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This study investigates influence of the anti-anthrax vaccination on blast transformation capacity of the leukocytes and also the possible in vitro influence of two, taxonomically different, autochthonous plants, Calendula officinalis and Vaccinium mirtyllus in promoting post-vaccinal proliferation. For that, blood was sampled on heparine (50 IU/ml) before and two weeks after the anti-anthrax vaccination (R 1190 Stamatin strain) of twenty-three dairy cows and 11 calves raised extensively. Blood samples were subjected to the in vitro blast transformation test, where glucose consumption was read after 72 hours of incubation at 37oC by the o-toluidine test. The experimental variants included samples treated with alcoholic extracts of the plants (7.5 µ/ml). There was a statistically significant increase in all the stimulation indices after the vaccination of the animals in cows, including those for Vaccinium myrtillus (48.57±13.27 to 53.53±12.49, p=0.02) and Calendulla officinalis (56.39±12.57 and 61.51±13.85, p=0.01). The increase induced in the growth of calf lymphocytes by the two plants was non-significant post vaccination. Both extracts could be used to enhance cell mediated immunity but there was an age dependent improvement of their in vitro response. [ABSTRACT FROM AUTHOR]

At higher order levels chromatin fibers in interphase nuclei are organized into loop domains. Gene regulatory elements (promoters and enhancers) are often located near the sites of loop attachments. Therefore, loop domains play a key role in regulation of cell transcriptional activity. We investigated the kinetics of DNA loop exit during single cell gel electrophoresis (the comet assay) of nucleoids obtained from two cell types that differ in their synthetic activity – human lymphocytes and lymphoblasts. Lymphocyte activation and transformation into lymphoblasts (blast transformation) was performed with interleukin 2. The results obtained suggest that a rearrangement of the loops occurs after lymphocyte activation. After blast transformation we observed an increase of the amount of loop domains on the surface of nucleoids against a decrease of the inner loop fraction. Therefore, the comet assay can be used for detection of large-scale changes in the cell nucleus that follow changes in cell functional state.

Objective: The experiment aimed at establishing the effects of an alcoholic Urtica dioica extract on the in vitro blastogenic response in antigen stimulated, immunologically mature chickens. Materials and Methods: Three equal groups (n=17) of 47 days old, Rock x Cornish chickens were subjected to oral administration of: a) 0.5 ml/chicken/ day of an alcoholic stinging nettle extract (I), b) 0.5 ml/chicken/day alcohol (II, solvent control), and c) 0.5 ml chicken/day water (III-environment control) for seven days. All birds were injected with 0.5 ml/bird of a 5% SRBC suspension (days 0 and 7). Leukocyte numbers (Burker Turk method) and their blastogenic capacity (blast transformation test) were monitored on days 0, 7 and 14. The specific response was evaluated against a SRBC lysate. Student's t-test was used to evaluate the statistical significance of the differences. Results and Discussion: The leukocyte numbers increased to from 18,083.33 ±4,879.81/mm3 to 42,833.33±7,547.99/mm3 by day 14. In vitro responses to C. officinalis and E. angustifolia decreased while SRBC response increased in the nettle treated group. Conclusion: The results did not validate the implemented protocol for the alcoholic stinging nettle treatment. Other administration routes, schemes or dosages should be tested to improve the functional capacity of the leukocytes. [ABSTRACT FROM AUTHOR]

AIM: To evaluate the clinical and hematologic efficiency of splenectomy (SE) in patients with myelofibrosis (MF) resistant to conventional traditional treatment/MATERIAL AND METHODS: Case histories were retrospectively analyzed in 52 MF patients who had been followed up at the Hematology Research Center, Ministry of Health of the Russian Federation, in 2004 to 2012 and undergone therapeutic SE (47 patients with primary myelofibrosis, 4 with postpolycythemia myelofibrosis, and 1 with postthrombocythemia myelofibrosis). The mean age was 47 years at diagnosis and 53 years before surgery. The patients younger than 50 years of age constituted 60%. Massive and giant splenomegaly was detected in 37 (71%) patients. The spleen weighing 0.9 to 2.9 and 3 to 7 kg was removed in 15 (29%) and 35 (67%) patients, respectively. In 2 cases, the weight of the removed spleen was as much as 10 and 11 kg/RESULTS: By the moment of SE, the disease duration averaged 76 (from 1 to 240) months. Twenty-one (40%) patients developed perioperative complications, including bleeding (15%), thrombosis (11.5%), and infectious complications (13.5%). There were no deaths from surgical interventions in the intra- and early postoperative periods. In more than 80% of the patients after SE, their general condition improved and the symptoms of intoxication disappeared; in the majority of patients, the therapeutic effect lasted about 2 years. In the follow-up period, 33 (63%) patients died; the time to death averaged 27 (1-84) months following SE. The causes of death were blast transformation in 27 (82%) patients and comorbidity in 6 (18%); 19 (37%) patients with an average post-SE follow-up of 37 (4-72) months continued hydroxyurea treatment. The median survival after SE was equal to 3 years; the median overall survival was 11 years/CONCLUSION: SE is effective palliative care with an acceptable level of occurring complications for individual patients with MF. Contraindications to SE as blast crisis and severe comorbidities should be strictly taken into account.

The purpose of this study was to monitor abundance and activation of local CD8β-expressing T-cell populations during Eimeria tenella infections of naïve chickens and chickens immune by previous infections. Chickens were infected with E. tenella up to three times. Caecal T-cell receptor ( TCR) γ/δ- CD8β+ cells (cytotoxic T lymphocytes; CTL) and TCRγ/δ+ CD8β+ cells were characterized with respect to activation markers (blast transformation, CD25 and cell surface CD107a). Cells were also induced to degranulate in vitro as a measure of activation potential. Major findings included a prominent long-lasting, up to 6 weeks, increase in the proportion of CTL among caecal CD45+ cells in the later stages after primary E. tenella infection. These CTL also showed clear signs of activation, that is blast transformation and increased in vitro induced degranulation. At second and third E. tenella infection, chickens showed strong protective immunity but discrete signs of cellular activation were observed, for example increased in vitro induced degranulation of CTL. Thus, primary E. tenella infection induced clear recruitment and activation of local CTL. Upon subsequent infections of strongly immune chickens cellular changes were less prominent, possibly due to lower overall numbers of cells being activated because of the severe restriction of parasite replication. [ABSTRACT FROM AUTHOR]

Introduction. Coccidiosis, caused by protozoan parasites of genus Eimeria , is a disease with large impact on poultry production worldwide. It is well known that Eimeria immunity is dependent on Th1-type responses. Gap Statement. In vitro assessment of Eimeria -specific T-cell activity would therefore be a valuable research tool but has so far proven difficult to establish. Aim. induced blast transformation and CD25 expression in defined chicken T-cell populations as a measure of in vitro induced blast transformation and CD25 expression in defined chicken T-cell populations as a measure of Eimeria immunity. Methodology. Three E. tenella infection experiments were performed and PBMC and/or spleen cells were collected between 6 and 16 days after infection of chickens. Cells were stimulated in vitro with E. tenella antigens and T-cell activation was assessed by immunofluorescence labelling and flow cytometry. Results. The results consistently showed statistically significant E. tenella induced blast transformation and/or CD25 expression albeit not always in concert on the same days after infection indicating complex kinetics of T-cell responses. In general, responses were higher for spleen cells compared to PBMC for all responding T-cell populations. E. tenella induced blast transformation and/or CD25 expression albeit not always in concert on the same days after infection indicating complex kinetics of T-cell responses. In general, responses were higher for spleen cells compared to PBMC for all responding T-cell populations. Conclusions. This methodology shows promise to study Eimeria -specific T-cells, e.g. to evaluate vaccine responses. Results indicated that a Th1-type response was induced and suggested a role for CD4+CD8αα+ cells in Eimeria immunity.

BACKGROUND: Isolated isochromosome (17q) is a rare cytogenetic abnormality in Philadelphia chromosome-negative myeloid neoplasms, usually myelodysplastic and/or myeloproliferative neoplasms (MDS/MPN). De novo acute myeloid leukemia (AML) with isochromosome 17q has rarely been reported. The frequency of genetic mutations is unknown. METHODS: The authors assessed clinicopathologic, immunophenotypic, and molecular genetic features of 22 myeloid neoplasms with isolated isochromosome 17q. RESULTS: Fourteen patients presented as MDS/MPN; 8 as de novo AML. Most presented with leukocytosis, anemia, thrombocytopenia, and splenomegaly. Morphologically, all showed myelodysplastic and myeloproliferative features, including pseudo-Pelger-Huet-like neutrophils, micromegakaryocytic hyperplasia, hypercellularity, fibrosis, and osteosclerosis. Blasts were increased (median, 40% in de novo AML; 9% in MDS/MPN). Immunohistochemical assessment of proliferation and apoptosis rates in AML were similar to a matched group without isochromosome 17q. In most patients, isochromosome 17q occurred at time of blast transformation or disease progression. DNA sequencing revealed no mutation in the uninvolved TP53 allele. Mutational analyses showed rare mutations in NRAS (3 of 10), FLT3 (2 of 16), and JAK2 (1 of 18), and no mutations in NPM1 (0 of 15), KIT (0 of 4), and CEBPA (0 of 4). The median overall survival was 14.5 months for de novo AML, and 11.0 months for MDS/MPN. With a median follow-up of 8.5 months (range, 1.5-107 months), 15 died of disease, 6 had persistent disease, and 1 was in remission after bone marrow transplantation. CONCLUSIONS: The authors conclude that myeloid neoplasms with isolated isochromosome 17q represent a distinct clinicopathologic entity with myelodysplastic and myeloproliferative features, high risk of leukemic transformation, and wild-type TP53. Cancer 2011;. © 2011 American Cancer Society. [ABSTRACT FROM AUTHOR]

We report a case of T cell prolymphocytic leukemia (T-PLL) involving blast transformation. At the initial diagnosis, most peripheral blood cells demonstrated proliferation of indolent T cell small cell variants, i.e., small to medium prolymphocytes with inconspicuous nucleoli and a normal karyotype. These cells were positive for surface CD4, CD5, and CD7, and cytoplasmic CD3, but negative for surface CD3 and CD8 and cytoplasmic terminal deoxynucleotidyl transferase (TdT). The T cell receptor (TCR) Cβ1 gene was rearranged in the cells. Large prolymphocytes with prominent nucleoli, irregular nuclei, and cytoplasmic vacuoles that exhibited chromosome 8 trisomy were observed about 1.5 years later. The CD4+CD8- single positive effector memory T cells transformed into surface CD4+CD8+ double positive precursor T cells. The clonal TCR gene rearrangement patterns of these cells were identical throughout the clinical course, suggesting clonal blast transformation. The CD4+CD8+ cells demonstrated increased chromosome 8 trisomy combined with complex chromosome abnormalities with t(14;14)(q11.2;q32) containing a 14q32 chromosome after transformation. T cell leukemia 1a (TCL1a) (14q32.1) may be implicated in this case. The TCL1a oncoprotein is expressed in approximately 70% of T-PLL cases. The disease gradually developed resistance to chemotherapy, and the patient died of the disease. It is known that indolent T-PLL can become aggressive. Therefore, similar transformations may occur in other aggressive T-PLL cases, particularly those involving trisomy 8 and TCL1a. [ABSTRACT FROM AUTHOR]

Testing the effect of a mesogenic virus strain on lymphocytes from broiler chickens, vaccinated against infectious bursal disease was performed by in vitro blast transformation of lymphocytes. Peripheral blood was collected on anticoagulant 24 hours prior to vaccination, as well as 7 and 14 days after the vaccination. Blood was diluted 1:4 with RPMI 1640, then distributed in 96 plates in duplicate for each experimental variant (control, phytohemagglutinin - PHA, concanavalin A - ConA, lipopolysaccharide - LPS). After 48 hours of incubation at 5% CO2, cell growth was estimated by spcetrophotometry of residual glucose (ortho-toluidin method). Blastization indices were calculated (%). The used vaccine strain decreased the ability of post vaccination in vitro lymphocytes transformation. Thus, the first week after vaccination, the inhibitory effect was higher for B lymphocytes when compared to I lymphocytes, while after two weeks following the vaccination, the degree of inhibition was non significant. [ABSTRACT FROM AUTHOR]

Marek's disease provides excellent opportunities for the study of a herpesvirus-induced tumour both experimentally and under natural conditions in the field. The mechanism of protection against this disease is mainly through an antiviral cell mediated immune response. Over the last decades, Marek's disease viruses have been evolving greater oncogenicity, some of which are not adequately controlled by the vaccines that are currently available (Biggs, 1997; 2001). The experiment aimed to monitor during a naturally occuring episode of Marek's disease, the changes in phagocytosis and blast transformation capacity of 65 weeks old Rosso hens (n=20) under field conditions. The influence of certain vegetal extracts on these two functions was observed in order to evaluate their restoring potential. Heparinized blood samples were taken on a farm where the disease appeared spontaneously. The birds in the experimental group were selected based on the changes of the iris and pupilla, characteristic to the eye affection during the disease as well as their progressive weight loss, worsening of general condition and abdominal distension. Total leukocytes, with a dominance of lymphoblasts and myeloid cells, varried from 35,000 to 66.000/mm3 . The birds were subjected to blod sampling twice, 11 days apart. Heparinized blood was used to perfom the in vitro carbon particle inclusion and blast transformation tests. Spontaneous phagocytosis and blast transformation were recorded and afterwards the effects of Calendula officinalis, Symphitum officinale, Echinaceea angustifolia and E. purpureea was monitored by the same techniques. The phagocytosis was diminished at the begining of the experiment and increased as a compensatory mechanism towrds the end of the experiment. The spontaneous blast transformation capacity was slightly increased at the first sampling in infected birds, when compared to that of healthy birds of the same category, but it degraded to day 11 as a consequence of the progressive disease. While during the first phases of the infection, the active principles from Echinaceea stimulated the phagocytosis, as the disease evolved, the stimulating effects became weaker. E. purpureea extract increased the in vitro growt of leukocytes from Marek's disease virus infected birds when compared to controls. Nevertheless, by day 11 none of the extracts acted stimnulating, pleeding for the lack of responsiveness of the lymphocytes to restoring treatments. [ABSTRACT FROM AUTHOR]

The phagocytic function represents one of the important aspects of the innate immunity, while the blast transformation is a measure of the specific cell-mediated reactivity. In the present study we investigated the alterations of the cell-mediated immune response induced by equine infectious anemia virus (EIAV) in eight EIAV positive animals compared to eight EIAV negative horses. Carbon particle inclusion test was used to monitor spontaneous innate phagocytosis, and the in vitro blast transformation test was performed with two experimental variants: untreated control and phytohaemaglutinin (PHA) M supplemented cultures (1 microliter/ml of culture). RPMI 1640 was the medium used to cultivate white blood cells. The spontaneous phagocytic activity was decreased in EIAV positive horses when compared to the EIAV negative ones. The spontaneous blast transformation capacity was slightly increased in infected horses (67.55 ± 13.59 %) compared to the healthy ones (68.99 ± 10.06 %), while the response to PHA M was decreased in positive horses (82.77± 7.19 %) versus the healthy individuals (80.37 ± 5.34%). The infection with equine anemia virus led to a decreased innate response while the specific cell-mediated reactivity was stronger, suggesting the activation of lymphocytes versus phagocytes subsequent to the infection. [ABSTRACT FROM AUTHOR]

One of the important aspects of the innate immunity is represented by the phagocytic function. The blast transformation is a measure of the specific cell-mediated reactivity. In our study we investigated the alterations of the cell-mediated immune response induced by equine infectious anemia virus (EIAV) in eight EIAV positive animals compared to eight EIAV negative horses to monitor spontaneous innate phagocytosis we used carbon particle inclusion test. The in vitro blast transformation test was performed with two experimental variants: untreated control (UC) and phytohaemaglutinin (PHA) M supplemented cultures (I microliter/ml of culture). RPMI 1640 was the medium used to cultivate white blood cells. The spontaneous blast transformation capacity was slightly increased in infected horses (67.96 ± 8.66 %) compared to the healthy ones (69.38 ± 9.93 %), while the response to PHA M was decreased in positive horses (67.01 ± 8.38%) versus the healthy individuals (62.42 ± 10.23 %). The spontaneous phagocytic activity has decreased in EIAV positive horses when compared to the EIAV negative ones. EIAV infection led to a decreased innate response while the specific cell-mediated reactivity was stronger, suggesting the activation of lymphocytes versus phagocytes subsequent to the infection. [ABSTRACT FROM AUTHOR]

The biological effect of visible light of low energy density was investigated in this study. The effects of diffuse (DL) and linearly polarized (LPL) light were compared on models in vitro and in vivo. Experiments in vitro were performed on human lymphocytes to study their blast-transformation and rosette-formation abilities. Both DL and LPL increased the number of blast-transformed cells even in a lymphocyte culture without PHA, and reduced rosette-formation of T lymphocytes. LPL had a more pronounced effect. In vivo exposure to DL and LPL of the spleens of tumour-bearing mice caused the appearance of factor(s) in their serum, inhibiting the incorporation in vitro of [3H)-thymidine into the tumour cells obtained from non-exposed animals. In the other series of experiments serum samples were taken from tumorous animals after the exposure of their spleens to LPL Following the daily administration of these sera to another group of non-exposed tumorous mice a decreasing tendency of the mitotic kinetics of ascites tumour was observed. The application of visible (preferably linearly polarized) light for the stimulation of human immune competent cells, and clinical trials with extracorporeal irradiation of blood for the promotion of natural defences of an immune-repressed organism are suggested. [ABSTRACT FROM AUTHOR]

Plasma γ-globulin fraction proteins, copper and zinc cations, and metal complexes formed by these cations and human serum γ-globulin induce blast transformation of splenocytes from BALB/c mice at a level comparable to that induced by concanavalin A. Zinc bound to γ-globulin reduces by 25% and copper in complex with this protein stimulates by 1.6 times its capacity to induce blast transformation. Combinations with concanavalin A reproduce the effects of γ-globulin-metal complex under conditions of mitogen induction. Incorporation of H-thymidine in splenocytes incubated with combinations of γ-globulin-copper metalcomplex, copper cations, and control protein with concanavalin A was by 1.4, 1.3 ( p<0.1), and 1.25 times higher ( p<0.05), respectively, than after incubation with concanavalin A alone. [ABSTRACT FROM AUTHOR]

Immunomodulating effects of synthetic peptides Vilon (Lys-Glu), Epithalon (Ala-Glu-Asp-Gly), and Cortagen (Ala-Glu-Asp-Pro) and possible involvement of the sphingomyelin signal transduction pathway in their effects in mouse thymocytes were studied. Vilon produced the most potent comitogenic effect on thymocyte proliferation and modulated comitogenic activity of interleukin-1b. Epithalon was less potent, while Cortagen produced no such effects. Vilon produced a more pronounced stimulatory effect on sphingomyelinase activity in mouse thymocyte membranes compared to Epithalon and Cortagen. [ABSTRACT FROM AUTHOR]

Blast transformation of peripheral blood lymphocytes (PBL) in dilated cardiomyopathy (DCM) patients and normal controls was examined to assess the cell mediated immune response of these patients. Uptake ofH-thymidine by PBL of DCM patients incubated for 72 hrs with phytohaemagglutinin (PHA) was 17787±9513 CPM and without PHA was 1475±529 CPM showing a stimulation index (SI) by PHA of 12.95±6.35. The corresponding values in healthy controls were 55078±12282 and 1868±792 respectively with a SI of 38.0±26.0. Blast transformation of PBL by PHA was significantly suppressed in DCM patients as compared to normal controls. This suppression was found to be highly significant and possibly indicates the defective cell medicated immune response in DCM patients. [ABSTRACT FROM AUTHOR]

Bilineal blast transformation of myeloid and T lymphoid type is a rare event in chronic myeloid leukemia. Here, we report a case in which an adult presented with high white cell counts and lymphadenopathy. Bone marrow studies confirmed the presence of 9 and 22 chromosomal translocation, and a diagnosis of chronic myeloid leukemia in chronic phase was made. Examination of a lymph node showed both myeloid and T lymphoblastoid blast crisis. Molecular studies demonstrated the presence of BCR-ABL fusion transcripts in both the myeloid and the T lymphoblastic component, indicating that the myeloid and T lymphoid blast crisis components shared common progenitors. TP53 deletion was demonstrated by fluorescence in situ hybridization.