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80 o E o .c E60 ~ Q. ~ o B> 20 '0 ~ o RAV -I , 51 birds 6. RAV-60s, 87 birds o RAV-O,58 birds • NTRE - 7,27 birds 8 16 24 32 Weeks after infec1ion 40 48 Fig. 6. Pathogenicity of NTRE-7, RAV-O, and RA V -60s. The figure shows cumulative disease incidence in the same birds as in Fig. 5 as a function of time after injection with 10 6 infectous units of RAV-l (0), RAV-O (0), NTRE-7 (e) and pooled data for the four RAV-60s shown in Fig. 3 (t:,.). All diagnoses were histologically confirmed. A small additional incidence of other neoplasms in the RAV-60 and RAV-1 groups is not included. Data taken in part from Robinson et al. (to be published) in the latter three groups at 1 month of age. Although there was substantial variation from bird to bird, the growth of the virus es in vivo seemed to mimic that in ceIl culture, with RA V -0 growing to a median value about lO-fold less than NTRE-7 or the RA V -60s. Figure 6 shows the incidence of leukosis in all the birds tested. The incidence and latent period for the RA V -60 strains were not significantly different from the RA V-I. Thus, the subgroup E host range does not contribute to the nonpathogenicity of endogenous viruses. Similar results were found by Crittenden et al. (1980). As previously reported (Motta et al. 1975), RAV-O was nonleukemogenic. Although the test of NTRE-7 is not yet complete, the results to date are quite surprising. In the first 34 weeks, there has been no disease whatever, compared with 40% in the RAV-60 infected birds. While we cannot yet conclude that NTRE-7 is completely nonpathogenic, it may weIl be so, and it is at least significantly slower than the RA V -60s. We must also point out that although td sarcoma viruses have been reported to be leukemogenic (Biggs et al. 1973 ; HalpernandHanafusa, personalcommunication), td Pr-B has not yet been tested. In any case, this result clearly separates growth rate from leukemogenicity, since RA V -60s and NTRE-7 show identical growth both in vitro and in vivo. This result is inconsistent with the simplest predictions of the downstream promotion model as weIl as models which invoke pathogenic side effects of the virus gene products. We have additional recombinants available for testing which should provide us with a suggestion of which region of the genome is responsible for these differences. A comparison of the structures of NTRE-7 and the RA V -60s gives a hint of where to look (Fig. 3). The only exogenous virus region besides U 3 consistently inherited by these virus es is immediately to the left of U 3 • This region is at present undefined. There are a number of reasons why such a sequence might have a role in pathogenicity, the most interesting of which is the regulation of downstream promotion by either the sequence itself or its product. We believe that the nonpathogenicity of the endogenous viruses is an important adaptive feature to their quiescent lifestyle, for which it is quite important that they not harm the host. Experiments such as we have presented he re should be useful in identifying and characterizing the regions of the virus genomes relevant to this adaptation and also in exploiting the differences in these regions in different viruses to probe the molecular mechanisms of viral leukemogenesis. Acknowledgements We thank M. Champion and L. Lieberman for expert technical assistance. This work was supported by grants CA17659, CA24530, and CA23086 from the National Cancer Institute. J. Coffin was a recipient of a Faculty Research Award from the American Cancer Society, and P. Tsichlis was a postdoctoral fellow of the National Cancer Institute. 437
References Astrin S (1978) Endogenous viral genes of white leghorn chickens: A common site of residence as weIl as sites associated with specific phenotypes of viral gene expression. Proc Natl Acad Sci USA 75:5941-5945 - Biggs P, Milne B, Graf T, Bauer H (1973) Oncogenicity of non-transforming mutants of avian sarcoma viruses. J Gen Virol 18:399-403 - Coffin JM, Champion MA, Chabot F (1978a) Genome structure of avian RNA tumor viruses: Relationships between exogenous and endogenous viruses. In: Barlati C, deGiuli-Morghen C (eds) Avian RNA tumorviruses. Piccin, Padua, pp 68-87 - Coffin J, Champion M, Chabot F (1978b) Nucleotide sequence relationships between the genomes of an endogenous and an exogenous avian tumor virus. J Virol 28: 972-991 - Crittenden LB, Hayward WS, Hanafusa H, Fadley AM (1980) Induction of neoplasms by subgroup E recombinants of exogenous and endogenous avian retroviruses (Rous-associated virus type 60). J Viro133: 915-919 - Czernilofsky A, Delorbe W, Swanstrom R, Varmus H, Bishop JM (to be published) The nucleotide sequence of "C" an untranslated but conserved domain in the genome of avian sarcoma virus. Nucleic Acids Res - Hanafusa T, Hanafusa H, Miyamoto T (1970) Recovery of a new virus from apparently normal cells by infection with avian tumor viruses. Proc Natl Acad Sci USA 67: 1797-1803 - Hayward WS, Hanafusa H (1975) Recombination between endogenous and exogenous RNA tumor virus genes as analyzed by nucleic acid hybridization. J Virol 15: 1367-1377 - Hsu J, Sabran J, Mark G, Guntaka R, Taylor J (1978) Analysis of unintegrated avian RNA tumor virus double-stranded DNA intermediates. J Virol 29:819-818 - Hughes S, Vogt PR, Shank PR, Spector PH, Kung HJ, Breitman ML, Bishop JM, Varmus HE (1978) Proviruses of avian sarcoma viruses are terminally redundant, co extensive with unintegrated linear DNA, and integrated at many sites in rat cell DNA. CeIlI5: 1397-1410-LiniaIM, Neiman PE (1976) Infection of chick cells by subgroup E virus. Virology 73: 508-520 - Moscovici C, Moscovici MG, Jimenez H, Lai MMG, Hayman MJ, Vogt PK (1977) Continuous tissue culture cell lines derived from chemically induced tumors of J apanese quail. Cell 11: 95-103 - Motta J, Crittenden L, Purchase H, Stone H, Okazaki W, Witter R (1975) Low oncogenic potential of avian endogenous RNA tumor virus infection or expression. J Natl Cancer Inst 55: 685-689 - Neiman PE, Das S, MacDonnel D, McMillan-Helsel C (1977) Organization of shared and unshared sequences in the genomes of chicken endogenous and sarcoma viruses. Ce II 11:321-329 - Robinson HL (1976) Intracellular restriction on the growth of induced subgroup E avian type C viruses in chicken cells. J Virol 18:856-866 - Robinson H (1978) Inheritance and expression of chicken genes which are related to avian-sarcoma viruses. Curr Top Microbiol Immunol 83:1-36 - Robinson HL, Pearson MN, Desimone DW, Tsichlis PN, Coffin JM (to be published) Subgroup E avian leukosis virus associated disease in chickens. Cold Spring Harbor Symp Quant Biol 44 - Sabran J, Hsu T, Yeater C, Kaji A, Mason W, Taylor J (1979) Analysis of integrated avian RNA tumor virus DNA in transformed chicken, duck, and quail fibroblasts. J Virol 29: 170-178 - Shank PR, Hughes S, Kung HJ, Majors JE, Quintrell N, Guntaka RV, Bishop JM, Varmus HE (1978) Mapping unintegrated forms of avian sarcoma virus DNA: Both termini of linear DNA bear a 300 nucleotide sequence present once or twice in two species of circularDNA. Ce1l15: 1383-1395 -Spector DH, Smith K, Padgett T, McCombe P, Roulland Dussoix D, Moscovici C, Varmus HE, Bishop JM (1978) Uninfected avian cells contain RNA unrelated to the transforming gene of avian sarcoma viruses. CeIl13: 371-379 - Stehelin D, Varmus HE, Bishop JM, Vogt PK (1976) DNA related to the transforming gene(s) of avian sarcoma virus is present in normal avian DNA. Nature 260: 170-172 - Stehelin D, Soule S, Roussel M, Sergeant A, Lagrou C, Rommens C, Raes MB (to be published) Three new types of viral oncogenes in defective avian leukemia viruses. Cold Spring Harbor Symp Quant Biol 44 - Tsichlis PN, Coffin JM (1980) Recombinants between endogenous and exogenous avian tumor virus es : Role of the C region and other portions of the genome in the control of replication and transformation. J Virol 33: 238-249 - Vogt PK (1971) Spontanous segregation of nontransforming viruses from cloned sarcoma viruses. Virology 46:939-946 - Vogt PK, Friis RR (1971) An avian leukosis virus related to RSV(O): Properties and evidence for helper activity. Virology 34:223-234 - Wang SY, Hayward WS, Hanafusa H (1977) Genetic variation in the RNA transcripts of endogenous virus genes in uninfected chicken cells. J Virol 24:64-73 - Yamamoto T, Jay G, Pastan 1(1980) Unusual features in the nucleotide sequence of a cDNA clone derived from the common region of avian sarcoma virus messenger RNA. Proc Natl AcadSci USA 77:176-180 438
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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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Page 307 and 308:
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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
Page 395 and 396: also Fig. 1). It followed that the
Page 397 and 398: whether this sequence is related to
Page 399 and 400: zed in infected cells argue that th
Page 401 and 402: a specific viral transforming prote
Page 403 and 404: detect viral RNA as the only charac
Page 405 and 406: Eisen H (1980) Myeloproliferate vir
Page 407 and 408: p,60- -34K - 1 2 3 4 Fig. 1. In vit
Page 409 and 410: .. p - • a'T ... I .t'l Fig. 4. D
Page 411 and 412: Haematology and Blood Transfusion V
Page 413 and 414: Fig. 2. Photomicrographs of NY68 in
Page 415 and 416: DEAE (o(lJmn 'liI GI -2 2 o e s 1
Page 417 and 418: vity was deterrnined. For endogenou
Page 419 and 420: pr 180 - - - P 12/15 - A B c D E F
Page 421 and 422: 85,- 66- -27 - -66 40- 27- -1'9 19-
Page 425 and 426: = 0.5 o E Q. o ILI 0.4 (J) « ILI .
Page 427 and 428: Table 1. Oncogenic potential of avi
Page 429 and 430: 100 III GI c: o "8 80 > w
Page 431 and 432: Expt. No. Infecting virus Proportio
Page 433 and 434: Haematology and Blood 'fransfusion
Page 435 and 436: sequences, provided its gag portion
Page 437 and 438: 77:3009-3013 - Hu SSF, Moscoviei C,
Page 439 and 440: RI 2.2 md and Hind III 2.6 md leuke
Page 443 and 444: RELATIONSHIP OF ENDOGENOUS AND EXOG
Page 445: E ........ E Cl. U E :::l .... Q) (
Page 449 and 450: the same animal were examined (e.g.
Page 451 and 452: I) 'IQ a \0 - CL -- Fig. 2. Restrie
Page 453 and 454: ~ ..................... ~ Cell 3' 5
Page 455 and 456: a :Sa,C! + . ~ 111 i' ( " '~ .J. b
Page 457 and 458: Table 1 Sampie Tissue TS fragment"
Page 459 and 460: a b Sac I / rep ._-_.-_ K J nl rep
Page 461 and 462: Haematology and Blood 'fiansfusion
Page 463 and 464: Osteopetrosis and osteosarcomas occ
Page 465 and 466: Lymphoma cellline Table 1. T and B
Page 467 and 468: Table 2. In vivo growth and oneogen
Page 471 and 472: Table 1. Transforming aetivity of c
Page 473 and 474: Table 3. Transformation aetivity of
Page 475 and 476: - 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 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 497 and 498:
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le of reacting in sensitive radioim
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gic approach. In: Hiatt HH, Watson
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12 Eco R [ Fig. 1. Hybridization pa
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Page 509 and 510:
10 - '1~ )( -~ S:! E A U I o 1 2
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Haematology and Blood 'fransfusion
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cultures. These cells did not conta
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indication of malignant T-cells in
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Fig. 3. Thin-section electron micro
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Table 3. Lack of detectable related
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J. HTLV Was Present in the Primary
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specific signals into nonspecific s
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Table 2. Distribution of HTLV-relat
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u 10 ~ 20 ~ :c 30 a 40 o Fig. 1. Ki
Page 529 and 530:
Page 531 and 532:
SSV TrS-gp labeled effectively with
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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 555 and 556:
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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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