Recombinant MHC Tetramers for Isolation of Virus-Specific CD8+ Cells from Healthy Donors: Potential Approach for Cell Therapy of Posttransplant Cytomegalovirus Infection.

Patients undergoing allogeneic hematopoietic stem cell transplantation have a high risk of cytomegalovirus reactivation, which in the absence of T-cell immunity can result in the development of an acute inflammatory reaction and damage of internal organs. Transfusion of the virus-specific donor T-lymphocytes represents an alternative to a highly toxic and often ineffective antiviral therapy. Potentially promising cell therapy approach comprises transfusion of cytotoxic T-lymphocytes, specific to the viral antigens, immediately after their isolation from the donor's blood circulation without any in vitro expansion. Specific T-cells could be separated from potentially alloreactive lymphocytes using recombinant major histocompatibility complex (MHC) multimers, carrying synthetic viral peptides. Rapid transfusion of virus-specific T-cells to patients has several crucial advantages in comparison with methods based on the in vitro expansion of the cells. About 30% of hematopoietic stem cell donors and 46% of transplant recipients at the National Research Center for Hematology were carriers of the HLA-A*02 allele. Moreover, 94% of Russian donors have an immune response against the cytomegalovirus (CMV). Using recombinant HLA-A*02 multimers carrying an immunodominant cytomegalovirus peptide (NLV), we have shown that the majority of healthy donors have pronounced T-cell immunity against this antigen, whereas shortly after the transplantation the patients do not have specific T-lymphocytes. The donor cells have the immune phenotype of memory cells and can be activated and proliferate after stimulation with the specific antigen. Donor lymphocytes can be substantially enriched to significant purity by magnetic separation with recombinant MHC multimers and are not activated upon cocultivation with the antigen-presenting cells from HLA-incompatible donors without addition of the specific antigen. This study demonstrated that strong immune response to CMV of healthy donors and prevalence of HLA-A*02 allele in the Russian population make it possible to isolate a significant number of virus-specific cells using HLA-A*02-NLV multimers. After the transfusion, these cells should protect patients from CMV without development of allogeneic immune response.

Identification of a novel allele HLA-C*12:138 in Russian patient by haplotype-specific sequence-based typing.

HLA-C*12:138 differs from HLA-C*12:03:01:01 by a single change, resulting in an amino acid substitution.

[Genes of killer cell immunoglobulin-like receptors and their HLA ligands after allogeneic hematopoietic stem cell transplantation in myeloid leukemia patients].

AIM: To study the impact of the genes of donor killer cell immunoglobulin-like receptors (KIR) and HLA-KIR ligands on overall (OS) and event-free survival (EFS) rates in patients with myeloid leukemia after transplantation with allogeneic hematopoietic stem cells (allo-HSCT) from HLA-identical related and HLA-compatible unrelated donors. SUBJECTS AND METHODS: The investigation enrolled 29 patients who had undergone allo-HSCT from KIR-genotyped donors at the Department of Bone Marrow Transplantation, Hematology Research Center (see symbol) in 2010-2013. OS and EFS rates after allo-HSCT were calculated using the Kaplan-Meier method. RESULTS: The main predictor of recurrence and survival in patients after allo-HSCT was a recurrence-risk group the patient belonged to before transplantation. The standard-risk group patients whose donors had telomeric gene-content motifs of KIR-B haplotypes had higher EFS rates than those whose donors lacked these genes. The standard-risk patients homozygous for HLA-1 alleles (i.e. without HLA-C2 ligand) tended to have higher EFS rates, so did the patients without HLA-Bw4 ligand. CONCLUSION: The donors having telomeric gene-content motifs of KIR-B haplotypes are more preferred for allo-HSCT for patients with myeloid leukemia as the presence of donor telomeric KIR-B genes increases EFS rates in standard-risk patients.

[Selection of an unrelated donor for hematopoietic stem cell transplantation. HLA haplotypes in patients with blood system diseases].

AIM: To study the distribution of HLA-A*-B*-C*-DRB1 *-DQB1 * haplotypes in patients with blood system diseases, to establish the most common HLA haplotypes, and to compare the findings with the data on the frequency and distribution of the highest-frequency HLA haplotypes in donors of a number of leading registries. SUBJECTS AND METHODS: In 2008-2012, the Hematology Research Center, Ministry of Health of the Russian Federation, examined 203 patients with blood system diseases who needed allogeneic hematopoietic stem cell transplantation (allo-HSCT) and their 386 blood relatives. Typing ascertained the kind of HLA haplotype in all the patients. Among the patients, there were 97 men who were aged 17 to 64 years (median 38 years) and 106 women who were aged 18 to 59 years (median 40 years). RESULTS: The examinees were found to have 265 different HLA haplotypes. There were 21 high-frequency HLA haplotypes; of them 7 belonged to 10 HLA haplotypes that are most frequent in the representatives of the Caucasoid race. Nearly 30% of the patients who needed allo-HSCT and had no HLA-identical siblings had HLA haplotypes out of the 10 ones that are most common in the representatives of the Caucasoids and thus could expect to find a compatible unrelated donor for a short time. The examinees were found to have a wide variety of HLA haplotypes (265 types in 203 persons). This variety, as well as the extreme polymorphism of HLA alleles, shows that there should be large registries of HLA-typed bone marrow donors in the country. These registries increase the chance to find a HLA-compatible unrelated donor for a short time for a patient with blood disease who has an indication for hematopoietic stem cell transplantation. The performed study supported that there were regional features in the distribution of HLA haplotypes within the same ethnic group. CONCLUSION: The chance to find a HLA-compatible donor for Russian patients in the large national registry that accumulates donors from different regional populations is substantially higher than that in the foreign registries. To create large cohorts of HLA-typic bone marrow donors from different regions of the country will substantially increase the chance of patients with blood system diseases to find a HLA-compatible unrelated donor.

[Search for non-relative donor by the Russian register of bone marrow donors]. / Poisk nerodstvennogo donora Rossiiskim registrom donorov kostnogo mozga.

AIM: To select maximally HLA compatible donor for hematological patients who need transplantation of bone marrow from non-relative donor. MATERIALS AND METHODS: 75 patients with hematological malignancy were observed. All of them have indications to non-relative transplantation of the bone marrow. Methods of polymerase chain reaction with sequence-specific primers and classic microlymphocytotoxic test were used. RESULTS: Typing of HLA antigens of class I and alleles of class II loci enabled search for non-relative donor for transplantation of bone marrow in accordance with the requirements of the European Federation of Immunogenetics. Most of the patients (86.6%) had at least one potential HLA-A, -B, -DR compatible donor. Half of the patients had potential donors typed at the allele level by class II loci. This diminishes time of HLA compatible donor selection. CONCLUSION: DNA typing enables the search for the non-relative donors meeting modern requirements. This allowed 5 non-relative bone marrow transplantations.

[Immunogenetic markers in hemoblastosis]. / Immunogeneticheskie markery gemoblastozov.

AIM: To establish for hemoblastoses general and individual associations between HLA genes and predisposition or resistance to malignant transformation of hemopoiesis. MATERIAL AND METHODS: Antigens of class I and class II genes were determined by microlymphocytotoxic test and DNA typing (PCR-SSP) in 30 patients with acute myeloblastic leukemia (AML), 47 patients with acute lymphoblastic leukemia (ALL), 52 patients with chronic myeloblastic leukemia (CML), 58 patients with lymphogranulomatosis (LGM) and 224 donors. RESULTS: HLA specificities were elucidated frequency of which was high in all the above nosological entities: Cw7 (RR 2.5 to 5.22), DRB1*11 (RR 2.3 to 4.88). Frequency of antigen B5 was high in three diseases (RR 2.14-2.6). A number of specificities have distribution deviations typical for only one or two diseases (A19--AML and CML, B27--ALL, DRB1*08--AML). The HLA-gene was detected the frequency of which was low in three diseases--DRB1*07 (RR 0.39 to 0.11). Individual preventive HLA genes for AML, ALL, LGM were A2, B8; A1, B8, B40; DRB1*01, respectively. CONCLUSION: Cw7, DRB1*11, B5 are HLA-specificities marking hemopoiesis predisposition to malignant transformation in general. Development into a certain nosological entity depends on combination of general HLA-genes with markers of individual hemoblastosis. DRB1*07 is a general preventive gene. Preventive HLA genes can be also individual for the above hematological diseases.

[The factors of hereditary predisposition to lymphogranulomatosis]. / Faktory nasledstvennoi predraspolozhennosti k limfogranulematozu.

AIM: To determine genetic factors of predisposition marked HLA with reference to serological and molecular characteristics. MATERIALS AND METHODS: Four groups of patients were included in the study: 51 patients with lymphogranulomatosis (LGM), 33 healthy relatives of these patients, 37 patients with chronic myeloid leukemia (CML), 24 healthy relatives of these CML patients. 224 donors served control. HLA-antigens were identified with the lymphocytoxic test and PSR-MSSR. Results of typing of class II antigens were taken into consideration in coincidence of serological and DNA typing. The significance of the differences was estimated according to the chi-square criterion. RESULTS: Differences in frequency of distribution of HLA-antigens (subloci A and B) were not found. Antigen CW7 was present in 70, DR5 in 60, DR6 in 50% of LGM patients. This frequency was much higher than in control groups. Carriers of CW7 are at 7 times higher risk to develop LGM. Among LGM patients the number of homozygous individuals is higher than in healthy ones (50 and 15.6%) while the number of individuals with a complete set of HLA-A.B antigens is significantly less. CONCLUSION: Genetic predisposition to LGM is predetermined by HLA antigens CW7, DR5, DR6. Genes HLA-DR1 and HLA-DR7 protect carriers from factors provoking LGM. Common HLA genes in the parents predispose their children to LGM. Insufficiency of the phenotype is a factor predisposing to LGM.

[The effect of endovascular helium-neon laser therapy on the immune status of patients with acute calculous pyelonephritis]. / Vliianie éndovaskuliarnoi gelii-neonovoi lazernoi terapii na immunnyi status bol'nykh ostrym kal'kuleznym pielonefritom.

Cellular immunity was assessed in 48 patients with acute calculous pyelonephritis exposed to intravenous He-Ne laser therapy. It was found that endovascular He-Ne laser therapy in the study regimens corrects immunological abnormalities arising in acute calculous pyelonephritis.

[Beta 2-microglobulin kinetics during hemodialysis with different membranes]. / Kinetika beta 2-mikroglobulina vo vremia gemodializa s razlichnymi membranami.

Dialysers F6 (polysulphon membrane) and Altra Nova-170 (acetate cellulose-H membrane) were tested for effects on concentration of beta 2-microglobulin (beta-2 MG) in the course of hemodialysis. F6 produced a rise in beta-2 MG levels (50.0 +/- 7.7 to 59.2 +/- 9.5 mg/l). Altra Nova-170 induced no significant changes in these levels. Dialyser F6 seems to stimulate generation or release of beta-2 MG the input and output amounts of which indicate that the process may be confined to dialyser interior. On hemodialysis minute 15 a sharp fall in beta-2 MG concentrations was registered in the use of either device. This phenomenon resembles leukopenic effect of cellulose membranes. Hemodialysis kinetics of beta-2 MG is likely to depend on leukocyte activation.