The role of H-2-linked genes in helper T cell function. V. I-region control of helper T cell interaction with antigen-presenting macrophages.
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Animals↗
Biomedical subjects
P Marrack
Publications and source records attributed to P Marrack.
The role of H-2 linked genes in helper T-cell function. IV. Importance of T-cell genotype and host environment in I-region and Ir gene expression.
We have studied the properties of helper T cells specific for sheep erythrocytes (SRBC), keyhole limpet hemocyanin (KLH), or poly-L-(Tyr,Glu)-poly-DL-Ala-poly-L-Lys [(T,G)-A--L]. These T cells differentiated and were primed in vivo in irradiation chimeras constructed of various combinations of F1 and parental bone marrow donors and irradiated recipients. Primed T cells were then tested for helper activity in the in vitro response of B cells and macrophages (Mphi) of parental or F1 origin to the hapten trinitrophenol coupled to the priming antigen. When testing either SRBC or KLH-specific T cells of parental H-2 type which had differentiated in F1 hosts, we found that they cooperated equally well with B cells and Mphi of either parental H-2 type. On the other hand, when testing F1 T cells which had differentiated in parental hosts, we found that they cooperated well only with B cells and Mphi which had the K-IA region type of the parental host. In similar experiments we found that (T,G)-A--L-specific T cells of low responder H-2 type which had differentiated in (high responder X low responder) F1 hosts induced high responses in high responder B cells and Mphi (T,G)-A--L-specific F1 T cells which differentiated in high responder but not those which differentiated in low responder hosts induced high responses in high responder B cells and Mphi. Low responder B cells and Mphi yielded low responses in all cases regardless of the source of (T,G)-A--L-specific T cells with what they were tested. Our results support the conclusion that I-region and Ir genes function via their expression in B cells and Mphi and in the host environment during helper T-cell differentiation, but not, at least under the conditions of these experiments, via their expression in the helper T cell itself. These findings place constraints upon models which attempt to explain the apparent dual recognition of antigen and I-region gene products by helper T cells.
Animals↗
The role of H-2-linked genes in helper T-cell function. III. Expression of immune response genes for trinitrophenyl conjugates of poly-L(Tyr, Glu)-poly-D,L-Ala--poly-L-Lys in B cells and macrophages.
Using lymph node T cells from poly-L(Tyr,Glu)-poly-D,L-Ala--poly-L-Lys[(TG)-A--L]-primed animals and B cells from animals primed with trinitrophenylated (TNP) protein or lipopolysaccharide, we have obtained anti-TNP-(TG)-A--L direct plaque-forming responses in vitro. Response to this antigen was shown to be controlled by the H-2 haplotype of the animal studied. The strain distribution of in vitro response was very similar to that previously reported by others for in vivo secondary IgG responses to (TG)-A--L. We investigated the cell types expressing the Ir gene(s) for (TG)-A--L in our cultures. F1, high responder x low responder mice were primed with (TG)-A--L. Their T cells were active in stimulating anti-TNP-(TG)-A--L responses of high responder but not low responder B cells and macrophages (MPHI), even though both preparations of B cells and Mphi were obtained from mice congenic at H-2 with one of the parents of the F1. For three low responder strains tested, of the H-2h2, H-2k, and H-2f haplotypes, the anti-TNP-(TG)-A--L response of low responder B cells and Mphis in the presence of high responder, F1 T cells could not be improved by the addition of high responder, antigen-bearing Mphis to the cultures. In one strain of the H-2a haplotype, it was shown that neither the B cells nor Mphis could be functional in anti-TNP-(TG)-A--L responses. Our results therefore suggested the Ir genes for anti-TNP-(TG)-A--L responses were expressed at least in B cells in all the low responder strains we studied, and, in mice of the H-2a haplotype, in Mphis too.
Animals↗
The role of H-2 linked genes in helper T-cell function. II. Isolation on antigen-pulsed macrophages of two separate populations of F1 helper T cells each specific for antigen and one set of parental H-2 products.
A method was established for isolating antigen-specific murine helper T cells by selective binding to antigen-pulsed macrophage (Mphi) monolayers. Sheep erythrocyte (SRBC)-primed T cells, which remained strongly adherent to SRBC-pulsed syngeneic Mphi after 20 h in culture, were markedly enriched for helper activity when tested in the in vitro antitrinitrophenol (TNP) response to TNP-SRBC. Successful binding and enrichment occurred only if the Mphi were pulsed with the specific antigen to which the T-cell donors had been primed. The genetic control governing helper function in this system was then examined by using primed F1 T cells isolated on Mphi monolayers from congenic strains bearing parental H-2 haplotypes. SRBC-primed BDF1 (H-2b X H-2d) T cells, which bound to SRBC-pulsed H-2d Mphi, subsequently functioned as helper cells in cultures containing H-2d B cells and Mphi, but not in those containing H-2b B cells and Mphi. They remained unable to collaborate with B cells of the H-2B haplotype even in the presence of additional H-2d Mphi, indicating that H-2 restriction occurs at least at the level of the B cell. Similary, primed BDF1 T cells isolated on H-2b Mphi cooperated preferentially with H-2b B cells and Mphi. In both cases, the haplotype preference of the T cell was not due to alloreactive suppressor activity. These results suggest that primed F1 mice contain individual populations of helper T cells, each of which recognize antigen in association with a parental H-2 gene product(s) expressed during both Mphi-T cell and T cell-B cell interactions.
Animals↗
Effects of warfarin administration on the immune response of mice.
Administration of warfarin to mice is shown to have little effect on the humoral immune response of these animals to sheep red blood cells (a thymus dependent antigen) or the trinitrophenyl hapten coupled to E. coli lipopolysaccharide (a thymus independent antigen). The ability of low doses of sheep red blood cells to prime T-cells in vivo for helper activity in the in vitro antibody-forming response to trinitrophenyl coupled to sheep red blood cells is also apparently unaffected. By contrast, delayed hypersensitivity responses, measured by foot pad swelling, are significantly stimulated by administration of the drug.
Animals↗
The role of H-2-linked genes in helper T-cell function. I. In vitro expression in B cells of immune response genes controlling helper T-cell activity.
The ability of murine helper T cells primed to the antigen, sheep erythrocytes (SRBC) to cross-react with burro erythrocytes (BRBC) in the in vitro anti-trinitrophenol (TNP) response to TNP-RBC was shown to be under genetic control. Although non-H-2 genes were shown to influence the absolute level of helper activity assayed after SRBC priming, the extent of cross-reaction of SRBC-primed helpers with BRBC was shown to be controlled by an H-2-1inked Ir gene(s). H-2 haplotypes were identified which determined high, intermediate, or low response to the cross- reacting determinants and the gene(s) controlling the cross-reaction tentatively mapped to the K through I-E end of the H-2 complex. Helpers primed in F(1) mice of high x intermediate or high x low responder parents were tested for cross-reaction using B cells and macrophages (Mphi) of parental haplotypes. In each case the extent of cross-reaction was predicted by the H-2 haplotype of the B cells and Mphi, establishing the expression of the Ir gene(s) in B cells and/or Mphi a t least, but not ruling out its expression in T cells as well. The low cross-reaction seen when T cells from F(1) mice of high x low responder parents were tested on low responder B cells and Mphi was not increased by the presence of high responder Mphi, indicating the Ir gene(s) is expressed in the B cell a t least although it may be expressed in Mphi as well. These and our previously reported experiments are consistent with the hypothesis that helper T cells recognize antigen bound to the surface of B cells and Mphi in association with the product(s) of Ir gene(s) expressed on the B cell and Mphi.
Antigens↗
Suppressor T-cell inactivation of a helper T-cell factor.
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Animals↗
Antigen-specific and nonspecific mediators of T-cell/B-cell cooperation. VI. Distinct patterns of cross-reactivity by two human gamma-globulin-primed helper T-cell subpopulations.
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Antigen-Antibody Reactions↗
Functional heterogeneity among the T-derived lymphocytes of the mouse. VII. Conversion of T1 cells to T2 cells by antigen.
The T1 subpopulation of peripheral T cells was defined in mice by its short half life, insensitivity to anti-thymocyte sera (ATS) in vivo, and slow kinetics of response to antigen. The T2 subpopulation was defined by its long life time, elimination by ATS in vivo, and rapid response to antigen. Mice containing only T1-type T cells were constructed by adult thymectomy (ATx) followed immediately by the elimination of T2 cells by ATS treatment. Immunization of these mice with SRBC led to the production of memory helper cells in the T2 subpopulation. This process depended on the presence of T1 cells and for the most part required SRBC immunization, although a few SRBC-specific T2 cells reappeared in the mice in the absence of antigen. We conclude that T1 cells can give rise to T2 cells in an antigen-driven step and that the two populations correspond to virgin and memory T cells, respectively.
Animals↗
Antigen-specific and nonspecific mediatiors of T cell/B cell cooperation. II. Two helper T cells distinguished by their antigen sensitivities.
Further evidence is presented for two types of helper T cells in the mouse specific for the protein antigen, keyhole limpet hemocyanin (KLH). The first cell helps B cells respond to the trinitrophenyl hapten (TNP) coupled to KLH, is primed by relatively high doses of antigen in vivo, and yet the effector cell is stimulated by very low doses of antigen in vitro. The second cell helps B cells respond to a non-cross-reacting antigen, sheep red blood cells, presumably via production of a nonspecific factor. This cell is primed by relatively low doses of antigen in vivo, but the effector cell requires relatively high doses of antigen in vitro. Thus, the two T cell types are differently sensitive to antigen dose, both in priming and challenge. The properties of T cells responding to KLH by proliferation in vitro were also studied. These cells showed the same antigen-sensitivity in vitro, as cells producing nonspecific B cell-stimulating factors.
Animals↗
Antigen-specific and nonspecific mediators of T cell/B cell cooperation. III. Characterization of the nonspecific mediator(s) from different sources.
T cell-containing lymphoid populations produce a nonantigen-specific mediator(s) (NSM) which can replace T cell helper function in vitro in the response of B cells to sheep red blood cells (SRBC), but not to the hapten-protein conjugate, trinitrophenyl-keyhole limpet hemocyanin, (TNP-KLH). NSM produced under three conditions: 1) stimulation of KLH-primed cells with KLH; 2) allogeneic stimulation of normal spleen cells; and 3) stimulation of normal spleen cells with Con A (but not PHA) are indistinguishable on the basis of their biologic activity and m.w., estimated as 30 to 40,000 daltons by G-200 chromatography. Production of NSM is dependent on the presence of T cells. The action of NSM on B cells responding to SRBC in the presence of 2-mercaptoethanol is unaffected by severe macrophage depletion. Extensive absorption of NSM with SRBC failed to remove its activity, confirming its nonantigen-specific nature.
Animals↗
Antigen-specific and nonspecific mediators of T cell/B cell cooperation. IV. Development of a model system demonstrating responsiveness of two T cell functions to HGG in vitro.
Previous work in this laboratory has defined two functionally distinct T cell subpopulations (viz., antigen-specific helper) and characerized some of the parameters of these subpopulations derived from the keyhole limpet hemocyanin (KLH)-primed mouse spleen. Characerization of further parameters such as relative susceptibility to tolerance induction and relative degree of specificity was not possible with the use of KLH as the antigen. In order to overcome this impediment, an experimental protocol was developed, as herewith described, to permit the study of these two T cell subpopulations in response to the antigen human gamma-globulin (HCG). This protocol qualitatively duplicates the parameters defined for the KLH system and provides an extremely useful model for the study of the response to serum proteins in the Mishell-Dutton culture system.
Animals↗
Antigen-specific and nonspecific mediators of T cell/B cell cooperation. V. Unresponsiveness to HGG in vitro of these two T cell subpopulations.
In previously published experiments we have distinguished two subpopulations of helper T cells (specific helper and nonspecific helper) in the spleens of antigen-primed mice. These subpopulations can be differentiated by their modes of action, and their activities and dose-response characteristics in mouse spleens primed with human gamma-globulin (HGG). In experiments designed to test shether the two subpopulations were derived from the same pool of precursors, or different pools, mice were injected with limiting doses of a tolerogenic form of HGG. They were subsequently challenged with an immunogeneic dose of HGG and their spleens assayed for specific helper and nonspecific helper activities in response to HGG in vitro. Over the dose range of tolerogenic HGG studied, both the helper activities were found to be equally tolerized. The question of the possible involvement of suppressor T cells in the maintenance of the state of unresponsiveness was also addressed quantitatively by the use of this model system. No active suppression was demonstrable in the maintenance of tolerance in either T cell subpopulation. These results suggest that the two types of helper T cells are derived from a common precursor pool, or from different pools with identical susceptibilities to tolerogen. Suppressor cells do not appear to play a role in this form of tolerance.
Animals↗
L3T4 interacts with class II molecules on target cells.
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Animals↗
Role of gamma-interferon in antibody-producing responses.
Interferon preparations, especially those containing gamma-interferon (IFN-gamma), have long been known to modulate immune responses. However, because many studies used only partially purified interferons, it has been difficult to separate the immunoregulatory effects of the interferons from those of other biologically active molecules contaminating the preparations. Recently, with the cloning of the interferon genes in mouse and man, it has become possible to use these cloned interferons directly to test their effects in assays other than those involving the protection of cells from viruses. For example, cloned IFN-gamma has been shown to be a potent inducer of Ia antigen expression on macrophages. Similarly, cloned IFN-gamma has been reported to act as a macrophage activation factors, as judged by the ability of activated macrophages to kill tumour cells in vitro. We demonstrate here that cloned murine IFN-gamma can also substitute for a late-acting helper factor which acts synergistically with other helper factors in the stimulation of B-cell antibody responses in vitro.
Animals↗
Immunoglobulin-like nature of the alpha-chain of a human T-cell antigen/MHC receptor.
Although the receptor with which T cells bind specific antigen can, like immunoglobulin, distinguish between antigens which differ only slightly in structure, it is unique in recognizing antigen only in conjunction with one of the self proteins of the major histocompatibility complex (MHC restriction). The receptor was identified and characterized in mouse and man by using monoclonal antibodies to receptor idiotypes, and consists of two disulphide-linked polypeptides, and acidic alpha-chain and a neutral to slightly basic beta-chain. Peptide maps have shown that, like immunoglobulin, both chains vary for receptors of different specificities. T-cell-derived cDNA clones have recently been identified in mouse and man encoding immunoglobulin-like molecules. These were identified as derived from beta-chain genes through a partial N-terminal protein sequence of the beta-chain isolated from a human T-cell tumour. We have now purified the alpha- and beta-chains of the receptor of the human T-cell leukaemia line HPB-MLT, and have determined the amino acid sequence of several tryptic peptides derived from each chain. Our results further confirm that the previously reported cDNA clones encode beta-chains. The sequence of the alpha-chain peptides identify this as another immunoglobulin-like polypeptide chain. Particularly striking was an alpha-chain peptide with high homology to the conserved portion of the immunoglobulin J segment and T-cell receptor beta-chains. Surprisingly, the alpha-chain peptides show little similarity to the sequence predicted by two overlapping putative murine alpha-chain cDNA clones.
Amino Acid Sequence↗
Primary structure of human T-cell receptor alpha-chain.
The T-cell receptor has been studied intensely over the past 10 years in an effort to understand the molecular basis for major histocompatibility complex (MHC) restricted antigen recognition. The use of anti-receptor monoclonal antibodies to isolate and characterize the receptor from human and murine T-cell clones has shown that the protein consists of two disulphide-linked glycopeptides, alpha and beta, distinct from known immunoglobulin light and heavy chains. Like immunoglobulin light and heavy chains, however, both the alpha- and beta-chains are composed of variable and constant regions. Molecular cloning has revealed that the beta-chain is evolutionarily related to immunoglobulins, and is encoded in separate V (variable), D (diversity), J (joining) and C (constant) segments that are rearranged in T cells to produce a functional gene. We report here cDNA clones encoding the alpha-chain of the receptor of the human T-cell leukaemia line HPB-MLT. Using these cDNA probes, we find that expression of alpha-chain mRNA and rearrangement of an alpha-chain V-gene segment occur only in T cells. The protein sequence predicted by these cDNAs is homologous to T-cell receptor beta-chains and to immunoglobulin heavy and light chains, particularly in the V and J segments.
Amino Acid Sequence↗