We have conducted a study of a novel monoclonal A3 antibody raised by means of hybridoma biotechnology. The study was performed with malignant cells of the patients with various lymphoproliferative disorders, and persons with nonmalignant diseases, as compared with intact lymphocytes from healthy people,using a method of immunocytochemical staining and indirect immunofluorescence technique. It was found that in cases of lymphoproliferative diseases with low proliferation rates, as based on the numbers of Ki-67 positive cells, as well as in non-malignant blood diseases, the A3 antigen was localized in nucleoli, and it was visualized as focal fluorescence. In malignant lymphoproliferative diseases with high proliferation indexes, the number of brightly fluorescent foci is observed, with formation of necklace-like structures within the nucleolar structures. The obtained data point to a diagnostic significance of A3 Mab in assessment of cellular proliferative rates in patients with various lymphoproliferative diseases. It was established that, in contrast to Ki-67, the proliferation stage could be determined for each cell, according to the number of fuorescent foci in nucleoli. This specific property of the A3 antigen points to its significance for diagnostics and malignancy staging of lymphoproliferative disorders.

The article proposes new (immune cytochemical) mode of evaluation of reaction of blast-cell transformation of human lymphocytes under their stimulation with phytohemagglutinin instead of standard radiometric method using 3H-thymidine as a marker. The reaction was assessed by means of luminescent microscope under indirect immunofluorescence using nuclear antigen Ki-67 as a marker of proliferating cells. The Ki-67 was detected using monoclonal antibodies Ki-67. The functional capacity of lymphocytes was evaluated by amount of visualized (luminescent) Ki-67 positive cells appearing after three day stimulation of proliferation with phytohemagglutinin. The comparison was made between immune cytochemical (with Ki-67 marker) and radiometric (with marking by 3H-thymidine) methods of phytohemagglutinin stimulation of lymphocytes. The article presents substantiation of advantage of immune cytochemical method of evaluation of reaction of blast-cell transformation of human lymphocytes as compared with radiometric method concerning possibility of its broader implementation in laboratory practice.

Incorporation of H-3-thymidine in the lymphocytes of short-term cultures was evaluated by MicroBeta Plus and 1409 DSA beta-scintillation counters ("Wallac", Finland). Comparable values of lymphocyte proliferative activity were obtained by both counters in specimens from 16 individuals in response to phytohemagglutinin (PHA) stimulation and in 8 donor-recipient pairs in mixed lymphocyte culture (MLC). MicroBeta Plus counter detected a trend to reduction of proliferative responses to stimulation by PHA and by allogeneic cells in MLC (p < 0.05); an increase of lymphocyte proliferation was detected in cultures without stimulation (p = 0.05). The advantages of the desktop MicroBeta Plus counter are multidetector system, convenient 96-pipe cell harvester and solid scintillator and rapid data processing. This recommends the counter for evaluation of proliferative activity of cultured lymphocytes.

At present time in spite of great achievements in modern chemotherapy of acute leukemia (AL) the issue of eradication of residual leukemic cells (MRD) is still relevant. Since 1971 we included specific immunotherapy in the treatment of children with acute lymphoblastic leukemia in remission using viable cryopreserved allogeneic leukemic cells. 67 children in remission were divided into 2 groups: 27 constituted the control group (only continued standard-for-that-time chemotherapy) and 40 children - the treatment which received immunotherapy in addition to standard chemotherapy. In 3 years all children in the control group relapsed. The median length of remission was 15 months. In the treatment group we observed stabilization of remission only in children over 7 years of age when immunization was initiated after 6 or more months of remission and in children younger than 7 if it was initiated after 1-1,5 years of remission. The median length of remission was 60 months which significantly exceeded (4 times) that parameter in the control group of children. Cytotoxic antibodies against leukemic cells appeared in the serum of effectively immunized children at a higher titer than against donor lymphocytes. Intrathecal administration of this hyperimmune serum to patients with neuroleukemia resistant to chemotherapy led to a sharp decrease in the amount of leukemic cells in the spinal fluid. After 5 years of remission (and 3-5 years of immunotherapy) all treatment in these patients was stopped. Out of 19 patients who received immunotherapy on time, 8 patients (42%) have been in event-free remission for 37 to 41 years (median - 38 years) through the present time and enjoy high quality of life. Our results indicate that immunotherapy initiated during remission period of AL can lead to creation of anti-leukemic immunity with subsequent eradication of MRD and complete recovery.

Трансплантация аллогенного костного мозга (ТКМ) до сих пор остается единственным методом, который может вылечить больных хроническим миелолейкозом (ХМЛ). Это доказано наблюдением за пациентами, которые после ТКМ уже десятки лет находятся в полной молекулярной ремиссии без какой-либо поддерживающей терапии. Представленный в статье 20-летний опыт выполнения ТКМ от HLA-идентичных сиблингов больным ХМЛ свидетельствует о возможности биологического выздоровления 70—80% больных в 1-й хронической фазе ХМЛ. Впервые обсуждаются вопросы, касающиеся современных взглядов на место ТКМ в терапии больных ХМЛ в эпоху широкого применения ингибиторов тирозинкиназ.

This review presents the history of development of mixed lymphocyte culture (MLC) method and its use for evaluation of the donor-recipient HLA-D histocompatibility before bone marrow transplantation. The relationship between the proliferative response in MLC reaction and donor sibling and recipient's HLA class II alleles and genes is shown. The informative value of MLC reaction is demonstrated and its important role (in addition with HLA genotyping in polymerase chain reaction) in search of HLA genoidentical donor siblings for bone marrow transplantation is defined.

The results of monitoring of qualitative content of 8 onco-antigens (CK, CA-242, CA-19-9, CA-125, REA, AFP, SCC, NSE) in serum of 414 patients are presented. The examination of 62 patients with tumors of different localization was carried our in dynamics before and after treatment. The monitoring of concentration of onco-antigens in serum permitted to apply early diagnostics of malignant process to 9 out of 28 patients with unclear clinical symptomatic. The technique informativeness is demonstrated in pre-clinical diagnostic and prognosis of course of oncological diseases. The possibility of transitory increase of concentration of certain onco-antigens under non-oncologic diseases returning to normal values after application of corresponding pharmaceutical treatment is revealed. The broader implementation of this technique into practice of public health is recommended.

We describe the reaction of nuclei in cultured human cells from different tissues to inhibition of total protein synthesis with anisomycin – ribotoxin, which is now considered as a potential antitumor drug. It was shown that nucleoli in sensitive cells demonstrate typical reaction: under the action of the inhibitor, labile nucleolar protein, a component of RNA polymerase I transcription complex (A3 antigen), rapidly migrates from the nucleolus to numerous discrete foci in the nucleoplasm. These changes are specific for translation suppression and are not induced by other influences on the cells. Migration of A3 antigen into the nucleoplasm manifests primarily in cells at the stage of DNA replication and is absent in resting cells. These results suggest that localization of A3 antigen can be a marker of artificial suppression of translation in proliferating human cells in vitro.

Immunocytochemical staining with specific antibodies was used to study the expression of three nucleolar proteins (fibrillarin, B23/nucleofozmin, and SURF6), which were involved in pRNA maturation, in the lymphoid cells of healthy individuals and patients with lymphoproliferative diseases and to compare it with the expression of the known proliferation marker Ki-67 protein. The results indicated that fibrillarin was detectable at the comparable level in the lymphoid cells of the patients and in the peripheral blood lymphocytes of the healthy individuals. In one fourth of the patients, the proportion of cells containing B23/nucleofozmin was noticeably higher than that in the lymphocytes of donors; however, there was no great difference in patients with different types of the disease. The number of SURF6-positive cells was directly correlated with that of Ki-67-positive cells. The maximum level (47-67%) of SUR6-positive lymphoid cells was found in splenic lymphosarcomas and mantle cell lymphoma. The findings suggest that SURG6 protein may be of additional diagnostic and prognostic value.