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J L Binet

Publications and source records attributed to J L Binet.

At least 19 recordsLinked to original sources

Comparative quantitative study of Ki-67 antibody staining in 78 B and T cell malignant lymphoma (ML) using two image analyser systems.

Total Ki-67 stained area percentage was studied in 32 B and 46 T malignant lymphomas (ML) using two different image analyser systems (TAS, Leitz; SAMBA TM 2005, TITN) respectively. The total Ki-67 area percentage was highly correlated to the number of Ki-67 positive cellular profiles (B-ML, r = 0.93; T-ML, r = 0.88), indicating that area percentage is a reliable alternative method to the manual cell counting. Image analysis allows quicker measurements, appropriate to large and strictly lymphomatous regions. The cell image processor (SAMBA TM 2005, TITN) linked to a color video camera was more suitable for immunohistochemical sections and allowed more automated and faster measurements than the texture analyser (TAS, Leitz) linked with a black and white camera. Alkaline phosphatase technique with fast red as chromogen was more suitable for the detection of Ki-67 stained area by thresholding than peroxidase technique with aminoethylcarbazol or with diaminobenzidine as chromogens. Significant differences were found between low and high grade in B and T ML according to the Kiel classification (mean values +/- SD of 7.7 +/- 3.8% and 16.6 +/- 6.2% in B-ML and of 10.2 +/- 7.9% and 25.6 +/- 16.3% in T-ML respectively). In follicular B-ML, considering follicular areas only, values were comparable to high grade ML; angioimmunoblastic-lymphadenopathy-like (AILD-type) T-ML belonging to low grade ML showed similar values to pleomorphic T-ML with medium and/or large cells belonging to high grade ML.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase

Evaluation of residual disease in B-cell chronic lymphocytic leukemia patients in clinical and bone-marrow remission using CD5-CD19 markers and PCR study of gene rearrangements.

We evaluated minimal residual disease (MRD) in 23 CD5 + B-chronic lymphocytic leukemia (CLL) patients who achieved clinico-hematological remission confirmed by bone-marrow biopsy. MRD was evaluated by dual marker analysis flow-cytometry using CD5 and CD19 markers, and by the study of Ig heavy chain gene rearrangements using the fast polymerase chain reaction (PCR). According to our laboratory conditions patients were considered to be in complete phenotypic remission when total CD19+ cells were < 25% and the ratio of CD5 + CD19 + /CD19 + cells was < 25%. According to these strict criteria only 9 of the 23 patients were in complete phenotypic remission. In order to evaluate the sensitivity of the above method, PCR analysis of the configuration of the Ig heavy chain gene region was performed in 12 of these patients. Five of 7 patients in complete phenotypic remission retained a detectable monoclonal rearrangement of the Ig heavy chain gene. For the remaining 5 patients in partial phenotypic remission, only one failed to show a monoclonal band and this is probably explained by the presence of an unusual gene rearrangement. In conclusion, this study suggests that PCR is more sensitive than dual marker flow-cytometry for evaluation of residual disease and that it is indeed possible to achieve complete remission at the molecular level, in B-CLL. Nevertheless, we suggest a word of caution as this was a retrospective study, and samples were not assessed before treatment. Thus the possibility that apparent molecular remission might correspond to unusual gene rearrangements cannot be completely excluded in these cases.

Adult

[Non-Hodgkin's lymphoma and AIDS: histopathologic features].

HIV-related non-hodgkin lymphomas currently occur in 5 to 8% of AIDS patients. AIDS-related lymphomas are high-grade tumors with the morphologic characteristics of either small noncleaved cell lymphomas of the Burkitt type or large cell centroblastic and immunoblastic lymphomas. Mixed features may be found, making classification difficult. Useful methods for characterizing AIDS-related non-hodgkin's lymphomas include immunophenotypic studies using B-cell differentiation and activation antigens (HLA-DR, CD10, CD19, CD20, CD21, CD22, CD23, CD38), evaluation of expression of surface immunoglobulins (IgS), activation and proliferation (CD25, CD30, CD71, Ki67), and identification of T-cell markers (CD1, CD2, CD3, CD4, CD5, CD7, CD8). Cases studied were of the B-cell type. Comparison with morphologic features revealed that Burkitt's lymphomas were monoclonal and expressed B-cell markers (CD10, CD19, CD20, CD22, CD38) and surface immunoglobulins, especially IgM kappa. This immunophenotype is similar to that of large cell or centroblastic non-hodgkin's lymphomas, suggesting that Burkitt lymphomas originate from centrofollicular cells. Immunoblastic non-hodgkin's lymphomas were monotypic or polytypic and expressed CD10 and CD38 antigens but not the other B-cell antigens Furthermore, a very large number of cells stained positively with the Ki67 antibody demonstrating that most lymphoma cells were undergoing cycling.

Burkitt Lymphoma

A soluble factor released by CD8+CD57+ lymphocytes from bone marrow transplanted patients inhibits cell-mediated cytolysis.

We report a new inhibitory activity of CD8+CD57+ peripheral blood lymphocytes from allo-bone marrow transplant patients. Positively selected CD8+CD57+ lymphocytes act as potent inhibitors of allospecific cytolytic T lymphocyte and lymphokine activated killer cell cytolytic activities. These suppressor cells are mature T cells expressing the CD2, 5, 7, CD3-TcR alpha beta complex, and lack natural killer cell markers as well as cytolytic function. Their inhibitory activity on both cytolytic processes and T-cell proliferation is mediated by a non-antigen-specific soluble factor released by CD8+CD57+ cells in culture supernatants. Preliminary characterization suggests that the CD8+CD57+ cells' inhibitory activity is mediated by a low molecular weight, glycosylated factor as indicated by its less than 1S coefficient of sedimentation and its binding to concanavalin A lectin.

Antigens, Differentiation, T-Lymphocyte

Antithrombin III activity is not related to plasma homocysteine concentrations.

Arterial and venous thromboembolic events represent frequent and life-threatening complications in homocystinuric patients and are responsible for their early deaths. Reduced levels of antithrombin III activity in homocystinuric patients have recently been reported. So, high plasma L-homocysteine concentration could play a role in the low antithrombin III activity level. In the present study, we have studied the relationship between total plasma homocysteine and inhibitors of blood coagulation levels in 16 patients with malignancies who received bone marrow grafts. There were no correlations between homocysteine values and inhibitors of blood coagulation levels. So, while the defect in amino acid transsulfuration that is responsible for homocystinuria can directly affect the synthesis or activity of some clotting factors, homocysteine concentration is not responsible for this effect.

Adult

Quantitative study of KI-67 antibody staining in non-Hodgkin's lymphomas using image analysis.

In sections from 32 B malignant lymphomas (ML), the total KI-67 stained area was compared to the number of KI-67 positive cells in order to demonstrate the reliability of using image analysis to quantify the proliferative activity. The total KI-67 area percentage correlated highly with the number of KI-67 positive cellular profiles (r = .93). Significant differences were found between low- and high-grade ML according to the Kiel classification (mean values +/- SD, respectively, of 7.7 +/- 3.81% and 16.6 +/- 6.23%), and between low-, or intermediate- and high-grade ML only, according to the International Working Formulation. Within the Working Formulation, the statistical analysis grouped the diffuse large cell subtype of intermediate grade with the immunoblastic high-grade subtype. A wide range of KI-67 area percentage values was noted, particularly in follicular ML; for these follicular ML, considering follicular areas only, values were comparable to high-grade ML (14.8 +/- 6.60%). In conclusion, the KI-67 area percentage is a reliable alternative method to manual cell counting, and image analysis allows quicker measurements appropriate to large and strictly lymphomatous areas, using a greater number of cells than in manual cell counting.

Antibodies, Monoclonal

Characterization of normal human bone marrow by flow cytometry.

Fifteen bone marrow samples were obtained from consenting donors for allogenic bone marrow transplantation and were studied by flow cytometry. The simultaneous analysis, cell by cell, of size (forward angle light scattering), structure (right angle light scattering) and surface antigenic expression with 20 monoclonal antibodies allowed selection of ten reproducible cell subsets which were sorted and studied by microscopic examination in order to confirm their distinct characteristics. This very accurate analysis of a bone marrow cytogram should find applications in view of a better characterization of normal and pathological human bone marrow.

Adult

Evidence that chronic lymphocytic leukemia B lymphocytes are frequently committed to production of natural autoantibodies.

CD5+ B lymphocytes have been postulated to be primarily involved in autoantibody production. To investigate whether CD5 B lymphocytes from chronic lymphocytic leukemia (CLL) patients display the same function, we fused B lymphocytes from 23 CD5+ B-CLL with nonsecreting murine myeloma X-63 cells. Hybrids were derived from 18 of the 23 patients, but only hybrids from 13 patients were found to secrete immunoglobulin (Ig) (IgM kappa 4, IgM lambda 8, and only kappa light chain 1). Supernatants of these hybrids were tested against an extensive panel of antigens by enzyme-linked immunosorbent assay, indirect immunofluorescence, and hemagglutination. At the same time, peripheral blood mononuclear cells (PBMC) from 15 of these patients, including most patients from whom secreting hybrids had not been obtained, were stimulated with phorbol myristate acetate (PMA). Our results indicated that secreting heterohybrids from CLL B lymphocytes are frequently obtained; however, this is highly dependent on the light chain phenotype, since 8 of 9 lambda-expressing CLL produced hybrids secreting complete Igs, as compared with 4 of 12 kappa-expressing CLL. In addition, B lymphocytes of most patients from whom we failed to derive secreting hybrids also failed to secrete Ig on PMA stimulation. Secondly, CD5+ B-CLL lymphocytes were frequently committed to the production of natural autoantibodies, since in 8 of 15 cases (including hybrids and PMA stimulation) anti-Fc activity was found; three of these also displayed a multispecific binding pattern.

Aged

Nucleolar topography of nuclei in histologic sections. Application of a nonparametric approach to the study of breast cancer and non-Hodgkin's lymphoma.

The position of the nucleolus within a near-convex nuclear profile was defined, without taking stereology into account, in terms of two distances, P and G, where P = pi p2/SN, G = pi 2/SN, p and g being, respectively, the smallest and greatest distances between the center of the nucleolus (considered as a disk) and the superficial nuclear membrane, and SN being the surface area of the nucleus. Nuclear elongation (ND) was defined by the ratio ND = SN/A, where A is the area of the largest inscribed disk. The nucleolus-nucleus ratio RS = Sn/SN (i.e., the surface of the nucleolus over that of the nucleus) describes the size of the nucleolus. A four-class classification of nucleolar topography was then developed from the model of an ellipsoid nuclear profile (1 less than ND less than 2.5), for which the probability that a randomly located nucleolus (with RS less than 0.23) will be classified as central (G less than 2.5 and 0.40 less than or equal to P less than 1) is less than 0.14, as paracentral (G less than 2.5 and 0.23 less than or equal to P less than 0.40) is less than 0.13, as "transversely eccentric" (G less than 2.5 and P less than 0.23) is less than 0.10 and as "longitudinally eccentric" (G greater than or equal to 2.5) is greater than 0.66. This theoretical distribution into four classes was compared to that of nuclear profiles in four cases of breast cancer and in typical cases of immunoblastic, centroblastic and Burkitt's non-Hodgkin's lymphomas. The findings illustrate the ability of this nonparametric method to indicate characteristic nucleolar locations in relation to the number of nucleoli, their size and their nuclear profile elongation.

Breast Neoplasms

Two statistical approaches to nuclei shape and size in a morphometric description of lymph node sections in non-Hodgkin's lymphoma.

An attempt is made to evaluate more clearly the potential contribution of quantitative nuclear profile shape and size measurements to lymph node section histologic description in 70 cases of non-Hodgkin's lymphoma (NHL). The area of nuclear profiles and five nonredundant and size-free shape indices were measured using the THECLA program on a Leitz Texture Analysis System (Leitz-TAS). Two statistical approaches were applied, known respectively as "parametric," (first statistical moments of variable distributions over samples of 200 nuclear profiles) and "nonparametric," which are percentages of nuclear profiles distributed into five "cytological" classes that are defined by shape: round (A), elongated (B), kidney shaped (C), irregular (D) and cleaved (E) nuclear profiles. Both statistical approaches provide proper overall discrimination of the eight histological categories identified with reasonable reliability by pathologists. Above all, the present report discusses the ability of a set of parametric and nonparametric variables to describe NHL cell populations, in an objective and meaningful way, according to nuclei shape. A method of synthesizing multidimensional correlations (CORICO program) is proposed in support of the discussion. Also, the specific descriptive power of each of the variables is described; in particular, it is concluded that there is a close link between the shape and size of the nuclear profiles of the cells.

Cell Nucleus

Separation of mononuclear bone marrow cells using the Cobe 2997 blood cell separator.

In the present study, we report the results of our evaluation of the use of the continuous-flow cell separator Cobe 2997 to isolate from human bone marrow (BM) aspirates the mononuclear cell (MNC) fraction containing hematopoietic stem cells. This MNC concentrate is isolated in 15% of the original BM volume and contains 23% of the initial nucleated cells. It is enriched as concerns the BM MNC fraction (lymphocytes + monocytes recovery; 80%), whereas the contamination with granulocytes, red blood cells and platelets is reduced to 7.2, 1.5% and 41%, respectively, of the cells initially present in the BM suspensions. Furthermore, it is demonstrated that this MNC concentrate is highly enriched in granulocyte-macrophage-colony-forming cells (CFU-GM; recovery 83%). The method is simple, inexpensive, efficient and reproducible. It allows rapid processing of a large volume of BM without substantial loss of hematopoietic progenitor cells. It represents a valuable method of BM MNC concentration prior to further in vitro manipulations such as T cell or tumor cell depletion or cryopreservation.

Adolescent

[Plasma exchange with very low molecular weight heparin CY 222. Biological profile and therapeutic value].

In a clinical, between-patient study we investigated the effects of a VLMW Heparin fragment (CY 222) versus standard heparin (SH) in plasma exchanges (n = 10) on coagulation factors (CF). Fibrinogen (FGN), II, V, VIIF + X, IX, XI, XII, VIIIc, VIIIRag, VIIIvwf and ATIII, ProtC, ProtS, Plasminogen (PGN), Activated Thromboplastin time (APTT), Prothrombin time (PT), Thrombin time (TT), anti factor Xa (Axa), DDimer, Platelet count. Heparin was administered as a bolus and by infusion during the session, CY 222 as a bolus dose only; 1 to 1.5 plasma volume was exchanged with substitution by 5% albumin. Results (mean, s.d.) at the end of the session (End) and 4 hours later (T4) were analyzed and showed no differences between groups (with CY 222 and with SH) for technical and clinical findings. Biologically, CF were similar in both groups except for Factor XII levels at the end of the session, and Factors II, V, XII at T4. Prolonged APTT in all samples appear related to low FGN. Significant differences in Axa activities were found for each treatment when compared with its own standard, suggesting different ranges of activities of both drugs. Changes in D-dimer levels differed during the session and four hours after the session with the drug tested, and could be related to their mode of administration. Clinical efficiency and tolerance were excellent both with CY 222 and SH.

Adult

Alternating chemotherapy and radiation therapy in the treatment of advanced Hodgkin's disease.

During the period from 1982 to 1987, 19 patients with advanced Hodgkin's disease, clinical stages IIIB and IV underwent treatment with combination chemotherapy and radiotherapy. Alternating chemotherapy (3 cycles of CVCPP/3 cycles of ABVD) was sandwiched with low-dose total nodal irradiation including the splenic area; at the end of the chemotherapy, a supplemental 20 Gy radiotherapy was sometimes delivered to bulky previously involved sites. The complete remission rate was 84.2% (16/19 patients) and actuarial survival was 85% at 48 months. These preliminary results show that the alternating technique for combining chemotherapy and sandwiched irradiation is feasible and improves the outcome in advanced Hodgkin's disease. Patients with mediastinal masses associated with pulmonary and/or pleural involvement may need more aggressive chemotherapy at an earlier phase of the treatment program.

Adolescent