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possesses a fairly small protein backbone. Preliminary results indicate that the protein moiety is important for the integrity of the NP p65 SSAV ga Staph Prot,ease' iP SSAV p6.5 gagl Papain P SSAV pos, gag moleeule on a 5 % SDS gel as wen as for its antigenicity (unpublished). The SSV TrS-gp is probably coded for by SSV, because it was demonstrated in cells from three different species transformed by SSV and not by other transforming viruses. Our studies indicate that the SSV TrS-gp is easily shed or secreted by the transformed cell, and since the go at was immunized with live autologous transformed cells, the SSV TrS-gp might serve as an important target antigen for tumor cell recognition in vivo. Elucidation of the role of this glycoprotein in transformation by SSV awaits its purification and characterization. References Fig. 4. After immune precipitation of 3H-Ieucine labeled p65 from SSV-NP cells and the gag-precursor from SSAV infected cells, the precipitates were electrophoresed on a preparative SDS gel and the respective bands cut from the gel. The slices were applied to a second SDS gel (15%) and electrophoresed in the presence of either no enzyme, Staphylococcus aureus protease (5 Ilg), or papain (40 ng) as described (Cleveland et al. 1976) Aaronson SA, Stephenson, JR, Hino S, Tronick SR (1975) Differential expression of helper viral structural polypeptides in cells transformed by donal isolates of woolly monkey sarcoma virus. J Virol 16:1117-1123 - Aaronson SA, Stephenson JR, Tronick SR, Hino S (1976) Immunological analysis of structural polypeptides of woolly monkey - gibbon ape type-C viruses. In: Clemmesen J, Y ohn DS (eds) Comparative leukemia research, vo143. Karger, Basel, pp 102-109 - Benveniste RE, Callahan R, Sherr Cl, Chapman V, Todaro GI (1977) Two distinct endogenous type C viruses isolated from the Asian rode nt Mus cervicolor: Conservation of virogene sequences in related rodent species. J Virol 21 :849-862 - Bergholz CM, Wolfe LG, Deinhardt F (197'7) Establishment of simian sarcoma virus, type 1 (SSV-1)-transformed non-producer marmoset celllines. Int J Cancer 20: 104-111 - Cleveland DW, Fischer SG Kirschner MW, Laemmli UK (1976) Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem 252: 1102-1106 - Deinhardt F, Bergholz C, Hunsmann G, Schneider J, Thiel HJ, Beug H, Schäfer W (1978) Studies of simian sarcoma and simian sarcoma-associated virus. I Analysis of viral structural proteins, and preparation and characterization of antiserum specific for viral envelope components. Z Naturforsch [Cl 33: 969-980 - Theilen GH, Gould D, Fowler M, Dungworth DL (1971) C-type virus in tumor tissue of a woolly monkey (Lagothrix spp.) with fibrosarcoma. J Natl Cancer Inst 47:881-889 523
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 Viral Gene Expression in CeUs Transformed by Simian Sarcoma Virus, an Infectious Primate Type C Retrovirus C. M. Bergholz A. Introduction Simian sareoma virus (SSV) is a replieationdefeetive transforming virus isolated in the presenee of exeess nontransforming associated virus (SSAV) from a spontaneous sareoma of a woolly monkey. Previous studies have shown that the struetural proteins of SSV(SSAV) are similar to the eorresponding eomponents of murine leukemia virus (MuLV). Studies reported he re foeus on viral gene expression in SSV-transformed virus producing marmoset monkey eells [HF/SSV(SSAV)] and SSVtransformed nonprodueer marmoset eell lines (HF/SSV-NP). I. Precursor Polypeptides in Virus Producing Cells Antiserum specifie for SSV(SSAV) p30 (kindly provided by Dr. H. J. Thiel) and another specifie for SSAV envelope proteins were reaeted with 35S-methionine-Iabeled extraets of HF ISSV(SSAV) eells. Normal HF and HF ISSAV eells were tested in parallel. Analysis of radioimmunopreeipitates (RIP) by SDS-PAGE and fluorography demonstrated that anti-SSAV p30 precipitated predominantly polypeptides of approximately 30,000 and 60,000-62,000 daltons and, in addition, polypeptides of approximately 72,000 and 52,000 daltons (Fig. 1). Results of pulse-eh ase experiments indieated that the 60,000 dalton pro tein (pr60gag) is proeessed to yield p30, the major viral eore protein (Fig. 1). Using antiserum speeific for gp70 and p15E/p12E large amounts of an 82,000 dalton polypeptide were precipitated, and results of pulse-ehase experiments indicated that it is an envelope preeursor protein proeessed to yield p15E and p12E (Fig. 2). Further experiments are aimed at analyzing glyeosylation of the env and gag gene produets. A gag polyprotein or nonstruetural polypeptide unique to transformed eells was not deteetable by RIP SDS-PAGE with antisera for viral structural pro teins or serum from tumor bearing marmosets. 11. Phosphorylated Proteins Two virus-specific phosphoproteins, the pr60gag and p12, were revealed by SDS-PAGE of 32P-Iabeled proteins. No phosphorylated pro teins were precipitated by antiserum specific for SSA V envelope proteins, providing further evidence that SSAV has a phosphorylated core p12 and a nonphosphorylated 12,000 dalton envelope protein (pI2E). It was further noted that IgG was phosphorylated in extraets of both HF/SSV(SSAV) and HF/SSAV eells, but not in extracts of HF/SSV-NP cells. 111. SSV Gene Expression Establishment of HF/SSV-NP cells has been deseribed previously (Bergholz et al. 1977). Presenee of the SSV genome was demonstrated by rescue of transforming virus following superinfeetion with SSAV, GALV, or MuLV. Expression of the SSAV -related sequenees by the SSV provirus was investigated by reaeting an SSAV cDNA with increasing eoneentrations of RNA extracted from two HF ISSV -NP eell lines. The results (Fig. 3) indicated that SSAV-related sequences were transcribed but represented a smaller proportion of the total eell RNA relative to virus producing cells. Comparison of maximum levels of hybridization indicated that less of the SSAV genome is 524
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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
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indieate a shifting of target eell
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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 and 528: u 10 ~ 20 ~ :c 30 a 40 o Fig. 1. Ki
Page 531: 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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