Thalassemia and qualitative hemoglobinopathy are hereditary disorders of Hb synthesis that lead to change in the Hb conformation or a decrease in the synthesis of structurally normal Hb, and consequently, to erythron pathology. Many variants of Hb are unstable or have altered affinity for oxygen, and, in heterozygous form can be associated with clinical and hematological manifestations (hemolytic anemia, hypochromic microcytic anemia, erythrocytosis). HbD-Punjab [β121 (GH4) Glu → Gln; HBB: C.364G> C] is variant of Hb carrying the amino acid substitution in the 121 position of β-globin chain. In all cases reported so far, patients with HbD-Punjab/β+-thalassemia (IVSI+5 G-C) combination experienced typical thalassemia with hypochromic microcytosis. HbD-Punjab was detected by electrophoresis from 37 to 94% of total Hb. The article describes rare clinical case of the cohabitation of HbD-Punjab/β+-thalassemia (IVSI+5 G-C) in a patient with homozygous variant of Gilbert's syndrome observed in AS Loginov Moscow Clinical Scientific Center.

Background. Pure red cell aplasia (PRCA) is a rare syndrome characterized by a decrease of erythroid progenitor cell count in the bone marrow. M-gradient with both a light and a heavy chain types in PRCA patients is a rare phenomenon which is considered to be a specific form of the disease. Aim. To review a clinical presentation, diagnostic capabilities, and treatment outcomes of PRCA with M-gradient. Materials & Methods. The analysis included 10 patients. The most effective empirically established treatment program was 200-400 g of cyclophosphamide 2-3 times a week to a total dose of 6-10 g and loading courses of 100-120 mg of oral and 180-240 mg of intravenous prednisone daily within 5 days. On the 6th day prednisone injections were discontinued, and from the 7th day the oral dose of prednisone was gradually reduced to permanent discontinuation in 2-3 days. This treatment course was repeated 1-3 times at intervals of a week. Targeted enzyme immunoassay of M-gradient was performed in 4 patients in order to determine whether M-gradient is the sum of two antibody types, i.e. erythrokaryocyte antibodies and secondary anti-idiotype antibodies against primary antibodies. Results. The total of 7 out of 10 PRCA patients reached complete remission within the period from 9 months to 22 years of follow-up, in 3 patients no remission was achieved. M-gradient contained IgG (n = 9) and IgA (n = 1) oligoclones. In typing it consisted of IgGA (n = 4), IgGK (n = 5) and IgAK (n = 1). M-gradient enzyme immunoassay showed no primary and secondary anti-idiotype antibodies. Conclusion. The obtained results allow to regard gammopathy in PRCA as an effect of oligoclonal hyper-immunoglobulin without any pathogenetic connection between M-gradient and PRCA.

AIM:To study the clinical manifestations, diagnosis, and treatment of lymphoproliferative diseases (LPD) concurrent with tuberculosis.SUBJECTS AND METHODS:In 1990 to 2013, the Hematology Research Center, Ministry of Health of Russia, followed up 4422 patients with LPD. Lymphomas and leukemias were diagnosed using the universally protocols. Tuberculosis was verified by the results of a comprehensive examination involving the histological study of biopsy specimens.RESULTS:Tuberculosis was identified in 85 (2%) patients with LPD. According to the nosological entity, the tuberculosis detection rates were 3% (40/1350) in Hodgkin lymphoma (HL), 1.2% (20/1627) in aggressive lymphomas, 1.4% (16/1136) in mature cell lymphomas and chronic lymphocytic leukemia, and 2.9% (9/309) in hairy cell leukemia. In accordance with its site, pulmonary tuberculosis was 73%; extrapulmonary tuberculosis, 14%; generalized tuberculosis, 12%. In pulmonary tuberculosis, its disseminated and focal involvements were found in 71 and 18% of cases, respectively. Tuberculosis was detected in 43% of the patients with HL in remission; it occurred only in other hemoblastoses in its active phase. When tuberculosis and LPD were simultaneously found, both diseases were concurrently treated. If the chemotherapy of LPD was effective, tuberculosis was cured in all the patients.CONCLUSION:Patients with LPD are a group at increased risk for tuberculosis. The diagnosis of recurrent LPD must be histologically proven. When tuberculosis and LPD are simultaneously found, both diseases should be concurrently treated.

Summary. M ale infertility remains one of the main side eff ects of chemotherapy for Hodgkin’s lympho-ma (HL). Aim of the study was to assess the fertility of adolescents and young adults treated for HL. Sixty-three patients aged 19—37 years (median 27 years), treated in 1993—2011, were examined. Spermatogenesis, plasma sex hormone levels, and number of children born after therapy were studied. The patients were divided into 4 groups, depending on schemes of chemotherapy: 1) (n = 11) ABVD; 2) (n = 30) BEACOPP-14; 3) (n = 15) MOPP + ABVD; 4) (n = 7) BEACOPP-Esc (strong). No azospermia was recorded in group 1; in group 2 it was recorded in 43% (13/30), in group 3 in 40% (6/15), and in group 4 in 57% (4/7). Seventeen healthy babies were born after chemotherapy: 3 after assist reproductive technology, 14 without it. Testosterone level depletion was detected in 39% (25/63). High incidence of spermatogenesis disorders makes it necessary to regular con-sult endocrinologist or andrologist. Cryopreservation of the semen in liquid nitrogen should be recommended to all young patients with HL before any program of chemotherapy. Key words:

Male infertility remains one of the main side effects of chemotherapy for Hodgkin's lymphoma (HL). Aim of the study was to assess the fertility of adolescents and young adults treated for HL. Sixty-three patients aged 19-37 years (median 27 years), treated in 1993-2011, were examined. Spermatogenesis, plasma sex hormone levels, and number of children born after therapy were studied. The patients were divided into 4 groups, depending on schemes of chemotherapy: 1) (n = 11) ABVD; 2) (n = 30) BEACOPP-14; 3) (n = 15) MOPP + ABVD; 4) (n = 7) BEACOPP-Esc (strong). No azospermia was recorded in group 1; in group 2 it was recorded in 43% (13/30), in group 3 in 40% (6/15), and in group 4 in 57% (4/7). Seventeen healthy babies were born after chemotherapy: 3 after assist reproductive technology, 14 without it. Testosterone level depletion was detected in 39% (25/63). High incidence of spermatogenesis disorders makes it necessary to regular consult endocrinologist or andrologist. Cryopreservation of the semen in liquid nitrogen should be recommended to all young patients with HL before any program of chemotherapy.

AIMTo give data on the frequency of recurrent hairy cell leukemia (HCL) and to characterize the immediate and late results of its treatment in this group of patients.MATERIALS AND METHODSThe data on the frequency of recurrences were analyzed in 165 patients with HCL after remission achieved by the purine analogue cladribin in the period 1995 to 2011. The treatment of recurrent HCL included splenectomy, interferon-a, cladribin, and rituximab.RESULTSAfter a course of cladribin therapy, the total frequency of recurrent HCL was 22%. The high (47%) frequency of recurrences was found in young patients (less than 45 years) as compared to that (9%) in older patients. A combination of cladribin and rituximab showed a high efficacy in treating the early recurrence of HCL.CONCLUSIONThe differences found in the frequency of recurrences give grounds to incorporate rituximab into the standard therapy regimen for HCL in young patients and in patients with early disease recurrence.

Infectious mononucleosis (IM) runs with a definite clonal response of T-lymphocytes to Epstein-Barr virus. The authors tried to utilize standard molecular method PCR-SSCP-TCRgamma for detection of some T-lymphocyte clones in IM. Determination of T-cell clonality by rearrangements of T-cell receptor gamma-chain genes was conducted with polymerase chain reaction (PCR) followed by analysis of conformation polymorphism of single chain products (SSCP). Of 20 examinees with IM, 19 ones demonstrated monoclonal or oligoclonal picture while out of 20 control patients with acute respiratory viral infections or lacunar tonsillitis, 18 had polyclonal picture. Thus, immune response in IM is accompanied with emergence of one or some clones of T-cells the size of which is within sensitivity of molecular methods of T-cell clonality identification.

Aim. To distinguish T-cell lymphomas and reactive T-cell proliferation it is important to confirm the. ability of T-cells to be cloned. Conventional histological and immunophenotypic methods fail to deter-cells to be cloned. An experience in the use of. detection of T-cell receptor gene mine the ability of T gamma-chain (TCRy) rearrangement for determining T-cellular clonality is described.Material and methods. Polymerase chain reaction (PCR) and single strand conformational polymorphism (SSCP) were used to determine T-cell clonality. Twenty healthy donors; 28 patients with T-lymphomas, and 26 patients with various non-T-cell lymphoproliferative disorders or reactive processes were studied.Results. T-cell monoclonality was detected in 23128 (8276) T-cell lymphoma cases, whereas in all the samples from normal subjects a polyclonal pattern of rearrangements TCRy was found. The sensitivity of the method was estimated as 2,5%, 7%, and 10% was demonstrated for bone marrow, spleen, and peripheral blood, respectively.Conclusion. PCR-SSCP for TCRy was found to be a useful supplement to routine histological and immunophenotypic methods in the diagnosis of T-cell lymphomas.

Objective. Pts with chronic clonal proliferation of large granular lymphocytes (LGL leukemia) often have neutropenia, splenomegaly, and rheumatoid arthritis (RA), thereby resembling the manifestations observed in pts with Felty’s syndrome. The present study sought to indicate that pts with these disorders represent two distinct subsets. We compare clinical, hematological, immunophenotiping and immunogenetic features in Felty’s syndrome pts with and without the LGL leukemia. Material and methods 10 pts with T-LGL leukemia were studied. Surface phenotype was estimated using monoclonal antibodies CD8-PE and CD3-FITC/CD16-PE (two-color) (Caltag, USA) by the flow cytometric analysis (Partec, Daco). Analysis of TCR gene rearrangement was performed by using PCR-LIS SSCP (low ionic strength single strand conformational polymorphism). Comparison with Felty s syndrome and RA pts based on the review of literature. Results. LGL leukemia is a distinct clinicopathologic entity often associated with RA. LGL leukemia pts with RA showed the same immunogenetis associations seen in RA/Felty’s syndrome, while LGL leukemia pts without arthritis did not. Conclusion. Hematologic, immunophenotyping and molecular genetic analysis are very important and highly representative tools in differential diagnosis of neutropenia in RA, and propose that Felty’s syndrome and LGL leukemia represent different variants of broader syndrome comprising RA, neutropenia, LGL expansions, and splenomegaly.

Anthracyclines are basic cytostatic drugs for the treatment of acute leukemias. Pharmacokinetics of these drugs determins their efficacy. Peak concentration (PC) correlates with response rate, for example in acute myeloid leukemia (AML) [1]. Area under the curve (AUC) parameters of daunorubicin are not optimal due to very fast phase of distribution and slow phase of elimination of the anthracycline. The usage of drug carriers -liposomes and erythrocytes—provides a new tool for overcoming these negative points. It was already proved that liposomal daunorubicin creates very high PC and large AUC [2]. It’s also supposed that liposomal formulation of daunorubicin can overcome P-glycoprotein related resistance of leukemic cells [3]. Erythrocytes as carriers of anthracyclines (doxorubicin) were already tested in a small cohort of lymphoma patients, and it was shown that this drug formulation did not create high PC but enlarge AUC [4].

AIM:Determination of the importance of serum cytokines (sCD30, sIL-2R, IL-10, IL-6) for diagnosis, response to chemotherapy and remission in patients with lymphogranulomatosis and lymphosarcoma.MATERIAL AND METHODS:Cytokine concentrations were measured in 87 samples of serum (plasma) from 54 patients by ELISA. Diagnosis of Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) was made histologically in 24 and 30 patients, respectively.RESULTS:The threshold concentrations of sCD30 (and less specific sIL-2R) for HD and NHL patients allowed to estimate sensitivity to chemotherapy after the second course. The threshold concentration of IL-10 can distinguish HD from NHL. Changes in IL-6 concentrations were nonspecific.CONCLUSION:On the basis of the threshold concentrations of sCD30 and IL-10 we offer the scheme of lymphoma diagnosis and prediction of the disease sensitivity to chemotherapy which reduce the duration of lymphoma restaging.

Aim. Determination of the importance of serum cytokines (sCD30, sIL-2R, IL-10, IL-6) for diagnosis, response to chemotherapy and remission in patients with lymphogranulomatosis and lymphosarcoma. Material and methods. Cytokine concentrations were measured in 87 samples of serum (plasma) from 54 patients by ELISA. Diagnosis of Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) was made histologically in 24 and 30 patients, respectively. Results. The threshold concentrations of sCD30 (and less specific sIL-2R) for HD and NHL patients allowed to estimate sensitivity to chemotherapy after the second course. The threshold concentration of IL-10 can distinguish HD fi-om NHL. Changes in IL-6 concentrations were nonspecific. Conclusion. On the basis of the threshold concentrations of sCD30 and IL-10 we offer the scheme of lymphoma diagnosis and prediction of the disease sensitivity to chemotherapy which reduce the duration of lymphoma restaging.

AIMTo assess diagnosis of parvovirus B19 infection (PI) in patients with aplastic crises by combined use of polymerase chain reaction (PCR) and enzyme immunoassay (EIA) of specific IgM and IgG.MATERIAL AND METHODSA total of 159 serum samples from 77 PI suspects were examined. The examination for virus DNA was conducted with modified "net" PCR in 108 samples, for specific IgM and IgG with EIA in 110 samples.RESULTSThe percentage of patients infected with parvovirus detected by PCR or EIA reached 60%. 21 of 77 patients with hemolytic anemias were infected with parvovirus B19, the virus persisting in 8 cases (40%). persistence of the virus was registered if viremia occurred in immunodeficiency due to the disease or immunosuppressive therapy. Immunity to parvovirus has not developed: IgM expression was the same as in patients without hemopoietic abnormalities, while IgG was not detected. The absence of specific immunity to parvovirus B19 occurred in patients treated with immunosuppressive drugs early after the end of viremia period in high IgM level and at the initial phase of IgG synthesis. IgM levels also remained unchanged; the level of IgG declined and was not identified furthermore. There were cases of reinfection.CONCLUSIONCombined use of PCR and EIA is optimal for diagnosis of parvovirus B19 infection in patients with hemolytic anemias. It was found that there are correlations between defects in specific immunity, persistence and immunodeficiency onset regarding viremia. Abnormal for the disease course levels of IgM and IgG indicate the persisting virus, the condition of specific immune response to parvovirus B19 and feasibility of reinfection. Reliable diagnosis of parvovirus infection is possible only in simultaneous use of PCR and EIA.