bluuebunny/crossref_metadata_2025 huggingface dataset:
THE ANEMIA OF INFECTION, VI. THE INFLUENCE OF COBALT ON THE ANEMIA ASSOCIATED WITH INFLAMMATION
Authors: WINTROBE M. M. , GRINSTEIN M. , DUBASH J. J. , HUMPHREYS S. R. , ASHENBRUCKER HELEN , WORTH WANDA
Published: October 2019

The influence of cobalt on the anemia associated with inflammation has been studied in three experiments involving observations in 108 rats.

It was found that by the simultaneous administration of cobalt the anemia associated with inflammation, as produced by the injection of turpentine, could be prevented from developing and polycythemia appeared instead.

This effect was accompanied by hypoferremia and an increase in erythrocyte protoporphyrin values similar to those encountered when anemia develops in association with inflammation.

Similar, though less marked, chemical changes were observed when only cobalt was given and polycythemia was produced.

A decrease in plasma albumin was noted in rats injected with cobalt or turpentine, or both, but this was not accompanied by an increased excretion of urinary nitrogen as measured by the urine urea and ammonia.

The observations cited are consistent with the hypothesis that cobalt favorably influences the utilization of iron for the synthesis of hemoglobin.
THE INFLUENCE OF ANEMIA ON PHAGOCYTIC FUNCTIONS AND RESISTANCE TO INFECTION IN MICE
Authors: BERRY L. JOE , HALLER EVELYN C.
Published: October 2019

The increase in phagocytic function of neutrophiles of mice was measured during the progressive development of anemia due to blood loss. The animals were bled from the tail, 2 per cent of total body weight being removed twice weekly for nine bleedings. Phagocytosis was 161 per cent of normal at the end of this period, and the erythrocyte totals and hemoglobin values were approximately two thirds of normal. The anemic mice were infected intraperitoneally with a suspension of Salmonella typhimurium and their survival rate was compared with normal mice similarly infected. Twenty-eight of 63 anemic mice survived and 14 of 63 normal mice survived.

In a second experiment, mice were subjected to bleeding every third day for nine bleedings, and were then infected along with a control group. One half of the anemic animals were bled for three additional bleedings after the infection. Two of 48 control animals survived, 5 of 47 anemic mice, additionally bled, survived, and 19 of 47 anemic mice, not bled after infection, survived. This suggests that the increase in phagocytosis that accompanies anemia may be, at least in part, responsible for the increased resistance to infection.
CERTAIN CHARACTERISTICS OF THE LEUKOCYTES OF GUINEA PIG BLOOD WITH PARTICULAR REFERENCE TO THE KURLOFF BODY
Authors: SMITH EMILY
Published: October 2019

A critical review of the literature on the leukocytes of the guinea pig has been presented, with added observations from studies of the blood cells of more than 500 guinea pigs.

Special studies of the Kurloff inclusion body were made on these animals to obtain information regarding its origin and significance because this entity occurs in the mononuclear leukocyte, which is a cell of great interest in the pathogenesis of tuberculosis.

The normal blood picture of the guinea pig was discussed and summarized in terms of the blood picture of the normal human being.
THROMBOCYTOPENIC PURPURA COMPLICATING GOLD THERAPY FOR RHEUMATOID ARTHRITIS
Authors: METTIER STACY R. , Mc BRIDE ALICE , LI JONAH
Published: October 2019

Of 160 patients treated with gold salt therapy, four developed thrombocytopenia with mild purpuric manifestations. In three patients there was a spontaneous remission and the hemorrhagic tendency disappeared in about one week’s time.

In the fourth patient there was a second occurrence of thrombocytopenia which persisted over a period of four months, and finally failed to respond to transfusions. Splenectomy was followed by a dramatic rise in the blood platelets and recovery of the patient.
THE HEREDITARY MECHANISM OF GAUCHER’S DISEASE
Authors: GROEN J.
Published: October 2019

The author reviews the literature of the familial incidence of Gaucher’s disease. Almost all the familial cases which have been described occurred in the members of one generation (siblings or cousins) only. To these cases reported in the literature the author adds 25, of which 4 were "sporadic" cases. The other 21 cases occurred in 6 families. The pedigrees of these families are presented. After an analysis of the available data the author presents the following hypothesis for the hereditary mechanism in these families:

Gaucher’s disease is a mutation which, once established, is transmitted as a simple dominant hereditary trait. In the affected individuals this trait gives rise to a disturbance of lipid metabolism which results in the accumulation of kerasin in the reticulum cells throughout the body. The severity of the disease may vary considerably. It can be present in such a slight degree that the amount of kerasin accumulated during life is too small to give rise to clinical manifestations. In other cases the progression may be so slow that the disease becomes manifest only in old age, provided the affected individual lives long enough. In these "subclinical cases," a diagnosis of Gaucher’s disease can sometimes be made by the detection of "early Gaucher cells" in the sternal marrow. Individuals thus affected suffer from Gaucher’s trait rather than from the actual disease. However, they can transmit the disease to 50 per cent of their offspring, and thus function as (almost) normal "carriers." In the family trees presented, it appeared that the disease tended to become more severe in every succeeding generation until after two or three generations it became clinically manifest in the affected individuals early in life. In the next generation it would then establish itself during fetal life so as to give rise to abortion, stillbirth or early death of the affected infant. In this way the mutation extinguishes itself, by permitting only the unaffected offspring of the affected individuals to persist. As a practical conclusion it is urged that a sternal marrow examination be included in every genetic investigation of Gaucher’s disease as the best method available at present for the detection of subclinical cases or "carriers."
A METHOD FOR DETERMINING THE FORM OF THE DISTRIBUTION OF RED CELL RESISTANCES TO SIMPLE HEMOLYSINS
Authors: PONDER ERIC
Published: October 2019

A method is described for determining quantitatively the form of the resistance distributions of red cell resistances to simple hemolysins. The normal range of variation is given for the resistance distributions to saponin, digitonin, and hypotonic NaCl, all at pH 7.0, and examples of departures from the normal, which may take the form of bimodalities after hemorrhagic or hemolytic episodes or of more persistent changes in the characteristics of the distributions, are provided.
THE Rh CHROMOSOME FREQUENCIES IN ENGLAND
Authors: RACE R. R. , MOURANT A. E. , LAWLER SYLVIA D. , SANGER RUTH
Published: October 2019

The results are reported of testing 1073 English bloods with the Rh antibodies anti-C, anti-Cw, anti-c, anti-D, anti-E and anti-e. The results of another series of 927 bloods, already published, are here reproduced. The total of 2000 bloods has been used by Fisher to estimate, by his method of maximum likelihood, the Rh chromosome frequencies in England. The estimates are: CDe 40.75 per cent, cde 38.86 per cent, cDE 14.11 per cent, cDe 2.57 percent, CwDe 1.29 per cent, cdE 1.19 per cent, Cde 0.98 per cent, and CDE 0.24 per cent.

A brief account is given of the three pairs of alternative antigens shown by Fisher to be the basis of the Rh blood groups. Fisher’s interpretation must now be considered as established beyond doubt. A possible genetic basis of these related antigens is discussed.
PARA-AMINOBENZOIC ACID IN LEUKEMIA
Authors: ZARAFONETIS C. J. D. , ANDREWS G. A. , MEYERS M. C. , BETHELL F. H.
Published: October 2019

Para-aminobenzoic acid, administered in large doses as sodium para-aminobenzoate caused a striking lowering of the leukocyte counts in five patients with chronic myelogenous leukemia and in one patient with subacute myelogenous leukemia.

NaPAB caused less definite decreases of the white cell count in two patients with chronic lymphatic leukemia, but the periods of administration may have been too short to obtain maximal effects.

In every instance, there was a prompt rise in the number of leukocytes when the administration of NaPAB was stopped.

Although there was decrease in spleen size in some of the patients, the objective clinical improvement was but slight and temporary.

All patients receiving NaPAB in large doses had concomitant glycosuria, apparently on a renal basis.

It is to be emphasized that NaPAB is not considered a practical adjunct to the therapy of leukemia at this time. Rather, it is hoped that studies of the cellular chemistry involved in the apparent inhibitory action of NaPAB may yield information concerning the disordered metabolism of leukemic cells.
CHRONIC INFECTIOUS MONONUCLEOSIS
Authors: ISAACS RAPHAEL
Published: October 2019

A group of patients is described in whom ease of fatigue, fever, splenomegaly, low blood pressure, low blood sugar, low specific gravity of the urine and the presence of infectious mononucleosis cells in the blood persisted for from three months to longer than four years after the initial attack.

Three of the group developed characteristics of lymphoblastoma and two showed the features of Banti’s congestive splenomegaly.

The symptoms responded to treatment with a preparation of adrenal cortical extract.

The syndrome is apparently not uncommon and the intense, prolonged debility, together with the marked improvement after therapy with adrenal cortical extract, makes its recognition of great practical importance.
THE USE OF URETHANE (ETHYL CARBAMATE) IN THE TREATMENT OF LEUKEMIA
Authors: WATKINS CHARLES H. , COOPER TALBERT , GIFFIN HERBERT Z.
Published: October 2019

Limited experience with the use of urethane in the treatment of leukemia indicates that this substance presents a considerable promise as a palliative agent in chronic myelogenous leukemia. It has no apparent value in the treatment of acute leukemia.

Further extensive observation will be necessary to provide a true measure of the clinical usefulness of this preparation.
URETHANE THERAPY IN LEUKEMIA
Authors: CRESKOFF ADOLPH J. , FITZ-HUGH THOMAS , FROST JOHN W.
Published: October 2019

1. The results of urethane therapy in 24 cases of leukemia are described.

2. The average daily dose is 4 Gm. orally or intravenously.

3. The drug is irregularly effective. Chronic myelogenous leukemia appears more responsive than the lymphatic variety.

4. Acute leukemias are not significantly altered in course by urethane.

5. Urethane produces a fall in the total leukocyte count in a majority of all types of leukemia. Clinical improvement does not necessarily follow.

6. Nausea is the most frequent side effect of urethane therapy. Possible marrow aplasia is the most dangerous toxic effect.

7. Urethane is of definite, but of limited value in the treatment of chronic leukemia. In some instances, it compares favorably with x-ray therapy, but in general, it is less dependable, particularly in its frequent failure to induce optimum return of normal red cell and platelet values, and optimum regression of organ infiltration.
A HEMATOPOIETIC PERIFOLLICULAR ENVELOPE IN THE RAT SPLEEN
Authors: KRUMBHAAR E. B.
Published: October 2019

About the Malpighian follicles of several species of rodents, and especially prominent in rats, is a perifollicular envelope composed of hematopoietic cells that takes active part in hemolytopoietic changes.

The identity of the mononuclear cell comprising the greater part of this tissue has not been positively determined; it is probably a young lymphocyte. It is possibly the homologue of the pale centers of the Malpighian follicles in man and other mammals, though it has been found in rabbit spleens which also have pale centers. It should be possible to determine this identity by the use of a greater variety of stains, and of test poisons, and, if possible, of dynamic methods such as tissue culture and moving pictures.

The envelope is separated from the Malpighian follicle by a thin rind of collagenous connective tissue, but on its outer margin it merges gradually with the red pulp. It often contains a scattering of erythrocytes, normoblasts, polymorphonuclear neutrophils, and rarely eosinophils and pigment-bearing macrophages. Some of these cells were so greatly increased under the pathologic conditions first studied that colonization was suggested; they were later thought to have probably wandered in. The collar never contains megakaryocytes or sinuses or blood vessels of any noteworthy size.
BARTONELLA BACILLIFORMIS ANEMIA (OROYA FEVER) A STUDY OF THIRTY CASES
Authors: RICKETTS WILLIAM E.
Published: October 2019

Bartonella bacilliformis anemia (Oroya fever) is a febrile hemolytic anemia with distinguishing clinical and hematologic characteristics. It occurs as an infrequent clinical form during the invasive stage. The onset is variable with or without chills, followed by a moderate temperature which does not parallel the intensity of the anemia. Hemorrhages, petechial spots, epistaxis may occur and are due to thrombocytopenia. Clouding of the sensorium and delirium are rather uncommon. There is a generalized lymphadenopathy but no splenomegaly.

The anemia is macrocytic and frequently hypochromic with signs of intense blood formation: young granulocytes, polychromatophilia, reticulocytosis and nucleated red cells. The reticulocytosis may increase to 50 per cent. The pathognomonic sign of the disease is the presence of bartonella bacilliformis on the erythrocytes. The leukocyte count varies; slight leukocytosis is not uncommon but marked leukocytosis is extremely rare in cases without intercurrent infections. There is a shift of the polymorphonuclear series to the left, characterized by the presence of myeloblasts, myelocytes and metamyelocytes. The anemia is normoblastic and not megaloblastic. It is hemolytic and the destruction of erythrocytes is dependent on the presence of bartonella bacilliformis on the erythrocytes. There is no spherocytosis and the fragility of the erythrocytes is normal. Histologically there is evidence of erythrocyte phagocytosis in the cells of the reticulo-endothelial system; the Kupffer cells, the littoral cells of the spleen and the lymph nodes.

The disappearance of the bartonellae from the erythrocytes occurs in a very few days and is called "critical stage" of the anemia. The hematologic changes of this transition are as follows: a change in the shape of the bartonellae from the "bacilliform" to "coccoid" form before complete clearance will take place, an increase in the erythrocyte count, reduction in the indirect hyperbilirubinemia to normal, increase in the number of reticulocytes, reversion to normocytosis, lymphocytosis, reappearance of monocytes and eosinophiles, a "shift to the right" of the polymorphonuclear series. Corresponding with the clearance of bartonella from the erythrocytes, the symptoms dependent on the anemia as well as the fever disappear. Clinical improvement, however, may not parallel the clearance of bartonellae because of intercurrent infections or an atypical course of the bartonellosis itself.

Bartonella bacilliformis anemia has a very severe prognosis, due largely to the occurrence of intercurrent infection by enteric organisms. The treatment today is largely symptomatic as there is no specific agent against the bartonellae bacilliformis infection.

It is suggested that the prognosis may be improved by the use of adequate blood transfusions and the prophylactic use of antibiotics to control intercurrent infection. At present there is no specific agent against bartonella bacilliformis anemia.
Rh ANTIBODIES; CORRELATION WITH CLINICAL FINDINGS
Authors: DAVIDSOHN I.
Published: October 2019

Rh antibodies were studied during pregnancy at frequent intervals in the blood of 73 mothers of babies with fetal erythroblastosis. The serums containing antibodies were divided in 2 groups : (1) serums with blocking antibodies without saline agglutinins; (2) serums with saline agglutinins and serum-albumin agglutinins, but without blocking antibodies. The correlation of these 2 groups with clinical findings in the newborn showed striking differences: blocking antibodies present in 85 per cent of mothers of babies with hydrops or stillborn, and only in 9 per cent of mothers of babies with icterus gravis.

Predominance of saline agglutinins over serum-albumin favored survival, and vice versa.

In a small group of women with a history of transfusions who gave birth to erythroblastotic babies blocking antibodies were found in 92 per cent.

The number of cases used for this study is admittedly small, and only very guarded conclusions may be drawn. The results, however, are in general agreement with recent studies of Wiener on correlation of antibodies with clinical manifestations, and with results of Levine and others regarding the sensitizing influence of blood transfusions.

Periodic tests for antibodies during pregnancy may permit prognostic conclusions.

An anamnestic Rh antibody reaction is described.
THE EFFECT OF HUMAN PLASMA TRANSFUSIONS ON THE FECAL UROBILINOGEN EXCRETION IN SICKLE CELL ANEMIA
Authors: KAPLAN EUGENE , LEWIS S. ROBERT
Published: October 2019

1. The results of transfusions of whole plasma and blood on the fecal urobilinogen excretion of 5 children with sickle cell anemia have been reported.

2. The phenomenon observed by Josephs—a reduction in urobilinogen output following plasma transfusions—has been confirmed.

3. It has been observed further that in certain patients the continued use of plasma transfusions induces a reversal of this phenomenon, namely an increase in urobilinogen output after each transfusion.

4. The possible significance of these phenomena is discussed and attention is called to the limitations inherent in accepting urobilinogen excretion as a valid index of hemoglobin destruction in certain disorders in which a defect of hemoglobin metabolism may be present.
THE EFFECT OF QUANTITATIVE AND QUALITATIVE PROTEIN DEFICIENCY ON BLOOD REGENERATION. II. HEMOGLOBIN
Authors: BUECHLER EDITH , GUGGENHEIM K.
Published: October 2019

1. The effect of diets, varying in quantity or quality of protein, on hemopoiesis was studied in protein depleted and anemic adult rats.

2. In experiments with diets containing different amounts of casein (0, 3, 9 and 18 per cent, respectively), and fed ad libitum, it was found that with a protein-free diet a further decrease of hemoglobin occurred, whereas the other three diets initiated a regeneration of hemoglobin; its degree being more or less proportional to the casein content.

3. In experiments, in which diets with 9 and 18 per cent of casein, respectively, were given in restricted amounts, it was found that the degree of hemoglobin formation was similar to that with the same diets when given ad libitum, whereas the weight gain was considerably less. It is concluded, therefore, that in caloric deficiency hemoglobin formation has a priority over weight recovery.

4. A diet containing 30 per cent casein and given in restricted amounts induced a further weight loss, whereas the concentration of hemoglobin showed a marked increase. Comparing the results obtained by this diet with those observed with 18 per cent casein diets, given either ad libitum or in controlled amounts, it was evident that restriction of the quantity of food protein interferes more seriously with hemopoiesis than restriction of calories.

5. Diets containing nutritionally inferior proteins fed at 9 per cent level, also impaired normal hemopoiesis. Hemoglobin regeneration induced by the proteins investigated was found to decrease in the following order: eggs, meat, processed soya, casein, peanut, maize, wheat, gelatin.

6. Comparing the nutritive value of various proteins for regeneration of hemoglobin and of granulocytes it was found, that casein and soya and maize proteins are considerably more efficient for hemoglobin formation than for production of granulocytes, whereas wheat protein and gelatin have a higher granulocytopoietic capacity.
A new form of hereditary persistence of fetal hemoglobin in blacks and its association with sickle cell trait
Authors: Stamatoyannopoulos G , Wood WG , Papayannopoulou T , Nute PE
Published: October 2019

A new form of hereditary persistence of fetal hemoglobin (HPFH) producing 3%-8% Hb F in heterozygotes and an elevation of F-cell counts as measured by both the Kleihauer test and an antibody fluorescent procedure was found during the study of a black family. Individuals with this anomaly also had sickle cell trait. A sickle cell homozygote who had apparently inherited the HPFH determinant had 20.3% Hb F. Both types of gamma-chains were present in equal proportions in the Hb F of these individuals. A population study revealed other AS individuals with increased Hb F synthesis, three of whom were sibs. The presence of this previously unrecognized form of HPFH might explain the mild clinical manifestations and the hemoglobin phenotypes of sickle cell homozygotes with unusual elevations of Hb F.
Use of cell separation and short-term culture techniques to study erythroid cell development
Authors: Glass J , Lavidor LM , Robinson SH
Published: October 2019

Cell populations highly enriched for the different stages of erythroid cell maturation were obtained by three sequential operations: harvesting of erythroid cells after induction of erythroid hyperplasia in the spleens of mice, elimination of the more mature erythrocytes by immunologic techniques, and separation of the residual nucleated erythroid cells as a function of size by the velocity sedimentation technique. The resulting cell fractions were studied both directly and after overnight incubation in the presence or absence of erythropoietin. In short-term culture, erythropoietin stimulated proliferation of pronormoblasts and basophilic normoblasts but probably not cells at later stages of differentiation. Erythropoietin also appeared to recruit increased numbers of pronormoblasts. In this experimental system, erythroid cell differentiation was able to proceed in the absence of erythropoietin, but without proliferation of these early erythroid cells. These techniques have provided a model system for the study of erythroid cells at different stages of maturation isolated from a uniform source at one point in time. The morphologic observations indicated that erythropoietin stimulates erythroid cell proliferation at several early stages of the maturation pathway.
Deoxycytidine kinase: properties of the enzyme from human leukemic granulocytes
Authors: Coleman CN , Stoller RG , Drake JC , Chabner BA
Published: October 2019

Deoxycytidine kinase, which phosphorylates deoxycytidine (CdR) and its analog, cytosine arabinoside (ara-C), has been purified 71-fold from human leukemic cells. Biochemical properties of the partially purified enzyme included a molecular weight of 68,000, Kms of 7.8 muM for CdR and 25.6 muM for ara-C, and optimal activity with ATP and GTP as phosphate donors. Ara-C phosphorylation was strongly inhibited by CdR (Ki = 0.17 muM) and dCTP (Ki = 7.3 muM) and was weakly inhibited by ara- CTP (Ki = 0.13 mM). Purification by calcium phosphate gel elution and DEAE chromatography effectively separated this enzyme from cytidine deaminase, which deaminates both CdR and ara-C, and from uridine- cytidine kinase, the enzyme which phosphorylates 5-azacytidine. CdR kinase activity was found to decrease and cytidine deaminase to increase with maturation of normal and leukemic granulocytes. Myeloblasts purified by Ficoll sedimentation revealed an average kinase activity of 15.4 U/mg protein in acute myelocytic leukemia and 12.3 U/mg protein in blastic crisis of chronic myelocytic leukemia (CML). The average ratio of CdR kinase to deaminase activity in crude cell extracts varied from 0.197 in AML and 0.089 in blastic crisis to 0.0004 in normal granulocytes, reflecting the changes which take place with cellular maturation. The absolute levels of kinase and deaminase and the ratio of these two enzymes varied considerably among patients with AML, indicating that quantitative differences may be found in the metabolism of CdR and its analogs in leukemic cells.
Five novel mutations located in exons III and IX of the protein C gene in patients presenting with defective protein C anticoagulant activity
Authors: Gandrille S , Alhenc-Gelas M , Gaussem P , Aillaud MF , Dupuy E , Juhan-Vague I , Aiach M
Published: October 2019

We describe five families presenting with type II hereditary protein C deficiency characterized by normal antigen and amidolytic activity levels but low anticoagulant activity. All the exons and intron/exon junctions of the protein C gene were studied using a strategy combining amplification by the polymerase chain reaction (PCR), denaturing gradient gel electrophoresis of the amplified fragments, and direct sequencing of fragments displaying altered melting behavior. We detected five novel mutations. Three were located in the C-terminal part of the propeptide encoded by exon III: Arginine (Arg)-5 to tryptophan (Trp), Arg-1 to histidine (His), and Arg-1 to cysteine (Cys) mutations. The two others, located in exon IX, affected Arg 229 and serine (Ser) 252, which were respectively replaced by glutamine (Gln) and asparagine (Asn). DNA studies of the other exons from affected individuals showed no other abnormalities. These novel mutations provide further insight into the importance of the affected amino acids located close to the active site, near Asp 257, one of the three amino acids of the catalytic triad. The low anticoagulant activity of the abnormal protein C indicated that Arg 229 and Ser 252 play a key role during the interaction between protein C and its cofactor protein S, phospholipids, or factors Va and VIIIa. The Arg-1 to Cys mutation led to the dimerization of protein C with another plasmatic component, as evidenced by the presence in the plasma of a high molecular weight form of protein C that disappeared after reduction. No molecular mass abnormalities were observed in heavy and light chains of all other protein C mutants. In the five families explored, 9 (64%) of the 14 subjects bearing the mutations reported thrombotic events. This suggests that the protein C amino acids affected by the mutations are very important for the in vivo expression of the antithrombotic properties of protein C.
Functional abnormalities of CD8+ T cells define a unique subset of patients with common variable immunodeficiency
Authors: Jaffe JS , Strober W , Sneller MC
Published: October 2019

A substantial subgroup of patients with common variable immunodeficiency (CVI) exhibit an abnormal T-cell phenotype characterized by a low CD4/CD8 ratio associated with a significant increase in the absolute number of CD8+ T cells (CVI4/8low patients). In the present study, we examined the phenotypic and functional properties of purified T-cell subsets in this group of CVI patients. CD8+ T cells from CVI4/8low patients manifested increased expression of HLA-DR and CD57 and decreased expression of CD45RA as compared with CD8+ T cells from normal controls. When stimulated with anti-CD3 and phorbol 12-myristate 13-acetate, purified patient CD8+ T cells exhibited significantly decreased proliferation, c-myc expression, and interleukin-2 (IL-2) production compared with that of normal CD8+ T cells. Nevertheless, mitogen-activated patient CD8+ T cells secreted elevated amounts of gamma-interferon and IL-5 and normal amounts of IL- 4. This abnormal pattern of proliferation and cytokine production was limited to the CD8+ T-cell subset as CD4+ T cells from these patients exhibited normal proliferation and cytokine production. In further functional studies, purified CD8+ T cells from CVI4/8low patients manifested increased cytotoxic T-lymphocyte activity and suppressor activity, as compared with normal CD8+ T cells, when they were tested in (1) an anti-CD3 “redirected” cytotoxicity assay and (2) a suppressor assay consisting of CD8+ T cells and Staphylococcus aureus Cowan I (SAC) plus IL-2-stimulated normal (allogeneic) B cells. In the latter case, patient CD8+ T cells suppressed IgG production, but not IgM production. Finally, in studies to evaluate the role of patient CD8+ T cells in the pathogenesis of hypogammaglobulinemia, we determined the capacity of SAC and IL-2 to induce Ig production in highly purified patient B cells, ie, in the absence of patient CD8+ T cells. We found that, whereas B cells from one patient produced normal amounts of IgG, B cells from three patients were unable to produce normal amounts of IgG under these conditions. These data establish the phenotypic and functional characteristics of CD8+ T cells in CVI4/8low and clearly distinguish CVI4/8low patients from other patients with this syndrome. The data do not support the contention that hypogammaglobulinemia in CVI4/8low patients is due to a direct effect of CD8+ T cells on terminal B-cell differentiation, except in the occasional patient. The abnormal CD8+ T cells may, nevertheless, have more subtle effects of lymphoid function that play a role in disease pathogenesis.
B-cell lymphoma after angioimmunoblastic lymphadenopathy: a case with oligoclonal gene rearrangements associated with Epstein-Barr virus
Authors: Abruzzo LV , Schmidt K , Weiss LM , Jaffe ES , Medeiros LJ , Sander CA , Raffeld M
Published: October 2019

We describe a patient with angioimmunoblastic lymphadenopathy with dysproteinemia (AILD), who subsequently developed large-cell immunoblastic lymphoma of B-cell immunophenotype. At the time of the initial diagnosis, histologic examination of an inguinal lymph node showed typical features of AILD, and there was no evidence of a monoclonal B-cell population by immunohistochemical analysis. In situ hybridization and Southern blot analysis for Epstein-Barr virus (EBV) were negative. At autopsy 2 years later, the patient had widespread lymph node and organ involvement by large-cell immunoblastic lymphoma of B-cell immunophenotype. Southern blot analysis performed on DNA extracted from lymph nodes, liver, and spleen showed two patterns of Ig heavy chain and kappa light chain gene rearrangements. The T-cell receptor beta chain gene was in the germline configuration. Analysis with an EBV terminal repeat region probe showed two clonal populations that paralleled the Ig gene rearrangement studies. Double-labeling immunohistochemistry and in situ hybridization confirmed the presence of EBV within the neoplastic B cells. The data support the hypothesis that EBV was not etiologically related to AILD in this case, and that EBV proliferation may occur after the onset of the disease. Further, the data suggest that some B-cell lymphomas that arise in the setting of AILD resemble EBV-associated B-cell lymphomas that arise in other immunodeficiency states.
Anti-erythropoietin receptor (EPO-R) monoclonal antibodies inhibit erythropoietin binding and neutralize bioactivity
Authors: D'Andrea AD , Rup BJ , Fisher MJ , Jones S
Published: October 2019

We have generated six high affinity monoclonal antibodies (MoAbs) to the human erythropoietin receptor (hEPO-R) polypeptide. All six MoAbs bind to the extracytoplasmic domain of the hEPO-R, and all immunoprecipitate 35S-labeled hEPO-R from metabolically labeled Ba/F3- hEPO-R cells. Four of the MoAbs neutralize the EPO-dependent growth of Ba/F3-hEPO-R cells, whereas two MoAbs are non-neutralizing. None of the MoAbs inhibit the EPO-dependent growth of Ba/F3 cells expressing the murine EPO-R (mEPO-R), even though the hEPO-R and mEPO-R share 82% amino acid identity. All six of the anti-EPO-R MoAbs bind to the cell surface human EPO-R but none bind to the cell surface murine EPO-R. Of the four neutralizing MoAbs, the one-half maximal inhibition occurs at MoAb concentrations ranging from 1 nmol/L to 50 nmol/L. These MoAbs also compete with radiolabeled EPO for hEPO-R binding. The two non- neutralizing MoAbs fail to inhibit EPO-dependent growth or compete with EPO-binding, even at antibody concentrations as high as 500 nmol/L. The four neutralizing MoAbs, designated group I, compete with each other for an epitope of the hEPO-R polypeptide required for EPO-binding. The two non-neutralizing MoAbs recognize discrete epitopes, and are designated group II and group III MoAbs. In conclusion, this is the first description of MoAbs specific for the hEPO-R. The MoAbs, which recognize three discrete epitopes, may be useful in characterizing the spectrum of cells that display the hEPO-R and in further defining the role of EPO in hematopoiesis.
Stage-specific oligonucleotide uptake in murine bone marrow B-cell precursors
Authors: Zhao Q , Waldschmidt T , Fisher E , Herrera CJ , Krieg AM
Published: October 2019

Fluorescein isothiocyanate (FITC)-conjugated phosphodiester and phosphorothioate oligonucleotides were used in four-color flow cytometry with murine bone marrow cells stained with monoclonal antibody specific for the differentiation markers B220, S7 (CD43), and BP-1 to show possible stage-specific oligonucleotide uptake. Relatively low uptake was observed among pre-Pro- and early Pro-B cells. Late Pro- B- and pre-B cells had increased oligonucleotide uptake, whereas B cells had a lower level. Cell membrane binding of oligonucleotides varied during B-cell differentiation in parallel with internalization, which was documented by confocal microscopy. An FITC-conjugated polyanionic dextran sulfate also showed differentiation-related B-cell association, suggesting the presence of cell membrane binding sites specific for polyanions as opposed to a unique feature of the DNA backbone. Interpretation of antisense experiments in murine bone marrow cells will need to account for the heterogeneous oligonucleotide uptake among differentiating B cells.
Augmentation of gamma-globin gene promoter activity by carboxylic acids and components of the human beta-globin locus control region
Authors: Safaya S , Ibrahim A , Rieder RF
Published: October 2019

Butyric acid increases fetal hemoglobin synthesis in adult animals and in erythroid cells in culture and induces the gamma-globin gene promoter in transient expression experiments in K562 cells (McDonagh KT, Nienhuis AW, Blood 78:255a, 1992 [abstr, suppl 1]). We compared the effect of butyrate and other short-chain carboxylic acids in transient expression studies with K562 cells using an expression plasmid bearing a luciferase reporter gene driven by the normal human A gamma-globin gene promoter. Butyrate (4 carbons) increased the activity of the human A gamma-globin gene promoter up to 123 times. Marked augmentation of the normal gamma-promoter activity was also noted with 5-carbon valeric acid (up to 394 times) and 3-carbon propionic acid (up to 129 times). The branched isobutyric acid as well as phenylacetate showed less ability to increase promoter activity. Addition of the tandemly repeated AP-1/NF-E2 (AP) enhancer sequences from hypersensitive site 2 (HS2) of the locus control region (LCR) increased gamma-promoter activity up to 24 times. Addition of a nearby 16-bp conserved motif (CM) in HS2 (Safaya S, Rieder RF, Blood 78:146a, 1992 [abstr, suppl 1]) to the AP-containing plasmid construct further increased gamma-promoter activity. In the presence of butyrate, the plasmid bearing both the AP and CM sequences showed gene expression up to 477 times greater than that of the basal gamma-promoter-driven luciferase plasmid in the absence of inducer. A plasmid bearing the herpes simplex thymidine kinase promoter was also tested and gene expression was markedly increased by the same organic acids. MEL cells responded to butyrate, valerate, and propionate with induction of hemoglobin synthesis. Responses to isobutyrate and 6-carbon caproate required higher concentrations of the compounds. Thus, other short-chain organic acids as well as butyrate increase gamma-promoter activity in the transient expression system, and this activity can be further augmented by incorporating LCR elements into the expression vector. Nonglobin promoters also respond to the same carboxylic acids.
Nonrandom cytogenetic changes accompany malignant progression in clonal lines abelson virus-infected lymphocytes
Authors: Clark SS , Liang Y , Reedstrom CK , Wu SQ
Published: October 2019

Initially, lymphoid cells transformed by v-abl or BCR/ABL oncogenes are poorly oncogenic but progress to full transformation over time. Although expression of the oncogene is necessary to initiate and maintain transformation, other molecular mechanisms are thought to be required for full transformation. To determine whether tumor progression in ABL oncogene-transformed lymphoid cells has a genetic basis, we examined whether progression of the malignant phenotype of transformed clones correlates with particular cytogenetic abnormalities. A modified in vitro bone marrow transformation model was used to obtain clonal Abelson murine leukemia virus-transformed B lymphoid cells that were poorly oncogenic. Multiple subclones were then derived from each clone and maintained over a marrow-derived stromal cell line for several weeks. Over time, clonally related Abelson murine leukemia virus-transformed subclones progressed asynchronously to full transformation. The data show that tumor progression can occur in the absence of detectable cytogenetic changes but, more importantly, that certain cytogenetic abnormalities appear reproducibly in highly malignant subclones. Therefore, three independent subclones showed deletion in a common region of chromosome 13. Other highly malignant cells carried a common breakpoint in the X chromosome, and, finally, two subclones carried an additional chromosome 5. These results are consistent with the hypothesis that ABL oncogenes are sufficient for the initial transformation of cells but that additional genetic events can drive oncogenic progression. These observations further suggest that diverse genetic mechanisms may be able to drive tumor progression in cells transformed with ABL oncogenes.
The murine interleukin-3 receptor alpha subunit gene: chromosomal localization, genomic structure, and promoter function
Authors: Miyajima I , Levitt L , Hara T , Bedell MA , Copeland NG , Jenkins NA , Miyajima A
Published: October 2019

The interleukin-3 receptor (IL-3R) is composed of alpha and beta subunits, members of the class I cytokine receptor family. Here we describe isolation and characterization of the chromosomal gene for the mouse IL-3R alpha subunit (mIL-3R alpha). Whereas the human IL-3R alpha gene is tightly linked with the granulocyte-macrophage colony- stimulating factor receptor alpha subunit (GM-CSFR alpha) gene in the pseudoautosomal region of the X and Y chromosomes, the mIL-3R alpha gene (II3ra) is located in the proximal region of mouse chromosome 14, separated from the mouse GM-CSFR alpha gene, which is on chromosome 19. The mIL-3R alpha gene spans about 10 kb and is divided into 12 exons. All the exon-intron boundaries possess the splicing junction consensus sequences (52GT-AG352), and the whole genomic structure is similar to those of the previously reported class I cytokine receptor genes. There are two major transcription initiation sites that are located at 215 and 188 nucleotides upstream of the initiator codon. The promoter region is GC-rich and contains potential binding sites for GATA, Ets, c- myb,, Sp1, Ap-2, and G-C boxes, but not a typical TATA or CAAT sequence. A fusion gene containing 0.8 kb of the 52 noncoding sequence linked to the firefly luciferase gene directed the transcription in mouse mast cells but not in fibroblasts or T cells, suggesting that this promoter functions in a cell type-specific manner. Further sequential deletion of the 52 region suggests two potential regulatory regions for transcription of the mIL-3R alpha gene.
The antineoplastic bryostatins affect human basophils and mast cells differently
Authors: Patella V , Casolaro V , Ciccarelli A , Pettit GR , Columbo M , Marone G
Published: October 2019

Bryostatins, macrocyclic lactones from the marine bryozoan Bugula neritina, are potent antineoplastic agents and multi-potential stimulators of immune cells. We have examined the effects of bryostatins on mediator release from human basophilic leukocytes and human tissue mast cells. Bryostatins 1, 2, and 5 (10 to 3,000 nmol/L) induced histamine secretion from purified and unpurified peripheral blood basophils, whereas they caused no release of peptide-leukotriene C4 from these cells. The rate of histamine release caused by bryostatin 1 was slower than that caused by anti-IgE (t1/2 +/- SEM = 38.2 +/- 4.7 minutes v 8.9 +/- 0.2 minutes; P < .01), whereas the temperature dependence was similar (optimum release at 37 degrees C, approximately 30% less at 30 degrees C, and no release at 22 degrees C or 4 degrees C). The addition of increasing concentrations of extracellular Ca2+ to the medium caused histamine release in the presence of bryostatins. Subeffective concentrations of bryostatins and anti-IgE produced a synergistic effect on histamine release from basophils. Staurosporine, chelerythrine, and calphostin C (0.1 to 10 nmol/L), which are protein kinase C inhibitors, inhibited the histamine secretion activated by bryostatin 1 and tetradecanoylphorbol-acetate (TPA). Preincubation with granulocyte-monocyte colony-stimulating factor (GM-CSF; 1 and 5 nmol/L) and interleukin-3 (IL-3; 10 ng/mL) potentiated the activation of human basophils induced by bryostatin 1. Neither bryostatin 1 nor bryostatin 2 induced the release of histamine from mast cells isolated from human lung or skin tissues. However, brief (10 minutes) preincubation with bryostatin 1 (3 to 300 nmol/L) potently inhibited the histamine secretion induced by anti-IgE from skin or lung mast cells. Bryostatin 1 was a more potent (by approximately 30 times) inhibitor of IgE- mediated histamine release than was TPA. The heterogeneous effects exerted by bryostatins on human basophils and mast cells can be of interest for those designing therapeutic trials using these agents.
Interleukin-9 is a major anti-apoptotic factor for thymic lymphomas
Authors: Renauld JC , Vink A , Louahed J , Van Snick J
Published: October 2019

We have recently shown that interleukin-9 (IL-9) strongly stimulates the proliferation of mouse thymic lymphomas in vitro. Here we report that this factor is also one of the most potent inhibitors of apoptosis induced by dexamethasone (DEX) in such cell lines, even if they do not depend on exogenous factors for growth. For the murine thymic lymphoma BW5147, protection against apoptosis was also obtained with IL-4 and less strongly with IL-6, whereas IL-2, IL-7, and IL-10 were inactive. Moreover, IL-4 and IL-9 maintained proliferation of these cells in the presence of DEX. Analysis of eight other factor-independent thymic lymphoma lines showed significant protection in seven and six cell lines with IL-9 and IL-4, respectively, whereas only three were protected by IL-7 and only two by IL-2. Comparison of the responses to IL-2 and IL-9 in a factor-dependent cell line that responds to both cytokines showed that IL-2 is a stronger inducer of proliferation, while IL-9 is more efficient in protecting the cells against apoptosis. Taken together, our observations suggest that, for thymic lymphomas, proliferation and apoptosis involve distinct regulatory mechanisms and can be differentially regulated by cytokines.
Eradication of polymerase chain reaction detectable immunoglobulin gene rearrangement in non-Hodgkin's lymphoma is associated with decreased relapse after autologous bone marrow transplantation
Authors: Zwicky CS , Maddocks AB , Andersen N , Gribben JG
Published: October 2019

In B-cell non-Hodgkin's lymphoma (NHL), as in other B-cell malignancies, clonal rearrangement of the third complementarity determining region (CDR III) of the immunoglobulin heavy chain gene (IgH) provides a useful marker for the detection of minimal residual disease (MRD) after treatment. To determine the clinical utility of IgH polymerase chain reaction (PCR), we analyzed peripheral blood (PB) and bone marrow (BM) samples from 25 patients with NHL with no PCR detectable chromosomal rearrangement who have undergone autologous bone marrow transplantation (ABMT). Patients with histologic bone marrow infiltration at the time of bone marrow harvest were selected for study since this provided us with diagnostic tissue samples. As an initial strategy DNA was amplified using consensus variable (VH) and joining (JH) region primers. In those cases failing to amplify using consensus region primers, PCR was performed using a panel of VH family-specific framework region 1 (FR1) primers. The clonal products were directly sequenced. From the V-N-D region nucleotide sequences, clone specific probes were constructed and used for subsequent detection of MRD. A clonal PCR product could be PCR amplified and directly sequenced in 18 (72%, 90% confidence intervals 54%-86%) of these 25 patients, 8 with diffuse and 10 with follicular NHL. Eight of these 18 patients have relapsed after ABMT. All had detectable lymphoma cells before relapse and the sequence of the CDR III region at the time of relapse was identical to that obtained at the time of ABMT. All 10 patients who remain in complete remission from 18 to 36 months after ABMT had eradication of PCR detectable lymphoma cells after ABMT, although in three patients PCR detectable MRD was detected early after ABMT. We conclude that sequencing and the use of patient specific IgH CDR III oligonucleotides probes provides a simple and highly reliable method to determine the specificity of the IgH PCR technique. The clinical utility of this technique is demonstrated by the finding that eradication of PCR detectable lymphoma cells in these patients is associated with decreased relapse after ABMT (P = .0002).
A cell proliferation-dependent multiprotein complex NC-3A positively regulates the CD34 promoter via a TCATTT-containing element
Authors: Perrotti D , Bellon T , Trotta R , Martinez R , Calabretta B
Published: October 2019

The CD34 cell surface antigen is a glycoprotein expressed by hematopoietic stem and progenitor cells and also by certain nonhematopoietic cell-types. Because CD34 expression is regulated both at the transcriptional and the posttranscriptional level, we attempted to identify factors that, by interacting with the 5′ flanking region of the human CD34 gene, may regulate its promoter activity in proliferating hematopoietic cells. By electrophoretic mobility shift assay, UV cross-linking and DNase I footprinting analyses, we identified a multiprotein complex, designated NC-3A, that specifically interacts with the CD34 promoter region from nucleotides -375 to -351. Sequence analysis of this region revealed the presence of a distinct motif, TCATTT. Chloramphenicol acetyl-transferase assays used to assess promoter activity in transiently transfected cells showed that this TCATTT-containing element, which is conserved in both the human and the murine CD34 genes, mediates positive regulatory activity in hematopoietic and nonhematopoietic cells, and acts as an enhancer when placed upstream of a heterologous promoter. Moreover, loss of CD34 promoter activity was caused by mutation of the TCATTT motif. In addition, the interaction of the nuclear multiprotein complex NC-3A with this enhancer element is proliferation-dependent. These data indicate that, although not cell-type specific, the formation of a multiprotein complex NC-3A interacting with the region from nucleotides -375 to 351 plays an important role in controlling CD34 promoter activity in proliferating hematopoietic cells.