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Table 1. Kinetic parameters derived from the least squares computer analysis applied to the hybridization values obtained with hnRNA from normal and leukemic cells CelIType First component Second component Third component % K Cot 1 / 2 % Normal promyelocytes 7.3 3.66125 0.273 9.4 Normal promyelocytes 5.7 1.97528 0.534 11.2 Normal lymphoblasts 5.1 0.60000 1.666 9.7 Normal lymphoblasts 4.4 0.76000 1.403 14.0 ALL 4.6 3.55457 0.280 29.0 AML1 10.7 0.59314 1.68 30.7 AML2 10.6 0.52518 1.90 38.9 HL60 5.9 0.43278 2.30 25.2 HL60 5.3 2.09849 0.47 24.6 K Cot1/2 % K Cot1/2 0.00992 100 26.8 0.00004 24870 0.00759 120 29.5 0.00004 25125 0.00473 211 28.8 0.00004 23192 0.00863 180 29.6 0.00006 17808 0.00317 315 0.00211 474 0.00206 484 0.00180 554 0.00261 382 hnRNA from different cell types ranging from 40% to 60%. Whereas some of them (Kline et al. 1974) assurne that the hnRNA which fails to hybridize belongs exclusively to the low frequency dass of sequences, other authors (Spradling et al. 1974) believe that the incomplete hybridization of RNA in vast DNA excess may affect all classes of sequences equally. Whatever interpretation finally proves to be correct, our results indicate that at least part of the hnRNA of leukemic cells is characterized by a sequence frequency higher than that of hnRNA of normal cells. Acknowledgments This research was supported by a grant from the National Research Council PFCCN contract no. 79.0068.396. References Davidson EH, Klein WH, Britten RJ (1977) Sequence organization in animal DNA and a speculation on hnRNA as a coordinate regulatory transcript. Dev Biol55 :69 - Kline WH, Murphy W, Attardi G, Britten RJ, Davidson EH (1974) Distribution of repetitive and non repetitive sequence transcripts in HeLa mRNA. Proc Natl Acad Sci USA 71: 1785 - Melli M, Whitefield DC, Rao KV, Richardson M, Bishop JO (1971) DNA-RNA hybridization in vast DNA excess. Nature New Biol 231 : 8 - Spradling A, Penman S, Campo MS, Bishop JD (1974) Repetition on hnRNA as a coordinate regulatory transcript. nuclear and cytoplasmic messenger RNA of mammalian cells. Cell 3 : 23 - Tobin AJ, Selvig SE, Lasky L (1978) RNA synthesis in avian erythroid cells. Dev Biol 67: 11 - Torelli G, Cadossi R, Ferrari S, Narni F, Ferrari S, Montagnani G, Torelli U, Bosi P (1979) Reassociation kinetics of the DNA of human acute leukemia cells Biochim Biophys Acta 561 :301 - Vogelstein B, Gillespie D (1977) RNA-DNA hybridization in solution without reannealing. Biochem Biophys Res Commun 75: 1127 519
Haematology and Blood Transfusion Vol. 26 <strong>Modern</strong> Trends in Human Leukemia IV Edited by Neth, Gallo, Graf, Mannweiler, Winkler © Springer-Verlag Berlin Heidelberg 1981 Characterization of Antigens in SSV Nonproducer Cells H.-J. Thiel, T. Matthews, E . Broughton, A. Butchko, andD. Bolognesi A.Summary 2 3 4 An autologous antiserum against simian sarcoma virus (SSV) nonproducer cells (SSV-NP celIs) was characterized by radioimmunoprecipitation. It reacts specifically with two different moleeules in SSV-NP cells, a SSV transformation-specific glycoprotein (SSV TrS-gp) and p65, which probably represents a modified gag-precursor . B. Introduction The SSV complex [SSV(SSAV)] contains a replication defective transforming virus (SSV) as weH as an associated helper virus (SSAV). This mixture of transforming pseudotype SSV(SSAV) and helper virus was originally isolated from a spontaneous fibrosarcoma of a New World primate, the woolly monkey (Theilen et al. 1971). While SSAV has been quite weIl characterized (Aaronson et al. 1976; Benveniste et al. 1977; Deinhardt et al. 1978), relatively Httle is known about the replication defective transforming virus, SSV (Aaronson et al. 1975; Bergholz et al. 1977). In an attempt to identify putatIve transformation specific information coded for by SSV, we isolated go at cells nonproductively transformed by SSV(SSV-NP) and inoculated these several tim es as live cells into the same goat from which they originated. The resulting hyperimrnune serum (SSV-NP serum) was reacted in radioimmunoprecipitation against different transformed and nontransformed cell lines. Fig. 1. Immunoprecipitation of SSV TrS-gp from tissue culture supernatants of NRK SSV-NP cells analyzed on a 5 % SDS gel. Cells were labeled by a 7 h pulse followed bya 14 h chase: 3H-glucosamine label with preimmune serum (Lane 1) and SSV-NP serum (Lane 2). 35S-cysteine label with SSV-NP serum (Lane 3) and preimmune serum (Lane 4) 520
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Haematology and Blood Transfusion 2
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Dr. Rolf Neth, Universitäts-Kinder
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Participants of the Meeting Aaronso
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Kurth, R, Friedrich-Miescher-Labora
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Wilsede Scholarship Holders (Grante
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Preface Gut ist eine Lehrart, wo ma
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Personal and scientific discussion
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Acknowledgement We should like to t
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Wilsede, June 21 1978 Memorial Trib
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limiting benign lymphoproliferative
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this most unlikely, however (for re
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longer subject to the same control
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Bregula U, Wiener F, Harris H (1971
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fresh lymph node biopsies from ten
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Fig. 3. Binucleate mitosis in an ob
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y Fig. 4. Schematic diagram of post
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than that among patients with Stage
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Tests of binding affinity, agglutin
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N Engl J Med 297: 963-968 - Green I
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Clinical Aspects
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"ring chromosome". "Mar" is marker,
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191 chromosomally abnormal patients
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tumors of both African and non-Afri
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Haematology and Blood Transfusion V
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Retrospective group 26/93 evolved l
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Treatment
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advantage in terms of duration of f
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leukaemia. An analysis of 168 patie
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11. Treatment Program Chemotherapy
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efractory congestive heart failure
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period (Sequence II) was gene rally
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children with a multiple drug proto
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DAYS 1 5 10 15 20 25 30 35 , I I I
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Biologically Drug Outcome fit" sens
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may counterbalance the potential be
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Ht (0--0) 50 40 30 20 10 0 Platelet
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10 7 ,-----------------------------
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11. Haematological and Biochemical
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0\ 0\ INTERFERON ... 10 3 rl E "'
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("pre-B" phenotype) has been descri
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human bone marrow cells exhibit the
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10 9.,0 MBKCC .. .,4 B prot:oool 1'
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demann W, Büehner T, Arlin Z, Gee
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-...l 00 a-tLDRENS CAt«:ER S TU>Y
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prognostic characteristics under in
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significantly higher in HR} (14.5%,
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normal or elevated immunoglobulin l
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References Andreeff M, Darzynkiewic
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R GROUP 1 ( 1970 - 1971 ) 17 PTS. R
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Table 3. Influence of initial featu
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LIJ (f) a.. < -1 LIJ Q: u. 0 >- I-
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me nt indicated that additional the
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42:2123-2134 - Chessells JM, Cornbl
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Baum et al. 1979). This study was b
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1. 0 '--'1--" 0.9 e: o .- VI VI E C
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e: o e: 1. 0 ,.--...-- 1'"'\ ......
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In conclusion, we can state that th
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Haematology and Blood Transfnsion V
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INDUCTION VCR + PRED + ASNASE (wk 3
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SCHED.INT 1 MTX • ! Adria MTX •
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~ looh---------ooooc>----o---c>----
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Consoll- Irra .tlon t) .tlon Indu
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Highrisk Standard risk Table 1. Cli
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Cumulo/ive comp/el9 remission 1.0 .
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10° 8 = 6 ö 4 Non T Cell .~ :ö
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% SURVIVAL 100 ~ ~II 0 80 60 40 - -
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a) Percentage of donor cell mitoses
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Table 5. Results of bone marrow tra
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...... w N Table 7. Clinical result
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agent had to be added and that in t
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C. Chronic Myelogenous Leukemia The
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Patients with a suitable donor rece
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No. Recurrent Leukaemia Other death
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followed by dialysis against isoton
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HS, HL- Heme >_ 40 0 HS I ;:. =====
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knowing at present. These genes may
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esistance to antifolates. In Vitro
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a DNA probe specific for the gene t
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translocation between chromosomes 9
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Table 1. Subdivision of 1827 Myelo-
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NORMAL CELL ®
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some change of the primary type, in
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F. Transformation of Mouse Bone Mar
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were carried out with 3H-MTX in the
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the lysozyme cDNA prepared from ovi
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et al. 1976). The BJAB cellline in
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dalton bands corresponding to light
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1979). The EBV-carrying cell lines
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producer lines (DAUDI and P3HR-1),
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G, Giovanella B, Westman A, Stehlin
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Dc;bt CT + i J.I9 Raji; 1. i .. 100
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c. Discussion During infectious mon
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C. Nonepisomal Viral DNA We have us
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A 260 3.0 A B 1670 70 N2 2.0 c: E .
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vivo. Nature 260:302-306 - Kirk JTO
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Fig. 1. a Polyacrylamide gelelectro
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1979). The lymphocyte subpopulation
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lysin 0 antigens are found, and mix
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Pope JH (1978) Long-term T cell-med
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Charaeteristie Morphologie maturati
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after TPA-induction (Table 3). The
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PA (1978) Reactivity of a rabbit an
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Blood Monocyte. B lood Monocytes 1-
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and nonhistone proteins can serve a
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The findings of the evaluation of s
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Fig. 3. Tumor grawth of L 428 cells
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Cell biological and Immunological A
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consideration. Sanel (1973) obtaine
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endotoxin serum, were used. A parti
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monocytic leukemia cell line in cul
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B. Isolation, Characterization and
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don al hemopathies generally displa
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nitors. Blood Cells 6:595-607 - Min
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\I) ....I ....I IU Jl 200 r 100 90
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their haemopoietic cells' ability t
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normal normal ALL ALL ALL persons p
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64.5 { V)
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References Biermann E, Neth R, Gros
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ehambers, the growth pattern observ
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References Anderssen LC, 10kinen M,
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REH CELL UNE TO CALLb~Ö ~q~ V.M. c
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the macrophage series in which the
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Table 1. Production of factor depen
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Table 1. Characteristics of the adh
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Fig. 3. A human marrow culture at 6
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300 250 • HYPERPROLIFERATIVE PATT
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Fig. 9. Nonadherent population from
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The cobblestone areas were often no
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L (1976) Induction of colony format
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B. Materials and Methods J. Tissue
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Fig. 2. Ultrastructural appearance
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Table 1. Monoclonal antibody reacti
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al. 1979; Janossy et al. 1979) is e
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to which this is an exact replica o
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inger JL (1976) Distribution of la-
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I region antigens were found to be
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have been reported to be present on
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V. FunctionalAssays Assays for PHA
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Functional studies of UCHT1 separat
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40 ,...... (5 L.. C o u 30 .9 ~ .~
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with immunohistochemieal methods (L
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can be dissected by cloning. Utiliz
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Table 1. Origin and marker profile
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complete identity of gp 100 from no
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Table 1. Reaction of single anti se
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media. Following 24-h incubation, c
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nonleukemic eells (Greaves 1979). L
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Fig. 2. Cytocentrifuged smears of b
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Modification of amino-groups of hum
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disturb this balance in the directi
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The majority of human blood lymphoc
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of the EBV infected B cells. Howeve
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and immunoglobulins. J Immunol 120:
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" K 10 • j ~ ALL RNlL AL wtfh les
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tion akuter Leukämiezellen in "spo
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D. Results and Discussion J. Acute
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selectively responsive to different
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l1J ~ 100 ~ Q. ::) l1J Z 0 ~ z >- .
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Table 2. Growth inhibition of S49 e
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Reverse Infectious ASC/2X1Q6 transc
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munized animals to 1316 tumor cells
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Fig. 3. Expression of retroviral gp
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pg70. J Exp Med 143: 151-166 - McGr
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Table 1. Expression of tumor antige
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Virological and Molecularbiological
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Table 1. Oncogenic properties of re
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also Fig. 1). It followed that the
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whether this sequence is related to
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zed in infected cells argue that th
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a specific viral transforming prote
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detect viral RNA as the only charac
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Eisen H (1980) Myeloproliferate vir
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p,60- -34K - 1 2 3 4 Fig. 1. In vit
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.. p - • a'T ... I .t'l Fig. 4. D
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Fig. 2. Photomicrographs of NY68 in
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DEAE (o(lJmn 'liI GI -2 2 o e s 1
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vity was deterrnined. For endogenou
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pr 180 - - - P 12/15 - A B c D E F
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85,- 66- -27 - -66 40- 27- -1'9 19-
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= 0.5 o E Q. o ILI 0.4 (J) « ILI .
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Table 1. Oncogenic potential of avi
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100 III GI c: o "8 80 > w
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Expt. No. Infecting virus Proportio
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Haematology and Blood 'fransfusion
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sequences, provided its gag portion
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77:3009-3013 - Hu SSF, Moscoviei C,
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RI 2.2 md and Hind III 2.6 md leuke
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RELATIONSHIP OF ENDOGENOUS AND EXOG
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E ........ E Cl. U E :::l .... Q) (
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References Astrin S (1978) Endogeno
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the same animal were examined (e.g.
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I) 'IQ a \0 - CL -- Fig. 2. Restrie
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~ ..................... ~ Cell 3' 5
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a :Sa,C! + . ~ 111 i' ( " '~ .J. b
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Table 1 Sampie Tissue TS fragment"
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a b Sac I / rep ._-_.-_ K J nl rep
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Haematology and Blood 'fiansfusion
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Osteopetrosis and osteosarcomas occ
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Lymphoma cellline Table 1. T and B
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Table 2. In vivo growth and oneogen
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Table 1. Transforming aetivity of c
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Table 3. Transformation aetivity of
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- Shoemaker C, Goff S, Gilboa E, Pa
Page 477 and 478: indieate a shifting of target eell
Page 479 and 480: Table 2. Expression of Jl and Sourc
Page 481 and 482: Haematology and Blood Transfusion V
Page 483 and 484: ~ i ,..1---------,,':::::::::::::::
Page 485 and 486: fragment of pBR322 DNA, a 3.0-kbp S
Page 487 and 488: Mak TW (1979) Proc Natl Acad Sci US
Page 489 and 490: Table 1. Transformation of 3T3 cell
Page 491 and 492: inserts in the genome of rapidly tr
Page 493 and 494: pulations, it seemed likely that th
Page 495 and 496: contains antigens which react with
Page 499 and 500: le of reacting in sensitive radioim
Page 503 and 504: gic approach. In: Hiatt HH, Watson
Page 505 and 506: 12 Eco R [ Fig. 1. Hybridization pa
Page 509 and 510: 10 - '1~ )( -~ S:! E A U I o 1 2
Page 511 and 512: Haematology and Blood 'fransfusion
Page 513 and 514: cultures. These cells did not conta
Page 515 and 516: indication of malignant T-cells in
Page 517 and 518: Fig. 3. Thin-section electron micro
Page 519 and 520: Table 3. Lack of detectable related
Page 521 and 522: J. HTLV Was Present in the Primary
Page 523 and 524: specific signals into nonspecific s
Page 525 and 526: Table 2. Distribution of HTLV-relat
Page 527: u 10 ~ 20 ~ :c 30 a 40 o Fig. 1. Ki
Page 531 and 532: SSV TrS-gp labeled effectively with
Page 535 and 536: 60 HFF/SSV cDNA CELL RNAs: 0 HF/SSV
Page 537 and 538: Antisera a Table 1. Immunofluoresce
Page 539 and 540: Haematology and Blood 'ftansfusion
Page 541 and 542: y competition RIA yielded virtually
Page 543 and 544: SiS'V' gp70: ANTIGEN, ANTI BODY, IM
Page 545 and 546: type-C virus p30 antigen in cells f
Page 547 and 548: c. Results The fractionated whole-b
Page 549 and 550: induced antibodies as well as exper
Page 551 and 552: - Fig. la-f. Retravirus particles p
Page 553 and 554: eplication of infecting murine leuk
Page 557 and 558: immunodeficiency, multic10nal lymph
Page 559 and 560: Bovine leukemia --, proteins of 499
Page 561 and 562: Human T- cell - -, characterization
Page 563 and 564: RNA -, of tumor virus 439 Rous sarc
Page 565 and 566: Haematology andBlood Transfusion Su
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