The role of nonlymphoid accessory cells in the immune response to different antigens.
01 Mar 1970-Journal of Experimental Medicine (The Rockefeller University Press)-Vol. 131, Iss: 3, pp 461-482
TL;DR: The results indicated that phagocytic cells are not required in the initiation of an immune response to POL, and some accessory cell, possibly a phagocytetic macrophage, is required for a response to SRC.
Abstract: Tissue culture techniques were combined with cell separation procedures to investigate the cellular requirements for a response to antigen, leading to the production of antibody-forming cells. Mouse spleen was dissociated, and the cells were separated into various groups on the basis of density, size, and active adherence. The ability of fractions to initiate a response in vivo, on transfer to an irradiated recipient, was compared to the response in vitro; and this ability was correlated with the presence or absence of phagocytic cells. Two different antigens were studied, sheep erythrocytes (SRC) and polymerized bacterial flagellin (POL). Density distribution analysis of spleen showed a wide density range of cells responding to both antigens in vivo. The same fractions responded to POL in vitro as in vivo. By contrast, only the light density regions responded in vitro to SRC. Response occurred in regions of overlap between lymphocytes and phagocytic macrophages. Separation by active adherence on columns of large glass beads gave a preparation containing large, medium, and small lymphocytes but no detectable phagocytic macrophages and very low levels of phagocytic polymorphs. This lymphocyte preparation responded to both antigens in vivo. In vitro it gave a full response to POL, but no response to SRC. Addition of a small quantity of the adherent fraction, enriched for phagocytic cells, restored response to SRC. The use of strain-specific antisera in a mixed culture containing a C57 phagocytic fraction and CBA lymphocytes showed that the lymphocyte fraction contributed the precursors of the final antibody-forming cells. The accessory cells from C57 spleen banded in the light regions of the density gradient where phagocytic macrophages were found. Irradiated spleen cells also activated the lymphocyte preparation, suggesting that the irradiated host provided the accessory cells for the in vivo response to SRC. Small lymphocytes were purified from spleen by the small glass bead size filtration technique. This sample of small lymphocytes responded less well to POL than the total lymphocyte population, but it responded as well in vitro as in vivo. The small lymphocyte preparation responded in vivo to SRC but not in vitro. Addition of a small quantity of the phagocyte-rich fraction from adherence columns restored the in vitro response to SRC. The results indicated that phagocytic cells are not required in the initiation of an immune response to POL. By contrast some accessory cell, possibly a phagocytic macrophage, is required for a response to SRC. The basis for this marked difference is discussed.
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TL;DR: It is concluded that LAF stimulates both central and peripheral T lymphocytes and enhances their responses to other stimulants.
Abstract: Effective supernatants (SUP), which potentiate mouse T-cell responses to phytohemagglutin (PHA), are obtained from cells of several species (human, rabbit, rat, mouse) and indeed from syngeneic spleen, thymus, or bone marrow cells. Unstimulated cells release some SUP activity but more is produced after stimulation. Lipopolysaccharide (LPS) produced very active SUP in all cultures tested. PHA was similarly active on human leukocytes only, whereas concanavalin A (Con A) gave highly efficient SUP only with mouse spleen cells. SUP production is not correlated with a mitotic response of the donor cells and is observed in cultures unable to respond mitotically to the stimulant. Adherent mouse spleen cell populations, consisting largely or entirely of macrophages, produce active SUP, while nonadherent cells do not. Similarly, purification of human peripheral leukocytes on nylon columns, with removal of macrophages and other adherent cells, destroys their ability to produce SUP. The importance of indirect effects in stimulating mitotic responses of T cells is emphasized by the fact that the mitotic response of mouse thymocytes to LPS and its ability to potentiate the response of these cells to PHA disappears with removal of adherent cells from the thymocyte population. Conversely the production of SUP from spleen cells stimulated by Con A requires the presence of T cells.
726 citations
TL;DR: The adoptive secondary response of mice to conjugates of NIP and DNP is used to elucidate the mechanism of cellular cooperation and shows that helper cells serve a role as handlers or concentrators of antigen, thus enabling AFCP which would otherwise be incapable of reacting to initiate antibody synthesis.
Abstract: The adoptive secondary response of mice to conjugates of NIP (4-hydroxy-5-iodo-3-nitro-phenacetyl-) and DNP (2,4-dinitrophenyl-) is here used to elucidate the mechanism of cellular cooperation. The framework into which the experiments fit can be formulated as follows. Priming immunization raises a crop not only of specific antibody-forming-cell-precursors (AFCP) but also of specific helper cells. Upon secondary stimulation the helper cells serve a role as handlers or concentrators of antigen, thus enabling AFCP which would otherwise be incapable of reacting to initiate antibody synthesis. In this act of cooperation both cells recognise antigen; in the system examined here the helpers recognise carrier determinants and the AFCP recognise either the hapten or other carrier determinants.
The first aim of the experiments was to raise populations of helpers and AFCP of distinguishable specificity. Mice were primed with NIP-Ovalbumin (OA) mixed with chicken γ-globulin (CGG) and bovine serum albumin (BSA); in comparison with controls primed with unmixed NIP-OA, their cells after transfer were relatively more sensitive to secondary stimulation with NIP-CGG or NIP-BSA and similar findings were obtained in cross-checks of these carriers. For reasons which are not entirely clear, non-transferred cells did not show the same effect. In further experiments cells primed with one conjugate (e. g. NIP-OA) were mixed with cells primed with another protein (e. g. BSA), transferred and challenged with the hapten conjugated to the second protein (i. e. NIP-BSA). In comparison with controls lacking the protein-primed cells, the mixture regularly showed greater sensitivity to stimulation. NIP and DNP were tested in many of the possible combinations with BSA, OA and CGG with the same result. The mixture system was used in the further analysis.
Tests with allotype-marked protein-primed cells showed that these cells did not participate in the production of the anti-hapten antibody and could therefore properly be regarded as helpers. Tests of specificity showed that physical union of the hapten and carrier were required: cells primed with BSA, for example, would not help NIP-OA-primed cells to make a response to NIP-HSA even when stimulated at the same time with BSA. Transfer of less than one-tenth of the spleen gives a maximum helper effect, whereas AFCP activity continues to rise as larger numbers of cells are transferred. Helper cells are therefore normally present in excess.
Helper activity is more resistant than AFCP activity to irradiation, drugs and semi-allogeneic cell transfer across an H-2 barrier. This suggests that helper cells play a relatively passive role in the immune response.
Several observations indicate that helper cells are thymus-derived mediators of cellular immunity. Passively transferred antibody did not substitute for helper cells. After immunization helper activity developed faster than AFCP activity. Spleen cells obtained from lethally-irradiated, thymocyte-repopulated, immunized donors provided help. Cells from the thymus-derived fraction of thymus/marrow chimeras also appear to provide help.
Thus, the hapten-carrier cooperative response maps onto the well-established synergy of thymus and marrow in the response to foreign erythrocytes.
642 citations
TL;DR: The positive immunogenic role of macrophages is related to their capacity to remove extracellular antigen, which might be capable of interacting with and eliminating isolated T or B lymphocytes and retain antigen in lymphoid tissues and promote its necessary meeting with both T and B cells.
Abstract: Publisher Summary This chapter focuses on the role of the macrophage in removing antigen from extracellular fluids, degrading the larger part of this antigen while presenting a small part of it in persisting immunogenic form. This handling of antigen is done without contributing to the specificity of the immune response, which is determined by the antigenreactive T and B lymphocytes. During the process of uptake of antigen, macrophages appear to retain a few molecules of antigen, undegraded or with few chemical changes. Macrophage-associated antigen becomes an effective immunogenic stimulus mainly in conditions that require two types of lymphocytes to meet with antigen molecules. These lymphocytes are specifically antigen-committed and few in number. The positive immunogenic role of macrophages is related to their capacity to (1) remove extracellular antigen, which might be capable of interacting with and eliminating isolated T or B lymphocytes and (2) retain antigen in lymphoid tissues and promote its necessary meeting with both T and B cells.
519 citations
TL;DR: The developments in macrophage biology over the past few years were not predicted till 1972, and Macrophages are definitely involved in some response to many of the conventional polyclonal stimuli.
Abstract: Publisher Summary Changes in antigen molecules that resulted in enhanced or decreased uptake by the phagocytes resulted in higher or lower immune responses, respectively. After the development of methodologies for obtaining live exudate cells rich in phagocytes and for pulsing these with antigen, the immune response to macrophage-associated antigens was possible to assay, using combinations of in vivo and in vitro methods. The presentation of antigen bound to live macrophages to the lymphocytes was a highly efficient mode of generating an immune response. In the marine system, the requirement for phagocytes or other acc|essory cells was such that as little as 1% contamination with phagocytes still enabled a T-cell proliferative response to develop. Thus, depletion procedures had to be extremely efficient. Macrophages are definitely involved in some response to many of the conventional polyclonal stimuli. Involvement of the macrophage may be by way of secreted active molecule. Fc fragments of Ig act as a polyclonal stimulant only after a processing of the fragment by the macrophages. The developments in macrophage biology over the past few years were not predicted till 1972.
467 citations
TL;DR: This chapter considers the interrelationships between radiation and the various components of the immune response from three perspectives: the effect of irradiation on normal lymphoid tissues and on isolated lymphocytes, the effect on antibody production, transplantation immunity, and other forms of cellular immunity.
Abstract: Publisher Summary This chapter illustrates several ways in which radiation may be employed to dissect several individual cellular components of the immune response. The chapter considers the interrelationships between radiation and the various components of the immune response from three perspectives: (1) the effect of irradiation on normal lymphoid tissues and on isolated lymphocytes, (2) the effect of irradiation on antibody production, transplantation immunity, and other forms of cellular immunity, and (3) the effect of irradiation upon tolerance with specific references to putative autoimmune consequences after radiation-induced alterations in normal immunological homeostasis. A wide variety of approaches are available to characterize and define distinct populations of lymphocytes. These include biophysical and functional methods and characterization of antigenic and cell surface receptor components. The mechanisms of radiation effects in biological systems, particularly in humans, have been derived from experiments utilizing cells exposed in vitro and maintained in tissue culture. Such cells can be examined for: (1) loss of viability, (2) alterations in biophysical structure, (3) loss of functional capabilities, (4) biochemical changes, and (5) evidence of injury to subcellular components.
408 citations
References
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Journal Article•
TL;DR: A simple technique for detecting plaque-forming cells that combines the sensitivity and improved optical conditions of the previously reported monolayer technique with the screening power and ease of quantification of the original agar-plate method is described.
Abstract: A simple technique for detecting plaque-forming cells is described. It combines the sensitivity and improved optical conditions of the previously reported monolayer technique with the screening power and ease of quantification of the original agar-plate method.
2,008 citations
"The role of nonlymphoid accessory c..." refers methods in this paper
...For SRC antigen , the assay was the formation, in the presence of complement, of plaques of lysis in a monolayer of SRC, using the modification of the Jerne technique introduced by Cunningham and Szenberg (15)....
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...For SRC antigen, the assay was the formation, in the presence of complement, of plaques of lysis in a monolayer of SRC, using the modification of the Jerne technique introduced by Cunningham and Szenberg (15)....
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TL;DR: It was found that both adherent and nonadherent cells were necessary for the induction of antibody formation to sheep red blood cells in vitro.
Abstract: A suspension of mouse spleen cells can be separated into two populations on the basis of their ability or inability to adhere to plastic dishes. It was found that both adherent and nonadherent cells were necessary for the induction of antibody formation to sheep red blood cells in vitro. Exposure of adherent cells to antigen for brief periods of time was sufficient to initiate a maximal in vitro response.
561 citations
"The role of nonlymphoid accessory c..." refers result in this paper
...For example, Mosier (9) has suggested the need for cell aggregate formation to obtain a response to SRC, and the accessory cell could have a "structural" role here....
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...Our results are in line with those of Mosier (29) who separated adherent and nonadherent components from mouse spleen by successive cultures in dishes and found these components did not respond in culture unless mixed together....
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...2) 108 (36-285) Adherent fraction 51 (27-155) 71 (18-195)...
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TL;DR: The viability of column-separated cells was shown by their non-staining with trypan blue, motility, phagocytic ability, oxygen consumption, and survival or development in tissue culture, whereas Nowell’s blast-like, dividing, phytohemagglutinin cells were produced only in cultures containing lymphocytes.
437 citations
"The role of nonlymphoid accessory c..." refers methods in this paper
...The technique has been modified from the Rabinowitz (6) procedure to give more efficient separation with various mouse lymphoid organs....
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...We have also developed methods of segregating different classes of cells on the basis of density (5), size (4), and active adherence (6), as modified by Shortman et al....
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TL;DR: Spleen cells from normal, unimmunised mice were grown on dialysis membranes above a reservoir of medium together with sheep or horse erythrocytes and antibody-producing cells appeared in significant numbers in the cultures within 3 to 5 days.
357 citations
"The role of nonlymphoid accessory c..." refers background or methods in this paper
...Following the work of Mischell and Dutton (1), Marbrook (2), and Diener and Armstrong (3) we have established tissue culture systems whereby dissociated mouse spleen cells develop an immune response to two different antigens, sheep erythrocytes (SRC) and polymerized bacterial flagellin (POL)....
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...The assay for the development of an immune response in vitro to both antigens was the technique of Marbrook (2) and Diener and Armstrong (3) as given in detail by Diener and Armstrong (20)....
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...--The assay for the development of an immune response in vitro to both antigens was the technique of Marbrook (2) and Diener and Armstrong (3) as given in detail by Diener and Armstrong (20)....
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...Our results also agree with those of Haskill, Byrt, and Marbrook (23) who found a radiation-resistant, light-density cell involved in the response to SRC in culture, but the detailed density profile of these two components does not agree well....
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...The activity profile parallels that obtained independently by Haskill, Byrt, and Marbrook (23)....
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TL;DR: The arrival directions of the solar particles aro such that they may first be trapped by the magnetic field of the Earth, then by the fields of the test magnets, and finally by this energy selection process exert a greater influence on the reproductive responses of flies in the Magnetic field than in control cultures.
Abstract: there exist both the deflexion effect of the magnetic fields on the cosmic ray particles as well as a possible genetically transferred conditioning effect of the magnetic field on the organism. During the periods of high solar activity, the incoming particles are of lower energy, and it can be shown by equation 8 that the radius of curvature of a charged particle in the magnetic field is considerably smaller. For example, a 1 Me V particle approaching the equatorial plane of the magnet at the medium surface would have approximately a 3-cm radius of curvature, and as a result of its lower energy there is a greater probability that it would be 'captured' by the magnetic field and spiral into the test region of the magnet. The situation in the magnetic field is quite different during high solar activity from that during the period of high cosmic radiation. This could explain the lack of correlation of the 1960 control data (Table 5) with the S' F parameter. Thus in the case of the control data, during the high solar flare activity the effects of both cosmic ray and solar flare radiation are acting on the organism. When encountering the magnetic field conditions, a portion of the cosmic ray component may be deflected out of the field region (as indicated for the 1962 data) and the magnetic field progeny are subjected to a lower cosmic ray flux but, by the same token, a higher solar flare flux. The arrival directions of the solar particles aro such that they may first be trapped by the magnetic field of the Earth, then by the fields of the test magnets, and finally by this energy selection process exert a greater influence on the reproductive responses of flies in the magnetic field than in control cultures. An example of the changing trends and correlations over the two periods of the solar cycle is indicated in a Table 6. SIMPLIFIED REPRESENTATION OF PROGENY TRENDS UNDER DIFFERENT ENVIRONMENTAL CONDITIONS
286 citations
"The role of nonlymphoid accessory c..." refers background in this paper
...There is considerable evidence suggesting this type of role (10-12)....
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