bluuebunny/crossref_metadata_2025 huggingface dataset:
Peripheral blood blast cell progenitors in human preleukemia
Authors: Senn JS , Messner HA , Pinkerton PH , Chang L , Nitsch B , McCulloch EA
Published: October 2019

Progenitors of blast cell colonies have been identified in acute leukemia. The peripheral blood of 18 of 25 patients with preleukemic states yielded low numbers of blast cell colonies, and the colony- forming cells were in an active proliferative state when assessed using short-term exposure to tritiated thymidine. The clinical significance of blast cell colonies is uncertain, but we suggest that further analysis of this cultural abnormality may lead to a better understanding of mechanisms and management in preleukemia.
The interaction of fibrinogen with human platelets in a plasma milieu
Authors: Marguerie GA , Thomas-Maison N , Larrieu MJ , Plow EF
Published: October 2019

Fibrinogen binds to specific receptors on human washed platelets and these sites are induced by adenosine diphosphate (ADP). This interaction is assumed to be the basis for the participation of the molecule in ADP-stimulated aggregation of platelets, but fibrinogen binding to platelets in plasma has not been directly demonstrated. In this study, we have characterized the interaction of 125I-fibrinogen to platelets in the platelet-rich plasma (PRP) of afibrinogenemic patients. In either citrated or heparinized PRP, association of fibrinogen with platelets was demonstrable and was dependent on ADP dose. This binding reached equilibrium in 10–15 min, and saturation was achieved at fibrinogen concentrations greater than 0.5 microM. A linear Scatchard plot was derived that indicated a single class of binding sites with an affinity constant of Ka = 1.8 X 10(6) M(-1), and 32,000 fibrinogen molecules were maximally bound per platelet. The kinetics of the platelet fibrinogen interaction in plasma were essentially the same at 37 degrees C and 22 degrees C, but fewer molecules were bound at 37 degrees C. The rate constants of association were k 22 degrees C on = 0.9 X 10(6) M-1 . min-1 and k 37 degrees C on = 0.4 X 10(6) M-1 . min- 1, respectively. Stabilization of the platelet-bound fibrinogen occurred only to a partial extent in both heparinized and citrated plasma. These results are similar to those obtained with washed platelets and establish that the previously defined steps in ADP- induced binding of fibrinogen to platelets occur in plasma, namely receptor induction by ADP, initial reversible binding, and irreversible binding.
Increase in the number of lymphocytes secreting IgG following blood transfusion
Authors: Shimizu K , Kitoh S
Published: October 2019

A sequential change in the number of circulating immunoglobulin (Ig) secreting cells of each Ig class following blood transfusion was studied using a reverse hemolytic plaque assay. The subjects studied were in two main groups, immunologically normal individuals and patients with malignant lymphoma or multiple myeloma who are, presumably, immune incompetent. A consistent increase in circulating IgG-secreting cells, along with either an earlier or simultaneous increase in IgM-secreting cells, was observed following blood transfusion in the immunologically normal individuals. An increase in IgA-secreting cells was also observed, but at a minimal magnitude. Such an increase was not apparent in patients with lymphoma or myeloma. The possible use of blood transfusion as a means of “challenging and checking” for the state of immune responsiveness in vivo is discussed.
Observations on the binding and interaction of radioiodinated colony- stimulating factor with murine bone marrow cells in vivo
Authors: Pigoli G , Waheed A , Shadduck RK
Published: October 2019

Radioiodinated L-cell-derived colony-stimulating factor (CSF) was used to characterize the binding reaction to murine bone marrow cells. The major increment in cell-associated radioactivity occurred over 24 hr incubation at 37 degrees C, but virtually no binding was observed at 4 degrees C. The reaction was saturable with approximately 1 ng/ml of purified CSF. Unlabeled CSF prevented the binding, whereas a number of other hormones and proteins did not compete for CSF uptake. Further specificity studies showed virtually no binding to human bone marrow, which is unresponsive to this form of murine CSF. Minimal CSF uptake was noted with murine peritoneal macrophages, but virtually no binding was detected with thymic, lymph node, liver, or kidney cells. The marrow cell interaction with tracer appeared to require a new protein synthesis, as the binding was prevented by cycloheximide or puromycin. Preincubation of marrow cells in medium devoid of CSF increased the degree of binding after 1 hr exposure to the tracer. This suggests that CSF binding sites may be occupied or perhaps decreased in response to ambient levels of CSF in vivo. Approximately 70% of the bound radioactivity was detected in the cytoplasm at 24 hr. This material was partially degraded as judged by a decrease in molecular weight from approximately 62,000 to 2 peaks of approximately 32,000 and approximately 49,000, but 72% of the binding activity was retained. After plateau binding was achieved, greater than 80% of the radioactivity released into the medium was degraded into biologically inactive peptides with molecular weights less than 10,000. These findings suggest that the interaction of CSF with marrow cells is characterized by binding with subsequent internalization and metabolic degradation into portions of the molecule that are devoid of biologic activity.
Absence of treatment-induced second neoplasms after ABVD in Hodgkin's disease
Authors: Valagussa P , Santoro A , Fossati Bellani F , Franchi F , Banfi A , Bonadonna G
Published: October 2019

One of the most serious complications of successful treatments for Hodgkin's disease is an increased incidence of acute nonlymphocytic leukemia (ANLL) and other malignancies. A retrospective analysis carried out on 1032 consecutive patients with Hodgkin's disease admitted to our Institute between 1965 and 1978 and treated with radiotherapy (RT) or chemotherapy or both modalities revealed that within 10 yr from initial therapy. ANLL was documented in 3% of patients, and over a comparable period of time 7.9% of patients developed other malignancies. ANLL was observed only in patients treated with chemotherapeutic regimens containing alkylating agents and/or procarbazine either alone (2.3%) or associated with RT (4%). Other second tumors were documented in patients given RT with or without chemotherapy. No second malignancies were observed in patients given ABVD (adriamycin, bleomycin, vinblastine, and dacarbazine) with or without RT. The incidence of ANLL was higher in patients given chemotherapy as salvage treatment upon relapse following primary irradiation (6.1%) compared to patients initially treated with combined modality (1.5%). The difference, however, failed to reach statistical significance. Since our analysis supports the evidence of a major role played by alkylating agents, procarbazine, and RT in inducing second malignancies, regimens not containing there drugs or their administration through treatments of different intensity warrant careful consideration.
The phenotype of the neoplastic cells of hairy cell leukemia studied with monoclonal antibodies
Authors: Jansen J , LeBien TW , Kersey JH
Published: October 2019

Eighteen cases of hairy cell leukemia were studied with a battery of polyclonal anti-Ig and nine monoclonal antibodies to determine the lineage of the hairy cells (HC) and the stage of their maturation arrest. Hairy cells tend to nonspecifically bind many antisera and precautions had to be taken to avoid nonspecific fluorescence of the cells. All but one case was reactive with one light chain type and one or more heavy chain isotype antisera as reported before. All cases studied were positive for monomorphic HLA-DR determinants, using monoclonal antibody 7.2. Most cases tested (6/7) were positive with the B-lineage related antibody PI 153/3. While most cases (15/18) were nonreactive with the B-lineage related antibody BA-1, they became positive (5/5) following in vitro culture. Seven out of nine cases were reactive with OKM1. Common acute lymphoblastic leukemia antigen (CALLA) was absent in all (15) cases tested and the ALL associated structure p24/BA-2 was absent from 16 of 18 cases. HC from none of the cases were clearly positive with the T-cell antibodies 9.6., or TA-1, whereas in only 1/18 the cells reacted with T101. The results of this study support the B cell lineage of most HC, and show the presence of multiple phenotypes. In combination with the surface Ib present on the cells, a hierarchy of phenotypes is postulated, with SIg-ormu delta, BA- 1+, PI 153/3+, HLA-DR+ being the most immature, and SIg(alpha) gamma, BA-1-, PI 153/3-, HLA-DR+ the most mature.
Evidence of the existence of a factor that induces Fc receptors on bone marrow cells
Authors: Calcagno M , Perez JR , Waldo MG , Cabrera G , Weiss-Steider B
Published: October 2019

The existence of a molecule responsible for the induction of Fc receptor (FcR) on bone marrow cells (FcR inducer, FcRI) is demonstrated in conditioned media from the macrophage-like cell line WR19M.1 activated by bacterial lipopolysaccharides. The molecular weight obtained from molecular sieving chromatography in gel and density gradient sedimentation is found to be 18,500 daltons and 16,000 daltons, respectively, with an isoelectric pH of 7.4. The factor is found to be thermolabile and trypsin sensitive. The macrophage and granulocyte inducer (MGI), also known as colony-stimulating factor (CSF) or colony-stimulating activity (CSA), is identified from the same source and found to have a molecular weight and an isoelectric pH different from FcRI. The fractions that contained the MGI did not induce FcR on bone marrow cells, while the fractions rich in FcRI did not induce colony formation.
Role of colony-stimulating activity in murine long-term bone marrow cultures: evidence for its production and consumption by the adherent cells
Authors: Heard JM , Fichelson S , Varet B
Published: October 2019

The involvement of colony-stimulating activity (CSA) in murine long- term bone marrow cultures (LTBMC) was studied using bilayer agar cultures. The supernatants of LTBMC were removed, a layer of dense agar was spread over the cells adherent to the bottom of the flask, and fresh myeloid cells were plated as source of CFU-C in an upper agar layer. Large numbers of granulocytic and macrophagic colonies developed regularly when target cells were plated over adherent cells of nonrecharged and greater than 12 wk old LTBMC that were hematopoietically inactive (i.e., producing a low number of nonadherent cells). The removal of adherent cells from the myeloid cells used as source of CFU-C did not decrease the number of colonies. This suggests that adherent cells of LTBMC release CSA that is directly active on CFU- C. This CSA was no longer detectable over adherent layers of hematopoietically active LTBMC. A close inverse relationship was demonstrated between the number of nonadherent cells harvested before the assay and the level of CSA. No inhibitor for CSA was demonstrated in the supernatant of hematopoietically active cultures. Murine exogenous CSA incubated over the adherent layer host its activity within 24 hr, whereas in the same conditions human CSA retained its activity. These data demonstrate the production of CSA by the adherent layer of LTBMC and strongly suggest its specific in situ consumption by differentiating myeloid cells.
Immunochemical studies of antisera to human fibrinopeptide-B
Authors: Butler VP Jr , Weber DA , Nossel HL , Tse-Eng D , LaGamma KS , Canfield RE
Published: October 2019

The immunochemical specificity of rabbit antisera to human fibrinopeptide-B (FPB) has been studied by comparing the relative abilities of FPB and of various proteins and peptides containing the NH2-terminal segment of the B beta-chain of human fibrinogen to inhibit the binding of a radioiodinated FPB derivative by each of seven anti- FPB sera. Anti-FBP sera varied in the extent to which they cross- reacted with fibrinogen, the NH2-terminal disulfide knot of fibrinogen (N-DSK), B beta 1(Pyr)-118(Met), B beta 1(Pyr)-42(Arg), and desarginyl- FPB. Anti-FPB sera have been identified that discriminate effectively between FPB and larger FBP-containing peptides; such antisera can be used to measure FPB in the absence of the larger peptides or to demonstrate the presence of larger peptides such as B beta 1(Pyr)- 42(Arg) in extracts of clinical plasma samples by means of an increase in FPB immunoreactivity following thrombin treatment. One anti-FPB serum has been identified that is capable of detecting desarginyl-FPB, and this antiserum has been used in the development of a radioimmunoassay for desarginyl-FPB. Thus, by precisely defining the specificity of anti-FPB sera, it has been possible to identify antisera that are useful, not only in the measurement of FPB, but also in the detection of other important related molecules, such as B beta 1(Pyr)- 42(Arg) and desarginyl-FPB. The immunochemical detection of these FPB- related peptides should provide useful information concerning the action of proteolytic enzymes, such as plasmin on the NH2-terminal segment of the B beta-chain of fibrinogen, and of carboxypeptidase-B on free FPB, in human plasma.
Alterations in the phenotype of hairy cells during culture in the presence of PHA: requirement for T cells
Authors: Worman CP , Beverley PC , Cawley JC
Published: October 2019

Culture studies of peripheral blood mononuclear cells from 7 entirely typical cases of hairy cell leukemia showed that after culture in the presence of PHA for 2--5 days, the predominant cell type changed from E- SIg+ CIg+ gamma FcR+ muFcR+ hairy cells to an E+ SIg- CIg- gamma FcR- muFcR- population of transformed cells derived from hairy cells. Depletion and readdition experiments demonstrated that cell-to-cell contact with T cells was necessary for the phenotypic change, while several observations indicated that the E+ population was not derived from T cells present before culture. The E positivity of the cultured cells was shown to be due to the possession of E receptor not acquired from the culture fluid, but the cells differed from true T cells in lacking both mature and immature T-cell antigens. The relevance of these in vitro observations to the continuing controversy concerning the nature of the hairy cell and to the in vivo fluctuations in immunologic phenotype not infrequently observed in hairy cell leukemia is briefly discussed.
Nonspecific esterase of B lymphocytes from a case of chronic lymhocytic leukemia and of normal T lymphocytes: similar constellations of isoenzymes
Authors: Yourno J , Burkart P , Mastropaolo W , Tartaglia A
Published: October 2019

Lymphocytes from a case of B-cell chronic lymphocytic leukemia (CLL) were obtained in a highly purified state from a therapeutic leukapheresis preparation. The CLL lymphocytes showed a fine, scattered, granular pattern of nonspecific esterase cytochemical reactivity with either alpha-naphthyl acetate (alpha NA) or alpha-naphthyl butyrate (alpha NB) substrate as opposed to the more focal pattern of control (T) lymphocytes. Nonspecific esterase of CLL lymphocytes and normal control lymphocytes was equally resistant to inhibition by fluoride ion. Extractable nonspecific esterases from the CLL lymphocytes and from purified normal T lymphocytes were indistinguishable in regard to specific activity, substrate specificity, pH optima, and zymogram profiles on polyacrylamide gel electrophoresis at pH 9.5 and pH 4.0. Zymograms of alpha NA esterase and alpha NB esterase prepared by isoelectric focusing were also similar, with no unequivocal differences. These results are consistent with recent reports that B lymphocytes contain detectable nonspecific esterase and suggest that the B lymphocytes from this case of CLL contained a constellation of isoenzymes similar to that of normal T lymphocytes. This is interpreted as a reflection of the close kinship of these cells.
Factor VIII/von Willebrand factor binding to von Willebrand's disease platelets
Authors: Gralnick HR , Williams SB , Shafer BC , Corash L
Published: October 2019

A form of von Willebrand's disease has been described with enhanced ristocetin-induced platelet aggregation and anodal migration of the factor VIII/von Willebrand factor protein (type IIb). We studied two families with this form of von Willebrand's disease and macrothrombocytopenia. We have found that these platelets bind more of the normal and intermediate-sized multimers of the factor VIII/von Willebrand factor than normal platelets. Analysis of the binding data show an increased affinity of these vWd platelets for the factor VIII/von Willebrand factor. These findings are consistent with an increased number of platelet receptors, which, either by their native topography or migration on the platelet surface, bind factor VIII/von Willebrand factor protein with greater affinity than normal platelets, platelets of other vWd patients, and large platelets of other etiologies.
Studies on the mechanism of the inhibition of platelet aggregation and release induced by high levels of arachidonate
Authors: Linder BL , Goodman DS
Published: October 2019

We have previously reported that arachidonic acid induced a biphasic pattern of platelet aggregation and the release of both dense and alpha- granule components. Low levels of arachidonate (0.025--0.1 mM) specifically induced aggregation and release, while high concentrations (0.15--0.35 mM) caused a progressive inhibition of these platelet responses in human gel-filtered platelets (GFP). We now report studies of the mechanism(s) responsible for this arachidonate-induced turn-off of platelet function. Electron micrographic studies demonstrated that there was no gross damage to the platelets during the turn-off. Active synthesis of malondialdehyde and thromboxane A2 was seen at the high arachidonate levels, despite the inhibition of aggregation. Furthermore, GFP inhibited by 0.25 mM arachidonate were capable of undergoing aggregation and serotonin release in response to other stimuli, such as collagen or thrombin. Thus, GFP appeared to be metabolically intact and functional during the inhibiton by high arachidonate levels. Thin-layer chromatographic studies revealed that prostaglandin metabolism was not changed at the high arachidonate levels. In addition, indomethacin (20 microM) did not abolish the arachidonate-induced inhibition of platelet function. Therefore, the inhibitory effect of high arachidonate did not depend on its conversion to other prostaglandin products. Platelet cyclic AMP levels increased twofold at the high arachidonate concentrations (1.3 +/- 0.3 pmole/10(8) platelets at peak aggregation, compared with 2.9 +/- 0.4 pmole/10(8) platelets at inhibition by 0.25 mM arachidonate, p less than 0.001). Prostaglandin-D2, a platelet inhibitor known to increase cyclic AMP, generated a similar rise (to 2.4 +/- 0.2 pmole/10(8) platelets). Thus, the magnitude of the arachidonate-induced increase in platelet cyclic AMP levels can account for the inhibition of aggregation and release.
Lactate dehydrogenase isoenzymes in normal and malignant human lymphoid cells
Authors: Blatt J , Spiegel RJ , Papadopoulos NM , Lazarou SA , Magrath IT , Poplack DG
Published: October 2019

Intracellular lactate dehydrogenase (LD) isoenzyme patterns were studied in the malignant cells of patients with a variety of lymphoid malignancies. These were compared with intracellular LD isoenzyme patterns of normal lymphoid cells and were also correlated with immunologic cell surface marker characteristics. Results showed that, in general, the malignant B cells of Burkitt's lymphoma and the lymphoblasts of T-cell acute lymphoblastic leukemia had isoenzyme patterns similar to those of normal B and T cells, respectively. The isoenzyme patterns of malignant lymphoid cells from patients with non- T, and non-B acute lymphoblastic leukemia, cutaneous T-cell lymphoma, and chronic lymphocytic leukemia were more heterogeneous. These data, although based on small numbers of patients, are consistent with the hypothesis that LD isoenzymes may reflect differences in the maturational status of cells within a single diagnostic category.
Extrarenal erythropoietin production by macrophages
Authors: Rich IN , Heit W , Kubanek B
Published: October 2019

Murine bone marrow and adherence-separated spleen cells cultured on hydrophobic, gas-permeable Teflon foils (petriperm dishes) can be shown to synthesize and secrete erythropoietin (Epo) and colony-stimulating activity (CSA) simultaneously into the surrounding medium. The Epo activity in the supernatants of primary cultures as measured by the fetal liver erythroid colony-forming technique, from adherent and nonadherent spleen cells, increases over the first 7 days in culture, followed by a plateau until 14 days. Use of the macrophage-specific cytotoxic agent, crystalline silica, as a tool to release residual Epo contained in these cells produces a similar time-Epo activity curve to that found in the primary supernatants. This, together with functional and morphological examination of the cells, indicates that macrophages are responsible for this activity. The total Epo activity released from adherent and nonadherent spleen cells at plateau levels was estimated to be 25 mU/ml culture/day. Weekly subcultivation of bone marrow and adherence-separated spleen cells initiated from primary cultures demonstrated a massive increase in both Epo activity and CSA above that obtained for the primary cultures. Subcultivation could be continued for at least 6 wk. These results, together with the reversible inhibition of Epo and CSA production by cycloheximide, demonstrate that these molecules are synthesized by the macrophage. The evidence supports the hypothesis that the macrophage is involved not only in extrarenal Epo production, but also in the possible short-range regulation of hemopoiesis.
Acidic isoferritins and E-type prostaglandins in sources of colony stimulatory factors mask detection of cycling granulocyte-macrophage progenitor cells
Authors: Broxmeyer HE
Published: October 2019

The effect of granulocyte-macrophage colony stimulatory factors (GM- CSF), acidic isoferritins, and E-type prostaglandins on the detection of the cycle status of human granulocyte-macrophage progenitor cells (CFU-GM) was investigated. Bone marrow cells were pulse-treated with control medium or high specific activity tritiated thymidine [3HTdr) and subsequently plated over feeder layers containing mononuclear blood leukocytes prepared in the absence or presence of anti-acidic isoferritins and/or indomethacin, or plated in the presence of medium conditioned by placental cell or GCT-conditioned media free of acidic isoferritins and prostaglandin-E. The presence of anti-acidic isoferritins and/or indomethacin in the blood leukocyte feeder layers increased the detectable stimulatory capacity of these cells and permitted detection of a larger proportion of marrow CFU-GM in cycle than in control cultures. The cycle status was not influenced by GM-CSF in conditioned medium regardless of the dilution of conditioned medium used to stimulate colony formation. This suggests that GM-CSF, supplied to the cells after treatment with 3HTdr, does not itself influence the detection of CFU-GM in cycle, but using sources of GM-CSF that contain acidic isoferritins or prostaglandin-E will underestimate the actual number of CFU-GM in S-phase.
A prospective study of the usefulness of the measurement of platelet- associated IgG for the diagnosis of idiopathic thrombocytopenic purpura
Authors: Kelton JG , Powers PJ , Carter CJ
Published: October 2019

The measurement of platelet-associated IgG (PAIgG) is a potentially useful diagnostic test for idiopathic thrombocytopenia purpura (ITP). However, the restricted application of PAIgG measurements to thrombocytopenic populations primarily comprised of ITP patients will artificially enhance its diagnosis specificity. For this reason, we performed a prospective study in which the results of a sensitive technique for quantitating PAIgG were related to the cause of the thrombocytopenia. Over a 1-yr period, clinicians were invited to submit patient blood samples encompassing as wide a spectrum of thrombocytopenic disorders as possible for PAIgG measurements. The physician was then contacted and requested to indicate the likeliest cause for the thrombocytopenia. The PAIgG was elevated in only 24 of 254 samples obtained from nonthrombocytopenic patients. In contrast, 134 (79%) of the 169 thrombocytopenic patients had elevated PAIgG results, and the increased levels were apparent in all diagnostic categories. The sensitivity of the PAIgG test for clinically diagnosed idiopathic thrombocytopenic purpura was 91% and the specificity was 27%. The positive predictive value for a raised PAIgG as a diagnostic test for ITP in a thrombocytopenic patient was only 46%, while the negative predictive value was 82%. This study indicates that the presence of increased PAIgG provides little additional information in the diagnosis of ITP. This study also suggests that immune mechanisms may mediate many more thrombocytopenic disorders than have been previously thought likely.
Elevated NAD(P) glycohydrolase activity: a possible enzymatic marker for malignancy in Burkitt's lymphoma cells
Authors: Skala H , Lenoir GM , Pichard AL , Vuillaume M , Dreyfus JC
Published: October 2019

A comparative analysis of enzymatic activities has been performed on 47 human continuous lymphoid lines: 22 tumors derived from Burkitt's lymphoma lines, 6 other lymphomatous long-term cultures, and 19 nonmalignant ties determined on the cell extracts. 4 showed no significant differences between the various lines. They included adenosine diphosphoribose incorporation, glucose-6-phosphate dehydrogenase, cyclic-AMP phosphodiesterase, and glutathione reductase. However, striking differences of activity were found for the enzyme, NAD(P) glycohydrolase (EC 3.2.2.6). Activity levels were, as a mean, four times higher in Burkitt's lymphoma-derived cell lines than in nonmalignant control lines, and the difference was highly significant (p less than 0.02). All Burkitt cell lines containing translocations of chromosome 8 with either chromosomes 2, 14 or 22 showed an increased activity. The specificity and significance of this possible enzymatic marker of Burkitt's lymphoma cells is discussed.
Acquired amegakaryocytic thrombocytopenic purpura: a syndrome of diverse etiologies
Authors: Hoffman R , Bruno E , Elwell J , Mazur E , Gewirtz AM , Dekker P , Denes AE
Published: October 2019

The possible pathogenetic mechanisms responsible for the production of acquired amegakaryocytic thrombocytopenic purpura (AATP) were investigated in a group of patients with this disorder. Absence of megakaryocytes and small platelet glycoprotein-bearing mononuclear cells, as determined by immunochemical staining of patient marrows with an antisera to platelet glycoproteins, suggested that the defect in AATP occurs in an early progenitor cell of the megakaryocytic lineage. Using an in vitro clonal assay system for negakaryocytic progenitor cells or megakaryocyte colony-forming units (CFU-M), the proliferative capacity of AATP marrow cells was then assessed. Bone marrow cells from three of four patients formed virtually no megakaryocyte colonies, suggesting that in these individuals the AATP was due to an intrinsic defect in the CFU-M. Bone marrow cells from an additional patient, however, formed 12% of the normal numbers of colonies, providing evidence for at least partial integrity of the CFU-M compartment in this patient. Serum specimens from all six patients were screened for their capacity to alter in vitro megakaryocyte colony formation. Five of six sera enhanced colony formation in a stepwise fashion, demonstrating appropriately elevated levels of megakaryocyte colony- stimulating activity. The serum of the patient with partial integrity of the CFU-M compartment, however, stimulated colony formation only at low concentrations. At higher concentrations, this patient's serum actually inhibited the number of colonies cloned, suggesting the presence of a humoral inhibitor to CFU-M. Serum samples from all patients were further screened for such humoral inhibitors of megakaryocyte colony formation using a cytotoxicity assay. The patient whose serum was inhibitory to CFU-M at high concentrations was indeed found to have a complement-dependent serum IgG inhibitor that was cytotoxic to allogeneic and autologous marrow CFU-M but did not alter erythroid colony formation. These-studies suggest that AATP can be due to at least two mechanisms: either an intrinsic effect at the level of the CFU-M or a circulating cytotoxic autoantibody directed against the CFU-M.
Colony-forming cell proliferation: a rapid and sensitive assay system for murine granulocyte and macrophage colony-stimulating factors
Authors: Burgess AW , Nicola NA , Johnson GR , Nice EC
Published: October 2019

A microculture assay for murine granulocyte-macrophage colony- stimulating factor (GM-CSF) has been developed using fetal liver GM colony-forming cells (CFC) isolated by fluorescence-activated cell sorting. These GM-CFC are free of mature hemopoietic cells, such as granulocytes and macrophages, which may interfere with direct assays for GM-CSF. The assay procedure allows the quantitation of GM-CSF within 48 hr by measuring the number of cells produced from 50 GM-CFC in microcultures (15 microliter). The assay is particularly simple to set up and score and yet, because of the reduced volumes, this assay is still capable of detecting 0.2 pg (i.e., 0.2 U) of GM-CSF within 48 hr, i.e., 100 times less GM-CSF than the conventional soft agar assay. By allowing the microcultures to develop for 7 days, the extra proliferation allows a further tenfold increase in the sensitivity of CSF detection. The time and cost of setting up hundreds of GM-CSF assays for fractions from chromatographic columns, e.g., reverse phase high performance liquid chromatography, is reduced by at least five- fold. Enough GM-CFC can be isolated and stored frozen in one afternoon to provide sufficient cells for the daily assay of 200 samples of GM- CSF for several months. Microassay results for several sources of GM- CSF at different stages of purification are compared to the results obtained from the soft agar assay.
A multivariate analysis of prognostic factors in chronic myeloid leukemia
Authors: Cervantes F , Rozman C
Published: October 2019

The prognostic value of different clinical and laboratory findings at diagnosis of chronic myeloid leukemia (CML) was analyzed in a series of 121 cytogenetically studied patients. From the univariate and multivariate analysis of the whole series it was apparent that the minority of Ph1-negative patients (11.5%) could be considered as a poor prognosis group. The analysis was then restricted to the Ph1-positive patients. From a multivariate survival analysis (Cox's regression model) of the latter group the following poor prognosis factors emerged: splenomegaly, hepatomegaly, presence of erythroid precursors in peripheral blood, and bone marrow myeloblasts over 5%. From the contribution of each one of these factors to the regression model, a clinical staging of Ph1-positive CML was derived: stage I (low risk, 32% of patients), including patients with one or no factors; stage II (intermediate risk, 38%), including cases with two factors; and stage III (high risk, 30%), including patients with three or four factors. The difference in survival of the patients at different stages was highly significant (p less than 0.001).
111In oxine-labeled rabbit platelets: in vivo distribution and sites of destruction
Authors: Hill-Zobel RL , Scheffel U , McIntyre PA , Tsan MF
Published: October 2019

We have studied the kinetics, biodistribution, and fate of autologous platelets labeled with 111In-oxine in rabbits. The initial recovery was 75% and mean survival time was 2.8 days when the data were analyzed by the multiple-hit gamma function model. Using a modified geometric mean for correction of attenuation, there was good correlation between the values obtained by in vivo quantification and those obtained by postmortem measurements of the radioactivity in the liver and the spleen (i.e., r = 0.854 and 0.899, respectively, n = 32). Using this method, it was shown that after infusion, the 111In-platelets rapidly accumulated in these two organs reaching 35% and 12% of the injected dose in the liver and spleen, respectively, by 1 day. Thereafter, there was little subsequent change. On the sixth day, when essentially all of the 111In-platelets had cleared from the circulation, a total of 82% of the injected dose was deposited in the three major reticuloendothelial organs: liver (40%), spleen (14%), and bone marrow (28%). Our results suggest that in addition to liver and spleen, bone marrow played an important role in sequestering platelets in rabbits.
A human granulocyte-specific antigen characterized by use of monoclonal antibodies
Authors: Skubitz KM , Zhen Y , August JT
Published: October 2019

Hybridoma cell lines secreting monoclonal antibodies that bind to a surface antigen of human neutrophils have been prepared by fusion of mouse myeloma cells with spleen cells from mice immunized with human neutrophils. Several of the monoclonal antibodies (AHN 1–6) were specific for a neutrophil surface antigen and did not bind lymphocytes, monocytes, red blood cells, platelets, or basophils. All of the granulocyte-specific antibodies immunoprecipitated a polypeptide of 145,000 daltons and an isoelectric point of about 4.5 and other heterogeneous polypeptides of 105,000 daltons. These same components were the major lactoperoxidase-labeled proteins precipitated by hyperimmune mouse serum. The antibodies were further characterized for binding to several human myeloid leukemia cell lines and cells from patients with myeloid or lymphoid leukemia. All antibodies bound the HL- 60, ML1, ML2, ML3, K562, and U937 myeloid leukemia cell lines. None of the antibodies bound the RPMI 6410 Raji, RPMI 8226, MOLT 4, or Daudi lymphoid cell lines. All of the hybridoma cell lines (AHN 1–6) produced IgM antibodies that were cytotoxic.
Glycosaminoglycans in human neutrophils and leukemic myeloblasts: ultrastructural, cytochemical, immunologic, and biochemical characterization
Authors: Parmley RT , Hurst RE , Takagi M , Spicer SS , Austin RL
Published: October 2019

Chondroitin sulfate is known to be present in normal and leukemic myeloid cells; however, its definitive subcellular location and association with other glycosaminoglycans (GAGs) has not been demonstrated. We have studied the type and distribution of GAGs in neutrophil granule subpopulations of normal and leukemic myeloid cells using ultrastructural, cytochemical, immunologic, and biochemical methods. At the ultrastructural level, high-iron diamine- thiocarbohydrazide-silver proteinate (HID-TCH-SP) stained sulfated glycoconjugates selectively in immature primary granules of normal promyelocytes and Auer rods and immature granules of leukemic myeloblasts. Staining was weak or absent in mature primary granules, whereas tertiary granules stained moderately. Primary granule staining with HID-TCH-SP was greatly diminished by prior treatment of the specimens with chondroitinase ABC and/or nitrous acid, indicating the presence of chondroitin sulfate and N-sulfated glycosaminoglycan. Immunostaining of myeloid cells with a rabbit antichondroitin 4-sulfate and ferritin-conjugated goat anti-rabbit IgG sequence resulted in staining of most primary granules. Biochemical analysis of GAGs from leukemic myeloblasts containing primary granules and Auer rods, but lacking secondary and tertiary granules, revealed 8 x 10(-17) mole of uronic acid/cell and electrophoretic and sulfaminohexose analysis showed 60%-70% chondroitin sulfate AC of heterogeneous molecular weight, 20%-30% of a GAG that most closely resembled heparan sulfate, and 10% dermatan sulfate. The lack of significant HID-TCH-SP staining of sulfate iin sites other than Auer rods and primary granules in leukemic myeloblasts indicates that these granules contain the chondroitin, dermatan, and heparan sulfate isolated from the same specimen. Similar GAGs are present in primary granules of normal cells as evidenced by their cytochemical and immunostaining properties. Thus, these studies demonstrate a heterogeneous population of GAGs not previously identified and localize these substances to the primary granule of leukemic and normal cells.
The significance of transferrin for intestinal iron absorption
Authors: Huebers HA , Huebers E , Csiba E , Rummel W , Finch CA
Published: October 2019

A mechanism is proposed by which apotransferrin is secreted from mucosal cells, loaded with iron in the intestinal lumen, and then the intact complex is taken into the cell. Within the cell, iron is released and transferred to the blood stream, whereas iron-free transferrin returns to the brush border to be recycled. We have investigated this hypothesis by measuring intestinal absorption of radioiron and 125I-labeled plasma transferrin using tied-off gut segments in normal and iron-deficient rats. There was no absorption of diferric transferrin from the ileum, but high absorption from the duodenum and jejunum segments. Jejunal absorption occurred as a function of the dose offered and showed saturation kinetics. In normal animals, 4 micrograms of the 50 micrograms of transferrin iron was absorbed over 1 hr. In iron-deficient animals, mean values as high as 13 micrograms were observed. Radioiron content of the jejunal mucosa bore a linear relationship to the dose administered and was inversely proportional to the amount of iron entering the plasma. Recycling of transferrin was indicated by the presence of labeled apotransferrin in the lumen, first observed between 15 and 60 min after the injection of diferric transferrin. A high resistance of diferric and apotransferrin to proteolytic degradation within the gut lumen was demonstrated. Comparative studies with lactoferrin and ferritin disclosed poor availability of their iron for absorption. The small amount that was absorbed did not relate to the iron status of the recipient animal. These studies support the role of mucosal transferrin as a shuttle protein for iron absorption.
Efficacy and morbidity of central nervous system "prophylaxis" in childhood acute lymphoblastic leukemia: eight years' experience with cranial irradiation and intrathecal methotrexate
Authors: Inati A , Sallan SE , Cassady JR , Hitchcock-Bryan S , Clavell LA , Belli JA , Sollee N
Published: October 2019

Between 1972 and 1979, 214 children with acute lymphoblastic leukemia and no evidence of central nervous system (CNS) disease prior to CNS prophylaxis were treated with 2400 rad cranial irradiation and concurrent intrathecal methotrexate. Only nine children developed CNS leukemia; five of them in the CNS only and four concurrently in the CNS and another site. Major acute effects of CNS prophylaxis were seizures in seven patients (3%). Sixty-nine children who had a minimum follow-up of 4 yr were evaluable for late effects of therapy. Small cataracts, incomplete regrowth of hair, and learning disabilities were noted. The latter occurred in 18% of patients, an incidence similar to that encountered in a normal community of school-age children. However, the incidence of learning disabilities in patients who were under 5 yr of age at the time of diagnosis was much higher, 35%. We conclude that the combination of cranial irradiation and intrathecal methotrexate was highly efficacious. The incidence and severity of neuropsychologic abnormalities, the principal late morbidity of this treatment program, varies among reporting institutions. Prospective longitudinal studies of neuropsychologic function are necessary to better define the incidence of abnormalities. Future programs should attempt to decrease late morbidity, but must also assure equal efficacy and improve overall disease-free survival.
Response to splenectomy in 65 patients with hairy cell leukemia: an evaluation of spleen weight and bone marrow involvement
Authors: Golomb HM , Vardiman JW
Published: October 2019

Sixty-five patients with hairy cell leukemia underwent splenectomy; 27 had a complete remission as defined by a return in WBC, RBC, and platelet counts to a defined level, and 38 had a partial remission with a return of only one or two of these parameters to the defined level. The 5-yr actuarial survival for all patients is 68%; for CR patients it is 76%, and for PR patients 62%. The response to splenectomy did not correlate with the spleen weight. Seventeen patients had a postsplenectomy platelet count of less than 200 x 10(9)/liter, and 34 patients had a postsplenectomy platelet count of 200 x 10(9)/liter or greater. A presplenectomy bone core biopsy hairy cell index (HCI) was calculated by multiplying the percent marrow cellularity by the percent of hairy cells in the marrow for 51 patients. The difference in the mean HCI between the two platelet response groups is statistically significant (p less than 0.05). Of the 15 patients with a presplenectomy HCI of 0.7 or greater, 9 (60%) did not have a satisfactory platelet response to splenectomy, whereas of 36 patients with an HCI of 0.7 or less, only 8(22%) did not have a satisfactory platelet response to splenectomy (p less than 0.01). The HCI appears to indicate the significance of underproduction of platelets as a result of marrow replacement by hairy cells.
Effect of aspirin and sodium salicylate on thrombosis, fibrinolysis, prothrombin time, and platelet survival In rabbits with indwelling aortic catheters
Authors: Cattaneo M , Chahil A , Somers D , Kinlough-Rathbone RL , Packham MA , Mustard JF
Published: October 2019

We have studied the effect of different doses of aspirin on platelet function, PGI2 formation, platelet survival, thrombosis, fibrinolysis, and prothrombin time in rabbits with indwelling aortic catheters. The thrombi formed around indwelling aortic catheters were found to have a large fibrin component, and their formation was inhibited by heparin administration. Thus, in these experiments we examined the effect of aspirin (a weak inhibitor of thrombin-mediated platelet aggregation) under conditions in which thrombin was a major factor in the initiation and growth of the thrombi. Only very high doses of aspirin tended to inhibit thrombus formation over the 5-day period of observation, and a statistically significant inhibition of thrombus formation was produced by equivalent concentrations of sodium salicylate. The failure of high doses of aspirin to achieve a significant inhibition of thrombosis under the conditions of these experiments (whereas an equivalent dose of sodium salicylate was inhibitory) could be due to aspirin inhibition of PGI2 formation. Shortened platelet survival was not affected by aspirin treatment or the dose sodium salicylate that inhibited thrombus formation. The tendency to inhibit thrombus formation appeared to be unrelated to an effect on platelets but was associated with prolongation of the one-stage prothrombin time and increased whole blood fibrinolytic activity; doses of aspirin that inhibited platelet aggregation in response to sodium arachidonate or collagen, and PGI2 formation by the vessel wall, did not have a significant effect on the amount of thrombus present at 5 days. However, the high doses of aspirin that inhibited PGI2 formation were associated with a tendency to increased thrombus formation during the first 3 hr after insertion of the catheter. The results of these experiments show that when thrombin is an important factor in the formation of thrombi, aspirin is a weak inhibitor of thrombosis unless doses are used that provide sufficient salicylate to interfere with blood coagulation and promote whole blood fibrinolytic activity. These results also show that thrombus formation can be inhibited without an apparent change in platelet survival.
The expression of myeloid-specific antigens on myeloid leukemia cells: correlations with leukemia subclasses and implications for normal myeloid differentiation
Authors: Ball ED , Fanger MW
Published: October 2019

The expression of three distinct myeloid-specific cell surface antigens detected by monoclonal antibodies (PMN 6, PMN 29, and AML-2–23) on acute and chronic myeloid leukemia cells is correlated with blast cell morphology and normal myeloid cell antigen display. In studies on normal peripheral blood cells, monoclonal antibodies PMN 6 and PMN 29 have previously been shown to react exclusively with neutrophils while AML-2–23 reacts with both neutrophils and monocytes. The present report demonstrates that these antigens are absent from blast cells of patients with acute myelocytic leukemia (AML) classified as M1 and M2 in the French-American-British system and chronic myelocytic leukemia in myeloid blast crisis. However, leukemia cells with myelomonocytic morphology (M4) expressed all three antigens, while cells with pure monocytic features (M5) were generally only positive for AML-2–23. Based on the absence of these antigens on both leukemic and normal myeloblasts and granulocyte-monocyte progenitors and their characteristic patterns of display on more differentiated leukemic and normal cells, we propose a modified concept of normal myelopoiesis. In this hypothesis, the myeloblast is an uncommitted cell that gives rise to a series of intermediate precursors that acquire committment to either the granulocytic or monocytic lineage marked by the acquisition of specific cell surface markers.
Extracellular matrix production by the adherent cells of long-term murine bone marrow cultures
Authors: Zuckerman KS , Wicha MS
Published: October 2019

We have studied the deposition of extracellular matrix proteins in the adherent stroma of long-term murine bone marrow cultures. Stable hematopoiesis was maintained for greater than 12 wk. At selected intervals, culture dishes were sacrificed by removing all nonadherent cells and air drying the dishes. The adherent stromal layer was analyzed for the presence of intracellular and extracellular collagen, fibronectin, and laminin using double immunofluorescent staining with specific antisera against these matrix components. In cultures examined during the first 2 wk, large numbers of stromal cells contained collagen, fibronectin, and laminin. Over the next 2 wk, an extensive extracellular network of fibronectin, laminin, and collagen was deposited on the dishes, which persisted throughout the life of the cultures. In contrast to a previous report, we detected substantial numbers of endothelial cells by means of immunofluorescent staining of stromal cells with antisera to type IV collagen, laminin, and factor VIII antigen. Although deposition of these extracellular matrix proteins coincides with onset of active hematopoietic cell production, the relative roles of the stromal cells and the extracellular matrix in supporting hematopoiesis in murine bone marrow cell cultures remain to be determined.
Circulating lymphocyte populations in Hodgkin's disease after mantle and paraaortic irradiation
Authors: Posner MR , Reinherz E , Lane H , Mauch P , Hellman S , Schlossman SF
Published: October 2019

The effect of mantle and paraaortic radiation on peripheral blood lymphocytes was studied in 11 previously untreated patients with early stage Hodgkin's disease using a series of monoclonal antibodies defining immunoregulatory lymphoid cells. Immediately following the completion of radiotherapy, there was a significant reduction in the number of lymphocytes and the percent of circulating T cells. This acute decrease in T cells was due to a marked diminution in the number of inducer T cells, while the fraction of suppressor T cells remained constant. These changes persisted for up to 12 mo and were accompanied by a later increase in the fraction of circulating B lymphocytes and cells bearing surface Ia. It thus appears that mantle and paraaortic radiotherapy causes a relatively selective reduction in the inducer T- cell population. The implications of a change in the ratio of inducer to suppressor T cells is discussed.