Bone Marrow Multipotent Mesenchymal Stromal Cells from Patients with Aplastic Anemia Retain Their Ability to Support Hematopoietic Precursors despite Pronounced Changes in Gene Expression.

The properties of bone marrow-derived multipotent mesenchymal stromal cells (MSC) of patients with aplastic anemia at the onset of the disease are studied insufficiently. The aim of this work was to test the ability of MSC from patients with aplastic anemia to maintain hematopoietic precursors and to analyze the expression of genes associated with hematopoiesis and immune response. The ability of MSC to maintain hematopoietic precursors was determined by counting cobblestone area-forming cells; gene expression was analyzed by quantitative PCR. It was shown that MSC of patients with aplastic anemia preserve their ability to maintain hematopoietic precursors. Pronounced changes in the expression of the VEGFA and ANGPT1 genes were found. MSC from aplastic anemia patients with PNH clone significantly differ from those from aplastic anemia patients without PNH clone in terms of the expression of the SDF1, IL1R, and VEGFA genes. Changes in gene expression can be associated with the pathogenesis of the disease.

[Detection of activating mutations in RAS/RAF/MEK/ERK and JAK/STAT signaling pathways].

ISSUE: The study of activating mutations (NRAS,KRAS,FLT3,JAK2,CRLF2genes) of RAS/RAF/MEK/ERK and JAK/STAT signaling pathways in B-cell acute lymphoblastic leukemia (B-ALL) in adult patients which are included in Russian multicenter clinical trials. MATERIALS AND METHODS: Within the multicenter study there were 119 adult patients included withde novoB-ALL. The study was considered as prospective and retrospective. The group withBCR-ABL1-negative B-ALL consisted of up to 93 patients (45 male and 48 female, at the age of 17 to 59, the median age 31), they were treated according to the protocols ALL-2009, ALL-2016. The median follow-up lasted for 19 months (1119). The group withBCR-ABL1-positive B-ALL with up to 26 patients (10 male and 16 female, at the age of 23 to 78, the median age 34 years) was included in the study as well. The treatment was carried out according to the protocols ALL-2009 and ALL-2012 in combination with tyrosine kinase inhibitors. The median follow-up lasted for 23 months (4120). The molecular analysis of activating mutations inNRAS,KRASgenes (RAS/RAF/MEK/ERK signaling pathway) andJAK2,CRLF2genes (JAK/STAT signaling cascade) was performed via Sanger sequencing. The internal tandem duplications (ITDs) inFLT3gene were studied by fragment analysis. The evaluation of CRLF2 expression was fulfilled via flow cytometry. RESULTS: Activating mutations inNRAS,KRAS,FLT3genes were found in 22 (23.6%) patients withBCR-ABL1-negative B-ALL. In total, 23 mutations were revealed in theNRAS(n=9),KRAS(n=12), andFLT3(n=2) genes, according to statistics that was significantly more frequent than withBCR-ABL1-positive B-ALL, these genes mutations were not identified in patients (p=0.007). The frequency of mutations detection inKRASandNRASgenes in patients withBCR-ABL1-negative B-ALL was comparable as 12.9% (12 of 93) to 9.7% (9 of 93), respectively (p=0.488). One patient was simultaneously revealed 2 mutations in theKRASgene (in codons 13 and 61).FLT3-ITD mutations were detected in 3.5% (2 of 57) cases ofBCR-ABL1-negative B-ALL. In patients withBCR-ABL1-positive B-ALLFLT3-ITD mutations were not assessed. Violations in the JAK/STAT signaling cascade were detected in 4 (4.3%) patients withBCR-ABL1-negative B-ALL. They were represented by the missense mutations ofJAK2gene (n=3) and the overexpression of CRLF2 (n=2); in one patient were detected the overexpression of CRLF2 and a mutation inJAK2gene simultaneously. No mutations were found inCRLF2gene. In patients withBCR-ABL1-positive B-ALL noJAK2mutations were detected. As long as analyzing demographic and clinical laboratory parameters between groups of patients with and without mutations, there were no statistically significant differences obtained. In the analyzed groups of patients, long-term therapy results did not differentiate according to the mutations presence inNRAS,KRAS,FLT3,JAK2genes. Also, substantive differences were not shown in the rate of the negative status achievement of the minimum residual disease between patients with and without activating mutations in the control points of the protocol (on the 70th, 133rd and 190th days). CONCLUSION: NRAS,KRAS,FLT3,JAK2activating mutations do not affect the long-term results of the therapy and the rate of the negative status achievement of the minimum residual disease in patients withBCR-ABL1-negative B-ALL treated by the Russian multicenter clinical trials.

Individual Differences of Multipotent Mesenchymal Stromal Cells Manifesting in during Interaction with Lymphocytes.

Analysis of changes in lymphocyte subpopulations during co-culturing with multipotent mesenchymal stromal cells (MSC) revealed two distinct MSC groups: one group (A) increased HLA-DR expression on lymphocytes during co-culturing and the other (B) did not change it in comparison with lymphocyte monoculture. In stromal cells interacting with lymphocytes, expression of HLA-DR molecules was initiated, but only in samples that induced enhanced expression on lymphocytes and irrespectively of whether allogeneic or autologous lymphocytes were used for co-culturing with MSC. In group A, the relative expression of IDO1 significantly increased in comparison with group B. The revealed individual differences in MSC can explain why not all MSC samples are effective in the treatment of autoimmune diseases, acute "graft-versus-host" disease, and other pathologies.

Co-Culturing of Multipotent Mesenchymal Stromal Cells with Autological and Allogenic Lymphocytes.

We studied the effect of autologous and allogeneic lymphocytes on multipotent mesenchymal stromal cells in co-culture. It is shown that changes in multipotent mesenchymal stromal cells and in lymphocytes did not depend on the source of lymphocytes. Contact with lymphocytes triggers expression of HLA-DR molecules on multipotent mesenchymal stromal cells and these cells lose their immune privilege. In multipotent mesenchymal stromal cells, the relative level of expression of factors involved in immunomodulation (IDO1, PTGES, and IL-6) and expression of adhesion molecule ICAM1 increased, while expression of genes involved in the differentiation of multipotent mesenchymal stromal cells remained unchanged. Priming of multipotent mesenchymal stromal cells with IFN did not affect these changes. In turn, lymphocytes underwent activation, expression of HLA-DR increased, subpopulation composition of lymphocytes changed towards the increase in the content of naïve T cells. These findings are important for cell therapy.

[Adult B-cell acute lymphoblastic leukemias: Conclusions of the Russian prospective multicenter study ALL-2009]. / Ostrye V-limfoblastnye leikozy vzroslykh: vyvody iz Rossiiskogo prospektivnogo mnogotsentrovogo issledovaniia OLL-2009.

AIM: To analyze the efficiency and reproducibility of the ALL-2009 protocol within the Russian prospective multicenter study based on different principles of cytostatic effects (non-intensive, but continuous cytotoxic treatment and a small number of allogeneic hematopoietic stem cells). SUBJECTS AND METHODS: The ALL-2009 (NCT01193933) study conducted in April 2009 to December 2016 included 194 patients (95 males and 99 females) aged 15 to 55 years (median age 28 years) with Ph-negative B-cell acute lymphoblastic leukemia (ALL). There was early pre-B-cell ALL in 54 patients, common ALL in 101, pre-B ALL in 39, initial leukocytosis in 9.4·109/l (0.4-899.0), lactate dehydrogenase in 901 IU (31-13 059), an initial central nervous system lesion in 17 (8.7%), mediastinal injury in 3 (1.5%), and splenomegaly in 111 (57.2%). The results of standard cytogenetic analysis are known in 113 (60.4%) patients. Normal karyotypes were detected in 49 (54.5%) out of the patients; t(4;11) in 9 (5.4%), t(1;19) in 2 (1.2%), and other karyotypic abnormalities in 53 (46.9%). Thirteen (7.8%) patients underwent allogeneic hematopoietic stem cell transplantation in first complete remission (CR); their proportion did not differ in the federal and regional centers. RESULTS: The frequency of CR achievement was the same in the federal and regional centers and generally amounted to 87.5%. Early (8.8%) and CR (9.6%) mortality rates remained high despite the low aggressiveness of cytotoxic action, necessitating the improvement of auxiliary treatment. The five-year overall survival (OS) rates vary considerably in the federal and regional centers (72.6 and 43.8%), the relapse-free survival (RFS) (70.2 and 53.4%) and recurrence risk (23.1 and 36.5%) are comparable. This suggests that the non-intensive, but continuous exposure principle built in the ALL-2009 protocol makes it possible to reproduce the envisaged treatment program and to achieve satisfactory results. CONCLUSION: The ALL-2009 protocol allows both the federal and regional centers to obtain the long-term results comparable with those of current foreign studies: OS (54.2%), RFS (56.5%); and relapse risk (35.4%). Multivariate analysis has identified age (over 30 years), initial leukocytosis (30·109/l and more) and t(4;11) among the main clinical prognostic factors. Gene mutation detection evaluated in a small number of patients (8/36) is not a poor prognostic sign. There is a need for further investigations with centralized evaluation of the mutation status of leukemic cells and the clearance of minimal residual disease.

Changing the Properties of Multipotent Mesenchymal Stromal Cells by IFNγ Administration.

We studied changes in the population of human multipotent mesenchymal stromal cells activated by IFNγ. The cells were cultured under standard conditions; IFNγ was added in various concentrations for 4 h or over 2 passages. It was shown that the total cell production significantly decreased after long-term culturing with IFNγ, but 4-h exposure did not affect this parameter. After 4-h culturing, the expression levels of IDO1, CSF1, and IL-6 increased by 300, 7, and 2.4 times, respectively, and this increase persisted 1 and 2 days after removal of IFNγ from the culture medium. The expression of class I and II MHC (HLA) on cell surface practically did not change immediately after exposure to IFNγ, but during further culturing, HLA-ABC (MHC I) and HLA-DR (MHC II) expression significantly increased, which abolished the immune privilege in these cells, the property allowing clinical use of allogenic multipotent mesenchymal stromal cells. Multipotent mesenchymal stromal cells can suppress proliferation of lymphocytes. The degree of this suppression depends on individual properties of multipotent mesenchymal stromal cell donor. Treatment with IFNγ did not significantly affect the intensity of inhibition of lymphocyte proliferation by these cells.

[Characteristics of stromal cell precursors in patients after allogeneic bone marrow transplantation].

UNLABELLED: AIM. To study the elements of the mesenchymal stromal cell compartment (multipotent mesenchymal stromal cells (MMSCs)) and their more mature progenies of fibroblast colony-forming units (CFU-F) in patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT). SUBJECTS AND METHODS. The total production of MMSCs after 5 passages, the time of their growth, and the concentration of CFU-F in the bone marrow from patients were determined using the control sections before transplantation and over time for 2 years after allo-HSCT. What is more, the genetic affiliation of the MMSCs from the patients after allo-HSCT and their immunophenotype were studied. RESULTS: The MMSCs from the patients after allo-HSCT belong to a recipient and have the immunophenotype that meets the international standard for these cells. The total production of MMSCs in the cultures obtained from the bone marrow of the patients with hematologic diseases was decreased. Not all the samples from the patients after allo-HSCT are able to undergo 5 passages. In addition, the time of growth substantially increases and the total production of cells decreases in all the analyzed cultures. These indicators are gradually restored; however, they never achieve the mean values in donors. The concentration of CFU-F in the bone marrow from the patients are reduced as compared to that in the donors prior to transplantation and decreased still further after allo-HSCT. These cell precursors are not restored for at least 2 years following allo-HSCT. CONCLUSION. Both examined categories of the cell precursors of the stromal environment suffer from both the disease itself and allo-HSCT in the patients with hematologic diseases.

[Effect of bortezomib on the efficiency of hematopoietic stem cell mobilization in patients with multiple myeloma].

AIM: To study the results of mobilizing and collecting autologous hematopoietic stem cells (HSC) in patients with multiple myeloma (MM) receiving bortezomib as part of induction therapy regimens. MATERIALS AND METHODS: In June 2001 to April 2010, the Department of Bone Marrow Transplantation, Hematology Research Center, Ministry of Health and Social Development of Russia, mobilized autologous HSC in 93 patients with MM, by using cyclophosphan (CF) and granulocyte colony-stimulating factor. The analysis covered 73 patients who received VAD and/or bortezomib-containing courses as induction therapy. Group 1 comprised 30 patients whose induction therapy was performed as 3-4 courses of VAD. Group 2 included 19 patients who had 2-4 courses of PAD or 4-8 courses of bortezomib + dexamethasone in addition to 1-3 courses of VAD. Group 3 combined 24 patients who used 6-8 courses of bortezomib + dexamethasone or 3-4 courses of PAD + 4-6 courses of bortezomib + dexamethasone. RESULTS: In Group 1 patients whose induction therapy was performed as 3-4 courses of VAD, baseline peripheral blood CD34+ cell counts were 3,575 +/- 631 in 1 ml, which was statistically significantly higher than those in Group 2 patients who had bortezomib-containing courses in addition to VAD courses. In Group 2 patients, premobilization CD34+ cell counts were 2,164 +/- 516 in 1 ml. The lowest blood CD34+ cell levels (1,586 -/+ 405 in 1 ml) were observed in Group 3 patients in whom bortezomib was used as first-line therapy. In Group 1 patients, the maximum peripheral blood counts of CD34+ cells were 322,287 +/- 73,994 in 1 ml, which was significantly higher than their maximum level in Groups 2 (231,624 +/- 39,708 in 1 ml) and 3 (161,007 +/- 44,266 in 1 ml) patients. The efficiency of mobilization proved to be high; more than 4.0.10(6)/kg of CD34+ cells were collected in all the patients with bortezomib-containing induction therapy, which allowed two autologous HSC transplantations to be carried out. CONCLUSION: Adding bortezomib at the stage of induction has no significant impact on the results of HSC mobilization and collection. By taking into account the possibility of achieving a complete or very good partial response in 40-60% of the patients using the bortezomib-containing regimens as first-line therapy, bortezomib should be considered as an essential drug as part of induction therapy.

[Dynamic study of Bcl-2, Bax, p53, and ACE expression in CD34+ cells of peripheral blood and bone marrow in acute leukemia patients in the course of induction chemotherapy].

AIM: To determine unbalance in the system of programmed cell death in the cells CD34+ of the bone marrow (BM) and peripheral blood (PB) before and after cytostatic impact in acute leukemia (AL). MATERIAL AND METHODS: Flow cytoflowmetry estimated expression of Bcl-2, Bax, p53 and ACE in the cells CD34+ of BM and PB from 10 AL (4 AML and 6 ALL) patients. PB and BM samples were studied before polychemotherapy (PCT) and in the course of induction treatment: on day +8, +21 (blood only), +36 - 38. Control group consisted of 4 BM donors. RESULTS: The number of CD34+ cells expressing Bcl-2 in AL patients was 46,5 +/- 9,35 % in BM and 39,4 + 10,8 % in PB, in healthy donors - 9 and 32,8 %, respectively. Bax expression in AL patients' cells CD34+ of BM versus this expression in donors was 3 times higher (36,7 +/- 8,1 and 14,8%, respectively), of PC - 2 times lower (40,7 +/- 6,59 and 75,8%, respectively). Expression of p53 in AL patients was 36,8 +/- 9 % in BM and 26 +/- 7,4 % in PB, in donors - 28,2 and 65 %, respectively. ACE expression on the cells CD34+ in AL patients in early disease was 62 +/- 7,57 % in BM and 48 +/- 8,1 % in PB, in donors - 40 and 85 %, respectively. Moreover, there were significant changes in expression of Bcl-2 in BM and Bax, ACE and p53 in PB in the cells CD34+ in AL patients during and after induction PCT. CONCLUSION: The above changes evidence for unbalance of pro- and antiapoptosis proteins of regulators in AL patients. PCT changes profile of expression of these proteins, but not to the level of healthy donors.

[Leukemic dendritic cells in patients with acute myeloid leukemia].

AIM: To determine surface and intracellular expression of ACE antigen and Bip shaperon on leukemic dendritic cells (LDC); to study expression of ACE genes and Bip, Calnexin, calreticulin shaperons in LDC at diagnosis of acute myeloid leukemia (AML) under standard and stress cultivation. MATERIAL AND METHODS: Expression of ACE antigens and Bip was studied with immunophenotyping and flow cytometry using monoclonal antibodies to shaperon Bip and to CD143), expression of genes of ACE and shaperons Bip, Calnexin, Calreticulin--with polymerase chain reaction (RT-PCR). Dendritic cells (DC) were obtained by culturing of a monoclonal fraction of donor peripheral blood and AML patients in the presence of 180 ng/ml calcium ionophor A23187 (Sigma) for 4 days in parallel at 37 degrees C and 33 degrees C in the atmosphere of 5% CO2. The trial included 9 patients (5 males and 4 females) aged 39-53 years (median 43 years). The control group consisted of 8 healthy donors. RESULTS: Lowering of cultivation temperature did not increase ACE expression. Intracellular shaperon Bip rose insignificantly (1.3-fold) in DC of the controls. ACE and Bip shaperon expression on LDC membrane increased 15- and 11-fold, respectively, while the level of intracellular ACE and Bip decreased 11- and 2-fold, respectively. Expression of the genes was investigated in cultivation temperature lowering from 37 to 33 degrees C and was presented as a logarithmic scale. Changes in expression of the genes Bip, Calnexin, Calreticulin in LDc and DC of the controls were insignificant. ACE expression in LDC significantly differed from ACE gene expression in DC (p = 0.05). CONCLUSION: LCD and DC of healthy donors are cells which differ by genetic and functional characteristics. Therefore, LDC may response inadequately in development of antitumor immune response. The phenomenon of ACE antigen expression normalization on cell membrane in stress open new opportunities for regulating functional activity of LDC.

[Transplantations of allogenic and autologous hemopoietic stem cells in acute leukemia (results of 20-year experience)].

AIM: To analyse results of transplantation of allogenic and autologous hemopoietic stem cells (allo-THSC and auto-THSC) with myeloablation preconditioning in patients with acute leukemia (AL) performed in 1987-2006. MATERIAL AND METHODS: A total of 71 allogenic and 45 autologous THSC were performed in 116 patients with different AL variants. Conditioning in all allo-THSC included busulfan (16 mg/kg) and cyclophosphamide (120 mg/kg). This regimen was used in 29 recipients of auto-HSC. Cyclophosphamide in a dose 120 mg/kg and total radiation of the body in a dose 12 Gy were given to 16 recipients. Overall, relapse-free and event-free survival of patients after THSC were analysed as well as early (first 100 days) and overall lethality. Auto-THSC in 15 patients was for the first time followed by immunomodulating therapy aimed at prevention of AL relapses: in acute myeloid leukemia ATRA in combination with alpha-interferon, in acute lymphoblastic leukemia (ALL)--ronkoleukin, interleukin-2 preparation. RESULTS: Overall survival of AL patients after allo-THSC for the observation period increased from 31 to 58%, early lethality fell from 44 to 4%. Results of allo-THSC conducted in the first complete remission were much better than in patients with other AL stages at the time of THSC. After auto-THSC 5-year survival rose from 22 to 60% while early lethality reduced from 33 to 4%. Administration of immunomodulating therapy after auto-THSC increases 5-year survival from 35 to 80%. CONCLUSION: Outcomes of THSC in AL has improved for the last 20 years. Outcomes of allo-THSC performed in the first complete remission are much higher. Immunomodulating therapy after auto-THSC promoted better results.

[Characteristics of T-cell immunity in patients with acute leukemia].

AIM: To study a cytokine profile of cytotoxic T-cells and T-helpers at diagnosis of acute myeloid and lymphoblastic leukemia (AML and ALL) and at different stages of its treatment. MATERIAL AND METHODS: T-cell population of peripheral blood lymphocytes was studied in 19 AML patients at diagnosis, 13 AML patients in remission, 5 AML patients in developing recurrence and 5 ALL patients at diagnosis. The control group consisted of 10 healthy donors. A mononuclear fraction of peripheral blood cells was stained with monoclonal antibodies. Surface expression of CD3, CD4, CD8 antigens was estimated with the use of flow cytofluorimeter FACS Calibur. Part of the cells were cultured for production of activated lymphocytes. The cytokine profile of T-cells was investigated according to the modified method of Rostaing et al. in the population of CD3+CD8+ and CD3+CD8- T-cells using the flow cytofluorimeter FACS Calibur. The results were processed with the programs CellQuest and WinMDI, Statistica Module Switcher. RESULTS: The percentage of CD3+CD8+ cells producing interferon-gamma (IF-g) was the same in AML patients and donors. The percentage of CD3+CD8- cells (T-helpers) producing IF-g was similar in the patients at diagnosis and in complete remission being higher than in healthy donors. The number of CD3+CD8- cells producing IF-g was reduced in recurrence while the number of CD3+CD8+ and CD3+CD8- cells producing interleukin-4 (IL-4) increased. In the onset of the disease, ALL patients had significantly increased percentage of CD3+CD8- IL-4 cells. CONCLUSION: The results obtained evidence for different polarization of T-cell immunity in AML and ALL patients at the time of diagnosis, remission and recurrence: in the onset of AML there was an increase in the production of Th1--proinflammatory cytokines, in ALL--in percent of Th-2 cytokines. In AML recurrence the balance of cytokines shifted in the direction of Th-2 response.

[Detection of minimal residual disease in patients with acute myeloid leukemia]. / Vyiavlenie minimal'noi ostatochnoi bolezni u bol'nykh ostrymi mieloidnymi leikozami.

AIM: Detection of minimal residual disease (MRD) in patients with acute myeloid leukemia (AML) and the study of its correlations with duration of recurrence-free interval. MATERIAL AND METHODS: Bone marrow samples obtained from 37 AML patients before treatment were studied at two-color flow cytometry. The panel of monoclonal antibodies to T- and B-cell, myeloid antigens was used. The residual cells were estimated in 20 patients in remission. RESULTS: 78% cases were diagnosed to have an anomalous immunophenotype including coexpression of lymphoid and myeloid antigens, asynchronous expression of myeloid antigens. In the first remission the residual cells were detected in 20 patients due to aberrant antigen expression. The presence of MRD was stated if bone marrow contained more than 0.12% leukemic cells. The duration of the first remission and MRD correlated. 8 patients with MRD had remission for 3 to 6 months (median 4.7 months). 12 patients free of MRD were in remission for more than 6 months (for 8 to 26 months, median 19.7 months). The threshold level of the residual cells (0.12%) was confirmed statistically using the three-parameter probability model. CONCLUSION: This study confirms feasibility of using flow cytometry for detection of residual cells. MRD and duration of the first remission correlate. Long-term observation of large groups of AML patients will try the validity of the above statistical model.

[Superhigh doses of dexamethasone in treatment of refractory forms of acute lymphoblast leukemia of adults]. / Sverkhvysokie dozy deksametazona v lechenii refrakternykh form ostrogo limfoblastnogo leikoza vzroslykh.

AIM: Assessment of high-dose dexamethasone efficacy in combination with standard drugs (adriablastin, vincristin, alpha-asparaginase) in patients with refractory acute lymphoblastic leukemia (ALL). MATERIAL AND METHODS: A pilot multicenter trial with participation of hematological departments of Hematological Research Center (Moscow), municipal hospital N 1 (Krasnoyarsk), municipal hospital N 8 (Yaroslavl), Research Institute of Hematology and Blood Transfusion (Kirov) included 34 patients (10 patients with late recurrences, 24--with primary resistant forms, early and secondary recurrences). RESULTS: In patients with late ALL recurrences a complete remission (CR) was achieved in 70% cases, the median being 10 months. In patients with primary resistant ALL, early and secondary recurrences CR reached 37.5%, the median was 14 months. CONCLUSION: The program HiDexa is highly effective: overall complete remission rate reached 47%, median of complete remission duration was 10 months. Dexamethasone in high doses must be used only intravenously.

[The efficacy of high-dose chemotherapy and the transplantation of autologous hemopoietic cells in lymphogranulomatosis]. / Effektivnost' vysokodoznoi khimioterapii i transplantatsii autologichnykh gemopoéticheskikh kletok pri limfogranulematoze.

AIM: To determine clinical effectiveness of high-dose polychemotherapy (PCT) and transplantation of autologous hemopoietic cells (TAHC) in patients with lymphogranulomatosis (LGM). MATERIAL AND METHODS: 27 LGM patients aged 16-42 years who have undergone TAHC after high-dose PCT (BEAM--17 patients or CBV--10 patients). 4 patients given high-dose PCT were in the first-second complete remission (CR), 7 patients--in the first partial remission (PR). Prior to TAHC, 8 patients had one, two and more relapses of LGM, and 8 patients had no remission at all. Bone marrow, hemopoietic blood cells and both were transplanted to 17, 2 and 8 patients, respectively. Mobilization of hemopoietic blood cells and stimulation of hemopoiesis after TAHC were achieved using colony-stimulating factors. RESULTS: The treatment resulted in CR or PR (from 6 to 95 months) in 70.4% of patients. The remission duration varied depending on the disease phase at transplantation. Four patients who underwent TAHC in PR maintained it for 13-95 months (median 47.5 months). Lasting remissions (29-59 months) were achieved in 42.9 and 37.5% of patients who underwent TAHC in the first PR or in recurrent LGM. None of the patients was in remission longer than 2 years after TAHC if high-dose PCT was conducted in advanced tumor process due to resistant LGM or inadequate previous treatment. Infectious complications lethality early after the transplantation reached 7.4%(2 patients). CONCLUSION: High-dose PCT followed by TAHC is effective in LGM if the tumor is chemosensitive.

[Effectiveness of allogeneic bone marrow transplantation in patients with acute leukemia in complete remission and in patients with chronic myeloid leukemia during the chronic phase]. / Effektivnost' transplantatsii allogennogo kostnogo mozga u bol'nykh ostrymi leikozami v faze polnoi remissii i u bol'nykh khronicheskim mieloleikozom v khronicheskoi faze.

AIM: To study clinical efficiency of allogenic bone marrow transplantation (ABMT) in patients with acute leukemia (AL) in the first remission and in patients with chronic myeloid leukemia (CML) in chronic phase; to analyse overall and recurrence-free survival in relation to the diagnosis and age. MATERIALS AND METHODS: 26 patients with AL and 24 ones with CML (29 males and 21 females) were subjected to ABMT within 10 last years. Median of age in AL and CML was 24.5 and 25.5 years, median of the time since the diagnosis was 9 and 13 months, respectively. Follow-up since the ABMT made up 67.5 months (31-107) and 38 months (6-108), respectively. Conditioning was made with cyclophosphamide (120 mg/kg) plus total radiation of the body (12 Gy) in 16 patients, myelosan (mileran) in a dose 16 mg/kg plus cyclophosphane (120 mg/kg) in 34 patients. The marrow was taken from HLA-identical sibs, enzygotic twins (5 recipients). Cytogenetic investigations were made in CML. The retention of the transplant was controlled by immunological and molecular tests. RESULTS: Among AL patients 50% are still alive. Probability of 80-month survival reached 55%, 110 months--42%. Probable recurrence-free survival was 78%. All the patients are in a complete clinico-hematological remission. Among CML patients 75% are still alive. Of them 89% had a complete hematological remission, 72% are in a complete hematological and cytogenetic remission. Probable 110 month survival equals 75%, probability to survive without recurrence--52%. Early lethality (100 days) of toxic and infectious complications was as low as 10 and 6%, respectively. Frequency of lethal acute secondary disease was under 8%. CONCLUSION: ABMT made in AL patients during the first complete remission and in CML patients in the chronic phase brings about very good results which are much better than after routine cytostatic chemotherapy.

[A trial of the clinical use of thienam in treating severe and life-threatening suppurative-septic complications]. / Opyt klinicheskogo primeneniia tienama v lechenii tiazhelykh i ugrozhaiushchikh zhizni gnoino-septicheskikh oslozhnenii.

The authors share their experiences with treatment of 12 patients using the antibiotic Tienam. No complications resulting from the administration of this medicine were noted. High therapeutic efficiency of Tienam has been shown. Recovery was noted in 11 out of 12 patients.