Thoracic duct lymphocytes obtained from mice 1-2 days after the injection of sheep erythrocytes and injected into thymectomized, irradiated, marrow-protected syngeneic hosts were deficient in adoptively transferring immune reactivity to sheep erythrocytes but normal with respect to horse erythrocytes. Cells collected at 3 days had normal reactivity, but cells collected at 5 days allowed their hosts to produce an enhanced response to sheep erythrocytes and a somewhat depressed response to horse erythrocytes. Thoracic duct lymphocytes obtained from CBA mice 2 days after injection of (CBA x C57BL) F-1 spleen cells produced splenomegaly in newborn (CBA X BALB/c)F-1 mice but not in (CBA x C57BL)F-1 recipients. When obtained 5 days after injection, they caused an increase in the splenic index of the (CBA x C57BL)F, recipients significantly above that given by control lymphocytes from saline-injected donors. These results are interpreted in terms of a selective recruitment from the circulation of specific antigen-sensitive cells, occurring soon after antigen administration and followed in turn by a rapid reentry of such cells into the pool in an increased proportion.

Small lymphocytes from the thoracic duct lymph of rats (1) and mice (2) can initiate immune responses to antigens such as sheep erythrocytes and histoincompatible cells when transferred to appropriate hosts. Recently, it was demonstrated that for 1-2 days after the injection of such antigens in mice (3) or rats (4, 5), there was a specific deficiency in thoracic duct lymph of cells able to mount adoptive immune responses to these antigens. The results were interpreted in terms of antigeninduced selective recruitment of specific lymphocytes from the recirculating pool to appropriate lymphoid organs. As a corollary, it would be expected that at this time such organs, e.g. the spleen, would be enriched in cells specific for these antigens.

Immunology is frequently the prisoner of its own semantics. Evolution designed an immune system equipped to recognize all foreign antigens, the structure of which it cannot know in advance. The solution involves repertoires of unique clonotypes for both B and T lymphocytes, resulting in a heterogeneity of binding avidities with respect to any given epitope. For the B cell, we now know that as many as 1 in 30 B cells can form antibody leading to lysis of haptenated erythrocytes, suggesting that there are many thousands of ways of forming an antihapten, antibody-combining site. So the question frequently is not, does this B cell recognize that antigen?, but, how well does this B cell recognize that antigen? This much our language can cope with, but when we come to issues like antigen- initiated clonal expansion and differentiation, it is too clumsy to say: This antigen in that dose will trigger two rounds of division in that B cell, with its low-affinity receptors, but ten in that other B cell with its high-affinity receptors. Therefore, we will pose the question and our experimental approaches to it in the simpler yes or no format.

T cells are an extremely heterogeneous population composed of various subsets which interact in a complex manner during an immune response. Furthermore each T cell expresses an antigen-specific receptor unique in its discriminating capacity and in its restriction by gene products of the major histocompatibility complex (MHC). The availability of defined T cell clones should help elucidate the nature of T cell specificity and the function of particular T cells in immune circuits. We have produced T cell lines either by growing T cells from immune mice with antigen, antigen-presenting cells (APC) and T cell growth factors (interleukin-2, IL-2), or by immortalizing T cells after fusing them to T lymphoma cell lines. Some of the results obtained in this work are summarized here.

Specific immune responses to a wide variety of antigens are under genetic control and many of these are linked to the species major histocompatibility complex (MHC) in the mouse, the rat and man [1]. Moreover, associations between histocompatibility type and susceptibility to disease have been observed in animals and man. Antigen-induced arthritis as a model of rheumatic inflammation was established in the mouse to define the role of humoral and cell-mediated immunity in the induction and persistence of dis ease and to investigate genetic aspects of susceptibility and resistance.

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Several studies have revealed that within 1 day of injecting mice or rats with antigens, such as heterologous erythrocytes, lymphocytes reactive to these antigens are specifically withdrawn from the recirculating lymphocyte pool and become sequestered in organs, such as the spleen. In mice injected with H2-incompatible spleen cells, the capacity of thoracic lymphocytes (TDL) to evoke a graft-versus-host (GVH) reaction against the injected determinants is found to be specifically abolished at 1–2 days, near normal on day 3, and above normal on day 5. This chapter describes studies that were designed to determine whether the unresponsiveness towards the GVH determinants also applies to cells producing a mixed-lymphocyte reaction (MLR), allograft rejection, and cell-mediated lympholysis (CML). The results from the studies showed that injecting parental strain mice with irradiated H2-incompatible spleen cells virtually abolished the capacity of TDL, collected 1 day later, to respond specifically to determinants controlling the GVH reaction, allograft rejection, and CML. The near normal reactivity of cells taken from the spleen at this time suggests that the injected irradiated cells carrying these various H2 determinants induced antigen-specific selective recruitment of recirculating lymphocytes (ASRL) reactive to those determinants to organs such as the spleen.

LYMPHOCYTES with the capacity to bind antibody-antigen complexes to their surface1–3 are probably bone marrow-derived, B, cells, not thymus-derived, T, cells3. We now have definite evidence that such lymphocytes are indeed B cells and will describe how this property can be utilized in a practical way for separating T cells from B cells.

MOUSE lymphocytes can be divided into two distinct populations according to the density of immunoglobulin determinants on their surface. Lymphocytes with a high density of immunoglobulin are marrow-derived, nonthymus-processed, B cells, whereas lymphocytes with little or no immunoglobulin are thymus-derived, T cells1–3. Since more than 95% of mouse immunoglobulin light chains are of the kappa type4, treatment of lymphocyte suspensions with an appropriate dilution of rabbit anti-mouse kappa serum and complement should be cytotoxic for only B lymphocytes. This prediction was tested by using lymphocyte populations enriched for either T or B cells or containing the two cell types in a known proportion.

Interaction between antigen and two separate classes of lymphocytes is a feature of many antibody responses. Thus, both thymus-derived “T” cells and non-thymus-derived “B” cells are involved. The T cells are required to initiate or facilitate antibody production by B cells in response to certain antigenic determinants. The exact nature of the interaction is not clear.

Experiments were designed to test the possibility that thymus-derived (T) cells cooperate with nonthymus derived (B) cells in antibody responses by acting as passive carriers of antigen. Thoracic duct lymphocytes (TDL) from fowl gammaG-tolerant mice were incubated in vitro with fowl anti-mouse lymphocyte globulin (FALG), which was shown not to be immunosuppressive in mice. On transfer into adult thymectomized, irradiated, and marrow protected (TxBM) hosts together with a control antigen, horse RBC, a response to horse RBC but not to fowl gammaG was obtained. By contrast, TxBM recipients of nontolerant, FALG-coated TDL responded to both antigens and the antibody-forming cells were shown to be derived from the host, not from the injected TDL. These findings suggested that, under the conditions of the experiment, triggering of unprimed B cells in the spleens of TxBM hosts was not achieved with antigen-coated tolerant lymphocytes. Another model utilized the ability of B cells to bind antibody-antigen complexes. Spleen cells from TxBM mice, incubated in vitro with anti-fowl gammaG-fowl gammaG.NIP, were injected with or without normal TDL (a source of T cells) into irradiated hosts. Only mice given both cell types could produce an anti-NIP antibody response. In a further experiment, spleen cells from HGG.NIP-primed mice were injected together with NIP-coated B cells (prepared as above) into irradiated hosts. A substantial anti-NIP antibody response occurred. If, however, the T cells in the spleens of HGG.NIP-primed mice were eliminated by treatment with anti-theta serum and complement, the NIP response was abolished. It was concluded that antigen-coated B cells could not substitute for T cells either in the primary or secondary response. Treatment of T cells from unprimed or primed mice with mitomycin C impaired their capacity to collaborate with B cells on transfer into irradiated hosts. Taken together these findings suggest that before collaboration can take place T cells must be activated by antigen to differentiate and in so doing may produce some factor essential for triggering of B cells.

THE mechanism, known as the carrier effect, whereby immunity to one or more determinant groups enhances the response to other determinants on the same multivalent antigen, was first recognized in delayed hypersensitivity to haptens, in which, for an appreciable response, the hapten must be coupled to the same protein carrier for priming and challenge1, 2. Carrier specificity has also been demonstrated in the secondary antibody responses to hapten protein conjugates3. Two alternative hypotheses have been advanced to explain this specificity. The “local environment” hypothesis supposes that the hapten-sensitive cell recognizes both the hapten and the carrier determinants. However, the antihapten antibodies produced do not distinguish details of the carrier molecule and so do not reflect the specificity of the cellular receptor. Furthermore, inert spacer molecules inserted between hapten and carrier do not interfere with carrier specificity in the antibody response3. Reflecting current views on the cooperation between thymus-derived (T) and bone marrow derived (B) lymphocytes in the antibody response to various antigens4, the second hypothesis invokes two or more cells, one with receptors directed towards the hapten (hapten-sensitive cell), the others specific for the carrier molecule proper (carrier-reactive cells). Supporting this is the observation that pre-immunization to a particular protein carrier alone could potentiate the primary or secondary antihapten response to a hapten conjugated to that protein5. In an adoptive transfer system, moreover, the efficiency of antihapten antibody production by cells primed to a particular hapten-protein conjugate and stimulated with the hapten conjugated on a heterologous protein, is significantly enhanced by the introduction of cells primed to the heterologous carrier alone. Anti-carrier serum antibody does not cause such enhancement6. The carrier-reactive cells must therefore cooperate in increasing the efficiency of the hapten-sensitive cells in some way other than by providing humoral anti-carrier antibody. Recent work strongly suggests that carrier reactive cells are thymus-derived6, 7.

“Educated” or “activated” thymus cells have been shown, by the use of “strong” histocompatibility antigens, to be immunocompetent only against the antigens by which they were originally activated.

An antigen “suicide” technique strongly suggests that both T and B cells can dictate the specificity of the antibody response.

Thymus lymphocytes injected into neo-natally thymectomized mice were identified in the thoracic duct lymph by their θ antigens and shown to form part of the recirculating lymphocyte pool.

Research Articles| July 21 2009 Assay for the Immunosuppressive Capacity of Antilymphocyte Serum Based on its Action on Thymus-Derived Cells Subject Area: Immunology and Allergy W.J. Martin; W.J. Martin The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Melbourne, Australia Search for other works by this author on: This Site PubMed Google Scholar J.F.A.P. Miller J.F.A.P. Miller The Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Melbourne, Australia Search for other works by this author on: This Site PubMed Google Scholar International Archives of Allergy and Applied Immunology (1969) 35 (2): 163–178. https://doi.org/10.1159/000230169 Article history Published Online: July 21 2009 Content Tools Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn Email Tools Icon Tools Get Permissions Cite Icon Cite Search Site Citation W.J. Martin, J.F.A.P. Miller; Assay for the Immunosuppressive Capacity of Antilymphocyte Serum Based on its Action on Thymus-Derived Cells. International Archives of Allergy and Applied Immunology 1 February 1969; 35 (2): 163–178. https://doi.org/10.1159/000230169 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsInternational Archives of Allergy and Applied Immunology Search Advanced Search This content is only available via PDF. 1969Copyright / Drug Dosage / DisclaimerCopyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements. Article PDF first page preview Close Modal You do not currently have access to this content.

In 1961, an immunological role for the thymus was revealed by experiments which indicated that thymectomy, at birth in mice, caused a severe depletion of lymphocytes in peripheral blood, lymph nodes, and spleen and a marked deficiency in the capacity to reject foreign skin grafts [1]. Since then numerous experiments have indicated that the thymus must perform a similar function in many species (reviewed by Miller and Osoba [2]). In spite of extensive research on the thymus in the last decade, there is much controversy regarding the fate and function of the thymus lymphocytes — the predominant cell type in thymus tissue. Recent work in our laboratory suggests that thymus lymphocytes are capable of recognizing and interacting with antigen by giving rise not to antibody-forming cells, but to a progeny of recirculating small lymphocytes through the intermediary of large pyroninophilic cells. As this work was performed using one antigenic system in mice, no generalization can yet be made with respect to other systems.