bluuebunny/crossref_metadata_2025 huggingface dataset:
Serologic characterization of a monkey antiserum to human leukemic myeloblasts
Authors: Mohanakumar T , Baker MA , Roncari DA , Taub RN
Published: October 2019

We have raised a monkey antiserum that is selectively reactive with human leukemic myeloblasts by immunization with a glycoprotein antigen (AMLSGA) released from myeloblasts in short-term culture. Antimyeloblast activity can be demonstrated using complement-dependent cytotoxicity or indirect immunofluorescence. Selective antimyeloblast activity is retained following absorption with leukemic lymphoblasts or lymphocytes, nonleukemic lymphocytes, neutrophils, or mononuclear cells from nonleukemic bone marrow. Anti-AMLSGA antisera are not reactive with B-cell-enriched cell populations, and antaimyeloblast activity is not reduced by absorption with Ia-positive cells. Anti-AMLSGA is a useful reagent for identification of human leukemic myeloblasts.
A murine model for the switch from fetal to adult hemoglobin during ontogeny
Authors: Alter BP , Goff SC
Published: October 2019

In murine erythroleukemia cells, the minor/major hemoglobin (Hb) ratio depends on the cell line and the inducing agent. To determine whether mouse minor hemoglobin is a “fetal” hemoglobin in vivo, globin chain composition and synthesis rates were determined in DBA/2 mice of various ages ranging from 14-day embryos to > 6-mo adults. Globin chains were separated by electrophoresis on polyacrylamide gels containing urea and Triton X-100. This method separates the embryonic (x,y,z) and the adult (alpha, beta ma, beta mi) globin chains. Fourteen day embryos had only 5%-10% adult globins but approximately 30% of the adult beta chains were beta mi. The % beta mi decreased with age and reached 20% in adult mice. Biosynthetic studies led to more pronounced differences: beta mi synthesis was 45% of total beta chain production in 14-day embryos and declined to 22% in adults. Thus beta minor/total beta globin synthesis declines during mouse ontogeny. This resembles qualitatively the human switch from fetal to adult hemoglobin and provides a murine model for studies of hemoglobin regulation.
Enhancement of hemopoietic recovery after bone marrow transplantation with the addition of nucleated blood cells in mice
Authors: Zander AR , Verma DS , Spitzer G , Huynh L , Johnston D , Beran M , Dicke KA
Published: October 2019

Recovery of bone marrow cellularity, CFU-C, and CFU-S were studied sequentially over 90 days time after syngeneic bone marrow transplantation in mice. A minimal cell dose of 2 X 10(5) bone marrow cells was given. At day 28 after transplantation, CFU-C reached more than 50% of the normal range whereas the CFU-S concentration was less than 15%. Normalization of CFU-S occurred at day 90. The effect of the addition of peripheral blood nucleated cells on bone marrow hemopoietic recovery was studied at day 28. The augmentation of CFU-C and CFU-S recoverey were dose dependent. Optimal enhancement was seen with bone marrow to blood ratios of 1:1 and 1:2.5. This enhancement effect was lost when nucleated blood cells in a ratio of 1:10 were administered.
The demonstration of antibody binding to platelet-associated antigens in patients with immune thrombocytopenic purpura
Authors: McMillan R , Tani P , Mason D
Published: October 2019

Platelet destruction in chronic immune thrombocytopenic purpura (ITP) is due either to antibody against platelet-associated antigen(s) that attaches by the antigen-specific Fab portion of the molecule or to platelet-bound immune complexes that bind nonspecifically to a platelet Fc receptor. Since pepsin digestion destroys the Fc fragment, the effect of this agent on platelet binding should allow differentiation betwen these two mechanisms. Normal serum IgG, aggregated normal serum IgG, and IgG produced in culture by splenic cells from control subjects and ITP patients were radiolabeled and tested for platelet binding before and after pepsin digestion. The binding to target platelets of both aggregated IgG and IgG produced in culture by ITP cells was increased when compared to controls. However, F(ab)2 fragments from the ITP samples retained their binding ability while those from the aggregated IgG did not. We conclude that these ITP patients produced antibody specific for platelet-associated antigens.
An anionic material at the advancing front of blood cells entering the bone marrow circulation
Authors: De Bruyn PP , Michelson S
Published: October 2019

The selective entry of mature blood cells into the peripheral circulation of the bone marrow is a transcellular process. The mature blood cells penetrate the cytoplasm of the endothelial cells lining the myeloid sinuses and form a migration pore in the cell body of the lining cell, which closes after the blood cell reaches the intravascular space. The changes at the surfaces of the cells involved in this selective transcellular process were studied by means of the cationic cell surface markers colloidal iron (CI) and polycationic (high isoelectric point) ferritin (PCF). The anionic cell surface charges resulting from the presence of sialated glycoproteins remain, as shown by the application of these markers at low pH (1.8), evenly distributed at the cell surfaces of both the blood cell and the sinus lining cell. However, there is at the advancing margin of the diapedesing blood cells a marked accumulation of a nonsialated anionic material binding PCF at high pH (7.2). This material first accumulates extravascularly beneath the endothelium at an area of the blood cell surface near the site of the migration pore formation, remains at the cell surface during the initial phases of transcellular passage and disappears, either through shedding in the vascular lumen or through redistribution on the cell surface, when the cell reaches the intravascular space. This anionic material, which is neuraminidase resistant and has a pKa higher than sialic acid, is a characteristic concommitant in the selective transcellular blood cell passage in bone marrow.
Adherent cell inhibition of human leukemic in vitro agar clonal growth by short-term cell-to-cell contact
Authors: Spitzer G , Verma DS , Beran M , Zander AR , Dicke KA , McCredie KB
Published: October 2019

The effects of peripheral blood adherent cells from normal donors on human myeloid leukemic cluster growth in agar were studied. A prior co- incubation of nonadherent leukemic cells with adherent cell monolayers from 9 out of 10 donors in liquid cultures over a 4-hr period was sufficient to reduce subsequent leukemic growth in semisolid agar cultures. Inhibition was seen with adherent to leukemic cell ratios of as low as 0.5:1. Conversely, identical numbers of adherent cells in agar cultures but separated from the leukemic cells enhanced growth more than the cultures containing human placental conditioned media alone. Because leukemic cell exposure to adherent cells was brief, a cytotoxic mechanism appeared likely; however, this could not be detected by 51Cr release. Human peripheral blood adherent cells not activated by any in vitro mechanism suppress clonal growth of human myeloid leukemic cells by a mechanism requiring cell to cell contact. Examination of the inhibition of clonal growth appears to be more sensitive than 51Cr release as an indicator of adherent cell effects on myeloid leukemia.
The kidneys in paroxysmal nocturnal hemoglobinuria
Authors: Clark DA , Butler SA , Braren V , Hartmann RC , Jenkins DE Jr
Published: October 2019

Long-term study of 21 PNH patients revealed an unexpectedly high incidence of functional and anatomic renal abnormalities. Most patients demonstrated varying degrees of hematuria and proteinuria distinct from hemoglobinuria. Evaluation of renal function revealed hyposthenuria, abnormal tubular function, and declining creatinine clearance. Radiologically these patients had enlarged kidneys, cortical infarcts, cortical thinning, and papillary necrosis which were confirmed by autopsy studies. Hypertension developed in eight patients. Urinary tract infection was uncommon. The renal findings bear striking similarity to those of sickle cell anemia. Contrary to the usual opinion, out studies clearly showed evidence of widespread renal pathology in PNH most likely due to repeated microvascular thrombosis similar to the venous thrombosis involving other organs in this disorder.
Acquired granular pool defect in stored platelets
Authors: Rao AK , Niewiarowski S , Murphy S
Published: October 2019

Platelets stored as concentrates (PC) for 72 h at 22 degrees C develop a functional defect. Alterations in adenine nucleotides of platelets have been shown to affect platelet function. Adenine nucleotide content of platelets was measured before and after storage and a decrease of 27.1 /+- 1.7% (mean /+- SE) in ATP and 39.1 /+- 2.6% in ADP were found in 34 PC stored with final volume of 50 ml. In 11 PC with 30 ml volume. ATP and ADP decreased by 39.4 /+- 3.2% and 49.4 /+- 2.1%, respectively. The mean ATP to ADP ratio of stored platelets was significantly higher than of fresh platelets in both groups, suggesting a relatively greater decrease in granular than metabolic pool nucleotides. Levels of low affinity platelet factor 4 measured by radioimmunoassay in plasma from 0.86 /+- 0.08 microgram/ml in the fresh PC to 8.59 /+- 0.39 microgram/ml in stored PC, indicating a concomitant alpha-granular secretion. Labeling of metabolic pool with 14C-adenine revealed a mean decrease in the adenylate energy charge of 2.0 /+- 0.4% in 12 of 16 stored PC, with a lower ATP and higher hypoxanthine labeling in stored as compared to fresh platelets. These observations suggest that stored platelets develop an acquired defect in both dense and alpha granules and in their ability to maintain ATP homeostasis.
Human antithrombin III heterogeneity
Authors: Williams L , Murano G
Published: October 2019

Crossed immunoelectrophoretic patterns were obtained on three patients with a congenital deficiency of anti-thrombin III (AT-III), and on normal single donor and “pooled” plasma controls. In the presence of heparin (incorporated into the agarose gel matrix), the AT-III of normal human plasma was separated into four components: two major, faster-moving components, and two minor, slower-moving components. The three patients with congenital deficiency of AT-III (levels approximately 50% of normal) appeared to possess only one of the faster- moving components, and one of the slower-moving components.
Cutaneous T cell lymphoma: characterization by monoclonal antibodies
Authors: Kung PC , Berger CL , Goldstein G , LoGerfo P , Edelson RL
Published: October 2019

Monoclonal antibodies to human T cells permit the characterization of the surface phenotype of cutaneous T cell lymphoma (CTCL). The majority of CTCL cells are reactive with OKT1 and OKT3 monoclonals, which identify peripheral T cells and mature thymocytes. The neoplastic cells also react with OKT4, which recognizes the inducer T cell subset; they are, however, unreactive with OKT5 monoclonal, which identifies cytotoxic/suppressor T cell subsets. These data are in agreement with previous functional studies demonstrating that CTCL is a neoplasm of inducer (helper) T cells.
Reed-Sternberg cells in Hodgkin's disease contain fibronectin
Authors: Resnick GD , Nachman RL
Published: October 2019

Reed-Sternberg cells in the lymph nodes from five patients with Hodgkin's disease were studied. Indirect immunofluorescence on fixed sections with a monospecific anti-serum to fibronectin revealed abundant cytoplasmic fibronectin in approximately 90% of the Reed- Sternberg cells. In addition, the cells were shown by immunofluorescence to contain polyclonal IgG; however, factor VIII antigen, albumin, fibrinogen, alpha-2-macroglobulin, anti-thrombin III, and ceruloplasmin were not present. The abundant cytoplasmic fibronectin suggests that Reed-Sternberg cells are derived from tissue macrophages.
Bone marrow transplantation for refractory acute leukemia in 34 patients with identical twins
Authors: Fefer A , Cheever MA , Thomas ED , Appelbaum FR , Buckner CD , Clift RA , Glucksberg H , Greenberg PD , Johnson FL , Kaplan HG , Sanders JE , Storb R , Weiden PL
Published: October 2019

Thirty-four patients aged 4–67 yr (median 17) with acute lymphocytic leukemia (ALL) (18 patients) or acute nonlymphocytic leukemia (ANL) (16 patients) who failed to enter complete remission (CR) or relapsed on conventional chemotherapy were treated with cyclophosphamide (CY), 60 mg/kg/day for 2 days, 1000 rad total body irradiation, and a marrow transplant from a genotypically identical normal twin. Sixteen of the patients received additional chemotherapy within the week before CY. After the transplant, 23 patients received immunotherapy consisting of killed autologous leukemic cells and/or normal twin peripheral blood lymphocytes, 16 as part of a prospectively randomized study. One moribund patient died before engraftment. Nine patients (6 ALL, 3 ANL) continued to have detectable leukemic cells. Twenty-four patients (70%) achieved CR. One of them died of viral hepatitis at 1 mo and another of viral interstitial pneumonitis at 4 mo in CR. Fourteen patients (7 ALL, 7 ANL) relapsed 2–16 mo (median 4) after transplantation. However, 8 patients (24%) (3 ALL, 5 ANL) remain in CR without any maintenance chemotherapy at 29–103 mo (median 80) after the transplant. The end results were not signficantly influenced by the type of leukemia, the immediated pre-CY chemotherapy, or the immunotherapy. The results show that this approach, even when applied to endstage patients with acute leukemia in relapse, causes tolerable morbidity, rare nonleukemic deaths, and frequent remissions, some of which represent cures.
Proliferation, differentiation, and cytogenetics of chronic leukemic B lymphocytes cultured with mitomycin-treated normal cells
Authors: Nowell P , Shankey TV , Finan J , Guerry D , Besa E
Published: October 2019

Lymphocytes from 6 patients with chronic lymphocytic leukemia of the B- cell variety (B-CLL) were cultured with equal numbers of mitomycin- treated mononuclear cells from normal blood. When stimulated with pokeweed mitogen (PWM), phytohemagglutinin (PHA), or the tumor- promoting agent, phorbol tetradecanoyl-acetate (TPA), the CLL cells proliferated actively by day 3 or 4 of culture, and in four cases, differentiated to significant numbers of immunoglobulin-containing cells. Chromosome studies on the proliferating lymphocytes demonstrated a cytogenetically abnormal clone in three patients, including two with a 14q+ marker chromosome and two with a translocation involving the short arm of chromosome 9. One patient had a translocation from 22q to 14q, producing a Philadelphia chromosome as well as the 14q+ marker. The results indicate that the neoplastic lymphocytes of B-CLL may proliferate and differentiate when appropriately stimulated in vitro, and that chromosomally abnormal clones are not uncommon. With several techniques now available for successful short-term culture of B-CLL lymphocytes, there is opportunity for better understanding of the cellular alterations in this disease.
Patterns of cell proliferation and cell migration in the Sezary syndrome
Authors: Bunn PA Jr , Edelson R , Ford SS , Shackney SE
Published: October 2019

The patterns of cell proliferation and cell migration were studied in three patients with the Sezary syndrome using autoradiographic techniques. Cell labeling patterns following pulse labeling with tritiated thymidine in vivo indicated that Sezary cells proliferate actively in skin and in lymph nodes but that few if any Sezary cells proliferate in the peripheral blood. In two of the patients serial samples were obtained. Label dilution patterns in skin and blood over time suggested that circulating Sezary cells originated in extracutaneous sites where cells were proliferating more rapidly than in the skin. Cells labeled in extracutaneous sites of proliferation appear rapidly in the blood, and their transit time through the peripheral blood compartment is short. Circulating Sezary cells may then be deposited in the skin where they resume proliferation at a low rate. Thus, while Sezary cells proliferate in both cutaneous and extracutaneous sites, proliferation appears to be more rapid in extracutaneous sites such as lymph nodes. This suggests that trials of systemic therapeutic approaches should be undertaken.
Progress report on chlorambucil therapy in postsplenectomy patients with progressive hairy cell leukemia
Authors: Golomb HM
Published: October 2019

Of eight patients with progressive hairy cell leukemia who were treated with daily doses of an alkylating agent (chlorambucil, 4 mg) for at least 6 mo, seven have had an objective response, as measured by improved blood counts. Two patients were pancytopenic and had almost total bone marrow replacement with hairy cells at the initiation of chemotherapy; approximatley 6 mo later, their blood counts and bone marrow had improved dramatically. The five other patients had the leukemic form of the disease, and all responded to therapy. It is important to identify postsplenectomy patients with progressive disease in order to initiate low-dose single-agent chemotherapy.
Timed sequential chemotherapy of cytoxan-refractory multiple myeloma with cytoxan and adriamycin based on induced tumor proliferation
Authors: Karp JE , Humphrey RL , Burke PJ
Published: October 2019

Malignant plasma cell proliferation and induced humoral stimulatory activity (HSA) occur in vivo at a predictable time following drug administration. Sequential sera from 11 patients with poor-risk multiple myeloma (MM) undergoing treatment with Cytoxan (CY) 2400 mq/sq m were assayed for their in vitro effects on malignant bone marrow plasma cell tritiated thymidine (3HTdR) incorporation. Peak HSA was detected day 9 following CY. Sequential changes in marrow malignant plasma cell 3HTdR-labeling indices (LI) paralleled changes in serum activity, with peak LI occurring at the time of peak HS. An in vitro model of chemotherapy demonstrated that malignant plasma cell proliferation was enhanced by HSA, as determined by 3HTdR incorporation assay, 3HTdR LI, and tumor cells counts, and that stimulated plasma cells were more sensitive to cytotoxic effects of adriamycin (ADR) than were cells cultured in autologous pretreatment serum. Based on these studies, we designed a clinical trial to treat 12 CY-refractory poor- risk patients with MM in which ADR (60 mg/sq m) was administered at the time of peak HSA and residual tumor cell LI (day 9) following initial CY, 2400 mg/m (CY1ADR9). Eight of 12 (67%) responded to timed sequential chemotherapy with a greater than 50% decrement in monoclonal protein marker and a median survival projected to be greater than 8 mo duration (range 4–21+ mo). These clinical results using timed sequential CY1ADR9 compare favorably with results obtained using ADR in nonsequential chemotherapeutic regimens.
Fibrinopeptide A in acute leukemia: relationship of activation of blood coagulation to disease activity
Authors: Myers TJ , Rickles FR , Barb C , Cronlund M
Published: October 2019

Plasma fibrinopeptide A (FPA) levels were determined in 20 unselected adult patients with acute nonlymphocytic and lymphocytic leukemia. The mean FPA level in patients with active disease (15.0 ng/ml) was significantly higher than during clinical remission (2.4 ng/ml, p less than 0.01). Elevated FPA levels were observed in patients with all morphological forms of acute leukemia. In the group of patients in clinical remission, 20/47 FPA values remained elevated beyond the normal range, suggesting that low-grade intravascular coagulation was present even when no leukemic cells were observed. Sequential studies revealed reduction of FPA levels to the normal range in five patients who entered clinical remission after chemotherapy and rapid elevation of the levels in eight patients who entered relapse after clinical remission. FPA levels rose significantly in five patients studied during induction chemotherapy. Thus, subclinical activation of blood coagulation, as defined by elevation of plasma FPA level, may occur commonly in acute leukemia. Plasma FPA generation may relate to leukemic disease activity.
Coincident chronic lymphocytic leukemia and osteosclerotic multiple myeloma
Authors: Jeha MT , Hamblin TJ , Smith JL
Published: October 2019

We report the first occurrence of chronic lymphatic leukemia (CLL) and osteosclerotic multiple myeloma (MM) in the same patient. The surface Ig of the CLL and the monoclonal serum Ig had different heavy and light chain classes, and CLL lymphocytes failed to secrete the monoclonal Ig in short-term culture. We conclude that they were entirely separate malignancies occuring together by chance.
Human T-cell lymphoma with suppressor effects on the mixed lymphocyte reaction (MLR). I. Morphological and cytogenetic analysis
Authors: Gaeke ME , Vardiman JW , Miller W , Medenica M , Hopper JE , Rowley JD
Published: October 2019

The aggressive clinical course and the distinctive histologic, cytochemical, and cytogenetic features of an adult non-Sezary T-cell lymphoma with suppressor activity have been investigated. Morphological and ultrastructural analysis of neoplastic cells from peripheral blood and involved lymph nodes revealed cells with convoluted nuclei, prominent cytoplasmic azurophilic granules, well developed Golgi apparatus, short strands of endoplasmic reticulum, and moderate numbers of ribosomes and mitochondria. Cytochemical reactions showed acid phosphatase (ACP) positivity in virtually all of the neoplastic cells; and a substantial percentage of the cells, the tartrate-resistant acid phosphatase (T-ACP) isoenzyme was observed. Granular naphthyl acetate esterase (A-EST) reactivity was not present. The histological and cytochemical features of these neoplastic suppressor cells were compared with those recently described for the suppressor T-cell fraction isolated from normal peripheral blood T-cell by Fc gamma- rosette formation. The aneuploid clone had 47 chromosomes with multiple complex abnormalities, including a 14q + chromosome formed by the tandem translocation of two no. 14 chromosomes and translocations involving the long arms of no. 2 and no. 9 at band 9q34. These latter changes are particularly common in T-cell disorders. The extensive analysis of this histologic, cytochemical, and cytogenetic features of this adult T-cell suppressor lymphoma should help to clarify the criteria for distinguishing among the subsets of T-cell neoplasms with definable immunologic function.
Reconstitution of normal megakaryocytopoiesis and immunologic functions in Wiskott-Aldrich syndrome by marrow transplantation following myeloablation and immunosuppression with busulfan and cyclophosphamide
Authors: Kapoor N , Kirkpatrick D , Blaese RM , Oleske J , Hilgartner MH , Chaganti RS , Good RA , O'Reilly RJ
Published: October 2019

Three patients with Wiskott-Aldrich syndrome received transplants of marrow from their HLA-A, B, C, D identical siblings after myeloablation with busulfan, 2 mg/kg/day x 4 days, followed by immunosuppression with cyclophosphamide, 50 mg/kg/day x 4. Sustained engraftment of lymphoid and hematopoietic elements was documented in each case. Platelet counts in excess of 100,000/cu mm were restored 20--50 days posttransplant and remain in the normal range 6--12 mo later. Platelets exhibit normal size and in vitro aggregation. The patients produce isoagglutinins and antibodies to other polysaccharides. The use of busulfan in moderate dosages as a myeloablative agent, coupled with cyclophosphamide, may offer an improved alternative to the use of lethal total body irradiation as a preparative regimen for complete correction of Wiskott- Aldrich syndrome by marrow transplantation.
The role of the nuclear protein matrix during development of rabbit gr anulocytes
Authors: Eastment CE , Scott RB , Shelton KR , Haar JL
Published: October 2019

A proteinaceous nuclear substructure (nuclear protein matrix or nuclear pore complex-lamina) has been described in a number of cells and may be a universal feature of cell nuclei. We have investigated the nuclear protein matrix (NPM) in the rabbit blood granulocyte and its precursor cells to determine (A) whether the NPM composition is similar to other cells that have been studied and (B) to determine whether the dramatic morphological changes that the granulocyte nucleus undergoes during cell maturation are related to changes in the composition or structure of the NPM. NPM preparations from rabbit granulocytes were similar but not identical to those found in HeLa cells or rat liver nuclei. The NPM structure of more mature cells retained more DNA during the isolation procedures than did immature cell NPMs, and the DNA was less accessible to DNAse in the mature cell nuclei. Scanning and transmission electron microscopy revealed a continuous outer covering of the NPM preparation and a lattice-like internal structure. Recognizable nuclear form persisted although the preparations represented less than 20% of the original nuclear protein. NPM preparations from mature cells were similar in overall dimensions and internal structure to NPM from immature cells, suggesting reexpansion during isolation of a previously compacted NPM structure in segmented neutrophils. NPM proteins are synthesized primarily in early stages of cell development. The NPM appears to play a major, but passive, structural role in the nuclear changes observed during maturation of granulocytes.
Human megakaryocyte stimulation of proliferation of bone marrow fibroblasts
Authors: Castro-Malaspina H , Rabellino EM , Yen A , Nachman RL , Moore MA
Published: October 2019

Human marrow cells were processed sequentially by density centrifugation and by velocity sedimentation in serum-free Percoll gradients in order to purify megakaryocytes and to determine if these cells are the source of the growth factor derived from platelets. Cell homogenates were made from the resulting fractions and tested for growth-promoting activity(ies) in 3T3 cells and in well characterized human marrow fibroblasts. Growth was evaluated by 3H-TdR incorporation and changes in DNA cell content, as measured by flow microfluorometry. The highest mitogenic activity was derived from homogenates of low density (less than 1.050 g/cu cm), rapidly sedimenting cells. This fraction contained the highest percentage of megakaryocytes. The assessment of growth-promoting activity(ies) derived from various megakaryocyte-enriched marrow cell homogenates containing different proportions of megakaryocytes demonstrated a positive correlation between the number of megakaryocytes and their stimulatory capacity as determined by 3H-TdR uptake. The growth-promoting activities elicited from homogenates of platelets and marrow fractions enriched for megakaryocytes were similar. The dose--response curves for both were parallel, and they were both temperature resistant and trypsin sensitive. These findings implicate megakaryocytes as a source of the growth factor derived from platelets and suggest that megakaryocytes may play a role in the pathogenesis of the marrow fibrosis observed in myeloproliferative disorders by stimulating fibroblast proliferation and collagen secretion.
Altered expression of histocompatibility antigens on "B" large cell lymphomas
Authors: Woda BA , Racklin B , Rappaport H
Published: October 2019

We describe three malignant lymphomas of the diffuse “histiocytic” (large noncleaved) type in which most of the large lymphoid cells express monoclonal surface immunoglobulin and HLA-DR antigens but do not express HLA-A,B,C, or beta-2-microglobulin as assessed by indirect immunofluorescence.
Filtration versus gravity leukapheresis in febrile granulocytopenic patients: a randomized prospective trial
Authors: Ambinder EP , Button GR , Cheung T , Goldberg JD , Holland JF
Published: October 2019

Forty-eight patients with fever greater than 38.3 degrees C for at least 24 hr despite broad spectrum antibiotics and an absolute granulocyte count less than 1000/microliter were randomly allocated to 4 days of granulocyte transfusions obtained by leukapheresis using filtration (n = 27) or gravity (n = 21) techniques, the latter permitting simultaneous nonmechanical collection of granulocytes and platelets utilizing hydroxyethyl starch as a sedimenting agent. Patient characteristics and dose of granulocytes obtained from both techniques were similar. Complete response to granulocyte transfusions was established by a reduction in temperature to less than 37.2 degrees C sustained for at least 48 hr after the fourth transfusion with sterilization of cultures where previously positive and diminution of measurable infection when present. This occurred in 6/21 (29%) for gravity leukapheresis and 9/27 (33%) for filtration leukapheresis. An additional group had diminution in temperature and clinical improvement during transfusions (6/21 gravity leukapheresis versus 10/27 filtration leukapheresis). Eighty-six percent of patients transfused with gravity leukapheresis cells were alive at day 20 compared with 81% for filtration leukapheresis cells. Transfusion reactions were comparable. Thus, gravity leukapheresis appears to be as efficacious as filtration leukapheresis for treating granulocytopenic febrile patients, with the added advantages of availability to any blood bank without new equipment, of having platelets as by-products, and of not requiring donor heparinization.
Proliferation kinetics of Sezary cells
Authors: Schwarzmeier JD , Paietta E , Radaszkiewicz T , Konrad K , Marosi L
Published: October 2019

The proliferation kinetics of neoplastic T cells arising in Sezary syndrome (Sezary cells) are still poorly understood. Kinetic studies with 3H-thymidine as a DNA precursor revealed a low incorporation rate of the nucleotide into Sezary cells obtained from the peripheral blood versus Sezary cells from the skin. Using the double-label autoradiography we determined the duration of single cell cycle phases of blood and cutaneous Sezary cells. The results indicate the almost complete lack of proliferative activity in the blood but a considerable portion of proliferatively active Sezary cells in skin infiltrates. The removal of a large mass of quiescent blood cells by leukapheresis did not affect the proliferative state of the residual peripheral cell population implying that the procedure did not induce the migration of proliferating skin cells towards the blood. Terminally, the disease underwent transition into immunoblastic lymphoma. At this time, the kinetic behavior of the peripheral immunoblasts showed great similarity to that of cutaneous Sezary cells. The findings point towards a common extravascular production site of Sezary cells and immunoblasts probably located in the lymphatic tissue.
Heterogeneous mechanisms of imparied lymphocyte responses in non- Hodgkin's lymphoma
Authors: Whisler RL , Balcerzak SP , Murray JL
Published: October 2019

Peripheral blood mononuclear cells (PBMC) from 18 untreated patients with non-Hodgkin's lymphoma (NHL) were studied to characterize the cellular mechanisms contributing to impaired in vitro lymphocyte responses after stimulation by the mitogen conconavalin A (Con-A). In vitro reactivity was quantitated by the 3H-thymidine incorporation in response to an optimal dose of Con-A. All patients demonstrated impaired in vitro reactivities compared to normal controls. These in vitro impairments were partially reversible since patient's cells precultured in media alone for 3 days demonstrated enhanced Con-A responses. In greater than half of the patients, the hyporeactive PBMC suppressed the enhanced reactivities of autologous precultured PBMC when assayed in cocultures. Suppressor activity was detected mainly in those untreated patients presenting with either constitutional symptoms or diffuse histology and in general was not marked compared to the severity of impairments. Adherent monocytes were shown to participate in the suppression of autologous lymphocyte reactivity but only appeared partially responsible for the in vitro impairments. In those patients lacking detectable suppressive activity, preculturing also enhanced Con-A reactivities and was compatible with the presence of a reversible, inhibitory mechanism differing from active suppression. Many patients' hyporeactive PBMC, however, failed to demonstrate normal responses after preculturing. This failure could not be directly attributed to aberrant regulatory populations, but rather appeared to possibly represent an additional intrinsic impairment of potentially reactive populations.
A new system of hemopoietic colony formation for permanent slides and medium changes: use of glass-fiber filters
Authors: Kodama F , Maruta A , Motomura S , Fukushima K , Umeda M
Published: October 2019

A new method for hemopoietic colony formation that allows the preparation of permanent slides and medium changes during the incubation period was developed in vitro. Bone marrow cells from mice were spread over glass-fiber filters, which were placed on agar medium and cultivated for 7 days. Hemopoietic colonies appeared on the glass- fibers filters. The glass-fiber filters with colonies were stained by a peroxidase and nonspecific esterase double-staining method and mounted as permanent slides. Each colony could be clearly identified and easily counted after the staining. The dose-response relationship between the number of seeded cells and the colony counts was a linear one, with the line very nearly passing through the origin on extrapolation. The colonies were classified into three types by the staining results: granuloid type, monocyte-macrophage type, and mixed type, the last containing both granuloid and monocyte-macrophage cells. Medium change during the incubation period was attempted in the experiment for phagocytic activity of the cultured cells and proved to be useful. This system appears to be useful and convenient in the study of hemopoietic colony formation.
Autonomous synthesis of alpha and beta hemoglobin chains in rabbit erythroid cells
Authors: Dembure PP , Garrick MD
Published: October 2019

Hemoglobin beta-chain synthesis by rabbit erythroid cells was tested for dependence on availability of complementary alpha-chains. Reticulocytes and bone marrow cells were obtained from variant rabbits that have hemoglobin with isoleucine in alpha-chains but not in beta- chains. This characteristic permits the use of L-O-methylthreonine, a specific isoleucine antagonist, to inhibit selectively the synthesis of hemoglobin alpha-chains without directly affecting that of beta-chains. Study of hemoglobin synthesis by bone marrow cells presents two problems that require careful management: (A) the fragility of the globin-synthesizing apparatus and (B) the isolation of globin from the various proteins made by the mixture of nucleated cells. Disruption of synthetic activity was minimized by collecting the bone marrow in autologous plasma then removing fat and connective tissue while the cells were suspended in this medium. Purification involved gel filtration of hemoglobin and globin then CM-cellulose chromatography of globin chains. Absence of radioactive isoleucine in beta-chains demonstrated the efficacy of this scheme in removing isoleucine- containing proteins that otherwise elute with beta-chains on CM- cellulose columns. In reticulocytes, when synthesis of alpha-chains is inhibited by 30%--80%, that of beta-chains is stimulated by 20%--60%, but in marrow cells, incorporation into beta-chains stays at control level when alpha incorporation is inhibited. The data indicate that beta-chain synthesis is independent of the availability of complementary alpha-chains.
Long-term disease-free survival in acute nonlymphocytic leukemia
Authors: Peterson BA , Bloomfield CD
Published: October 2019

Twenty-six of 45 adults (58%) with acute nonlymphocytic leukemia who were treated with intensive induction chemotherapy over 5 yr ago entered complete remission. All patients entering remission were placed on weekly maintenance chemotherapy consisting of cytosine arabinoside and 6-thioguanine. The median duration of complete remission was 17 mo and 7 patients (27%) remained in their initial remission for 62 + to 102 + mo. All but one of the patients in complete remission over 5 yr have had treatment discontinued. Only 1 of 7 patients in remission for more than 5 yr has relapsed. Median survival is 26.5 mo, and 8 patients (31%) currently remain alive without evidence of leukemia 63--105 mo from diagnosis. It is possible to achieve long-term disease-free survival with chemotherapy alone in acute nonlymphocytic leukemia.
Effect of leukocyte antibodies on the fate in vivo of indium-111- labeled granulocytes
Authors: McCullough J , Weiblen BJ , Clay ME , Forstrom L
Published: October 2019

The effect of different leukocyte antibodies on the fate in vivo of granulocytes is not known. Thus, the optimum in vitro serologic tests to determine a safe and effective granulocyte transfusion or to diagnose immune destruction of granulocytes in other clinical situations have not been identified. We have studied the effect of granulocyte agglutinating (GA), granulocytotoxic (GC), and lymphocytotoxic (LC) antibodies on the intravascular recovery and half- life (t 1/2) and the extravascular localization of Indium-111- granulocytes in 50 patients. GA antibodies caused reduced granulocyte recovery and t 1/2 in three of three non-neutropenic patients (one with anti-NB1), increased sequestration of cells in the liver, and failure of granulocytes to localize at sites of infection in two of two patients (one with anti-NA1). In contrast, GC antibodies in five patients and LC antibodies in one patient did not cause reduced intravascular recovery or t 1/2 of granulocytes. In nine patients with GC and six patients with LC antibodies, incompatible granulocytes localized at known sites of infection. It appears that GA, but not GC nor LC, antibodies alter the fate in vivo of granulocytes.
Regulation of in vitro erythropoiesis by normal T cells: evidence for two T-cell subsets with opposing function
Authors: Torok-Storb B , Martin PJ , Hansen JA
Published: October 2019

Cellular interactions responsible for regulating in vitro erythropoiesis were studied using murine monoclonal antibodies recognizing antigens expressed by human mononuclear cells. Cell populations of interest were negatively selected by complement- dependent cytotoxicity and then evaluated for their effect on in vitro growth of erythroid burst-forming units (BFU-E). The data suggest that normal peripheral blood T cells contain at least two functionally distinct subpopulations with opposing regulatory effects: one that enhances burst formation had one that limits burst formation. Whether these effects are mediated by direct interactions of T cells with BFU-E or with auxillary cells remains to be determined.
Factors predicting long-term survival in diffuse mixed, histiocytic, or undifferentiated lymphoma
Authors: Fisher RI , Hubbard SM , DeVita VT , Berard CW , Wesley R , Cossman J , Young RC
Published: October 2019

Clinical and histopathologic material from 151 cases of diffuse mixed, diffuse histiocytic, and diffuse undifferentiated non-Burkitt's lymphomas have been reviewed to determine the factors that predict long- term survival. Median survival of all patients was 34 mo with 43% alive at 70 mo. Factors associated with a poor prognosis include: male sex, constitutional symptoms, advanced stage, bone marrow involvement, huge (greater than 10 cm) abdominal masses with gastrointestinal involvement, hepatic involvement, hemoglobin greater than 12 g/dl, or serum LDH greater than 250 U. The best prediction of a given patient's survival was defined by a set of four variables, which includes sex, symptoms, bone marrow status, and the presence or absence of a huge abdominal mass with gastrointestinal involvement. In contrast, classification of these patients according to the histopathologic categories of Rappaport of Strauchen did not define patient groups with significant differences in survival, nor did these categories correlate with the previously described clinical factors. Knowledge of the distribution of these prognostic factors in any clinical trial is needed before therapeutic results can be compared. In addition, such data may define subsets of patients for whom current therapy is inadequate and conversely those patients for whom current therapy yields excellent long-term survival.
Biochemical analysis of specific histamine HI and H2 receptors on lymphocytes
Authors: Osband ME , Cohen EB , Miller BR , Shen YJ , Cohen L , Flescher L , Brown AE , McCaffrey RP
Published: October 2019

It is increasingly clear that histamine mediates a variety of lymphocyte functions. Further understanding of these mechanisms requires a method for the analysis of histamine membrane receptors on the lymphocyte surface. We report now a biochemical technique for the identification and quantitation of specific histamine H1 and H2 receptors of lymphocytes. The method can be performed on small numbers of formaldehyde-fixed cells. The data this assay yields, together with that resulting from the flow cytometric analysis of histamine receptor distribution (a technique we have previously described), will be a powerful tool in the study of histamine mediation of lymphocyte function.
Monocyte-mediated antibody-dependent cell-mediated cytotoxicity: the role of the metabolic burst
Authors: Koller CA , LoBuglio AF
Published: October 2019

Human monocytes respond to opsonized microorganisms with a “metabolic burst” composed of an increase in oxygen consumption, an increase in hexose monophosphate shunt (HMPS) activity, and the generation of reactive oxygen species (ROS). We investigated the role of the metabolic burst in antibody-dependent cell-mediated cytotoxicity (ADCC) by human monocytes toward anti-D coated erythrocyte target cells because recent studies suggested a role for oxygen-dependent bactericidal mechanisms in ADCC. In normal monocytes, we found that ADCC was nearly halved under hypoxic conditions. Several agents known to impair activation of the burst, such as vincristine, cation chelators, and a sulfhydryl reagent, all decreased cytotoxicity if added before initiation of contact between target and effector cells. Cytotoxicity was inhibited by 2-deoxyglucose but not fluoride, suggesting a nonglycolytic role for glucose in ADCC, perhaps in the HMPS pathway. Although these data suggested a role for the metabolic burst in ADCC, scavengers of ROS did not impair cytotoxicity, and monocytes from chronic granulomatous disease (CGD) patients who had a defective metabolic burst had normal levels of ADCC. We conclude that ADCC toward anti-D coated erythrocyte target cells was the result of at least two independent but closely related cytotoxic pathways. Although one of these pathways appeared to involve the metabolic burst, the potentially cytotoxic reactive oxygen species did not appear to play a role in this system.
Lymphoblast purine pathway enzymes in B-cell acute lymphoblastic leukemia
Authors: Reaman GH , Blatt J , Poplack DG
Published: October 2019

Activities of enzymes of the purine metabolic pathway, adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and 5′- nucleotidase (5′-N), were investigated in the lymphoblasts of a patient with B-cell acute lymphoblastic leukemia. These lymphoblasts exhibited increased ADA activity and diminished activities of both PNP and 5′N' as compared to normal lymphocytes as well as non-T, non-B leukemia cells. This enzymatic pattern is identical to that which has been described in T-cell leukemic lymphoblasts and differs from that which has been observed in the malignant cells of undifferentiated B-cell lymphomas. These data suggest that there is biochemical heterogeneity within the spectrum of B-cell malignancies. Furthermore, inhibitors of ADA may be of use in those B-cell lymphoid neoplasms that exhibit increased ADA activity.
Loss of 111Indium as indicator of platelet injury
Authors: Joist JH , Baker RK
Published: October 2019

We previously demonstrated that platelets can be labeled with 111Inoxine with high labeling efficiency and that 111In is not liberated from labeled platelets during the platelet release reaction or prolonged in vitro storage. In view of these findings, we examined the potential usefulness of loss of 111In from labeled platelets as an indicator or platelet damage by comparing the loss of 111In with that of 51Cr and LDH (in some experiments also with platelet factor 3 availability) under different conditions of platelet injury. When washed human platelets labeled with either 51Cr-chromate or 111In-oxine were exposed to increasing concentrations of detergents (Triton X-100, lysolecithin), threshold, rate, and extent of loss of 111In, 51Cr and, LDH were similar. In contrast, when labeled platelets were depleted of metabolic energy by incubation in glucose-free Tyrode albumin solution or glucose-depleted plasma in the presence of antimycin A and 2-deoxy-D- glucose, loss of 51Cr (and PF3a) occurred earlier and progressed at a faster rate than that of 111In or LDH. Similar results were obtained when platelets were exposed to increasing concentrations of PlA1 antibody, causing complement-mediated immune injury. The findings indicate that with certain agents that cause rapid platelet disruption (lysis), different platelet constituents are lost at similar rates. However, under conditions of more subtle or slowly progressive platelet injury, small molecules such as adenine nucleotides (51Cr) may escape earlier and at faster rates than larger molecules such as LDH or 111In- binding platelet protein. Thus, neither 111In loss nor LDH loss appear to be suitable indicators for sublytic or prelytic platelet injury.
Intensive chemotherapy is the treatment of choice for elderly patients with acute myelogenous leukemia
Authors: Foon KA , Zighelboim J , Yale C , Gale RP
Published: October 2019

One-hundred and seven patients with acute myelogenous leukemia (AML) ranging in age from 15 to 82 yr who were previously untreated, received a 70 day high-dose remission induction regimen consisting of daunorubicin, cytarabine, and thioguanine (TAD). Identical complete remission rates of 65% were observed for 33 patients 60 yr of age and older and for 74 patients age 15–59 yr. Median remission duration and survival were 14 mo and 22 mo for patients 60 yr and older, and 16 mo and 22 mo for patients 15–59 yr. These differences are not significant. These data indicate that older patients respond to intensive chemotherapy in a similar manner to younger patients with this disease.
Prostacyclin and beta-adrenergic catecholamines inhibit arachidonate release and PGI2 synthesis by vascular endothelium
Authors: Adler B , Gimbrone MA Jr , Schafer AI , Handin RI
Published: October 2019

We have investigated the mechanism by which cyclic AMP inhibits PGI2 synthesis in cultured bovine aortic endothelial cells. Inhibition of cyclic AMP phosphodiesterase activity by 3-isobutyl-1-methylxanthine (IBMX) blocks calcium ionophore-induced PGI2 production by 62%. The addition of 3 mM dibutyryl cyclic AMP, alone with IBMX, increases the inhibition to 96%. Release o 3H-arachidonate from membrane phospholipids was inhibited 25% by dibutyryl cyclic AMP, 48% by IBMX, and 76% by isoproterenol plus IBMX. Inhibition by isoproterenol was reversed by 10 micro M propranolol. Release of 3H-arachidonate was also reduced 75% by a combination of 10 micro M PGI2 and 3 mM IBMX. We conclude that hormones like isoproterenol and PGI2 may regulate endothelial cell PGI2 biosynthesis by increasing intracellular cyclic AMP, which then inhibits release of endogenous arachidonate from membrane phospholipids.