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y 3H-thymidine uptake during a 16-h period 48 or 72 h after the initiation of the cultures (Fathman et al. 1977). The initial bulk culture A(B6) exhibited identical stimulation indexes against C57BL/6 and (C57BL/6XA/J)F 1 stimulator cells, and the stimulation indexes against third party stimulator cells stayed constant even after the 20th restimulation. The bulk culture A(B6A) showed a stimulation index against (C57BL/6xA/J)F 1 stimulator cells which was twice as high as that against the C57BL/6 stimulator cells, suggesting a unique F 1 MLR determinant on the semiallogeneic stimulator cells. These proliferating T cells could be cloned in soft agar. The responder T cells were stimulated in suspension for 24 hand then seeded on soft agar in a petri dish. After 5 days colonies were picked from the ag ar and subsequently expanded in 0.2-ml, 2-ml, 15-ml and 45-ml cultures by restimulation with X-ray-irradiated stimulator spleen cells every 10 to 14 days. Between two restimulations the cloned alloreactive T cells undergo an activation, a proliferative, and a resting phase. The continous stimulation by allogeneic stimulator cells in vitro leads to this expansion of the alloreactive T cells. The analysis of such cultures clearly demonstrated the existence of an F 1 MLR determinant expressed on semiallogeneic stimulator cells which is absent on C57BL/6 cells. The analysis of such F 1 alloreactive proliferating T cell clones using different recombinant F 1 stimulator spleen cells further demonstrated the existence of at least four different clone types with different fine specificities (Fathman and Hengartner 1979). In the A(B6) bulk cultures we have demonstrated through cloning and subcloning the existence of a T cell clone type which can be stimulated only by C57BL/6 stimulator cells but not by the semiallogeneic (C57BL/6 X A/J)F 1 stimulator cells. This suggests the expression of a unique MLR determinant on the homozygous C57BL/6 stimulator cells (Hengartner and Fathman 1980). The alloreactive T cells can be thawed and restimulated after freezing and storage in liquid nitrogen. The chromosome number of the clones is 40 and the chromosomes do not show any obvious abnormalities. Fathman and Weissman (1980) injected 10 7 AI J cells of one of the F 1 specific clones (clone A [B6A) 1-1] intraperitoneally into A/J, C57BL/6, and (C57BL/6 XA/J)F 1 animals. After 4 weeks the F 1 animals exhibited enlarged lymph nodes, and the H-2 typing demonstrated the A/J origin of a large number of the lymph node cells. This clearly demonstrated the generation of a T cell lymphoma by exposing proliferating T cells to a constant stimulatory environment in the (C57BL/6 X A/J)F 1 mouse. C. Cytotoxic T Cells The MHC-restricted cytotoxic T cells against the H -Y antigen and the hapten sp were generated in secondary mixed lymphocyte cultures 14 days after the primary immunization in vivo. Analogous to the alloreactive T cells, the cytotoxic T cells were cloned in soft agar or und er conditions of limited dilution (Nabholz et al. 1980; Von Böhmer et al. 1979). In contrast to alloreactive proliferating T cells, cytotoxic T cells have to be kept in medium containing T cell growth factor. We routinely use 10% supernatant of ConA-activated (48 h) rat spleen cells in the culture medium as a source for T cell growth factor. The growth of cloned cytotoxic T cells is strictly dependent on T cell growth factors and cells usually die within hours in regular medium. Cytotoxic T cells can not be stimulated by the specific antigens, which are X-ray-irradia ted syngeneic male spleen cells or haptenated syngeneic spleen cells (Nabholz et al. 1980; Von Böhmer et al. 1979). The chromosome number of such cells is higher than 40 and occasionally even metacentric chromosomes can be demonstrated. D. Summary Using techniques of cloning, it became possible to generate alloreactively proliferating (Hengartner and Fathman (1980), cytotoxic (Von Böhmer et al. 1979), and helper (Schreier and Tees 1980; Von Böhmer et al. 1979; Watson 1979) T cells. The homogeneity of functional T cells makes it possible to start to tackle problems such as the chemical nature of the T cell receptor and the organization of genes wh ich lead to the T cell differentiation and specificity. The specific proliferation or killing observed in a complex mixture of cells after activation in a mixed lymphocyte culture 321
can be dissected by cloning. Utilizing such clones, it might also be possible to study the phenomenon of specific activation and the still obscure phenomenon of cytolytic activity. References Fathman CG, Hengartner H (1978) Clones of alloreactive T cells. Nature 272: 617 - Fathman CG, Hengartner H (1979) Cross-reactive mixed lymphocyte re action determinants recognized by cloned alloreactive T cells. Proc Natl Acad Sci USA 76:5863 - Fathman CG, Nabholz M (1977) In vitro secondary mixed leucocyte reaction H. Interaction MLR determinants expressed on F j cells. Eur J Immunol 7:370 - Fathman CG, Weissman IL (1980) Production of alloreactive Tcelllymphomas. Nature 283:404 - Gillis S, Smith KA (1977) Long term cultures of tumor specific cytotoxic T cells. Nature 268:154 - Hengartner H, Fathman CG (1980) Clones of alloreactive Tcells. Immunogenetics 10: 175 - T cell stimulating growth factors. Immunol Rev 51 (1980) -KontiainenS, SimpsonE, Bohrer E, Beverley PC, Herzenberg LA, Fitzpatrick WC, Vogt P, Torano A, McKenzie IFC, Feldman M (1978) T cell lines producing antigen-specific suppressor factor. Nature 274:477 - Nabholz M, Conzelmann A, Acuto 0, North M, Haas W, Pohlit H, von Böhmer H, Hengartner H, Mach JP, Engers H, Johnson JP (1980) Established murine cytolytic T celllines as tools for a somatic cell genetic analysis of T cell functions. Immunol Rev 51 : 125 - Schreier MH, Tees R (1980) Clonal induction of helper T cells: Conversion of specific signals into nonspecific signals. Int Arch Allergy Appl Immunol61 :227 - Schreier MH, Anderson J, Lernhardt W, Melchers F (1980) Antigen-specific T-helper cells stimulate H-2 compatible and incompatible B cell blasts polyclonally. J Exp Med 151: 194 - Taniguchi M, Saito T, Tada T (1979) Antigen-specific suppressive factor produced by a transplantable I -J bearing T cell hybridoma. Nature 274: 555 - Taussig MJ, CorvaUm JRF, Binns RM, Holliman A (1979) Production of H-2 related suppressor factor by a hybrid T cellline. Nature 277:305 - Von Böhmer H, Hengartner H, Nabholz M, Lernhardt W, Schreier MH, Haas W (1979) Fine specificity of a continously growing killer cell clone specific for H-Y antigen. Eur J Immunol 9:592 - Watanabe T, Kimoto M, Maruyama S, Kishimoto T, Yamamura Y (1978) Establishment of T cell hybrid cellline secreting IgE dass specific suppressor factor. J Immunol 121 :2113 - Watson J (1979) Continuous proliferation of murine antigen-specific helper T lymphocytes in culture. J Exp Med 150: 1510 322
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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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Page 53 and 54:
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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Page 281 and 282: the macrophage series in which the
Page 283 and 284: Haematology and Blood Transfusion V
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 and 330: with immunohistochemieal methods (L
Page 331: Haematology and Blood Transfusion V
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
Page 381 and 382: 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
Page 431 and 432:
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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Page 443 and 444:
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
Page 465 and 466:
Lymphoma cellline Table 1. T and B
Page 467 and 468:
Table 2. In vivo growth and oneogen
Page 469 and 470:
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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
Page 479 and 480:
Table 2. Expression of Jl and Sourc
Page 481 and 482:
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~ 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
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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
Page 507 and 508:
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
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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
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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
Page 543 and 544:
SiS'V' gp70: ANTIGEN, ANTI BODY, IM
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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:
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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