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CJ, Singh J, Taylor RB (1973) The effect of capping by antiimmunoglobulin antibody on the expression of cell surface immunoglobulin and on lymphocytic leukemic lymphocytes. Scand J Immunol2: 143-149 - Fialkow PJ (1976) Clonal origin of human tumors. Biochim Biophys Acta 458:283-321 - Godal T, Henriksen A, Iversen J-G, Landaas T0, Lindmo T (1978) Altered membrane-associated functions in chronic lymphocytie leukemia cells. Int J Cancer 21 :561-569 - Godal T, Lindmo T, Marton PF, Landaas T0, Langholm R, H0ie J, Abrahamsen AF (to be published) Immunologieal subsets in human B-celllymphomas - Henriksen A, Godal T, Landaas T0 (1980) Mitogenie effect on human lymphocytes of insolubilized anti-immunoglobulins. 1. Specificity of the stimulating agent. J Immunol 124:921-925 - Herzenberg LA, Black SJ, Tokuhisa T, Herzenberg LA (1980) Memory B cells at successive stages of differentiation. J Exp Med 151: 1071-1087 - Iversen J-G (1976) Unidirectional K+ fluxes in rat thymocytes stimulated by concanavalin A. J Cell Physiol 89:267-276 - Klaus GGB (1979) Generation of memory cells. IH. Antibody dass requirements for the generation of B-memory cells by antigen-antibody complexes. Immunology 37:345-351 - Landaas T0, Godal T, Marton PF, Kval0Y S, Langholm R, Lindmo T, J0rgensen OG, H0st H (1981) Cell-associated immunoglobulin in human non-Hodgkin lymphomas. A comparative study of surface immunoglobulin on cells in suspension and cytoplasmie immunoglobulin by immunohistochemistry. Acta path microbiol scand, sect A (in press) - Lennert K (ed) (1978) Malignant lymphomas other than Hodgkin's disease. Springer, Berlin Heidelberg New York - Levy R, Warnke R, Dorfman RF, Haimovich J (1977) The monoclonality of human B-cell lymphomas. J Exp Med 145: 1014-1028 -LindmoT, SteenHB (1977)Flow cytometric measurement of the polarization of fluorescence from intracellular fluorescein in mammalian cells. Biophys J 18: 173-187 - Mason DW (1976) The requirement for C3 receptors on the precursors of 19S and 7S antibody-forming cells. J Exp Med 143: 1111-1121-Rohrer JW, Lynch RG (1979) Immunoregulation of localized and disseminated murine myeloma: Antigen-specifie regulation of MOPC-315 stern cell proliferation and secretory cell differentiation. J Immunol 123: 1083-1087 - Steen HB, Lindmo T (1978) Cellular and nudear volume during the cell cyde of NHIK 3025 cells. Cell Tissue Kinet 11 :69-81 - Stein H (1978) The immunologie and immunochemieal basis for the Kiel dassification. In: Lennert K (ed) Malignant lymphomas other than Hodgkin's disease, part 6. Springer, Berlin Heidelberg New York - Stewart CC, Ingram M (1967) A method for counting phytohemagglutininstimulated lymphocytes. Blood 29:628-639 - Taylor CR (1978) Immunocytochemieal methods in the study of lymphoma and related conditions. J Histochem Cytochem 26:496-512 - White RG, Henderson DC, Eslami MB, Nielsen KH (1975) Localization of a protein antigen in the chieken spleen. Effect of various manipulative procedures on the morphogenesis of the germinal centre. Immunology 28: 1-21 - Zan-Bar I, Strober S, Vitetta ES (1979) The relationship between surface immunoglobulin isotype and immune function of murine B lymphocytes. IV. Role of IgD-bearing cells in the propagation of immunologie memory. J Immunol 123:925-930 319
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 Clones of Murine Functional T Cells H. Hengartner A. Introduction The serologie and biochemieal analysis of different classes of T cells, such as cytotoxic, helper , and suppressor T cells, has been impeded by the lack of pure populations of functional T celllines. None of the established weIl characterized AKR thymoma and lymphoma T celllines exhibit any T cell functions besides the expression of serologie T cell surface markers. To date all attempts to transform functional T cells using oncogenie viruses have failed. Although several groups reported T cell hybridomas which secrete specific and nonspecific hel per or suppressor factors (Kontiainen et al. 1978; Taniguchi et al. 1979; Taussig et al. 1979), most of the described hybridomas are not very stable and have to be subcloned and selected for activity over short periods. During the past 3 years, however, several groups have succeeded in establishing and cloning functional human and mouse T cells (reviewed in Schreier et al. 1980, Immunological Reviews, Vol. 51, Ed. Möller, G., Munksgaard, Kopenhagen). Functional T cells were expanded in vitro after in vive immunization and kept under optimal tissue culture conditions for the appropriate T cell type. Alloreactive proliferating T cells can be kept functional by periodieal re stimulation using irradiated spleen cells (Fathman and Nabholz 1977), whereas the growth of cytotoxic or helper T cells after initial antigen specific stimulation, in some cases in vivo and then in vitro, is strictly dependent on the presence of T cell growth factors in the culture medium (Gillis and Smith 1977). All attempts to establish long-term cultures of cloned or uncloned T cells appears to depend upon the following principles: 1. Expansion of the antigen-specifie T cells by in vivo immunization; 2. Secondary immunization in vitro followed by the biologie assay for the appropriate T cell specificity; and 3. Maintaining the functional T cells either by periodical restimulation or by supplementing the cultures with T cell growth factor. To prevent overgrowth by T cells whieh are not antigen specific, it is important to clone the cells at this point and select those clones which exhibit the biologie activity desired. Inour laboratory we have been dealing over the past 4 years with the cloning of T cells of the following types: alloreactively proliferating T cells (Fathman and Hengartner 1978, 1979; Hengartner and Fathman 1980), H-Y antigen (14) and hapten (9) specific H-2 restrieted cytotoxic T cells in the mouse. B. A1loreactive T CeUs Alloreactive T cells can be generated in an ordinary mixed lymphocyte culture where lymph node cells are stimulated by X-ray-irradiated spleen cells. Every 10 to 14 days the viable responder T cells are diluted into fresh medium and restimulated by X-ray-irradiated stimulator spleen cells. Fathman et al. (1977) described the following two long term culture systems: A/J anti-C57BL/6 (A[B6]) and A/J anti-(C57BL/6XA/J)F 1 (A[B6A]). The bulk cultures consisted mainly of haplotype-specifie proliferating T cells lacking any cytotoxic activity. After three to four restimulations, the specificity of the two systems may be analysed by stimulating 10 4 responder T cells with 10 6 X-ray-irradiated spleen cells of different haplotypes. The stimulatory effect is measured 320
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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
Page 279 and 280: Haematology and Blood Transfusion V
Page 281 and 282: the macrophage series in which the
Page 285 and 286: Table 1. Production of factor depen
Page 289 and 290: Table 1. Characteristics of the adh
Page 291 and 292: Fig. 3. A human marrow culture at 6
Page 293 and 294: 300 250 • HYPERPROLIFERATIVE PATT
Page 295 and 296: Fig. 9. Nonadherent population from
Page 297 and 298: The cobblestone areas were often no
Page 299 and 300: L (1976) Induction of colony format
Page 301 and 302: B. Materials and Methods J. Tissue
Page 303 and 304: Fig. 2. Ultrastructural appearance
Page 309 and 310: Table 1. Monoclonal antibody reacti
Page 311 and 312: al. 1979; Janossy et al. 1979) is e
Page 313 and 314: to which this is an exact replica o
Page 315 and 316: inger JL (1976) Distribution of la-
Page 317 and 318: I region antigens were found to be
Page 319 and 320: have been reported to be present on
Page 321 and 322: V. FunctionalAssays Assays for PHA
Page 323 and 324: Functional studies of UCHT1 separat
Page 327 and 328: 40 ,...... (5 L.. C o u 30 .9 ~ .~
Page 329: with immunohistochemieal methods (L
Page 333 and 334: can be dissected by cloning. Utiliz
Page 335 and 336: Table 1. Origin and marker profile
Page 339 and 340: complete identity of gp 100 from no
Page 341 and 342: Table 1. Reaction of single anti se
Page 345 and 346: media. Following 24-h incubation, c
Page 347 and 348: nonleukemic eells (Greaves 1979). L
Page 351 and 352: Fig. 2. Cytocentrifuged smears of b
Page 353 and 354: Modification of amino-groups of hum
Page 355 and 356: disturb this balance in the directi
Page 357 and 358: The majority of human blood lymphoc
Page 359 and 360: of the EBV infected B cells. Howeve
Page 361 and 362: and immunoglobulins. J Immunol 120:
Page 363 and 364: " K 10 • j ~ ALL RNlL AL wtfh les
Page 365 and 366: tion akuter Leukämiezellen in "spo
Page 367 and 368: D. Results and Discussion J. Acute
Page 369 and 370: selectively responsive to different
Page 373 and 374: l1J ~ 100 ~ Q. ::) l1J Z 0 ~ z >- .
Page 375 and 376: Table 2. Growth inhibition of S49 e
Page 377 and 378: 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
Page 435 and 436:
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.
Page 451 and 452:
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
Page 463 and 464:
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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Page 471 and 472:
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
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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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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
Page 533 and 534:
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
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SiS'V' gp70: ANTIGEN, ANTI BODY, IM
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type-C virus p30 antigen in cells f
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c. Results The fractionated whole-b
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induced antibodies as well as exper
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- Fig. la-f. Retravirus particles p
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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
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Haematology andBlood Transfusion Su
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