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After the induction treatment not only a striking increase in virus production but also an alteration in cell morphology can be recognized. While uninduced cultures of Tera-l show a monolayer of homogeneous ceIls, after induction several cell types arise within a matter of 3 to 4 days and dome and vesicle formation can be seen. The GH and HL ceIls, already selected for dome and vesicle formation, exhibit only limited morphologie alterations. In contrast, Tera-2 cells cannot be induced by chemical means to initiate virus production and cellular differentiation (see Table 1). In the differentiated tumor cell populations the proportion of cells producing virus-like particles as judged by electron microscopy varies from a few in HL to over 10% in Tera-l to almost 100 % in GH cultures. The latter cell line, subcultivated only by transferring vesicles to new culture flasks, exhibits in any case a low spontaneous production of retrovirus partie Ies. Thus it appears likely that virus replication is permitted only in a specific cell type of the differentiating culture. In placentas it is the syncytiotrophoblast cell layer from which retrovirus particles can occasionally be seen budding (Kalter et al. 1973; Vernon et al. 1974). Syncytiotrophoblasts at the same time produce human chorionic gonadotropin (HCG) (Morisada et al. 1972). As a first step in determining the type of the virus-producing ceIl, we tested the secretion of HCG into the culture medium before and after the induction procedure (Table 1). While in Tera-l, GH, and HL cells HCG production after induction is significantly increased, this hormone is not detectable in the supernatant of Tera-2 cell cultures. Induction or enhancement of virus synthesis is thus paralleled by induction of HCG productiön. The alteration in hormone release may well reflect cellular differentiation, but it is not possible at present to claim that in analogy to the findings in human placentas, it is a trophoblastic or at least a HCG-secreting cell that is permissive for virus replication. Attempts to further characterize the human teratocarcinoma derived viruses have so far been unsuccessful. Firstly, we were not able to demonstrate unambiguously reverse transcriptase (RT) activity either in uninduced or induced cultures. Transient small peaks of RT activity (Kurth et al. 1980) are observed but are difficult to interpret. The difficulty in detecting RT activity in the supernatant of teratocarcinoma cell cultures has also been reported by Bronson et al. (1978). Secondly, the teratocarcinoma viruses have not yet been demonstrated to be infectious. Cultivations between induced teratocarcinoma cells and a number of uninfected indicator celllines did not yield replicating virus even after repeated cocultivations and passages for over 20 weeks. The indicator cells were derived from dog thymus (8155), bat lung (Tb-I-Lu), marmoset fibroblasts (HF), tupeia embryo kidney (TEK) , mink lung (C58), and feHne embryo fibroblasts (FEF), as well as from human cells (rhabdomyosarcoma RD, amnion cells AV -3). c. Discussion Whereas in uninduced human teratocarcinoma cells expression of retrovirus particles is a rare event (Bronson et al. 1978, 1979; Kurth et al. 1980), the production of such particles in three out of four teratocarcinoma celliines can easily be demonstrated after treatment with IUdR, DXM, and DMSO. Individual tumor lines mayaiso produce virus particles spontaneously. The human teratocarcinoma derived particles differ morphologically from mammalian type-C viruses by the lack of a c1ear electron lucent space between core and envelope. In this regard they resemble the particles observed in human term placentas. The lack of easily demonstrable RT activity and infectivity as weIl as the absence of "mature" particles with condensed cores is also reminiscent of a number of retrovirus mutants (e.g., murine leukemia virus murants ts3 and ts24) (Witte and Baltimore 1978). These mutants are characterized by a defect in viral precursor pro tein processing. It is as yet largely unknown to what extent viral or host cell proteases contribute to the specifie c1eavage of virus precursor proteins to yield mature and infectious particles. Therefore, the question cannot yet be decided whether the human teratocarcinoma-derived viruses posses an intrinsic genetic defect or whether the host cell is lacking a factor which is necessary for an appropriate processing of viral structural components. The latter possibility is somewhat favored by the recent findings of Gautsch (1980) that murine teratocarcinoma cells lack a factor needed to support the 543
eplication of infecting murine leukemia (Teich et al. 1977) and papova (Swartzendruber and Lehman 1975) viruses. References Branson DL, Ritzi DM, Fraley EE, Dalton AJ (1978) Morphologic evidence for retrovirus production by epithelial cells derived from a human testicular tumor metastasis. J Natl Cancer Inst 60: 1305-1308 - Bronson DL, Fraley EE, Fogh J, Kalter SS (1979) Induction of retrovirus particles in human testicular tumor (Tera-l) cell cultures: an electron microscopic study. J Natl Cancer Inst 63:337-339 - Dalton AJ, Hellman A, Kalter SS, Helmke RJ (1974) Ultrastructural comparison of placental virus with several type-C oncogenic viruses. J Natl Cancer Inst 52:1379-1381 - Fogh J, Trempe G (1975) New human tumor celllines. In: Fogh I (ed) Human tumor cells in vitra. Plenum, New York, pp 115-159 - Gautsch JW (1980) Embryonal carcinoma stem cells lack a function required for virus replication. Nature 285: 11 0-112 - Kalter SS, Helmke RJ, Heberling RL, Panigel M, Fowler AK, Strickland JE, Hellman A (1973) C-type particles in normal human placentas. J Natl Cancer Inst 50:1081-1084 - Kurth R, Löwer R, Löwer J, Harzmann R, Pfeiffer R, Schmidt CG, Fogh J, Frank H (1980) Oncornavirus synthesis in human teratocarcinoma cultures and an increased antiviral immune reactivity in corresponding patients. In: Todaro G, Essex M, zur Hausen H (eds) Viruses in naturally occurring cancers. Cold Spring Harbor Laboratory, New York pp 835-846 - Morisada M, Yamaguchi H, Lizuka R (1972) Toxie action of anti-h.c.g. antibody to human trophoblasts. Int J Fertil 17:65-71 - Pimentel E (1979) Human oncoviralogy. Biochim Biophys Acta 560: 169-216 - Swartzendruber DE, Lehman JM (1975) Neoplastic differentiation: Interaction of simian virus 40 and Polyoma virus with murine teratocarcinoma cells in vitra. J Cell Physiol85 : 179-188 - Teich NM, Weiss RA, Martin GR, Lowy DR (1977) Virus infection of murine teratocarcinoma stern celllines. Cell 12:973-982 - Vernon ML, McMahon JM, Hackett JJ (1974) Additional evidence of type-C particles in human placentas. J Natl Cancer Inst 52:987-989 - Witte ON, Baltimore D (1978) Relationship of retrovirus Polypratein c1eavages to to virion maturation studied with temperature-sensitive murine leukemia virus mutants. J Viral 26:750-761 544
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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) (
Page 447 and 448:
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
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~ i ,..1---------,,':::::::::::::::
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fragment of pBR322 DNA, a 3.0-kbp S
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Mak TW (1979) Proc Natl Acad Sci US
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Table 1. Transformation of 3T3 cell
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inserts in the genome of rapidly tr
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pulations, it seemed likely that th
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contains antigens which react with
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le of reacting in sensitive radioim
Page 501 and 502: Haematology and Blood Transfusion V
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 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: - Fig. la-f. Retravirus particles p
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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