The Role of Lymphocytes and Macrophages in the Immunological Response XIII International Congress of Haematology, Munich August 2-8,1970 Edited by D. C. Dumonde With 37 Figures Springer -Verlag Berlin· Heidelberg. New York 1971 Dr. Dudley Cohen Dumonde, Kennedy Institute of Rheumatology, Division of Immunology, Bute Gardens, Hammersmith, London, W 6/Great Britain ISBN-13: 978-3-642-65135-9 e-ISBN-13: 978-3-642-65133-5 DOl: 10.1 007/978-3-642-65133-5 This work is subject to copyright. All rights are reserved, whether the whole or part of the material is con cerned, specifically those of translation, reprinting, re-use of illustrations, broadcasting, reproduction by photo copying mamine or similar means, and storage in data banks. Under § 54 of the German Copyright Law where copies are made for other than private use, a fee is payable to the publisher, the amount of the fee to be determined by agreement with the publisher. © by Springer Verlag Berlin' Heidelberg 1971. Library of Congress Catalog Card Num- ber 72-151103. Solkover reprint of the hardcover 1st edition 1971 The use of general descriptive names, trade names, trade marks, etc. in this publication, even if the former are not especially identified, is not be taken as a sign that sum names, as understood by the Trade Marks and Mermandise Marks act, may accordingly be used freely by anone. Preface: Editorial Introduction This volume contains papers presented at a symposium on "The Role of Lympho cytes and Macrophages in the Immunological Response" at the XIII International Congress of Haematology, Munich, August 1970. This symposium was designed to highlight current work submitted to the Haemato logy Congress which related to the role of cellular cooperation in induction and expression of the immune response. The symposium was divided into two parts. The morning session consisted of invited papers dealing with various general aspects of the field (papers 1 to 5) which were relevant to the afternoon session, which con sisted of free communications submitted to the Congress. In the discussion it was emphasized that whereas lymphocyte activIty ultimately underlies all immune induction, the cellular and humoral manifestations of the immune response are governed by distinct yet interacting compartments of the ly.mphoid system. Much interest centred on the role of thymus-derived lymphocytes as cooperator cells in antibody formation; and the mechanisms by which macro phages assist antibody production. The discussion brought together work on lympho cyte activation, the generation of lymphocyte activation products (lymphokine factors) and the cellular events underlying the induction and expression of the Immune response. The helpful cooperation of Springer-Verlag has enabled rapid publication of the papers presented at this symposium. London, February 1971 D. C. DUMONDE Symposium Moderator Contents The Spectrum of Thymus Dependency. E. LEUCHARS 1 The Flow of Information during Primary Antibody Synthesis. D. ]ACHERTS. With 3 Figures 5 Biochemical Evidence against a Physiological Role for Macrophage RNA- Antigen Complexes in the Immune Induction G. E. ROELANTS. . 15 In vitro Studies of Cellular Co-Operation during Immune Induction. K.-U. HARTMANN. 22 Lymphocyte Mitogenic Factor in Cell-Mediated Immunity. R. A. WOLSTEN CROFT, M. MATTHEW, C. OATES, R. N. MAINI, and D. C. DUMONDE. With 2 Figures 28 The Uptake of Antigens and Non-Antigenic Markers by Mouse Spleen and Peritoneal Cells. T. MANDEL and P. BYRT. With 3 Figures 38 Mechanisms by which Human Macrophages Resist Intracellular Growth of Listeria Monocytogenes: Studies with a Redox Reagent Active in Leprosy and Experimental Tuberculosis (B 663). M. ]. CLINE . 43 The Effect of Cyclophosphamide Treatment on Immunological Competence as Measured by the Mixed Lymphocyte Reaction. M. R. SCHWARZ. With 2 Figures 45 Influence of E. Coli L-Asparaginase (EC-2 A-se) on Phytohaemagglutinin (PHA)-Stimulated Human Lymphocytes: Inhibition of the Cytoaggressive Effect. H. OERKERMANN, W. D. HIRSCHMANN, K. SCHUMACHER, G. ALZER, G. UHLENBRUCK, G. WINTZER, and R. GROSS. With 1 Figure 49 Mitotic and Chromosomal Abnormalities Induced by Disrupted Lymphocytes Obtained from Patients with Autoimmune Disorders and Lymphoid Malig nancies. T. HOSHINO, S. KAWASAKI, S. ITANI, S. NAKAYAMA, and M. FUKASE. With 3 Figures. 52 Migration Inhibitory Factor and Mitogenic Factor Produced by Antigen Stimulated Human Lymphocytes. H.-D. FLAD and G. HOCHAPFEL. With 2 Figures . . . . 58 In vitro Studies on Human Lymphocyte Factors of Sensitized Lymphocytes. H. M. DOSCH, K. HAVEMANN, H. MALCHOW, C. P. SODOMANN, and M. SCHMIDT. With 5 Figures 62 The Effect of DEAE-Dextran on Stimulated Human Lymphocytes, with and without "Exogenous" RNA. P. G. RIGBY. 68 VI Contents Changes of Euchromatin Content in Human Peripheral Lymphocytes under the Influence of Phytohaemagglutinin. P. DRINGS. With 3 Figures. 73 Inhomogeneity of Peripheral Lymphocytes - Complement Receptor Sites on a Portion of Human Lymphocytes. H. HUBER, G. MICHLMAYR, C. HUBER, H. ASAMER, and S. D. DOUGLAS. With 3 Figures 78 Studies on the Biological Properties of the Isolated Active Fractions of Phyto haemagglutinin (PHA) from Phaseolus vulgaris. K. SCHUMACHER, G. WINTZER, H. OERKERMANN, G. UHLENBRUCK, G. ALZER, W. D. HIRSCH- MANN, and R. GROSS. With 3 Figures . 83 The Nature of the PHA Receptor on Red Cells. G. UHLENBRUCK, G. WINTZER, K. SCHUMACHER, H. OERKERMANN, G. I. PARDOE, W. D. HIRSCHMANN, G. ALzER, and R. GROSS. With 3 Figures. 87 Immunochemistry of Galactosyl Groups of Cell Membranes. G. I. PARDOE, G. W. G. BIRD, G. UHLENBRUCK, and D. J. ANSTEE. With 4 Figures 91 Subject Index . 99 Author Index D. C. DUMONDE, Division of Immunology, Kennedy Institute of Rheumatology, Hammersmith, London W 6/Great Britain D. J. ANSTEE, Bristol Regional Blood Transfusion Service, Southmead, Bristol/Great Britain G. ALZER, Medizinische Universit1itsklinik, D-5000 Koln-Lindenthal H. ASAMER, Medizinische Universit1itsklinik, lnnsbruck/Austria G. W. G. BIRD, Birmingham Regional Blood Transfusion Service, Birmingham 15/ Great Britain P. BYRT, The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Parkville 3050, Australia M. J. CLINE, University of California, San Francisco, CA 94122/USA P. DRINGS, Medizinische Universit1itsklinik, D-6900 Heidelberg H. M. DOSCH, Medizinische Klinik der Universit1it, D-355 Marburg S. D. DOUGLAs, Mount Sinai School of Medicine, New York, NY 10029/USA H.-D. FLAD, Abteilung fur klinische Physiologie des Zentrums fur klinische Grund lagenforschung der Universit1it Ulm, D-7900 Ulm/Donau, ParkstraBe 11 M. FUKAsE, The Second Division of Department of Internal Medicine, Faculty of Medicine, Kyoto University, Kyoto/Japan R. GROSS, Medizinische Universit1itsklinik, D-5000 Koln-Lindenthal K. U. HARTMANN, Max-Planck-Institut fUr Virusforschung, Abteilung Physikalische Biologie, D-7400 TUbingen K. HAVEMANN, Medizinische Klinik der Universit1it, D-355 Marburg W. D. HIRSCHMANN, Medizinische Universit1itsklinik, D-5000 Koln-Lindenthal G. HOCHAPFEL, Abteilung fur klinische Physiologie des Zentrums fur klinische Grund lagenforschung der Universit1it Ulm, D-7900 Ulm/Donau, ParkstraBe 11 VIII Author Index T. HOSHINO, The Second Division of Department of Internal Medicine, Faculty of Medicine, Kyoto University, Kyoto! Japan C. HUBER, Medizinische Universitatsklinik, Innsbruck/Austria H. HUBER, Medizinische Universitatsklinik, Innsbruck/Austria S. ITANI, The Second Division of Department of Internal Medicine, Faculty of Medi cine, Kyoto University, Kyoto! Japan D. JACHERTS, Institut fiir Hygiene und Medizinische Mikrobiologie, CH-3000 Bern, FriedbiihlstraBe 51 S. KAWASAKI, The Second Division of Department of Internal Medicine, Faculty of Medicine, Kyoto University, Kyoto/Japan E. LEUCHARS, Chester Beatty Research Institute, Institute of Cancer Research, Royal Cancer Hospital, Fulham Road, London, S.W.3!Great Britain R. N. MAINI, Division of Clinical Research, Kennedy Institute of Rheumatology, Hammersmith, London W 6/Great Britain H. MALCHOW, Medizinische Klinik der Universitat, D-355 Marburg T. MANDEL, The Walter and Eliza Hall Institute of Medical Research, Royal Mel bourne Hospital, Parkville 3050, Australia M. MATTHEW, Division of Immunology, Kennedy Institute of Rheumatology, Hammersmith, London W 6/Great Britain G. MICHLMAYR, Medizinische Universitatsklinik, Innsbruck/Austria S. NAKAYAMA, The Second Division of Department of Internal Medicine, Faculty of Medicine, Kyoto University, Kyoto/Japan C. OATES, Division of Immunology, Kennedy Institute of Rheumatology, Hammer smith, London W 6/Great Britain H. OERKERMANN, Medizinische Universitatsklinik, D-5000 Koln-Lindenthal G. I. PARDOE, Department of Experimental Pathology, Medical School, University of Birmingham, Birmingham 15!Great Britain P. G. RIGBY, University of Nebraska, College of Medicine, Omaha, NB 68105/USA G. E. ROELANTS, National Institute for Medical Research, Mill Hill, London NW 7/ Great Britain M. SCHMIDT, Medizinische Klinik der Universitat, D-355 Marburg Author Index IX K. SCHUMACHER, Medizinische Universitatsklinik, D-5000 Koln-Lindenthal M. R. SCHWARZ, Department of Biological Structure, University of Washington, Seattle, WA 98105/USA C. P. SODOMANN, Medizinische Klinik der Universitat, D-355 Marburg G. UHLENBRUCK, Medizinische Universitatsklinik, D-5000 Koln-Lindenthal G. WINTZER, Medizinische Universiditsklinik, D-5000 Koln-Lindenthal R. A. WOLSTENCROFT, Division of Immunology, Kennedy Institute of Rheumatology, Hammersmith, London W 6/Great Britain The Spectrum of Thymus Dependency E. LEUCHARS Introduction Studies on the thymus have revealed differences in its control of the development of immunological responsiveness in different species [1]. In the chicken, cell mediated immune responses have been demonstrated to be thymus-dependent but humoral antibody responses have been found to be affected by a different lymphoid organ, the bursa of Fabricius [2]. It has been argued that a similar system might operate in mammals [3, 4] but investigations in the mouse have so far failed to reveal unequivocal evidence of a dichotomy in the development of immunological responsiveness. Studying the responses to a wide variety of antigens, varying from skin homografts and oxazolone to serum proteins and bacterial antigens, in normal, thymus-deprived and thymus-reconstituted CBA mice, it has been show that all, with the possible exception of pneumococcal polysaccharide, are thymus dependent. This finding will be considered in relation to the numbers of residual thymus derived cells in deprived mice [5]. The significance of the demonstrated spectrum of thymus dependency in mice and its relation to the "bursal equivalent" theory will be discussed. Experimental Procedure A variety of responses have been tested using a standard experimental system. Details of this system have been described previously [6] but an outline of the method of preparation of the test mice is shown in Table 1. Bone marrow cells and thymus grafts were given on the day of irradiation. Mice were tested with the appropriate stimulus approximately 50 days later. Table 1. Groups of CBA/Lac male mice used to test the thymus dependency of various responses Group Thymus Irradiation Bone marrow Thymus graft in situ (850 r) (5 X 106 syngeneic (1 syngeneic neonatal cells LV.) thymus lobe under kidney capsule) Normal + Deprived + + Reconstituted + + + 2 E. LEucHARs Results The results which have been obtained are summarised in Table 2. The extent to which each response depended on the presence of thymus tissue is indicated. Table 2. Thymus dependency of a variety of responses Test material Response measured Thymus Reference dependent or not In vivo: Oxazolone Contact sensitivity + 7 8 Skin homograft Rejection + 9 Tumour homograft Control of growth + 10 Sheep red blood cells Haemagglutinin and + 11 haemolysin tit res Maia squinada Antibody titre + 12 haemocyanin (antigen binding capacity) NIP-BSA Antibody titre (modified + 13 phage inactivation) Bacteriophage fd Antibody titre + 14 (phage inactivation) Salmonellar flagellar antigen Agglutinin titre ± 15 Pneumococcal polysaccharide Agglutinin titre ?- 15 Leishmania tropica Healing of lesion + 16 promastigotes (ID) Trichinella spiralis Eosinophil count + 17 larvae(IV) Experimentally induced Neutrophil count 17 pyelonephritis In vitro: PHA Lymphocyte transformation + 5 Discussion The majority of the responses reviewed in Table 2 have proved to be thymus dependent. However, the mechanism by which the thymus exerts an effect in such a wide range of reactions is by no means fully understood. Of the responses listed above, the ability to make antibody to sheep red blood cells has been most fully investigated [6, 18, 19, 20]. It has been shown that the thymus provides cells, "T lymphocytes" [21], which are essential for the full expres sion of the antibody response. These cells react by mitosis to antigen [6] but are not the cells which secrete antibody [7]. The cells which co-operate with "T lymphocytes" and give rise to plasma cells have been called "B lymphocytes" [21] to indicate that they may be "bursal-equivalent cells"; but there is as yet no critical evidence that a "bursal-equivalent" exists in mammals, although gut-