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The factor of immunization in the rat. The effect of allogeneic immunization on graft-versus-host activity

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Abstract

Using a popliteal lymph node weight assay the graft-versus-host activity of lymphocytes from donors immunized with allogeneic tissue has been assayed by comparison with that of lymphocytes from nonimmune donors. When the donors were immunized against weak histocompatibility antigens (non-AgB) the specific GVH activity of its lymphocytes was increased. This increase was greater if spleen cells rather than thoracic duct lymphocytes were the source of the donor cells used for assay. The increase in GVH activity was also greater if the standard immunization procedure of two successive skin allografts was followed by three boosting injections of allogeneic lymphoid cells. When donors were immunized against strong histocompatibility antigens the specific GVH activity of the donors' lymphocytes was slightly increased, was unchanged, or was actually decreased depending on the experimental situation. In donors rendered incapable of a humoral alloantibody response by whole body X-irradiation, immunization across a strong barrier was followed by little or no increase in the specific GVH activity of TDL. In the rat, as in other species, the increase in GVH activity after immunization is inversely proportional to the strength of the antigenic barrier involved.

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... For the GVHR one must assume that the time-course of the lymph node enlargement is identical. This has not been tested, but the tempo of the enlargement is the same whether immune or nonimmune cells are used (12). Furthermore, one must assume that the percentage of lymphocytes in the spleen cell suspensions are identical. ...
... The F.I. after immunization against the weak H-ag AS cannot be estimated precisely, because only immune cells have been injected in single doses. About 25 X 106 nonimmune spleen cells are needed to give a lymph node weight of 10 mg (12). There is no difference between the GVH activity of conventional and germfree rats after immunization against the weak AS antigen. ...
Article
The reactivity of lymphoid cells from germfree and conventional CDF rats were compared in mixed leukocyte culture (MLC) with both allogeneic rat leukocytes and with human leukocytes as stimulating cells, and in a graft-versus-host (GVH) assay. There was no difference between germfree and conventional rats in any of these systems. Furthermore, the increase in GVH activity after specific immunization against a strong histocompatibility antigen was small in both cases. The findings are incompatible with the hypothesis that the high proportion of antigen-sensitive cells against strong histocompatibility antigens is caused by cross-reactions between microbial antigens and histocompatibility antigens.
... Assays of T cell alloreactivity include quantitative GVH (162,163), MLC (164,165) and cell mediated lympholysis assays (CML) (166). CD4 + T cells responding to Class II MHC are assayed in GVH (65,167) and MLC (168). ...
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The quest to understand how allogeneic transplanted tissue is not rejected and how tolerance is induced led to fundamental concepts in immunology. First, we review the research that led to the Clonal Deletion theory in the late 1950s that has since dominated the field of immunology and transplantation. At that time many basic mechanisms of immune response were unknown, including the role of lymphocytes and T cells in rejection. These original observations are reassessed by considering T regulatory cells that are produced by thymus of neonates to prevent autoimmunity. Second, we review “operational tolerance” induced in adult rodents and larger animals such as pigs. This can occur spontaneously especially with liver allografts, but also can develop after short courses of a variety of rejection inhibiting therapies. Over time these animals develop alloantigen specific tolerance to the graft but retain the capacity to reject third-party grafts. These animals have a “split tolerance” as peripheral lymphocytes from these animals respond to donor alloantigen in graft versus host assays and in mixed lymphocyte cultures, indicating there is no clonal deletion. Investigation of this phenomenon excludes many mechanisms, including anti-donor antibody blocking rejection as well as anti-idiotypic responses mediated by antibody or T cells. This split tolerance is transferred to a second immune-depleted host by T cells that retain the capacity to effect rejection of third-party grafts by the same host. Third, we review research on alloantigen specific inhibitory T cells that led to the first identification of the CD4+CD25+T regulatory cell. The key role of T cell derived cytokines, other than IL-2, in promoting survival and expansion of antigen specific T regulatory cells that mediate transplant tolerance is reviewed. The precise methods for inducing and diagnosing operational tolerance remain to be defined, but antigen specific T regulatory cells are key mediators.
... There have been repeated observations that long term memory of prior exposure to antigens of the MHC, in contrast to that for minor antigens, is not associated with a demonstrable increase in proliferative responses (16)(17)(18)(19). It is however associated with a change in the effector arm of the allograft response which can be demonstrated in vitro in CML assays (20)(21)(22). ...
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An adoptive transfer system was used to study the cellular basis of memory in animals immunized by grafting with major histocompatibility complex incompatible tissue. Memory was characterised by a large (greater than 100 fold) increase in the potency of lymphocytes to precure graft rejection. This increase in potency endured for at least 1 yr after sensitization. The memory cells were shown to be Ig-- small lymphocytes which were long lived and which did not recirculate from blood to lymph in normal recipients although they did home to lymphoid tissue from which they could be recovered several months later. The thymus was not required either for the generation of memory cells or their maintenance. Cells carrying memory for alloantibody synthesis did recirculate normally but alloantibody synthesis was shown not to be required for rejection.
... Generation of CTL (a) AGAINST MINOR HISTOCOMPATIBILITY ANTIGENS. Standard immunization procedure involved two sets of bilateral skin grafts spaced at least 3 wk apart followed by injection of 50-100 × 10 e allogeneic spleen and LNC approximately 1 mo later (15). One-half the immunization dose was injected i.p. and the rest was distributed among five subcutaneous sites. ...
Article
The regulatory influence of the rat major histocompatibility complex (MHC) (Ag-B complex) on the specificity of cytotoxic T lymphocytes was investigated. It was shown that the effector cells were specific for the original Ag-B phenotype in rat systems in which the responder and stimulator cell populations were unquestionably MHC identical but expressed different minor alloantigens of viral antigens. However, combined in vivo immunization and restimulation in culture of lymphocytes from rat strains previously thought to be MHC compatible resulted in the generation of cytotoxic T lymphocytes which effectively lyse not only target cells from the specific stimulating strains but also, to varying degrees, target cells from third party strains regardless of their Ag-B haplotypes. Genetic analysis indicates that expression of these cytotoxic T-cell-defined ("CT") antigens, found on both T and B lymphocytes, detectable thus far only with cytotoxic lymphocytes, is controlled by a single locus which segregates in backcross populations with the rat MHC. Discrepancies between the nature of CT antigens of the rat Ag-B and I-region specificities of the mouse H-2 are discussed.
... Although immunization across weak histocompatibility differences does lead to augmented GVH responsiveness, there is abundant evidence that it does not regularly do so across strong differences. A negative factor of immunization (Simonsen, 1970;Ford and Simonsen, 1971) or even complete loss of GVH activity (Simonsen, I960;Sprent and Miller, 1971) may sometimes result from prior contact \vith stiong allo-antigens. In contrast to the variability of their behaviour in GVH assays, the capacity of lymphocyte populations to mediate allograft responses in vitro and in vivo is in\'ariably increased by prior immunization with the appropriate strong allo-antigens. ...
Small lymphocytes of the recirculating pool are competent to mediate the rejection of first-set allografts. As a population, these cells are multicompetent, i.e. they can procure the rejection of a variety of antigenically distinct grafts. The basis of this multicompetence was shown to be the admixture, within the population, of separate, clones of allo-antigen-sensitive cells, each competent against one set of allo-antigens. This was done using supralethally irradiated rats as biological filters for large inocula of thoracic duct lymphocytes, and assaying the competence of those, cells which successfully recirculated from the blood to the lymph. When recovered from the lymph of allogeneic “passage” rats, these cells were specifically depleted of the clone of cells competent against “passage” allo-antigens. This clone of cells was recovered from the lymphoid tissues of the passage animal at the time when it was missing from the lymph. It was possible to exclude antibody mediated enhancement or the induction of conventional transplantation tolerance as explanations for the specific defect in reactivity suffered by passaged thoracic duct lymphocytes. The reproducible sequestration of specific allo-antigen-sensitive cells which occurred in supralethally irradiated passage rats with a variety of donor-host combinations is interpreted as confirming the clonal nature of the cells sensitive to Ag-B allo-antigenic specificities among the small lymphocytes in the recirculating pool of normal rats.
... generation of new responding units similar in quality to the primary responding units, but more abundant resembles the "quantal" immunological memory for strong transplantation antigens postulated by Brent and Medawar (17); it is likely that this accounts for the effects of in vivo priming on the normal lymphocyte transfer reaction in guinea pigs. Nevertheless, this phenomenon has proved impossible to demonstrate in the M L I or GVH reactions in mice or rats after in vivo immunization (4)(5)(6). The GVH studies reported here show T-cell populations with potencies, relative to normal, of from 5 to 20 with respect to a strong alloantigen system; these figures compare with the mean factor of immunization of 1.4 which was obtained in a variety of strain combinations with various in vivo immunization schedules done by Ford and Simonsen (5). ...
Article
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Selected populations of thymus-derived (T) rat lymphocytes having specific immunological reactivity to chosen histocompatibility (H) alloantigens are found among the cellular products of the mixed lymphocyte interaction (MLI). Such specific selection seems to depend on (a) the antigen-induced proliferation of specific H antigen reactive cells (HARC), and (b) the disappearance of nonreactive cells from the cultures. When the surviving cells from this lymphocyte-antigen interaction are transferred into thymectomized, X-irradiated, marrow-reconstituted syngeneic recipients (B rats) which lack detectable T-lymphocyte functions, the lymphocyte populations subsequently recovered from the hosts possess the capacity to react in the MLI and in the graft-vs.-host (GVH) reaction, and the reactions have specificity for the original priming alloantigens. In addition, these findings identify the cell that reacts in the MLI with the GVH reactive cell.
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Graft-vs.-host (GVH)-induced lymphadenopathy of the popliteal lymph node has been produced in C57BL/6 x A/J F1 (BAF1) mice by injecting A/J spleen cells into the rear footpads. By giving 51Cr-labeled BAF1 lymphoid cells intravenously to the hosts, 24 h before sacrifice, we have demonstrated that a large portion of the GVH-induced lymphadenopathy is due to the trapping of circuating lymphocytes in the challenged lymph nodes. Most of the remaining enlargement can be attributed to proliferation of host cells within the reacting lymph nodes. Conditions have been defined under which the weights and [14C]thymidine incorporation of the popliteal nodes can be plotted against the dose of injected A/J spleen cells on a double-log scale to give a linear dose-response. The popliteal lymph node GVH assay is a simple and effective means of quantitating immune reactivity to histocompatibility antigens in mice.
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Lethally irradiated (Lewis X Brown Norway)F1 hybrid (LBN) rats were grafted with either marrow or marrow and spleen cells from normal Lewis (L) donors. All recipients of inocula containing both marrow and spleen cells died of graft-versus-host (GVH) disease. In other irradiated and grafted rats the severity of the GVH disease was reduced by challenge of the donor with LBN spleen cells before or after treatment with cyclophosphamide (CY). The magnitude of this suppressed immunity depended on the interval between the administration of CY and the allogeneic spleen cells. Maximum suppression was obtained when donors received CY on day 0 and LBN spleen cells on day +1. In all experiments, the treated donors were killed on day +7. The observed immunosuppression was shown to result from the synergistic action of CY and cells bearing the alloantigons of the recipient. Furthermore, the phenomenon was demon-strated to be antigen specific.
Article
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The graft-versus-host reactivity of Lewis rat spleen cells was measured using a popliteal lymph node assay in Lewis-Brown Norway (LBN) recipient rats. Specific sensitization of donors with a single iv injection of Lewis-Brown Norway spleen cells 7–14 days prior to assay resulted in a significant decrease in graft-versus-host reactivity. Third party sensitization of donors with Lewis-Buffalo spleen cells did not alter graft-versus-host reactivity. Lewis spleen cells suspended in Lewis anti-LBN serum were equally as reactive in the popliteal lymph node assay as Lewis spleen cells suspended in normal Lewis serum.An association was observed between the degree of immunosuppression as a function of time after sensitization and the titer of cytotoxic antibody in the donor serum. This association was not shown in a study of graft-versus-host reactivity and serum cytotoxic antibody titer as a function of the number of cells used.Velocity sedimentation was used to separate spleen cells obtained from specifically sensitized Lewis rats into fractions either enriched or depleted of antibody-forming cells. The addition of 105 or 106 sensitized cells from the antibody-forming cells enriched fraction to 107 normal spleen cells resulted in a significant decrease in graft-versus-host reactivity. The addition of 106 sensitized cells depleted in antibody-forming cells did not significantly change the graft-versus-host reactivity of 107 normal Lewis spleen cells. From these data we conclude that the suppression in graft-versus-host reactivity produced by donor sensitization can be accounted for by immunoblocking antibodies secreted by cells transferred with the graft.
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In this study T cells isolated from isopregnant and virgin CBA/J mice were examined for reactivity to self antigen(s) in vitro and in vivo. The autoproliferative capacity of maternal versus virgin T cells was tested in vitro using autologous mixed lymphocyte reactions (AMLR). The popliteal lymph node (PLN) assay was used to compare the ability of maternal versus virgin T lymphocytes to mediate syngeneic graft-versus-host (SGvH) reactions in vivo. Splenic T cells obtained from pregnant animals near term were found to be approximately 10-fold more reactive towards syngeneic virgin non-T stimulator cells in AMLR than splenic T lymphocytes from age-matched virgin animals. In addition, T cells isolated from the spleens of gravid CBA/J mice displayed a significantly enhanced capacity to mediate SGvH reactions in virgin CBA/J females as measured by regional lymph node enlargement. These findings indicate that syngeneic murine pregnancy is accompanied by an increase in autoreactive T cell activity.
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By exploiting congenic rat strains (HO.B2 and PVG/c) cell-mediated immune responses against Ag-B antigens alone were measured and compared with responses against (1) non-Ag-B antigens and (2) Ag-B and non-Ag-B antigens in combination. It was confirmed that multiple non-Ag-B antigens provoke prompt first-set skin graft rejections, but are much weaker than Ag-B antigens in stimulating both graft-versus-host (GVH) and cytotoxic activity. No evidence of synergistic interaction was found between anti-Ag-B and anti-non-Ag-B responses either by GVH assay or in the generation of cytotoxic cells. Specific partitioning of cytotoxic cells on antigenic monolayers suggested that cytotoxic cells on antigenic monolayers suggested that cytotoxicity is predominantly directed against Ag-B antigens. The measurements of GVH activity consolidate previous work, which suggested that 4.5 to 12% of nonimmune T cells can respond to each Ag-B determined antigenic complex and eliminate the possibility that most of these cells were responding to non-Ag-B antigens. Two principles for measuring GVH activity were compared: (1) 3H-thymidine incorporation into donor lymphocytes at 24 hr after transfer to irradiated F1 hybrid recipients and (2) the popliteal lymph node assay, which depends on a secondary phase of host cell proliferation.
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The experimental use of the laboratory rat, Rattus norvegicus (Robinson 1965), resembles in general that of the laboratory mouse, Mus musculus. Both species belong to different genera in the family of Muridae.
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The in vitro generation of memory cells reactive to transplantation antigens is described. Blast cells, obtained from rat lymphocytes sensitized on xenogeneic or allogeneic fibroblast monolayers, reverted to secondary small lymphocytes after transfer from the foreign sensitizing to syngeneic monolayers. These secondary small lymphocytes had a limited in vitro life span of 4–6 wk. They manifested properties of memory cells: upon re-exposure to fibroblasts of the sensitizing phenotype, the secondary lymphocytes adhered to the fibroblast monolayer and transformed into blast cells with cytotoxic activity. The response of secondary lymphocytes was rapid, compared to that of normal lymphocytes, and directed specifically against the primary sensitizing antigens.
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Current concepts of the inflammatory process include the formation of oxidation products which have the ability to damage tissue. In a few selected inflammation models which included irritant and immunologic stimulus, some representative compounds with antioxidant activity were studied. A diverse set of responses were obtained, but consistently DPPD and ethoxyquin exhibited antiinflammatory activity. Although it cannot be categorically stated that antioxidant capability leads to antiinflammatory activity, it appears that these data provide a theoretical basis for modulating the inflammatory process.
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DNA synthesis and cell proliferation were measured, using [125I]iododeoxyuridine (125IUDR) incorporation and lymph node weight, respectively, in the popliteal nodes of rats undergoing local graft-versus-host (GVH) and host-versus-graft (HVG) reactions in response to a wide range of cell doses. The relationship between 125IUDR incorporation and lymph node weight was investigated at various times during the course of these reactions. Good correlation was demonstrated on linear-linear, log-log, and log-linear plots at early, peak, and late response times in both reactions. These results confirm the usefulness of 125IUDR incorporation as a measure of the local GVH reaction at peak response and show that its use extends to the local HVG reaction and to the measurement of both these reactions at early and late response times. There were no statistically significant quantitative differences in the correlations obtained with linear-linear, log-log, and log-linear plots at any time in either reaction, but comparison of the distributions of the residuals about the regression lines indicated that at peak and late response times in the GVH reaction the log-linear plot gave a qualitatively better fit of the results to the regression line. This could imply that at peak and late response times in the local GVH reaction more DNA synthesis is occurring than is required for cell proliferation alone. The theoretical implications of these findings are discussed.
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Lymph gland cells from sensitized and normal CBA/HT6T6 mice were injected into F1(CBA/HT6T6×C57BL/6) mice subjected to bilateral adrenalectomy 7 days before injection of the donor cells. In one of the experiments both adrenals were removed from the donors four weeks before the lymph gland cells were taken. Adrenalectomy on the recipients led to an increase in the percentage of donor mitoses in the spleen at all times of transfer, provided that the lymphocytes were taken from sensitized donors, whereas the number of donor mitoses in the lymph glands was indistinguishable from that in the control groups. An increase in the percentage of donor mitoses in the spleen of the adrenalectomized mice was observed only on the 5th day after transfer if lymphocytes from unsensitized donors were used; on the first day after injection of the cells an increase in mitotic activity of the donor cells in the lymph glands was observed. If the lymphocytes were taken from adrenalectomized and sensitized donors, the injected cells retained their power of increased mitotic activity in the spleen of the irradiated recipient on the first day after injection.
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Cell proliferation was investigated during local host-versus-graft (HVG) and graft-versus-host (GVH) reactions in rats by means of the popliteal lymph node weight assay. Dose-response regression lines were obtained at early, peak, and late response times for both reactions. Changes in the rate of cell proliferation were demonstrated by changes in the slopes of the dose-response regression lines and were confirmed statistically. In the local HVG reaction the rate of cell proliferation was constant from early (Day +2) to peak (Day +4) response times but possibly was reduced later (Day +8), when the response was dose dependent only at low doses. In the local GVH reaction the rate of cell proliferation increased markedly between early (Day +4) and peak (Day +8) response times and then remained constant. Examination of time-response curves following the injection of a fixed cell dose confirmed these findings. In the HVG reaction lymph node weight increased linearly with respect to time whereas in the GVH reaction the increase in lymph node weight was linear initially but became exponential before peak response. These results are consistent with the concept that, whereas HVG reactions involve proliferation of one principal population of cells (host cells), GVH reactions involve proliferation of two principal populations of cells (host and donor cells). The change in the rate of proliferation occurring between early and peak response in the GVH reaction probably reflects an initial proliferation of donor cells being joined by proliferation of host cells.
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Graft versus host reaction (GvHR) induced in 10-day-old F1 mice by in vitro allogeneically or mitogenically stimulated spleen cells is lower than that induced by unstimulated fresh spleen cells. In vitro stimulated lymphoblasts are unable or only slightly able to induce a GvHR. An active suppression by the blasts is not involved. Since lymphoblasts after in vivo stimulation show an increased ability to elicit a GvHR it is concluded that in vitro and in vivo stimulated lymphoblasts have different properties. A different homing cannot be excluded after transfer to the mouse.
Article
The graft-vs-host (G-v-H) reactivity of sensitized or nonsensitized mouse lymphoid cell populations was measured using a splenomegaly assay. Sensitized populations were obtained either from the local lymph nodes of alloimmunized animals or from the spleens of heavily irradiated mice previously infused iv with allogeneic lymphocytes (educated cells). Immunization of animals resulted in increased G-v-H responses of the cells in their local lymph nodes. This effect was more pronounced when the immunizing cells differed only at non-H-2 transplantation antigens than when H-2-disparate strain combinations were tested. There was no evidence of a changed doseresponse profile of lymphocytes obtained from immunized mice. The G-v-H reactivity of educated cell populations was complex. The slopes of the dose-response lines obtained for lymph node cells or thymic cells educated in an H-2-disparate strain were generally lower than those obtained for nonsensitized cells. This difference was particularly evident when testing educated thymocytes. By studying the G-v-H indices obtained in A/Sn × C57B1 hybrids after inoculation of nonsensitized C57B1 lymph node cells or specifically educated C57BL lymph node cells, it was observed that the latter cells were approximately 30 times more reactive when small cell inocula were compared. On the contrary, education of lymphocytes in H-2-compatible allogeneic hosts did not result in any increment of their G-v-H reactivity. The results indicate that different methods of sensitizing lymphocyte populations against alloantigens may lead to activation of different subclasses of T-cells which differ in their mode of antigen reactivity.
Article
The aim of the studies presented here has been to characterize the cells and elucidate the signals which comprise the flow of immune information leading to the generation of T cell mediated immunity. Rat or mouse lymphocytes were sensitized against fibroblasts in vitro, and were injected into syngeneic animals. The central phase of the reaction, the accumulation of lymphocytes in the draining popliteal lymph node (PLN), was measured as a PLN index. The development of specific effector lymphocytes was detected in two ways. Suspensions of lymphocytes from rat PLNs were tested for immunospecific cytotoxicity against target cells in vitro. In mice the effector phase was assayed by injecting allogeneic tumor cells to recipient animals which had been injected previously with lymphocytes sensitized against allogeneic fibroblasts. The results support the conclusion that a T cell mediated response requires the sequential interaction of at least two types of T lymphocytes. Initiator T lymphocytes (ITL), the agents of the afferent sensitization phase, migrate from the tissue spaces to the draining lymph node after their sensitization. The development of effector and memory lymphocytes requires the recruitment of the ITL of T lymphocytes of another type in the central lymph node phase of the response. The recruited T lymphocytes (RTL) appear to be immunospecific and are part of a circulating pool of lymphocytes trapped in the lymph node and activated by the double signal of syngeneic ITL plus immunogen. The RTL and the ITL are prevalent in different lymphatic organs, have distinct migration patterns, and can be physically separated according to their surface properties. This model system provides a conceptual framework for integrating a number of apparently diverse aspects of cell mediated immunity. (Tonder - Bergen)
Article
ALTHOUGH the phenomenon of memory is central to the adoptive immune response to antigenic stimulation, the magnitude of the memory effect in graft rejection has never been quantified. We have directly compared the potency of naive and memory cells in two quantitative adoptive transfer assays, one measuring graft rejection and the other graft-versus-host activity. While the well known lack of a significant factor of immunisation in the latter was confirmed, memory cells showed a dramatic, long term increase in their potency to reject heart grafts.
Chapter
Some types of graft, e.g. allogeneic bone marrow cells, are vulnerable to an antibody response by the host but this chapter is concerned with those allografts which are rejected by a cell-mediated response. Most attention is directed towards the rejection of skin grafts exchanged between inbred strains of mice or rats because much of the experimentation on the lymphocyte response to allografts has used these species. Small rodents have also been most favored for graft-versus-host studies. Several aspects of the lymphocytic response to allografts in regional lymph-nodes and in the spleen were investigated many years ago and these have been lucidly reviewed by Gowans and McGregor (1965) and by Wilson and Billingham (1967). In the past few years important progress has been made in consolidating areas which have been doubtful or speculative. This chapter is intended to set some of these advances within a matrix of long-established information.
Article
The alloantigen reactivity of spleen and lymph node cells from specifically sensitized (MHA anti-CB) hansters is analyzed in graft-versus-host reactions (GVHR) and mixed lymphocyte reactions (MLR). Analysis of GVHR in appropriate F1 hybrid animals revealed that parental MHA anti-CB hamster lymphoid cells elicit a significantly greater local GVHR than normal unsensitized cells. This pattern of response was observed for hyperimmune lymph node cells (LNC) and for spleen and LNC various times after a single s.c. inoculation of allogeneic (CB) lymphoid cells. This unrestricted clonal expansion was only partially reflected in the MLR. Seven days after a second inoculation of CB lymphoid cells, MHA anti-CB LNC demonstrate a dramatic increase in proliferative capacity over that evoked by unsensitized cells. This was also true for MHA anti-CB spleen and LNC 7 days after a single immunization regime. However, both spleen and LNC demonstrate a delayed onset of clonal proliferation. The immune cell response for both populations is less than that observed for unsensitized cells between 3 and 5 days postimmunization. The unrestricted clonal expansion of hamster lymphoid cells in GVHR and MLR to allogeneic antigens is discussed in terms of the current understanding of the immuno-genetics of the hamster major histocompatibility complex, and the capacity of this species to regulate conventional immune responses.
Article
The Field effect is the decreased biological response to a second challenge in the graft versus host reaction (GvHR). Treatment of recipeint rats or mice during the initial GvHR by methotrexate or cyclophosphamide effected the secondary unresponsivienss. Methotrexate and cyclophosphamide demonstrated different temporal responses indicating different mechanisms of their immunosuppresdive activities.
Article
A single intraperitoneal injection of 0.5 mg methotrexate (MTX) has been found to increase the immune reactivity of spleen cells from (C57B1/6 X DBA/2)F1 mice. Five days after injection, spleen cells from MTX-treated mice exhibited greater PHA responsiveness and GvH reactivity, and mice given SRBC at this time developed greater than normal direct PFC responses. This pattern of effects of MTX was particularly evident in mice that had been given high doses of BCG intravenously 14 days before testing, a treatment that generally depressed the measured activities. MTX enhancement of GvH was also observed in mice that had been depleted of short-lived T lymphocytes by adult thymectomy. We suggest that MTX-sensitive cells possible exert, particularly in BCG-treated mice, a suppressive action on the responding cells.
Article
The graft-versus-host (GVH) response of spleen cells from rats bearing either orthotopic skin grafts or allogeneic implants in the anterior chamber of the eye was evaluated using popliteal lymph node (PLN) assay. When a viable implant remained in the anterior chamber, the spleen cells of these rats produced a popliteal lymph node enlargement in F1 hybrids which was approximately 50% of that produced by a similar number of cells from a normal animal. Conversely, the GVH response of spleen cells from orthotopically skin-grafted rats was noted to be significantly increased over the response of spleen cells from normal animals. The decrease in the GVH response of implanted rat spleen cells was a specific reaction and not because of trauma or implantation, since spleen cells from rats bearing syngeneic implants had shown no reduction in their GVH-inducing ability. The PLN weights of rats receiving mixed population of normal and implanted rat spleen cells were always less than the weights observed with an equal number of normal spleen cells. These findings permit the assumption that implant-bearing rats may be lacking or low in cells that induce GVH reactions or that there is a delayed conversion of effector cells after early immune recognition.
Article
Graft-versus-host reactions (GVHRs) are one of the most intriguing facets of transplantation immunology. This chapter describes the GVHRs, and mechanisms of GVHRs. All GVHRs, both in vivo and in vitro, involve the interaction of lymphocytes as the pivotal event. The chapter discusses the role of donor lymphocytes in the induction and prosecution of GVHRs, the dual roles of host lymphoid cells that are both the primary source of antigenic stimulus and the primary target of donor antihost effector cells, and the probable consequences of donor-host interactions. The host participation in GVHRs involves immunogenic stimulus, development of lesions, and alteration of host immunologic capability. The implication to be drawn from a consideration of the cellular basis of GVHRs is that donor and host both bring to the response unique contributions. The chapter discusses the modification of GVHRs based on altered donors or donor cells and altered hosts after initiation of the GVHR. The chapter also discusses the cellular and molecular aspects of immunoregulation in GVHRs and highlights the imniunoregulatory role of the spleen.
Article
The regulatory influence of the rat major histocompatibility complex (MHC) (Ag-B complex) on the specificity of cytotoxic T lymphocytes was investigated. It was shown that the effector cells were specific for the original Ag-B phenotype in rat systems in which the responder and stimulator cell populations were unquestionably MHC identical but expressed different minor alloantigens of viral antigens. However, combined in vivo immunization and restimulation in culture of lymphocytes from rat strains previously thought to be MHC compatible resulted in the generation of cytotoxic T lymphocytes which effectively lyse not only target cells from the specific stimulating strains but also, to varying degrees, target cells from third party strains regardless of their Ag-B haplotypes. Genetic analysis indicates that expression of these cytotoxic T-cell-defined ("CT") antigens, found on both T and B lymphocytes, detectable thus far only with cytotoxic lymphocytes, is controlled by a single locus which segregates in backcross populations with the rat MHC. Discrepancies between the nature of CT antigens of the rat Ag-B and I-region specificities of the mouse H-2 are discussed.
Article
Rat lymph node cells taken at the peak of cytolytic activity following a skin allograft were separated into adherent and nonadherent fractions by incubation on monolayers of thoracic duct lymphocytes either of the same strain as the graft donor or of an Ag-B different strain. In the face of a 3-fold enrichment of cytolytic activity in the adherent cells and a 3-fold depletion in the nonadherent cells there was no detectable partition of graft-vs-host (GVH) activity. Supplementary experiments supported the simplest interpretation of this finding, namely that the antigen receptors on GVH-reactive cells did not influence their adherence in this system. Similarly, there was no partition of the GVH activity of nonimmune lymph node cells by adherence. Labeling lymph node cells with either radioactive uridine or thymidine in vitro, suggested that about 20% of DNA-synthesizing cells in the immune population adhered because of antigen recognition.
Article
The alloantigenic reactivity of hamster spleen cells from unsensitized animals was analyzed in vitro. When compared with normal MHA cells, MHA anti-CB spleen cells responded better or to the same degree in mixed lymphocyte reaction (MLR) during the first 3 days after alloantigenic exposure. However, by day 5 postsensitization there was a signigicant reduction in spleen cell MLR activity, which returned to normal or even an exaggerated response 7 days following immunization. We examined the possibility that the reduction in MLR responsiveness 5 days following immunization might be attributable to the generation splenic suppressor cells. Irradiated spleen cells from unsensitized and from MHA anti-CB donors were cocultured with normal responder MHA lymph node cells stimulated by (MHA x CB)F1 hybrid lymph node cells in MLR. No regulatory effect of either cell population was seen, indicating that even hamster spleen cells from alloimmune donors fail to suppress the mitotic response of normal lymph node cells to allogeneic targets.
Article
This paper describes a method for quantitating relative potencies of mixed lymphocyte interactions (MLI) in the rat by parallel line analysis of responder cell titrations. The MLI titration assay is based upon, and has certain advantages over, the popliteal lymph node graft-versus-host assay for investigations of allogeneic interactions determined by the major histocompatibility complex. Log-log plots of dose of responder cells versus cpm of [3H]thymidine incorporated were linear in the range 1 × 104 to 2 × 105 responder cells per microtiter well. Dilution experiments indicated that the overriding influence on the potency of any allogeneic interaction was the initial frequency of reactive cells. We have defined the limits within which the MLI titration assay behaved ideally and suggested explanations for the nonideal behavior seen when reactions of widely differing potency were compared.
Article
Lethally irradiated mice reconstituted with syngeneic bone marrow cells were grafted with allogeneic skin grafts 6-7 weeks after irradiation and reconstitution. Mice with intact thymuses rejected the grafts whereas the mice thymectomized before irradiation and reconstitution did not. Thymectomized irradiated mice (TIR mice) reconstituted with bone marrow cells from donors immune to the allografts rejected the grafts. Bone marrow cells from immunized donors, pretreated with Thy 1.2 antibody and C', did not confer immunity to TIR recipients. To determine the number of T lymphocytes necessary for the transfer of immunity by bone marrow cells from immunized donors, thymectomized irradiated mice were reconstituted with nonimmune bone marrow cells treated with Thy 1.2 antibody and C' and with various numbers of splenic T lymphocytes from nonimmune and immune donors. Allogeneic skin graft rejection was obtained with 10(6) nonimmune or 10(4) immune T cells. The effect of immune T cells was specific: i.e., immune T cells accelerated only rejection of the relevant skin grafts whereas against a third-party skin grafts acted as normal T lymphocytes.
Article
The representation of mouse alloantigens belonging to three systems, H-2, θ and TL, on the surface of cells from thymus, spleen, lymph nodes, and peritoneal cavity, was studied by electron microscopy with ferritin-labeled antibody. As expected from earlier serological data, TL was confined to thymocytes, θ was found on thymocytes and lymphocytes, and H-2 occurred to some extent on all cell types observed. On reticular cells, lymphocytes, plasma cells, and eosinophils, the majority of the cell surface was occupied by H-2; thymocytes had considerably less H-2, and erythrocytes and peritoneal macrophages least of all. In every instance the representation of antigen was discontinuous, the fraction of the cell surface covered being characteristic both of the antigen and of the type of cell. H-2 and θ provide a striking example of this; H-2 is present in far higher amounts on lymphocytes than on thymocytes, whereas the converse is true of θ. Within areas positive for H-2 or θ, protuberances of the surface membrane were often antigen-negative. A better definition of cell surface structure, gained from studies such as this, is necessary for further inquiry into how the cell surface is assembled, and into selective gene action in relation to cellular differentiation.
Article
Both primary and secondary responses to sheep erythrocytes and to Brucella abortus antigen have been obtained in cultures of dispersed rabbit spleen cells. Removal of adherent cells by repeated incubation of spleen cells on absorbent cotton diminished the ability of the spleen cell suspensions to give secondary as well as primary responses in vitro. When comparing cultures made in dishes and in tubes, the loss of responsiveness after incubation on cotton was much more evident in the dish cultures. It was concluded that the cell-to-cell interaction needed for immune responses to particulate antigens in vitro was more readily interfered with when the cells were spread over a larger surface area. The proliferative response to antigen, as measured by uptake of 3H-thymidine in tube cultures of the sensitive spleen cells, appeared particularly resistant to the depletion effect of adherent cell removal. Dispersed spleen cells from sensitized mice gave a secondary response to sheep erythrocytes. This response was readily abolished by one incubation on absorbent cotton when the cells were cultured in dishes.
The inoculation of blood leucocytes from an adult non-inbred fowl onto the dropped chorioallantoic membrane of non-inbred eggs results in the production of visible lesions after four days’ further incubation. This reaction is a graft versus host reaction of immunologically competent lymphocytes against major histocompatibility antigens present in the host membrane and is termed the Simonsen phenomenon.Although tolerance to this range of histocompatibility antigens can be induced in birds, previous studies had failed to show any increase in the proportion of reactive cells following immunisation of the donor bird with the same range of histocompatibility antigens.Results presented in this paper show that preimmunisation of donor non-inbred fowls with pooled spleen cells from embryos of the AA inbred line produced an increase in the relative number of reactive cells as assessed by the number of foci produced on subsequent inoculation of blood leucocytes onto membranes of AA eggs.It is concluded that this represents immunisation to minor histocompatibility antigens in the fowl, and parallels Simonsen's results in mice.
Article
The migration of lymphocytes from the blood into the splenic pulp and the release of lymphocytes from the spleen into the blood was studied by isolating the rat spleen and perfusing it with 15 ml of recirculating, oxygenated blood. When thoracic duct lymphocytes labelled with tritiated uridine were added to the initial perfusate the concentration of these cells fell exponentially for 2–3 hr and then rose to a flat secondary peak. From this pattern it was inferred that small lymphocytes entered the spleen at a rate proportional to their instantaneous concentration in the perfusate, traversed the splenic pulp and re-entered the perfusate with a minimum transit time of 2–3 hr. The rate of release of small lymphocytes from the spleen was not influenced by the prevailing concentration of small lymphocytes in the perfusate but probably reflected the rate of migration into the spleen over a period earlier than 2 hr before. The rate of exchange of small lymphocytes between the blood and the intact spleen in vivo was estimated to be about 84 × 106 cells/hr. The size of the intrasplenic pool of recirculating small lymphocytes was probably 400–500 × 106 cells. The rate of migration of small lymphocytes into the spleen was not affected by prior irradiation of the spleen donor. When either of two antigenic materials were added to the perfusate no inhibition of lymphocyte migration into the spleen was noted although the release of lymphocytes from the spleen was diminished by the addition of a large dose of sheep erythrocytes.
Chapter
Material and Methods Mouse strainsGeneral Experimental DesignTechnique of the Spleen AssayPreimmunization of DonorsFactor of Immunization in Response to St/A AntigenResultsComparison of F.I. in 4 Strain CombinationsAntigenic Strength in Terms of Runting PowerDiscussionThe Present Experiments Compared with Earlier FindingsThe Present Experiments in Relation to Acquired ToleranceThe Choice Between HypothesesSummaryReferences
Article
Popliteal lymph nodes were obtained from rabbits 4 days to 9 months after a primary injection of diphtheria toxoid or bovine γ-globulin into the footpad. The ability of cells from these nodes to proliferate upon reexposure to antigen in vitro was compared to the height of the secondary response produced by tissue fragments. In addition, a comparison was made between the responsiveness of draining and contralateral lymph nodes. While the secondary antibody response in vitro increased markedly with the time after immunization at which the lymph nodes were taken from the animals, the degree of proliferation induced by antigen was highest with cells from lymph nodes taken early after priming (peak day 7) and was very much lower with lymph node cells taken longer than 3 wk after priming. This striking difference between these two responses has been discussed. Contralateral lymph nodes were much inferior to draining nodes in their ability to give a secondary antibody response in vitro, and never gave a detectable proliferative response. This difference became less marked with time after priming, but could still be demonstrated after 4 months. These results suggest a concentration of primed cells in the lymphoid tissue draining the site of injection, and a slow release of these cells into the circulation, to be distributed to the remaining lymphoid tissue.
The normal lymphocyte transfer reaction (NLT reaction) is a cutaneous inflammatory episode of delayed onset that is aroused when living lymphocytes from one guinea-pig are injected into the skin of another, and it occurs only in those genetic situations that show it to be an immunological response of the transferred lymphocytes to antigens of the animals into which they are injected (section 4). When the recipient guinea-pig is exposed to 600 r whole-body irradiation before transfer, to delay the onset of an immunological counter-attack on the transferred cells, the NLT reaction evolves in three phases spread over about 6 days (section 3). These are: (a) the first inflammatory episode, of moderate intensity, which reaches its peak at 24 h and remains stationary for 2 further days; (b) the flare-up, starting between the third and fourth days and rising to a peak of violent intensity at about the sixth day; and (c) the fade-out, which is mainly due to an immunological recovery of the host. The same three components may be discerned when the transferred lymphocytes have been presensitized against the tissues of their future recipients, but the pitch of the reaction is much higher throughout and presensitized cells perform as strongly at 24 h as normal cells do when they reach the peak of their activity at 5 to 6 days (figure 2). In general, lymphocyte transfer reactions vary in intensity rather than in tempo: the weak reactions that occur where antigenic disparity is slight, or when relatively few cells are injected, differ from strong reactions-even the very strong reactions caused by presensitized cells-in their general pitch of intensity but not in the relative timing of the various episodes of the response. In terms of the power of a given number of cells to excite an NLT reaction, blood lymphocytes were two to five times more active, and thymocytes ten to twenty times less active, than lymph node cells (section 5). Cells from lymph nodes caused to enlarge greatly by stimulation with human gamma globulin emulsified in Freund's complete adjuvant were not more effective than normal cells in exciting an NLT reaction. Lymphoid cells from foetal or newborn mesenteric nodes gave bold and clear NLT reactions rising to a peak not lower than that achieved by the same number of adult lymphoid cells (section 5). The NLT reaction lent itself very well to a study of inhibitors of the immunological response. Immunosuppressive agents (section 6) were applied, as appropriate, to the cell donor before transfer, to the lymphoid cells in transit, or to the recipient before, during or after transfer. Immunosuppressive agents did not in general affect the first inflammatory episode; with varying degrees of effectiveness they did, however, eliminate the flare-up. The two most effective agents in this respect (Methotrexate and cyclophosphamide) did not, however, oppose the immunological performance of presensitized cells at concentrations more than sufficient to eliminate the flare-up. It is therefore reasoned that conventional immuno-suppressive agents do not affect any distinctively immunological activity of lymphoid cells: they merely prevent the multiplication of the cells activated in the first episode of the NLT reaction, and therefore the transformation of a 'normal' into a sensitized population. Reasons are given for thinking that the antigens which excite the NLT reaction belong to the homograft system and that the reaction as a whole can be construed as a homograft reaction in reverse (section 7). The first inflammatory episode is interpreted as the outward design of a 'recognition' event, i.e. of a distinctively immunological process, unaccompanied by cell division and in no way dependent on it, that occurs when a lymphocyte is first engaged by an antigen of the homograft system and is committed to the evolution revealed outwardly by the flare-up. A quantal theory of the reaction is proposed, according to which the violent response given by normal lymphoid cells at the peak of their flare-up, and by presensitized cells from the outset, is simply an arithmetic multiple of the recognition process, i.e. it simply consists in more cells doing what a relatively small number does in the first inflammatory episode. Immunosuppressive agents act by preventing this multiplication. This interpretation implies that any 'true' immunosuppressive agent which weakens or abolishes the first inflammatory episode must weaken the performance of a presensitized population to an exactly similar degree.
Article
When parental strain spleen cells or thoracic duct lymphocytes were injected into the feet of F1 hybrid recipients, the draining popliteal lymph nodes reached a weight which was up to 30 times greater than that of control lymph nodes. Seven days after injection, the mean lymph node weight was linearly related to the dose of cells injected on a double log scale. As a graft-versus-host assay, this method was found to be more sensitive as well as more convenient than other methods which have been used in the rat. The degree of lymph node enlargement produced by a given dose of cells was influenced by the volume in which they were injected, the precise site of injection, and the age of the recipient. On the other hand, the sex of the recipient and addition of F1 hybrid cells to the inoculum did not affect the lymph node weight. Graft-versus-host activity can be assayed in both Ag-B-disparate and Ag-B-identical strain combinations although, in the latter case, about 100 times as many cells are required to produce the same amount of lymph node enlargement. (C) Williams & Wilkins 1970. All Rights Reserved.
Article
The ferritin-labelled antibody technique has been applied to the study of the distribution of the H-2 antigen. Using a specific antiserum for the isoantigens determined by the H-2d allele (H-2b anti H-2d), the H-2 isoantigens were shown to be evenly distributed on the cell surface of lymphocytes, polymorphonuclear leukocytes, eosinophils, platelets, and erythrocytes. Lymphocytes were shown to possess the largest concentration of antigen at the cell surface. In preliminary experiments, nonspecific attachment of the ferritin-antibody marker to the intracellular components of dead or formalin-treated cells prevented the demonstration of an intracellular distribution of the H-2 antigen. The potential application of the ferritin-labelled antibody technique to the study of single H-2 isoantigenic specificities is discussed.
Article
The effect of pretreatment of C57BL mice with lyophilized tissues of RIII or Swiss mice on the capacity of their spleen cells to induce runt disease in newborn recipients of these strains was studied. Pretreatment with a transplantable RIII tumor (MMCIA) did not affect the induction of runt disease in RIII mice by large doses of C57BL spleen cells, but increased the incidence of the disease induced by small doses of cells. The mean survival time of mice injected with small doses of cells from either normal or pretreated C57BL donors was longer than that of mice receiving large doses of cells. Spleen cells from pretreated donors had a lower capacity than cells from normal donors to induce runt disease in 2- to 3-day-old RIII recipients. Pretreatment of C57BL donors with Swiss mouse tissues specifically decreased their capacity to induce runt disease in newborn Swiss mice. Surviving Swiss mice did not tolerate C57BL skin grafts and no C57BL antigens could be demonstrated in their spleens.
Article
The representation of mouse alloantigens belonging to three systems, H-2, theta and TL, on the surface of cells from thymus, spleen, lymph nodes, and peritoneal cavity, was studied by electron microscopy with ferritin-labeled antibody. As expected from earlier serological data, TL was confined to thymocytes, theta was found on thymocytes and lymphocytes, and H-2 occurred to some extent on all cell types observed. On reticular cells, lymphocytes, plasma cells, and eosinophils, the majority of the cell surface was occupied by H-2; thymocytes had considerably less H-2, and erythrocytes and peritoneal macrophages least of all. In every instance the representation of antigen was discontinuous, the fraction of the cell surface covered being characteristic both of the antigen and of the type of cell. H-2 and theta provide a striking example of this; H-2 is present in far higher amounts on lymphocytes than on thymocytes, whereas the converse is true of theta. Within areas positive for H-2 or theta, protuberances of the surface membrane were often antigen-negative. A better definition of cell surface structure, gained from studies such as this, is necessary for further inquiry into how the cell surface is assembled, and into selective gene action in relation to cellular differentiation.
The capacity of lymph nodes to show secondary reactivity has been investigated in sheep and in mice, using Swine Influenza virus and sheep red blood cells as antigens, respectively. Following a primary challenge to the political node of one leg, the node of the contralateral leg becomes primed and reacts with a secondary response to its first direct encounter of the antigen. This secondary reactivity is acquired in an indirect way and is not due to the transfer of antigen from the injection site in the opposite leg. The secondary response that occurs in “indirectly primed” nodes is not as vigorous or as prompt as mat occurring in “directly primed” nodes. This difference is thought to be due to the presence of both “residential” and “circulating” memory cells, the former class of cells being generated and remaining in lymph nodes where antigen becomes localized while the latter are free-floating cells of lymph and blood. Circulating memory cells are recruited to a lymph node when an antigen is taken up there, but this recruitment does not appear to be a specific phenomenon.
Article
Hybrid adult mice can be protected against wasting disease induced by spleen cells from parental origin, by utilizing the immunological enhancement-facilitation phenomenon. Passive enhancement-facilitation was applied using a "B anti-A" type of immune serum injected into (B X A)F1 hybrids before the injection of B lymphoid cells. Active enhancement-facilitation was used in similar strain combinations by means of appropriate immunization of the prospective B spleen cell donors; typically the immunizing material consisted of extracts prepared from lyophilized A tissues (3 mg) and immunizing injections were made 5 clays before the donor spleens were transferred. (1) Passive enhancement-facilitation of (B1OD2 X CBA)F1 hybrids receiving B1OD2 spleen cells, by means of an anti-CBA B1OD2 immune serum, increased survival rates from 1% to 13-40%. This effect of the immune serum was removed by specific absorption. (2) Active enhancement-facilitation displayed its effect clearly in the same strain combination and also in the C57BL6 -> (C57BL6 X CBA)F1 combination, where specificity of the phenomenon was controlled in a group of hybrids receiving cells from parental donors immunized with extracts of guinea pig lyophilized tissues. (3) Varying immunization procedures or strain combinations had sometimes an opposite effect, resulting in an aggravation of the disease (sometimes limited to an increased lymphocytopenia). (4) The spontaneous hemagglutination phenomenon in Gorer-Mikulska's medium is described as a sign of antibody fixation at the surface of mouse erythrocytes. In the present experiments, it results from a graft-versus-host reaction.
Article
The patterns of deoxyribonucleic acid synthesis and cell survival were studied in mixed spleen cell cultures from normal and crossimmunized inbred rats. In mixed cultures from nonsensitized animals, the peak of tritiated thymidine (3H-TdR) incorporation took place at the 7th day in culture, and there was a steady decrease of cell survival throughout the culture period. In mixed cultures from rats sensitized 4 days previously with spleen and lymph node cells of the opposite strain, the patterns were markedly different: there was an early high peak of 3H-TdR incorporation followed by a rapid decline to values lower than those observed in nonmixed control cultures. At the time of decline of 3H-TdR incorporation in mixed presensitized cultures, there was an abrupt decline of cell viability. The sera from the sensitized animals was not cytotoxic in the system, and the cultures from the sensitized rats responded to phytohemagglutinin stimulation as well as those from nonsensitized rats. It is proposed that both afferent and efferent arms of the cellular immune response develop in cultures of allogeneic lymphoid cells.
Article
Lymphocytes were obtained from the thoracic duct of rats 1½ to 15 months after primary immunization with a single dose of bacteriophage ϕX 174. An intravenous injection of these lymphocytes conferred on heavily X-irradiated rats the ability to form antibody in a secondary-type manner after a first injection of ϕX. Negligible responses were obtained after cell transfer if the recipients were not challenged with antigen. Thoracic duct cells from some immunized donors were incubated in vitro for 24 hr before transfer in order to destroy selectively the large, dividing lymphocytes. The responsiveness conferred on X-irradiated recipients by such "incubated" inocula was then compared with that given by equal numbers of "fresh" thoracic duct cells. In all such comparisons the recipients of the "incubated" cells gave higher and more rapid antibody responses. It was concluded that the cells in thoracic duct lymph which carried immunological memory were small lymphocytes.
Article
51Cr-labelled (CBA X A)F1 and A strain lymph node cells were injected by either the i.v. or the i.p. route into adult A strain mice, and the distribution of radioactivity in various organs was studied over a period of 48 hours. The extent to which radioactivity in an organ is a measure of the number of living donor cells is considered in the light of control experiments with frozen-thawed and intact heat-killed cells. From these and other experiments it is concluded that the elution of 51Cr and its reattachment to host cells plays little part over this period, and that the differences between the results obtained by i.v. and i.p. inoculation are largely attributable to differences in the distribution of living cells. Both with allogeneic (hybrid) and syngeneic (A strain) lymph node cells, the activity recovered in the organs was many times greater when the cells had been administered by the i.v., as opposed to the i.p., route. This was true for the liver, lungs, spleen, and lymph nodes, but not for the thymus and salivary glands, from which little activity was recovered by either route. Using both routes the greatest part of the recovered radioactivity was found in the liver and spleen, but relative to organ weight uptake was highest in the lymph nodes and spleen. It is suggested that the different distribution of cells, together with the fact that cells injected i.p. are exposed to peritoneal as well as to liver macrophages, accounts for the very different results obtained when attempting to induce sensitization or tolerance by these two routes. Adult A strain mice presensitized with CBA antigens showed what appeared to be immune elimination of hybrid lymph node cells-an observation suggesting that it might be possible to use this approach as a test of homograft sensitivity.
Article
When lymphoid cell suspensions from the spleen, lymph nodes, blood, and thoracic duct of parental strain adult rats were injected beneath the renal capsule of F(1) hybrid hosts, the transferred cells and/or their progeny invaded the underlying renal cortex and destroyed most of the tubules which they surrounded. The immunogenetic conditions under which this reaction was observed defined it as a graft vs. host reaction (GVHR). On the 7th day the GVHRs were histologically similar to primary renal homografts undergoing rejection. Lymphoid cells from donors tolerant to the other parental strain were inactive after transfer to the hybrid, whereas cells from either normal or sensitized donors consistently produced reactions of about equal severity. Lewis lymphoma cells displayed malignant, invasive activity but did not destroy either isologous or homologous tissue, showing that the presence of an infiltrate was not per se sufficient to damage the parenchyma. These observations indicate that the GVHRs were manifestations of the ability of the transferred lymphocytes to enter into a homograft reaction with consequent destruction of renal parenchyma, and support the hypothesis that at least some of the lymphocytes which are seen infiltrating primary homografts are the agents which effect their destruction.
Article
The immune responses of normal and x-irradiated mice were compared by ; studying the survival of skin homografts and by following changes of ; isohemagglutinin titers in host serum. Male CBA mice were presensitized by ; intraperitoneal injection of spleen cells from strain A mice, and 7 days later ; were irradiated with 200, 400, or 750 rads, given at a rate of 73 rads/min. They ; were then grafted with A-strain skin on the 7th, 29th, or 91st day after ; sensitization. Graft rejection and hemagglutinin formation were hoth depressed ; by radiation in proportion to the dose given. The radioresistance of the ; hemagglutinin response was greater in mice sensitized by spleen cells than in ; nonsensitized mice. The 400-rad dose suppressed this response completely in ; nonsensitized mice although it only slightly retarded the rejection of 2 ; successive skin homografts. These results confirm that circulating antibody is ; not essential for the rejection of skin grafts. They also suggest that plasma ; cells are not involved in graft rejection and that these cells are most ; radiosensitive before actively synthesizing antibody, since irradiation one week ; after presensitization has relatively little effect on hemagglutinin production. ; (H.H.D.);
In General Discussion
  • G Msller
MSller, G. 1970. In General Discussion. Transplant. Rev. 3:81.
Modification of runt disease by treatment of the donor with lyophilized tissues 95:323. on March 18, 2013 jem.rupress.org Downloaded from Published April 1 Protection against homologous disease in hybrid mice by passive and active immunological enhancementfacilitation
  • M Schlesinger
  • R Goitein
  • L Ford
Schlesinger, M., and R. Goitein. 1965. Modification of runt disease by treatment of the donor with lyophilized tissues. J. Immunol. 95:323. on March 18, 2013 jem.rupress.org Downloaded from Published April 1, 1971 WILLIAM L. FORD AND MORTEN SIMONSEN 949 12. Voisin, G. A., R. Kinsky, and J. Maillard. 1968. Protection against homologous disease in hybrid mice by passive and active immunological enhancementfacilitation. Transplantation. 6:187.
  • W L Ford
Ford, W. L. 1970. In General Discussion. Transplant. Rev. 3:83.