Biphenotypic acute leukemia with t(15;17).

Biphenotypic acute leukemias (BAL) represent 5% of all acute leukemias. The most frequent cytogenetic abnormalities described are Philadelphia chromosome and 11q23 involvement. Here we report a BAL case, with blasts showing lymphoblast morphology and positivity for myeloperoxidase (in 6% of the blast cells). Immunophenotype revealed the compromise of myeloid and B-lymphoid lineages. Cytogenetic analysis showed the t(15;17) and 8 trisomy. PML/RARa rearrangement was detected by fluorescent in situ hybridization (FISH) on interphase nuclei while PML/RARa fusion transcript was detected in the bone marrow and peripheral blood by molecular biology studies (RT-PCR). This report describes a BAL case with an unfrequent cytogenetic abnormality, and highlights the importance of correlating the results of multiple diagnostic methods in order to establish a correct diagnosis and treatment in BAL patients.

Decreased chemotaxis of neutrophils in acromegaly and hyperprolactinemia.

Both growth hormone (GH) and prolactin (PRL) modulate immune responses in vitro. We studied chemotaxis under agarose of polymorphonuclear cells from patients with acromegaly or hyperprolactinemia. Polymorphonuclear cells were purified by dextran sedimentation and subjected to stimulation with N-formylmethionyl-phenylalanine. The results showed a decrease in both directed migration (acromegaly: 971 +/- 155 microns; hyperprolactinemia: 1123 +/- 137 microns, expressed as mean +/- SEM) and spontaneous migration (acromegaly: 270 +/- 77 microns; hyperprolactinemia: 298 +/- 77 microns) when compared to similar features from normal controls (directed migration: 2019 +/- 99 microns; spontaneous migration: 590 +/- 49 microns) and from patients with non-GH/PRL-secreting pituitary tumours (directed migration: 1633 +/- 282 microns; spontaneous migration: 562 +/- 116 microns), suggesting that this defect is selective for acromegaly and hyperprolactinemia. Our results point to a putative direct or indirect effect of GH and PRL on polymorphonuclear cell chemotaxis.

Trilineage phenotypic compromise in acute leukemia.

The expression of three lineage specific antigens in the leukemic blasts is extremely infrequent. We here report a case of triphenotypic acute leukemia with involvement of the myeloid and B and T lineages. The morphology of the blasts showed promyelocytic features with agranular cytoplasm, suggesting a M3-variant of AML. The blasts were positive for myeloperoxidase PAS and Sudan Black. Immunophenotype and cytogenetics did not confirm M3-AML diagnosis, showing a trilineage compromise (myeloid and T and B lymphoid markers) and the cytogenetic alterations +8 and +11, respectively. This report highlights the importance of correlating the results of multiple diagnostic methods in order to establish a correct diagnosis of mixed lineage acute leukemias, and allows evaluation of the prognostic importance of this subgroup of patients.

Promyelocytic blast crisis of chronic myelogenous leukaemia with translocations (9;22) and (15;17).

The promyelocytic blast crisis is a rare form of transformation during the evolution of chronic myeloid leukaemia (CML). We report a case of promyelocytic blast crisis with t(15;17) in addition to t(9;22). The morphology and immunophenotype of the blasts were similar to those seen in acute promyelocytic leukaemia (APL). The t(15;17) was confirmed by FISH. The patient had evidence of coagulopathy with clinical and laboratory findings of disseminated intravascular coagulation (DIC). This report highlights the importance of correlating the results of multiple diagnostic methods in order to establish a correct diagnosis of the promyelocytic blast crisis of CML.

Effect of soybean variety and processing on growth performance of young chicks and pigs.

The objective of this study was to determine whether soybeans without the Kunitz trypsin inhibitor and lectins could be fed effectively to young chicks and pigs. Specifically, we compared the growth performance of chicks and pigs fed diets containing modified soybeans: Kunitz trypsin inhibitor-free (KF), lectin-free (LF), lectin and Kunitz trypsin inhibitor-free (LFKF), conventional soybeans (CSB), and commercially obtained, dehulled, solvent-extracted soybean meal (SBM). A 7-d chick experiment was conducted to evaluate the nutritional value of CSB, KF, LF, LFKF, and SBM. The experiment was conducted as a completely randomized design, with four replicates, five treatments, and six male chicks per pen (n = 120). The five treatments consisted of 23% CP dextrose-soybean-based diets containing KF, LF, LFKF, CSB, or SBM as the source of dietary protein. A 28-d pig experiment was conducted to evaluate the nutritional value of CSB, LF, LFKF, and SBM. Pens of four pigs were assigned randomly to a control, corn-SBM, or one of six corn-soybean diets containing raw or extruded soybean varieties as a 2 x 3 factorial arrangement of treatments in a randomized complete block design with five blocks per treatment (n = 140). Chicks fed diets containing any of the raw soybean varieties gained less weight (P < 0.05) than chicks fed SBM (22.81 g/d for SBM vs. 14.17 g/d for the raw soybeans combined). Among the raw soybean treatments, there was a greater effect on growth performance (P < 0.05) by removing both lectins and Kunitz trypsin inhibitor (ADG of 16.56 g for LFKF) than by removing each antinutritional factor separately (ADG of 14.38 and 14.11 g for KF and LF, respectively). Pig growth performance was different (P < 0.001) for SBM (ADG of 409 g) and all the varieties when extruded (ADG of 450 g for CSB, 417 g for LF, and 408 g for LFKF) compared with the raw soybean treatments (ADG of 101 g for CSB, 165 g for LF, and 266 g for LFKF). Among the raw soybean treatments, growth performance improved (P = 0.003) as the antinutritional factor, lectin, was removed from the soybean and improved further (P = 0.045) when both lectins and Kunitz trypsin inhibitor were removed. The growth-inhibiting effect of feeding modified soybeans to young animals was more detrimental for pigs than for chicks in our experiments. Soybeans without the Kunitz trypsin inhibitor and lectins cannot be fed successfully to young chicks and pigs without heating.

[Circulating lymphocyte populations and B-cell differentiation in a young female patient with multicentric giant lymph node hyperplasia]. / Poblaciones linfocitarias circulantes y diferenciación B en una paciente joven con hiperplasia gigante multicéntrica de los ganglios linfáticos.

Giant multicentric hyperplasia of the lymph nodes (GMHLN) is currently regarded as a disseminated form of angiofollicular lymphoid hyperplasia, or Castleman's disease. An immunologic study was carried out on a 24 year-old Caucasian woman who was admitted to hospital with generalised lymph node enlargement, jaundice and fever, and showed an excellent response to steroid therapy. No alterations were found in the lymphocytic subsets, the intrinsic B cell function or the T-B cooperative capability. These data disagree with the hypothesis than GMHLN is due to a deficiency of the T cell suppressive function with concomitant immunoglobulin over-production.

Dup(12)(q13-q22) and 13q14 deletion in a case of B-cell chronic lymphocytic leukemia.

Cases with partial trisomy 12 have rarely been found in B-cell chronic lymphocytic leukemia (CLL). We report our clinical, cytogenetic and fluorescence in situ hybridization (FISH) findings in a CLL patient with a duplication of part of the long arm of chromosome 12 between bands q13-q22. This patient was the only case with this duplication among the 112 cases (0.9%) of CLL cytogenetically analyzed in our laboratory. FISH studies using unique-sequence specific probes for the RB-1 (retinoblastoma) gene and the D13S319 locus at the 13q14 band showed a monoallelic loss for the D13S319 locus (20% of cells) with a diploid RB-1 gene. Our case showed an atypical morphology (35% prolymphocytes), a high proliferation rate and progression of the disease, indicating that the duplication of this region may be equivalent to complete trisomy 12 in CLL patients.

Growth hormone inhibits normal B-cell differentiation and neutrophils' chemotaxis in vitro.

In acromegalic patients we have previously described a low ability of B-lymphocytes to differentiate into plasma cells under PWM stimulation, and a decreased chemotaxis of polymorphonuclear leukocytes (PMN) towards N-formylmethionylphenilalanine (FMP). In this study we examined the effect of exogenous GH over these immune functions in normal cells. PMN were purified by dextran sedimentation, incubated with recombinant human GH (0 to 20 ng/ml) and subjected to stimulation with FMP. PBMC were cultured with or without PWM, in the presence of GH (between 0 and 100 ng/ml). Plasma cells were determined as hemolysis plaque forming cells and also by immunofluorescence. GH, in a dose-dependent way, decreased directed migration of PMN (5 ng/ml: 1.787 +/- 148 microns; 10 ng/ml: 1.581 +/- 221 microns; 20 ng/ml: 1.569 +/- 149 microns, all as mean +/- S.E.M.), when compared to similar values of untreated PMN (0 ng/ml 2.085 +/- 139 microns). GH treatment did not modify spontaneous migration. Net migration showed the same pattern as directed migration. GH decreased dose-dependently the PWM-driven differentiation of B-lymphocytes into plasma cells to 60% of the basal level. Although not significantly, GH tended to increase spontaneous B-cell differentiation. These results could account for the already described defect in B-cell differentiation and PWN chemotaxis in acromegaly, emphasizing the relationship between the endocrine and immune systems.

Autologous rosette forming cells in subjects at risk for the acquired immunodeficiency syndrome.

We investigated the distribution of autologous rosette forming cells (ARFC) in the peripheral blood from subjects at risk for the acquired immunodeficiency syndrome (AIDS). The mean percentage of ARFC with autologous plasma from 35 male homosexual individuals was significantly lower than that of 31 normal controls. The mean percentage of ARFC of SARA had a direct linear correlation with the percentage of T4+ cells (p less than 0.01). Within the SARA group, those with antibodies against HTLV-III/LAV had percentages of ARFC significantly lower than SARA with negative antibodies. Plasma from SARA decreased the percentage of ARFC of normal cells when compared to normal homologous plasma (p less than 0.005), whereas normal homologous plasma did not modify the low percentage of ARFC from SARA. These results indicate that SARA possess a depletion of this subpopulation of T cells. This is more pronounced in subjects with anti-HTLV-III/LAV antibodies. The low ARFC is related to a decrease in the number of cells that correlates with the number of T4+ cells. The inhibition of normal ARFC by plasma from SARA suggests the existence of plasma factors that could mask the lymphocyte receptors for their own red blood cells and/or coat erythrocytes preventing lymphocyte-erythrocyte binding.

[Neutrophil chemotactic dysfunction in multitransfused thalassemia patients]. / Disfunción quimiotáctica de los neutrófilos en pacientes talasémicos politransfundidos.

PURPOSE: To evaluate the chemotactic capability of neutrophils in thalassaemic patients under poly-transfusion regimens and in thalassaemia carriers. PATIENTS AND METHODS: Twenty-one patients in multi-transfusion regimen diagnosed in the Ricardo Gutiérrez Children Hospital were studied. Of them, 17 had thalassaemia major, 3 S/beta thalassaemia and one sickle-cell anaemia. Twenty-one normal subjects comprised a control group. Chemotaxis was evaluated by two methods, namely, migration under agarose layer and in microchemotaxis chamber under stimulation with N-formyl-methionyl-n-phenylalanine at optimal concentrations of 10(-5) M and 10(-6) M, respectively. RESULTS: In thalassaemia major patients, directed mobility of neutrophil assessed by both methods was significantly decreased with regard to the normal controls, whereas random mobility was preserved. The four patients under poly-transfusion who had not thalassaemia major showed the same neutrophil defect. On the contrary, chemotaxis and random mobility of the neutrophils from thalassaemia carriers (thalassaemia minor) were similar to those of the normal controls. CONCLUSIONS: These results suggest that the defect found in the patients might be caused by transfusion overload.

Lymphocyte subpopulations and cytokine production in paracoccidioidomycosis patients.

Despite the postulated role of the immune system in the control of the infection by Paracoccidioides brasiliensis, only a few studies have addressed this point in patients. The determination of total lymphocytes and their subpopulations in 6 untreated patients with the chronic form of paracoccidiodomycosis showed that half of them were lymphopenic, because of low number of CD4+ T-lymphocytes. All patients had low CD4/CD8 ratios. On the contrary, B-lymphocytes were normal in all patients. An additional patient, studied on treatment with ketoconazole, had normal lymphocyte counts in all subpopulations, as did one of the patients previously studied at diagnosis when he received specific antimycotic treatment. The production of interferon and tumor necrosis factor, determined by bioassay in supernatants of mononuclear blood cells of the patients, induced by interleukin 2 in vitro was significantly lower than that of normal subjects. These results show that patients with paracoccidioidomycosis have a defect in blood lymphocyte subsets as well as in the ability to produce regulatory cytokines.

Defect of NK regulation in HIV-infected patients.

PIP: In Buenos Aires, Argentina, health workers obtained peripheral blood samples from 22 HIV-infected people with either no symptoms or persistent lymphadenopathy to examine natural killer cytotoxicity (NKC) of asymptomatic HIV-infected (HIV+AS) cases and the effect of factors that regulate normal NKC. Researchers compared these results with those of 10 healthy heterosexual controls. Even though NK cells of the HIV+AS cases were present in the same numbers and functioned as well as those in the controls, an inducer of interferon (Concanavalin A or ConA) could not force the cell system in vitro which demonstrated NK defectiveness. NK response to ConA of HIV+AS cases was lower than that of the healthy controls (p.05). On the other hand, the NK response to a prostaglandin antagonist (Indomethacin or IM) matched that of the healthy controls. Cell supernatants from normal peripheral blood mononuclear cells increased normal NK cell function (p.001), but those from HIV+AS cases did not do so. Thus it appeared that the HIV+AS cases were unable to produce sufficient NK enhancer factors. Cell supernatants from HIV+AS reduced normal NKC below baseline (p.05) indicating the presence of NK suppressor factors. The researchers believed products of the arachidonic acid metabolism by the cyclooexgenase pathway, maybe PGE2, may have contributed to NK suppression since IM negated suppressor activity of cell supernatants from HIV+AS subjects on normal NK function. They concluded that reduced production of enhancing factors, additional release of inhibitory factors, and deficiency at the NK effector system are likely to be the underlying causes for NK deficient function in AIDS. They noted that these effects function synergistically.^ieng

Immortalized Epstein-Barr virus-positive B-cell lines obtained by prolonged culture of peripheral blood mononuclear cells from human immunodeficiency virus type 1-positive patients.

OBJECTIVES: To study the factors that determine malignant B cell growth in human immunodeficiency virus type 1 (HIV-1)-infected patients. STUDY DESIGN: B-cell lines (lymphocyte cell lines [LCL]) were developed after nonstimulated culture of peripheral blood mononuclear cells (PBMC) from HIV-1-positive (HIV-1(+)) patients. Human immunodeficiency virus type 1 replication in culture, Epstein-Barr virus (EBV) latent oncogene expression, and cell-to-cell interaction were studied after nonstimulated culture of HIV-1(+) PBMC, analyzing their contribution to LCL appearance. METHODS: Nonstimulated PBMC cultures of HIV-1(+) PBMC and controls (N-PBMC) were established. Lymphocyte cell lines were characterized. Epstein-Barr virus latent membrane protein 1 (LMP-1) and Epstein-Barr nuclear antigen 2 were detected by polymerase chain reaction (PCR). Clonality of LCL was determined by light chain restriction (flow cytometry) and immunoglobulin H chain rearrangement (semi-nested PCR). Peripheral blood mononuclear cell phenotypes were studied at different intervals of culture. RESULTS: Lymphocyte cell lines were obtained in 73% of HIV-1(+) PBMC cultures, compared with 6% in N-PBMC. All LCL were EBV-positive (EBV(+)). B-cell lineage was established, and up to 12 different B-cell clones were expanded from the same individual. Occurrence of LCL was more frequent in cultures with HIV-1 replication, high LMP-1 expression in viable B cells, and high CD4:CD8 ratio. Human immunodeficiency virus type 1 replication persisted in 53% of the LCL. CONCLUSIONS: In vitro HIV-1 replication and persistence of viable EBV(+) lymphoblasts favor spontaneous in vitro outgrowth of LCL in HIV-1(+) patients.

CD8 expression in a case of chronic lymphocytic leukemia with trisomy 12.

We report a case of B-cell chronic lymphocytic leukemia (B-CLL) with aberrant expression of the T-cell-associated antigen CD8, as revealed by two-color flow-cytometric analysis. DNA studies showed immunoglobulin heavy-chain gene rearrangement, but not of gamma-chain T-cell receptor, confirming the B-cell origin of the neoplastic cells. Ploidy analysis showed a tetraploid population and high S-phase fraction. B-CLL cells also carried trisomy 12, detected by fluorescence in situ hybridization. The identification of more cases with the same features would be necessary to establish the prognosis of this subtype of B-CLL.

Nonhepatosplenic gamma delta T-cell lymphoma with initial testicular compromise.

We report here a case of nonhepatosplenic gammadelta T-cell lymphoma with undescribed initial localization in testis, without hepatosplenomegaly or adenopathies, and subsequent development in the maxillary sinus. The maxillar mass biopsy revealed a T-cell infiltration, and its immunologic characterization by flow cytometry showed a gammadelta T-cell phenotype (CD45+, CD3+, CD2+, TCR gammadelta+), without expression of CD7, CD5, CD1a, TdT, CD4, CD8, TCR alphabeta, or NK antigens (CD16, CD56, and CD57). Clonal gamma-chain gene rearrangement by polymerase chain reaction (PCR) was detected in testicular and maxillar biopsies. Epstein-Barr virus type 1 (EBV) sequences were detected by molecular biology in the biopsy material, suggesting that this oncogenic virus may play a role in the genesis of the clonal expansion of gammadelta T-cells. The patient was initially treated with standard chemotherapeutic protocols, with poor response and aggressive course.

Evaluación clínica-inmunológica del tratamiento con Tp-5 en sujetos infectados con HIV / Clinical immunological evaluation of the treatment with tp-5 in subjects infected with HIV

We studied the effect of Tp-5 treatment on the immune system of 15 HIV+ males. The control groups consisted of 7 asymphtomatic HIV+ homosexual men and 33 health individuals. At the time of admission, the HIV+ subjects showed a significant decrease in the number of CD4+ T-cell and an increase in CD8+ cell when compared to normal controls. During Tp-5 treatment, 4 out of 15 of the HIV infected individuals expressed a transient increase of the CD4+ lymphocyte subpopulation. The number of CD8+ subset was not modified by the pentapepotide. The ability of pokeweed mitogen driven B lymphocytes to differentiate into plasma cell, already significantly low at admission, persisted without change at the end of the treatment. Thus, Tp-5 was not able to restore B-lynphocyte function. The T-lymphocyte PHA proliferative response of T-lymphocytes of HIV infected individuals did not differ from normal controls and no variation was observed after 3 months of the Tp-5 therapy. Nevertheless, the delayed skin reactivity to the 7 antigens tested, increased in 5 out of 9 patients, suggesting a partial restoration of T cell immunity by Tp-5. We conclude that Tp-5 in vivo can induce a transient partial variation at the T lymphocyte level in HIV infected patients (AU)

Evaluación clínica-inmunológica del tratamiento con Tp-5 en sujetos infectados con HIV / Clinical immunological evaluation of the treatment with tp-5 in subjects infected with HIV

We studied the effect of Tp-5 treatment on the immune system of 15 HIV+ males. The control groups consisted of 7 asymphtomatic HIV+ homosexual men and 33 health individuals. At the time of admission, the HIV+ subjects showed a significant decrease in the number of CD4+ T-cell and an increase in CD8+ cell when compared to normal controls. During Tp-5 treatment, 4 out of 15 of the HIV infected individuals expressed a transient increase of the CD4+ lymphocyte subpopulation. The number of CD8+ subset was not modified by the pentapepotide. The ability of pokeweed mitogen driven B lymphocytes to differentiate into plasma cell, already significantly low at admission, persisted without change at the end of the treatment. Thus, Tp-5 was not able to restore B-lynphocyte function. The T-lymphocyte PHA proliferative response of T-lymphocytes of HIV infected individuals did not differ from normal controls and no variation was observed after 3 months of the Tp-5 therapy. Nevertheless, the delayed skin reactivity to the 7 antigens tested, increased in 5 out of 9 patients, suggesting a partial restoration of T cell immunity by Tp-5. We conclude that Tp-5 in vivo can induce a transient partial variation at the T lymphocyte level in HIV infected patients