates which ensured their high stability. Mixing competitors and samples before RNA extraction eliminated problems with quantification of cDNA or RNA entering the competitive reaction and with checking for the RNA quality and reverse transcription (RT) efficiency. The bulk of the malignant clone was expressed as the number of leukemic cells in 10(6) leukocytes when the overproduction of the BCR/ABL mRNA in the cell lines we used as competitors was taken into account. It was found to be 82-fold and 14-fold in K562 and BV173, respectively, in comparison with 100% Ph-positive CML standard. The assay reliability was verified by comparison of results with the mathematical model of competitive PCR. The assay is highly reproducible and sensitive (10(-5)). Its accuracy was proved to be excellent in a wide range of malignant cell concentrations. The method is demonstrated on three CML patients suffering from MRD after BMT. In conclusion, this method fulfills all criteria of competitive Q-RT-PCR. Because of its simplicity it is suitable for clinical laboratories and due to the high stability of the lysates used it may serve in the standardization of results between different laboratories.

[Treatment of chronic myeloid leukemia with bone marrow transplantation at the Institute of Hematology and Blood Transfusion in Prague]. / Lécba chronické myeloidní leukémie transplantací kostní drene v UHKT Praha.

The outcomes of bone marrow transplantation (BMT) performed at the Institute of Haematology and Blood Transfusion from April 1988 to December 1994 in 31 patients with chronic myelogenous leukemia are presented. Age of the patients range from 18 to 49 years, median 34 years. Male:female ratio was 1.58:1. The conditioning regimen consisted of Cyclophosphamide and total body irradiation (TBI) or Busulfan and Cyclophosphamide. The results are evaluated as of January 1, 1995. Nineteen patients (61.3%) are alive, 12 patients (38.7%) died. The causes of death are discussed. The median time of follow up all patients is 10.4 months, range 0.3-81.5. The median time of follow up of surviving patients is 21.8 months, range 2.5-81.5. Probability of 2 years survival by Kaplan-Meier analysis is 58 +/- 10%. Of the 24 transplanted in the first chronic phase, 18 patients are alive. Of the 7 transplanted in advanced phase of the disease, 1 patient is alive. Of the 27 patients, who received bone marrow from an HLA identical sibling, 19 are alive. Of the 4 patients who received bone marrow from other donor than an HLA identical sibling, none is alive. Acute GvHD III.-IV. grade developed in 5 patients (16.1%), moderate and severe chronic GvHD developed in 11 patients (31.5%). Cytogenetic relapse was diagnosed in 1 patient, hematological relapse in 2 patients. Karnofsky scores of patients surviving after BMT range from 30% to 100%, median 90%.

[Results of induction therapy in newly diagnosed acute myeloid leukemias in study 911 at the Institute of Hematology and Blood Transfusion]. / Výsledky indukcní lécby nove zjistených akutních myeloidních leukémií ve studii UHKT-911.

Thirty-seven patients with de novo acute myeloid leukemias were admitted to the Institute of Hematology and Blood Transfusion in Prague in February 1991-December 1993. Their age was 18-85 years with a median of 46 years. Two patients died on the day of admission, chemotherapy was initiated in 35 patients. Altogether 27 patients (77%) achieved complete remission (CR), i.e. 18 (81%) of 22 patients younger than 55 years and 9 (70%) of 13 patients older than 55 years. Only 7 (35%) of 20 patients achieved CR after a single therapy course 3/7 consisting of 3 doses of daunorubicin 45 mg/m2 on days 1, 3, 5 and cytosine arabinoside 150-200 mg/m2 every 12 hours for 7 days. However, 8 (61%) of 13 patients achieved CR after a single treatment course 4/7 with 4 doses of daunorubicin 45 mg/m2 on days 1, 3, 5, 7 and identical doses of cytosine arabinoside as in the 3/7 treatment. We used the course with 10 high-doses of cytosine arabinoside 2000 mg/m2 every 12 hours and daunorubicin 45 mg/m2 on days 4 and 5 (treatment HDAC/DNR) as the 1st, 2nd or 3rd induction therapy in 12 patients and 9 (75%) of them achieved CR. The treatment was associated with a high toxicity. An intensified therapy 3/7h similar to the 3/7 one but with the doubled dose of cytosine arabinoside 300-400 mg/m2 on days 5-7 was given to 5 patients as the 2nd induction but it did not improve the CR rate and it was associated with a high toxicity similar to the HDAC/DNR therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Frequency analysis of cytotoxic T lymphocyte precursors in search for donors in bone marrow transplantation.

The usefulness of cytotoxic T lymphocytes precursors (CTLp) frequency analysis in the search for donors in bone marrow transplantation was studied. The frequency of anti-recipient CTLp was approached by limiting dilution assay in HLA matched unrelated, HLA partially matched related and HLA genotypically identical donors. The majority of patients examined were affected with different hematological malignancies. Alloreactive CTLp recognizing non-HLA gene products were not detected in pretransplant examination of two pairs of HLA identical siblings. However, an increased incidence of allospecific CTLp was identified in HLA matched MLC negative unrelated pairs. Thus, CTLp assay allowed to uncover the residual Class I incompatibilities that remained hidden in standard serotyping. In two matched unrelated pairs with high pretransplant CTLp frequency the severe acute graft-versus-host disease (GVHD) developed after bone marrow transplantation. Examination of other relatives in patients lacking an HLA identical sibling showed the importance of Class I incompatibility for CTLp generation as well. The lack of correlation between CTLp frequency and HLA-D disparity could suggest that Class II antigens do not participate in CTLp induction. With one exception we had good correlation between MLC and DNA analysis of Class II antigens demonstrating that MLC gives interpretable results even in unrelated pairs. Our results demonstrate the significance of CTLp frequency assay in detection of residual Class I incompatibilities in matched unrelated pairs and in assessment of Class I compatibility in related pairs. For that it should be used in the final selection of BMT donors.

Sequential cytogenetic study of patients after bone marrow transplantation.

Thirty-one patients (19 males and 12 females; mean age 23.9 years, range 4-41 years) were treated with bone marrow transplantation (BMT) after intensive chemoradiotherapy. Their diagnoses were as follows: chronic myeloid leukemia (CML) in 13, acute myeloid leukemia (AML) in seven, acute lymphocytic leukemia (ALL) in six, myelodysplastic syndrome (MDS) in two, aplastic anemia (AA) in two, and Fanconi anemia (FA) in one. Allogeneic BMT was performed in 28 cases (17 donors were of like sex, 11 were of unlike sex), one patient received syngenic transplant, and one received transplant of cells obtained from an unrelated donor through a computerized international registry in London. Autologous BMT was performed in three patients. BM cells were analyzed cytogenetically at diagnosis, before and serially after BMT (three to nine times). Follow-up ranged from 2 to 55.5 months. Cytogenetic examination was a very useful method for monitoring posttransplantation course in patients with CML or in those who received BM cells of unlike sex. Results of concomitant cytogenetic examinations are reported in detail.

[Monitoring lymphocyte activity in the peripheral blood of patients after bone marrow transplantation]. / Monitorování aktivních lymfocytu v periferní krvi pacientu po transplantaci kostní drene.

BACKGROUND: After bone marrow transplantation serious complications develop which may limit the success of this therapeutic method. One of the early complications is an acute graft versus host reaction. The objective of the investigation was to evaluate the relationship between the number of active lymphocytes in the patient's blood stream after bone marrow transplantation and the development of an acute graft versus host reaction or rejection of the graft, and thus contribute towards prediction or diagnosis of these complications. METHODS AND RESULTS: In 14 patients with acute myeloid leukaemia (3), acute lymphatic leukaemia (1), chronic myeloid leukaemia (6), myelodysplastic syndrome (2) and aplastic anaemia (2) bone marrow was transplanted: the donor was in all instances a HLA identical sibling. However, only 11 patients were evaluated. In the latter changes in the number of circulating active lymphocytes were assessed: their activity was evaluated from nucleolar characteristics expressed by RNA synthesis. Their values at the time of the acute graft versus host reaction (GVHD) varied between 7.4% and 17.3%; at the time when these patients were free from complications they were 2.2%-6.0% (the difference is at the borderline of significance). In 8 patients the rise of active lymphocytes preceded the manifestation of the graft versus host reaction by 3-7 days. At the time of infectious complications after bone marrow transplantation (temperatures of obscure origin, herpetic infections, varicella, adenoviral infections) the number of active lymphocytes did not increase (2.0%-10.0%), as compared with 3.4%-9.5% in the group without complications. CONCLUSIONS: The increased percentage of activated lymphocytes in the peripheral blood stream of patients with an acute graft versus host reaction (GVHD) after bone marrow transplantation results from specific immunological procedures. Their assessment could help with the differential diagnostic difficulties frequently associated with the diagnosis of the acute graft versus host reaction.

[Results of bone marrow transplantation at the Institute of Hematology and Blood Transfusion in Prague]. / Výsledky transplantace kostní drene v Ustavu hematologie a krevní transfúze v Praze.

The first allogenic bone marrow transplantation (TKD), when for the preparation whole body irradiation was used, was implemented in the Institute of Haematology and Blood Transfusion (UHKT) in Prague in 1986. Before June 1992 36 TKD were performed incl. 28 allogenic, 2 syngenic and 6 autologous. For the first time bone marrow from a non-related donor was transplanted. Of 30 allogenic and syngenic TKD to the present time 17 patients survive, i.e. 56.6% of the whole group. According to individual diagnoses 8 patients with the diagnosis of chronic myeloid leukaemia (CML) survive, 5 of 10 patients with the diagnosis of acute leukaemia (AL) and 3 of 4 patients with the diagnosis of severe aplastic anaemia (SAA) or with Fancon's anaemia (FA) resp. The survival period of the whole group is from 1-62 months since the transplantation. The main cause of death of 8 from 13 patients who died were infections associated with acute or chronic disease of the graft against the host (GVHD). In autologous TKD the bone marrow was treated with etoposide. Of the six transplanted patients with AL five survive 1.5-30 months after transplantation. The authors present some general information of pretransplantation preparation, prevention of GVHD, its incidence and results of TKD.

[Personal experience with treatment of acute myeloid leukemia using the DAT combination]. / Nase zkusenosti s lécbou akutních myeloidních leukémií kombinací DAT.

A combination of cytostatics--Rubomycin (Medexport), Alcysten (Spofa) and LANVIS (Wellcome) was used to treat 44 patients with the diagnosis AML. A total of 66% CR was achieved. The median duration of CR was 15 months and the median of survival of patients with CR was 25 months. 20% of the patients died in induction mostly from infectious complications. The authors assume that this combination with the use of the Czech preparation Alcysten and the Soviet preparation Rubomycin is suitable induction treatment in AML. Treatment, however, calls for perfect supporting therapy and therefore should by concentrated in selected departments.

Cytogenetic study of circulating blasts in leukemias.

A special cultivation technique of separated blasts of peripheral blood in suspension culture has been used for cytogenetic diagnosis of patients suffering from acute nonlymphocytic leukemia, chronic myeloid leukemia, and refractory anemia with excess of blasts. Twenty-three patients were examined; in ten cases isolation of blasts was performed on Ficoll-Verografin and in the remaining 13 patients further separation of T-lymphocyte precursors by means of sheep erythrocytes was performed. Remarkably, a 100% success rate was attained in all cultivations. The optimum harvesting time was 72-96 hours; the rate of cell division per cultivation was determined by means of bromodeoxyuridine incorporation; second mitoses were revealed only after 96-hour cultivation. In all patients, except one, abnormal karyotypic changes were ascertained in separated blasts of peripheral blood cultivations. In most cases the morphology of chromosomes obtained from separated blasts of peripheral blood cultivations was of excellent quality.

Auto anti-A1 and auto anti-NA1 after bone marrow transplantation.

The production of auto anti-A1 and auto anti-NA1 antibodies in patient with aplastic anemia has been described. The patient of group A1 received bone marrow from his brother of group A2. For immunosuppression cyclosporine A was administered.