No difference between cytochemical and immunological detection of myeloperoxidase in AML.
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Fluorescent Antibody Technique↗
Biomedical subjects
H C Kluin-Nelemans
Publications and source records attributed to H C Kluin-Nelemans.
Staging of patients with Hodgkin's disease: what should be done?
An overview of the place and use of diagnostic investigations necessary for staging of patients with Hodgkin's disease is given. Staging followed by specific treatment according to stage remains the corner stone in the management. New imaging techniques such as computed tomography and ultrasound cannot completely replace lymphangiography. Staging laparotomy should not be used routinely in clinical stage I and II patients. In the past, staging laparotomy was performed to identify those patients for whom management decisions would depend on the identification of abdominal disease. However, with the identification of prognostic factors and the subsequently selected therapy-regimens including treatment of possible spleen involvement, the role of laparotomy in patients management has largely decreased.
Bone Marrow Diseases↗
Inhibition of human macrophage colony formation by interleukin 4.
We investigated the effects of human rIL-4 on in vitro hematopoiesis. A profound inhibition of macrophage colony formation by IL-4 was observed, whereas colony growth of other lineages was not affected. Inhibition of macrophage colony growth was not restricted to GM-CSF-induced colony growth but was also present in cultures stimulated with M-CSF. This inhibition was not only observed in cultures of light density bone marrow cells, but also in cultures of monocyte- and T lymphocyte-depleted bone marrow cells. Since a similar inhibition was observed in cultures of CD34+HLA-DR+-enriched bone marrow cells, a direct action of IL-4 on monocyte-committed progenitor cells is suggested.
Antigen-Presenting Cells↗
Comparison between clonogenic and cytotoxic assays for measuring LAK cell activity.
The antiproliferative effect of lymphokine-activated killer (LAK) cells was studied using a clonogenic assay in an attempt to find a model for predicting this effect in vivo or ex vivo (in the case of purging) in cancer treatment. The results were compared with the standard 51Cr-release cytotoxic assay. Cells from clonogenic neoplastic cell lines (K562 and HL-60) were plated in methylcellulose with LAK cells obtained from ten different donors in various effector-to-target (E:T) ratios. At E:T ratios of 16:1, elimination of greater than 90% of the clonogenic cells was seen in 20 of 21 experiments, whereas such lysis was incidentally found in the 51Cr-release assay. In almost all paired combinations, clonogenic cells tested in a colony assay were more sensitive to kill by LAK cells than the whole tumor cell suspensions measured in the 51Cr-release assay.
Cell Line↗
Haematopoietic and immunologic abnormalities in severe aplastic anaemia patients treated with anti-thymocyte globulin.
Thirty-five patients with severe aplastic anaemia (SAA) were extensively evaluated 0.3-12.4 years (median 3.8) after anti-thymocyte globulin (ATG) treatment. All but one were transfusion independent. Most patients revealed a normal Hb level and a granulocyte count over 1.5 x 10(9)/l but were still thrombocytopenic due to decreased platelet production. Lymphocytopenia and/or monocytopenia was found in about 30%. Two patients had a monocytosis. Although there was a great range in degree of recovery at various time intervals after ATG, patients tested more than 4 years after ATG tended to have higher cell counts. Lymphocyte counts correlated with the interval between ATG and evaluation, and with haematopoietic recovery. Qualitative abnormalities were found in all cell lines. Most patients showed a homogeneous macrocytic RBC population, and almost 50% a positive sucrose lysis test; only three patients showed evidence of haemolysis and only two of these showed a positive Ham test. Mean platelet volumes were reduced out of proportion to their number. Platelet function, determined by bleeding time and aggregometry, was impaired in over 30%. The granulocytic series showed a shift to the left in about 30%. Hypersegmentation and pseudo Pelger-Huet anomaly were seen in some patients. Lymphocyte subset distribution in blood and bone marrow was within the normal range but absolute blood levels of CD4 cells in particular were slightly decreased, and tended to increase gradually with time after ATG. IgG and IgA levels were significantly decreased. In only one patient cytogenetic analysis of unstimulated bone marrow cells revealed an abnormal karyotype, but in eight of eight patients an increased sensitivity of lymphocytes to X-rays was found. These data suggest impairment at the level of the very early haematopoietic progenitor cell in all patients up to 10 years after ATG. Since similar findings have been reported in clonal (pre-)malignant disease, SAA, improved after ATG treatment, might be prone to clonal (malignant) evolution.
Adolescent↗
Bone marrow histopathology of patients with severe aplastic anaemia before treatment and at follow-up.
Pretreatment bone marrow biopsies of 63 patients with severe aplastic anaemia (SAA) who were not transplanted and of whom 55 received ATG, were evaluated according to the amount and character of residual haematopoiesis ('genuine' aplasia/intermediate/hypoplastic myelodysplasia (MD], inflammatory infiltrate (Te Velde & Haak, 1977, grade I/II/III), and number of mast cells (normal or slightly increased/increased). Of 61 evaluable biopsies, 47 were 'genuine' aplastic, 11 intermediate and three hypoplastic MD. Inflammatory infiltrates were graded as III in 36/60 evaluable biopsies, as II in 21 and I in three. A moderate to marked increase of mast cells was seen in 19/61. Of grade III patients, 86% had a less than 90 d interval between diagnosis and administration of ATG, versus 50% of grade I/II patients (P less than 0.01). No other correlations with pretreatment characteristics were found. No significant prognostic value for survival or response to ATG of any of these three criteria has been identified. More patients with grade III inflammation tended to show adequate recovery at 4 and 6 months after ATG. Stem cell damage, not identifiable morphologically, and/or impairment of accessory cells might play a major role in eventual outcome of SAA patients. Thirty-five patients are currently alive, median 3.8 years (up to 12.4) after ATG. Follow-up bone marrow aspirates and biopsies of 32 patients were evaluable and none showed normal haematopoiesis. One patient revealed persistent aplasia. Of the remaining 31, haematopoiesis was decreased in 14 and increased in eight. All had dyserythropoiesis, 28 dysplastic myelopoiesis and in 16/29 with evaluable megakaryocytes, dysmegakaryopoiesis was found. Sixteen patients had normo- to hypercellular bone marrows with two dysplastic cell lines (consistent with myelodysplastic syndrome (MDS) according to the FAB-group). The prognostic impact of the dysplastic abnormalities found in these patients needs longer follow-up. Close observation is indicated in view of the previously recognized, albeit uncommon, evolution of SAA to MDS/acute non-lymphocytic leukaemia.
Adolescent↗
CD3+, 4+ and/or 8+ T cells and CD3+, 4-, 8- T cells repopulate at different rates after allogeneic bone marrow transplantation.
The antigen receptors of the majority of peripheral blood T lymphocytes are constituted of alpha- and beta-chains in association with CD3. The phenotype of those T cell receptor-alpha, beta cells is CD3+, 4+ and/or 8+. The small subset of CD3+, 4-, 8- T cells includes TCR-gamma, delta cells. These two T cell subsets have different TCR gene rearrangement patterns, tissue distributions and mechanisms of antigen recognition. We studied the repopulation of both T cell subsets in 20 allogeneic marrow graft recipients in relation to the type of graft (T cell-depleted versus non-depleted) and the occurrence of active cytomegalovirus (CMV) infection, using three-color immunofluorescence and flow cytometry. The CD3+, 4+ and/or 8+ and CD3+, 4-, 8- T cells had clearly different repopulation patterns. At 1 month post-BMT, they had repopulated the blood to similar levels. Thereafter, the CD3+, 4+ and/or 8+ T cells increased further in number, whereas the CD3+, 4-, 8- T cells stabilized on average between 100 and 200 x 10(6)/l. The nine recipients of T cell-depleted marrow grafts showed a relatively delayed repopulation of their CD3+, 4+ and/or 8+ T cells compared with the 11 recipients of non-depleted marrow. In contrast, the repopulation rate of the CD3+, 4-, 8- T cells was similar in both groups. The occurrence of active CMV infection post-BMT was associated with an increased rate of repopulation of the CD3+, 4+ and/or 8+ T cells, particularly those expressing HNK1, but did not affect the repopulation of the CD3+, 4-, 8- T cells.
Adolescent↗
Circulating buttock cells in non-Hodgkin's lymphoma.
Leukemic B cells with a characteristically sharp nuclear cleft seemingly dividing the nucleus into two or more parts have been entitled "buttock cells" and are subject of this study. These cells were found in leukemic non-Hodgkin's lymphomas (NHL) and usually have been related to follicular center cell lymphomas. However, buttock cells also closely resemble cells present in intermediate lymphocytic lymphoma (ILL) and mantle zone cells of reactive lymphoid tissues. Ultrastructurally, it became apparent that the separate nuclear lobes of buttock cells were connected by chromatin bridges. Immunophenotypically, circulating buttock cells had a variable phenotype, which may indicate either a follicle center or mantle zone origin. The use of CD5 and FMC7 monoclonal antibodies might be of discriminative help. These leukemic NHL have to be differentiated from classical chronic lymphocytic leukemia (CLL) with help of cytomorphology and immunophenotyping, since the former usually have a worse prognosis and generally will require a different treatment.
Adult↗
Residual T lymphocytes, and not malignant B cells, proliferate upon mitogenic stimulation.
Malignant B cells, derived from hairy cell leukemia (HCL), non-Hodgkin's lymphoma (NHL) or prolymphocytic leukemia were stimulated with mitogens and interleukin 2 after careful depletion of contaminating T cells resulting in final residual percentages of less than 0.1. No proliferation was found as measured by 3H-thymidine incorporation. Subsequently, to the non-T B cell cultures very small amounts of autologous or allogeneic T cells were added, ending up in final concentrations of 0.1, 0.5, 1, or 2% T cells. It appeared that a marked proliferation occurred which had in various coculture combinations to be ascribed to the T cells alone. Moreover, most HCL and other B cell NHL additionally stimulated the T cells, resulting in a further increase in proliferation. We conclude that 3H-thymidine incorporation by malignant B cells such as HCL and B-NHL stimulated with mitogens and IL-2 will in most cases wrongly be attributed to proliferation by the B cells themselves, and instead has to be ascribed to contaminating T cells.
B-Lymphocytes↗
Evolution of acquired severe aplastic anaemia to myelodysplasia and subsequent leukaemia in adults.
Myelodysplasia (MDS) and leukaemia following acquired aplastic anaemia has been reported as a rare event occurring in about 5% of patients. Improved results in survival of patients with severe aplastic anaemia (SAA) and subsequent prolonged follow-up created the possibility of evaluating the occurrence of MDS and leukaemia in 38 adult patients with acquired SAA surviving two or more years without bone marrow transplantation. Five patients, age 22, 35, 47, 56, 72 years, two females, three males, all with idiopathic SAA and normal cytogenetic analysis developed a refractory anaemia (RA) 7, 30, 48, 56, 142 months after diagnosis of SAA. In 3/5 RA evolved into an acute myeloid leukaemia (AML) either via a chronic myelomonocytic leukaemia (CMML) (2/3) or via RA with excess of blasts (RAEB) (1/3). Three patients revealed a monosomy 7 during MDS and/or leukaemic phase. One patient died during RA phase without cytogenetic abnormalities. A pattern of evolution could be identified in these patients revealing well-documented SAA - improvement of bone marrow haematopoiesis - dyshaematopoietic features of one or more cell lines with predominance of dyserythropoiesis - RA - RAEB or CMML - AML. These five patients represent more than 10% of all patients surviving at least 2 years. This implies that the risk of developing MDS and leukaemia in SAA patients surviving with autologous marrow, might increase with longer follow-up.
Adult↗
Flow cytometric and morphologic studies of HNK1+ (Leu 7+) lymphocytes in relation to cytomegalovirus carrier status.
CMV carrier status of healthy individuals causes a persistent increase in the numbers of lymphocytes expressing the HNK1 (Leu 7) antigen. We investigated this phenomenon by correlating HNK1 fluorescence intensity of the CD3+, CD4+, CD8+ and CD16+ lymphocytes with their morphology. The study was performed on 112 healthy individuals as a function of their CMV carrier status. To this end, the CD3+, CD4+, CD8+ and CD16+ lymphocytes were stratified into subsets with undetectable (-), weak (+) and strong (++) levels of HNK1 fluorescence intensity, as well as into subsets with low, intermediate or high perpendicular light scatter (PLS) signals. Among the CD3+, CD4+ and CD8+ lymphocytes. CMV carrier status was associated with significantly increased numbers of HNK1(+) and particularly HNK1(++) cells. Among the CD16+ lymphocytes, CMV carrier status increased only the HNK1(++) ones significantly. There were clear positive correlations between the PLS characteristics of the CD3+, CD4+, CD8+ and CD16+ lymphocytes and their HNK1 fluorescence intensity. Microscopic analysis of CD3+ lymphocytes sorted into HNK1(-), HNK1(+) and HNK1(++) fractions revealed that the percentage of those cells containing azurophilic granules and the numbers of such granules per cell increased in proportion to their HNK1 fluorescence intensity. Thus, CMV also interacts with the HNK1+ lymphocytes of its healthy carriers by increasing the level of expression of that antigen which is accompanied by increased granularity. Those granular lymphocytes may include cytotoxic cells which are involved in the control of viral replication.
Adolescent↗
Colony growth of normal and neoplastic cells in various concentrations of methylcellulose.
To assess the semisolid character of methylcellulose (MC) and its ability to prevent cell migration and aggregation in clonogenic assays, we studied the influence of various concentrations of MC (0.7%-1.26%) on colony growth of neoplastic cell lines, normal bone marrow cells, and hairy cell leukemia (HCL). All cell lines (K562, HL-60, JOK-1, Daudi, and BB3, an IgM-kappa B-cell line) showed a prominent decrease in colony numbers and remarkable changes in colony morphology at rising MC concentrations, whereas no such influence could be demonstrated for HCL, mixed lineage colony-forming units (CFU-GEMM), granulocyte-macrophage CFU (CFU-GM), erythroid burst-forming units (BFU-E), and erythroid CFU (CFU-E). Despite a decrease in colony numbers at high MC concentrations, some cell lines showed a sustained proliferation as measured by growth index calculations and bromodeoxyuridine (BrdUrd) incorporation. This indicates that at certain MC concentrations colony formation is not always a reflection of proliferation. BrdUrd incorporation yielded an extremely low proliferation capacity for HCL. It is likely that HCL cells, which strongly aggregate, formed pseudo-colonies in spite of high MC concentrations.
Bone Marrow Cells↗
T-cell prolymphocytic leukemia with an unusual phenotype CD4+ CD8+.
A patient with T-cell prolymphocytic leukemia (T-PLL) is described. The outcome was poor, with death 8 months after diagnosis, despite several therapeutic interventions. The cells carried both CD4 and CD8 epitopes, but other thymocytic markers were absent. The spleen showed infiltration of CD4+ CD8+ prolymphocytes in the red pulp and in T-cell-dependent areas of the white pulp. Immunologic function studies revealed proliferation after stimulation with mitogens and even several antigens. However, in the mixed lymphocyte culture the T-PLL cells did not proliferate. Cytotoxic T-cells could not be induced. In T-non-T recombination experiments neither helper nor suppressor cell function was found for pokeweed mitogen-dependent plasmablast generation of normal B-cells. Cytogenetically, many abnormalities were found. Among them, 14q+; absence of chromosomes 8, 11, and 22; and the presence of large marker chromosomes and fragments.
Aged↗
Effects of herpes virus carrier status on peripheral T lymphocyte subsets.
We studied the effects of herpes virus carrier status on peripheral blood T lymphocyte subsets in 334 healthy individuals. IgG-class antibodies against cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus (HSV), and varicella-zoster virus (VZV) were used as markers for the carrier status of those viruses. CMV carrier status was associated with significant increases in the numbers of some T cell subsets, whereas the carrier status of EBV, HSV, and VZV had no significant effects. The 159 CMV-seropositive individuals had higher numbers of HNK1+ T cells than did the 175 CMV-seronegative individuals [mean (SD), 292 (196)/microL v 164 (89)/microL, respectively], including the CD4+HNK1+ T cells [38 (48)/microL v 9 (13)/microL, respectively] and the CD8+HNK1+ T cells [166 (146)/microL v 73 (54)/microL, respectively]. Morphological and cytochemical studies showed that the expression of HNK1 by the CD4+ and CD8+ T cells was associated with the occurrence of azurophilic cytoplasmatic granules and a loss of nonspecific esterase activity. The numbers of CD4+HNK1+ and CD8+HNK1+ T cells increased proportionally to the levels of the IgG-class CMV antibody titers. We suggest that the increased numbers of CD4+HNK1+ and CD8+HNK1+ granular T cells in CMV carriers reflect the persistent interaction between CMV and the immune system of its hosts.
Age Factors↗
Thalidomide in the treatment of neuro-Behçet's syndrome.
No relapses of symptoms in a patient with neuro-Behçet's syndrome were observed after the addition of thalidomide to chlorambucil and prednisolone therapy. At present, the patient has been free of symptoms for 3 years. This observation, together with those reported from the literature, strongly support a re-evaluation of thalidomide therapy in certain life-threatening diseases such as neuro-Behçet's syndrome.
Adult↗
Characterization of normal and regenerating bone marrow cells with a panel of monoclonal antibodies.
To obtain baseline data for the diagnosis of residual leukaemia or lymphoma in bone marrow (BM), BM samples of 12 normal adults, 6 patients treated with chemotherapy for solid tumours and 6 patients treated with chemotherapy for leukaemia or lymphoma during remission were assessed with a panel of monoclonal antibodies by fluorescence microscopy immediately following chemotherapy. A large variability between normal individuals was found concerning the percentages of T lymphocytes and T cell subsets, B lymphocytes, HLA-DR+ and OKT10+ cells, due to variation in BM composition and, presumably, peripheral blood contamination. The regenerating BM cells also showed a large variability and, in addition, sometimes very high percentages (up to 19%) of CALLA+, c mu + s mu-, c mu + kappa lambda- and TdT+ cells were found. The interpretation of data on individual BM samples of patients assessed for leukaemia or lymphoma should therefore be related to: the amount of contaminating peripheral blood cells; the cellular composition of the BM samples as determined by May-Grünwald Giemsa staining; baseline data of age-matched normal controls and of regenerating BM of patients without haematological diseases.
Adolescent↗
No preferential sensitivity of clonogenic AML cells to ASTA-Z-7557.
ASTA-Z-7557, an in vitro active metabolite of cyclophosphamide, has recently been introduced to purge autologous bone marrow grafts of patients with AML. The rationale of this approach assumes a relatively higher sensitivity of leukemic cells to the drug as compared to that of normal marrow precursors. We have investigated in direct comparison the sensitivity to ASTA-Z-7557 of normal bone marrow progenitors (GM-CFC and BFU-E) and clonogenic leukemic cells (L-CFC). Normal bone marrow cells and purified leukemic blast cells were exposed to varying concentrations of the drug. Dose-response relationships did not indicate a selective cytotoxic susceptibility of L-CFC to ASTA-Z-7557. The recovery of bone marrow precursors following exposure to ASTA-Z-7557 depended on the cell concentration during exposure and was higher for 2 X 10(7) cells/ml than for 1 X 10(6)/ml. To mimic minimal residual leukemia cell mixtures of 95% irradiated normal bone marrow cells with 5% leukemic blast cells were exposed to ASTA-Z-7557. In this mixture killing of L-CFC was largely decreased. These data suggest that in vitro incubation of autologous bone marrow grafts of patients with minimal residual leukemia with ASTA-Z-7557 might not offer a therapeutic advantage.
Cell Survival↗
Functional deficiency of complement factor D in a monozygous twin.
An adult twin with recurrent bacterial infections was found to have a partial functional deficiency of complement factor D. Full restoration of alternative pathway activity and zymosan- or cobra venom factor-induced consumption of C3 and B was found after reconstitution of patient's serum with purified D. Family studies revealed normal D levels in the mother, a brother and another sister. After gel filtration of patient's sera only little D activity could be detected in the fractions, and trypsin activation of the fractions also did not uncover detectable precursor D activity.
Adult↗