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Biomedical subjects

C W Pierce

Publications and source records attributed to C W Pierce.

At least 37 records · Page 2Linked to original sources
PubMed1988

Antigen presentation to T cell lines and clones by peritoneal macrophages, peritoneal B cells and antigen-specific B cell hybridomas.

The antigen presenting cell (APC) activity of uninduced, resident peritoneal macrophages and B cells was compared to that of antigen-specific B cell hybridomas by measuring proliferative responses of antigen-specific, MHC-restricted T cell clones. The results demonstrate that peritoneal macrophages and B cells are much more efficient APC than irradiated splenic filler cells, and that unirradiated B cells were as good as, if not better than, macrophages. Both B cells and macrophages can be pulsed with antigen, although pulsed B cells were always found to be more efficient than pulsed macrophages. However, the APC activity of B cells was exquisitely sensitive to irradiation. The relative contribution of macrophages and B cells to the APC activity of mixed populations was easily distinguished by complement dependent lysis with monoclonal antibodies specific for unique differentiation antigens expressed by these cells. Normal peritoneal macrophages and B cells present the synthetic terpolymer of L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) to GAT-specific T cells clones and beef insulin to insulin-specific T cell lines nonspecifically. The APC activity of antigen-specific B cells was also examined by using novel GAT-specific, nonsecretor B cell hybridomas produced by fusing GAT-primed spleen cells to the HAT sensitive Balb/c lymphoma, M12.4.5. The hybridomas selected for these studies were GAT-specific, sIg+, Ia+ cells. These hybridomas presented GAT to GAT-specific T cells more efficiently than heterogeneous B cells suggesting that interaction with surface Ig receptors facilitated the uptake and/or processing of antigen. GAT-specific B cell hybridomas, like normal B cells, presented soluble beef insulin to an insulin-specific T cell clone nonspecifically. However, after pulsing with antigen overnight, the GAT-specific B cell hybridoma could activate only GAT-specific T cells.

Animals↗
PubMed1988

A comparison of the conventional and the rosette method of determination of interleukin 1 (IL-1) activity. The advantages of the rosette method.

In this report we compare the conventional method of determination of IL-1 activity with a new rosette assay. Determination of IL-1 activity in the rosette assay is well correlated with the conventional method and equally sensitive. However, the rosette method is able to dissect the activity of IL-1 from IL-2 and to determine the immunotropic, optimal dose of IL-1 more accurately.

Animals↗
PubMed1988

Secretion of interleukin 1 (IL-1) by peritoneal macrophages upon contact with syngeneic T cells is Ia-restricted and antigen-independent process.

We deliver an evidence that secretion of interleukin 1 by macrophages upon contact with T cells is an Ia-restricted and antigen-independent event. Antigen-specific T cell lines or quiescent thymocytes induce comparable quantities of IL-1 from Ia-compatible macrophages. Addition of antigen together with antigen-specific T cells does not increase production of IL-1. Con A-activated thymocyte-derived blasts induce less IL-1 than unstimulated thymocytes. Peritoneal B cells do not secrete IL-1 although they present GAT effeciently to GAT-specific T cell clones. Lastly, the production of IL-1 can be inhibited by a treatment of macrophages with monoclonal antibodies against Ia but not H-2D antigens. The results indicate that the release of IL-1 is solely a result of an interaction between Ia molecules on macrophages and the receptor for Ia on interacting T cells. This process does not require antigen/lectin bridge between the interacting cells.

Animals↗
PubMed1987

Receptor diversity of insulin-specific T cell lines from C57BL (H-2b) mice.

To characterize the T cell receptor repertoire in an immune response in which the Ia and nominal antigenic determinants are defined and limited, we have cloned and sequenced the expressed receptors from four independent, beef insulin-specific T cell lines from C57BL mice. Each of these lines responded to beef but not to the pork insulin, thus defining the nominal antigenic determinant recognized. Furthermore, each of these lines could only be presented antigen by B6 but not mutant B6.C-H-2bm12 antigen-presenting cells, thus defining the requisite Ia recognition or antigen-association site. In spite of this functional similarity in ligand specificity, each of these T cell lines was found to use different V alpha and V beta gene segments. Moreover, structural comparisons of implied protein sequences of each of these receptors showed no stretches of conserved amino acid residues that could be implicated in ligand interaction. However, the V alpha genes used by these four clones appeared considerably more homologous to each other than were their V beta genes.

Animals↗
PubMed1987

Molecular genetic characterization of the mRNA coding for an inducible suppressor factor specific for L-glutamic acid60-L-alanine30-L-tyrosine10.

The suppressor T-cell hybridoma 1556A2.1 can be induced by the monoclonal L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT)-specific suppressor inducer 372B3.5 and soluble GAT to synthesize a disulfide-linked heterodimeric protein (GAT-TsF2), which directly suppresses a primary in vitro immune response to GAT. Induction and synthesis of the GAT-TsF2 protein is correlated with the appearance of specific mRNA, as detected by translation in vitro in a wheat germ cell-free extract of RNA isolated at various times after induction. The mRNA coding for the polypeptide chain that bears a serologically defined I-J determinant (I-J+ chain) appeared 8 hr after induction, whereas the mRNA coding for the antigen-binding chain (AB+ chain) was not detected until 16 hr after induction. The mRNAs coding for the individual chains sedimented as different species, suggesting that the two-chain factor is the product of two genes. The AB+ chain of the 1556A2.1 GAT-TsF2 was synthesized on membrane-bound polysomes, whereas the I-J+ chain was translated on free polysomes. The AB+ chain was synthesized from two independent mRNA species sedimenting at 10 S and 28 S, whereas a single 16S mRNA encoded the I-J+ chain. The in vitro translated I-J+ chain was bound by a monoclonal antibody against the I-J+ determinant of only the appropriate H-2 haplotype. These results suggest that posttranslational modification, including glycosylation, is not required for biological activity or for expression of the I-J epitope on the GAT-TsF2 molecule.

Animals↗
PubMed1987

The role of suppressor T cells in the expression of immune response gene function.

Mechanisms underlying major histocompatibility complex (MHC)-linked immune response (Ir) gene regulation of immune responses have been the subject of considerable interest and debate in recent years. Two general mechanisms have been proposed to account for antigen-specific, Ir gene-mediated unresponsiveness. In one, defective antigen presentation resulted from the failure of processed nominal antigen and Ia antigen to associate on the antigen presenting cell membrane in a manner sufficient for helper T cell (Th cell) activation. By contrast, it has been proposed that selected Th cell clones were deleted from the repertoire during ontogeny or otherwise rendered unresponsive to the antigen-Ia complex, i.e., functionally deleted. Either of these mechanisms would account for the deficient activation of antigen-specific, Th cells observed in genetic low or nonresponder mice. In addition, the failure of mice to respond to certain antigens under Ir gene control has been attributed to the activation of specific suppressor T (Ts) cells. The latter mechanism might be considered a corollary or subset of the clonal deletion model. However, an important distinction exists. In the case of active, Ts cell-mediated Ir gene regulation, genetic low responder animals should retain the capacity for antigen-induced activation of Th cells, or Th cell activity should be demonstrable in these mice. In this communication, experiments are described which are designed to evaluate the possibility that active Ts cell-mediated regulatory mechanisms were of general importance in mediating Ir gene-related unresponsiveness.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Proliferative response of T lymphocytes to a proline-rich polypeptide (PRP): PRP mimics mitogenic activity of Il-1.

Mitogenic properties of a proline-rich polypeptide were investigated. The mitogenic action of PRP was compared with the mitogenic action of Il-1. PRP was not mitogenic for thymocytes at doses 0.01-50 micrograms/ml. PRP, at doses 0.1-50 micrograms/ml, augmented the proliferative response of thymocytes to Con A in a similar fashion as Il-1. At doses higher than 10 micrograms/ml, PRP induced proliferation of lymph node cells and splenocytes as well as T cells from the lymph nodes. It did not, however, cause significant proliferation of B cells from the lymph nodes, at the doses used. PRP did not induce proliferation of an antigen specific Lyt 1+ T cell clone. Il-1 behaved in a similar way as PRP in all the tests described. We consider a possibility that under physiological conditions, at a very early stage of postneonatal life, PRP may replace some functions of Il-1.

Animals↗
PubMed1986

Characterization of two monoclonal idiotype-binding suppressor T cell factors specific for the antibody response to L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT).

We have previously identified and described two distinct types of suppressor T cell factors specific for the PFC response to L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) or L-glutamic acid50-L-tyrosine50 (GT). Both of these factors are antigen-specific and I-J+. GAT-TsF1 is not MHC-restricted and is composed of a single polypeptide chain, whereas GAT-TsF2 is MHC-restricted and is composed of two different polypeptide chains. We have not previously found evidence for an obligatory, idiotype-specific suppressor T cell in this suppressor pathway. However, we now report that idiotype-specific suppressor T cells can be elicited by exposing normal spleen cells to GAT-TsF1 or GAT-TsF2 in the absence of antigen in vitro. These factor (TsF1/TsF2)-induced cells have been fused with the AKR thymoma, BW5147, and hybridomas were selected for production of suppressor factors that inhibit GAT-specific antibody responses in vitro. In this report, we characterize one monoclonal factor from each fusion. Neither factor binds GAT or the related co-polymer, GT; both factors have binding sites for GAT-specific idiotypes but not for unrelated idiotypes either in the form of antibody immobilized on Sepharose or as cell surface determinants expressed by B cell hybridomas. Moreover, their reactivities for a panel of monoclonal anti-GAT antibodies are overlapping but not identical. Both factors are composed of two polypeptide chains, and both chains are required for suppressive activity; one chain bears the I-J determinant, whereas the other possesses the idiotype-binding activity of the intact molecule. Both idiotype-binding factors are restricted by MHC- and Igh-linked genes, and transcomplementation is observed in the F1 mice between MHC-congenic and Igh-congenic parents. Both factors are active late in Mishell-Dutton cultures. These data support the contention that these two factors are similar but nonidentical members of an anti-idiotypic class of GAT-specific suppressor factors.

Animals↗
PubMed1986

Identification and characterization of a suppressor T cell hybridoma specifically inducible by L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT).

In vitro activation of naive spleen cells from C57BL/10 mice with GAT and the monoclonal GAT-TsF1, 372B3.5, followed by fusion with BW5147 resulted in generation of a hybridoma that fails to produce GAT-TsF constitutively, but upon reexposure to GAT and 372B3.5 is induced to secrete GAT-TsF2. The induction is GAT specific and requires de novo RNA, protein synthesis, and DNA synthesis. Although both GAT and 372B3.5 are required for induction, they may be added sequentially, provided the GAT is added first. The GAT-TsF produced by the induced cell is antigen specific and composed of two polypeptide chains: one capable of binding antigen, the other bearing determinants encoded by the I-J region of the MHC. The utility of this inducible GAT-TsF2 cell line for molecular biology and other studies is discussed.

Animals↗
PubMed1986

Regulation of immune responses by T cell subsets. Role of helper and suppressor T cells in the development and expression of MHC-restricted antibody responses to L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) by (responder X responder)F1 spleen cells.

The roles of helper and suppressor T cells in the development and expression of antibody responses to GAT were studied in (responder X responder)F1 mice immunized with parental GAT-M phi. Spleen cells from (B10 X B10.D2)F1 mice primed in vivo with B10 or B10.D2 GAT-M phi developed secondary in vitro plaque-forming cell (PFC) responses only when stimulated by GAT-M phi syngeneic with the GAT-M phi used for in vivo priming. By contrast, virgin F1 spleen cells developed comparable primary PFC responses to both parental GAT-M phi Co-culture of T cells from (B10 X B10.D2)F1 mice primed in vivo by B10 GAT-M phi with virgin (B10 X B10.D2)F1 spleen cells demonstrated the presence of suppressor cells that inhibited the primary response of virgin spleen cells stimulated by B10.D2 GAT-M phi. Spleen cells from (B10 X B10.D2)F1 mice primed in vivo with B10.D2 GAT-M phi had suppressor T cells that suppressed primary responses stimulated by B10 GAT-M phi. The suppressor T cell mechanism was composed of at least two regulatory T cell subsets. Suppressor-inducer T cells were Lyt-2-, I-J+ and must be derived from immune spleen cells. Suppressor-effector T cells can be derived from virgin or immune spleens and were Lyt-2+ cells. When the suppressor mechanism was disabled by treatment with 1000 rad gamma irradiation or removal of Lyt-2+ cells, Lyt-2-helper T cells from (B10 X B10.D2)F1 mice primed with B10 GAT-M phi provided radioresistant help to virgin F1 B cells stimulated by B10 but not B10.D2 GAT-M phi. Suppressor inducer Lyt-2-,I-J+ cells from B10 GAT-M phi-primed (B10 X B10.D2)F1 mice were separated from the primed Lyt-2-,I-J-helper T cells. In the presence of Lyt-2+ suppressor effector cells, the Lyt-2-,I-J+ suppressor-inducer suppressed the primary response of virgin spleen or virgin T plus B cells stimulated by both B10 and B10.D2 GAT-M phi. Therefore, suppressor T cells were able to suppress primary but not secondary GAT-specific PFC responses stimulated by either parental GAT-M phi. These results showed that immunization of (responder X responder)F1 mice with parental GAT-M phi results in the development of antigen-specific helper and suppressor T cells. The primed helper T cells were radioresistant and were genetically restricted to interact with GAT in association with the major histocompatibility complex antigens of the M phi used for in vivo priming.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Cellular interactions of L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT)-specific suppressor factors. I. Inhibition of the activity of GAT-specific helper T cell clones by monoclonal GAT-specific suppressor T cell factors.

Considerable information concerning the serology and biochemistry of antigen-specific, T cell-derived suppressor factors has been obtained with the use of T cell hybridomas as a source of homogeneous material. Similarly, knowledge of helper T cell products and receptors is accumulating from studies of helper T cell clones and hybridomas. Our strategy for studying the mechanisms by which suppressor factors inhibit responses was to determine whether monoclonal suppressor factors could inhibit antibody responses specific for L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT) in cultures containing unprimed splenic B cells, macrophages, and GAT-specific T cell clones as a source of helper activity. The MHC-restricted, two chain suppressor factors, GAT-TsF2, inhibited these responses if the helper T cell clones and suppressor factor were derived from H-2-compatible mice. Furthermore, responses were inhibited by briefly pulsing T cell clones with GAT-TsF2 in the presence of GAT, indicating that suppressor factors need not be present continuously. In addition, helper T cell clones adsorbed syngeneic, but not allogeneic, GAT-TsF2 in the presence of GAT. Adsorption also requires a shared antigenic specificity between the H-2b-derived helper T cells and TsF2 factor. Thus, helper T cells can serve as the cellular target of antigen-specific, MHC-restricted GAT-TsF2, and cloned helper T cells can be used as a homogeneous target population for analysis of the molecular mechanisms of T cell suppression.

Adsorption↗
PubMed1986

Genetic restrictions in the development of antibody responses to L-glutamic acid60-L-alanine30-L-tyrosine10 by nude mice implanted with semiallogeneic thymus glands.

Athymic nude mice implanted with F1 thymus glands were used to investigate genetic restrictions regulating T cell-macrophage (M phi) interactions in the development of antibody responses to GAT. Spleen cells from conventional mice developed comparable primary plaque-forming cell (PFC) responses when stimulated by syngeneic and allogeneic GAT-M phi. However, spleen cells from strain A nude mice implanted with (A X B)F1 thymus glands were tolerant of strain B alloantigens and developed GAT-specific PFC responses to strain A GAT-M phi and allogeneic strain C GAT-M phi, but failed to respond to strain B GAT-M phi. The lack of primary GAT-specific PFC responses by spleen cells from (A X B)thy----A nude mice stimulated by strain B GAT-M phi was not due to detectable suppressor mechanisms. However, an allogeneic effect stimulated by H-2- or non-H-2-disparate GAT-pulsed or unpulsed M phi was able to overcome the inability of spleen cells from (A X B)F1 thy----A nude mice to respond to strain B GAT-M phi. Furthermore, the inability to respond to strain B GAT-M phi was overcome by the addition of supernatant fluids from independent cultures of H-2-disparate cells. These results 1) demonstrate that T cells from A nude mice implanted with (A X B)F1 thymus glands did not recognize nominal antigen in the context of B MHC antigens, and 2) suggested that the T cell repertoire was altered in strain A nude mice implanted with (A X B)F1 thymus glands, such that T cells that could recognize GAT in association with strain B MHC antigens were functionally deleted.

Animals↗
PubMed1985

Identification of Igh-C-linked determinants on suppressor T cell hybrids and factors specific for L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT).

Hyperimmunization of BALB/c mice with concanavalin A-stimulated blasts from the Ig allotype-congenic strain, C.B20, results in the production of antibodies reactive with T cells in an allotype-restricted manner. Spleen cells from these hyperimmune BALB/c mice were used to generate a panel of hybridomas that secrete monoclonal antibodies, reactive, in an allotype-restricted manner, exclusively with T cells subpopulations, and in particular, reactive with suppressor T cell hybridomas and their secreted soluble factors. Two functional classes of antibodies were identified: those that react with single polypeptide-chain suppressor T cell factors (TsF1) and the suppressor T cell hybridomas that produce such factors, and those that react with two polypeptide-chain suppressor T cell factors (TsF2) and their corresponding suppressor T cell hybridomas. These two classes of antibody were used to isolate molecules from the membranes of the respective suppressor T cell hybrids that are functionally and structurally related to the secreted suppressor T cell factors, suggesting a receptor function for these molecules.

Animals↗
PubMed1985

Haplotype-specific suppression of antibody responses in vitro. III. Haplotype-specific suppressor factor (TsF-H) binds to but fails to suppress responses by the I-A mutant strain B6.C-H-2bm12.

Spleen cells from C57BL/6 and B6.C-H-2bm12 mice, both responder strains to GAT, differ in their ability to be suppressed by the monoclonal I-A-restricted, nonantigen-specific, but haplotype-specific suppressor factor, TsF-H, from the hybridoma 266A4.5. Whereas GAT-specific responses by C57BL/6 spleen cells are susceptible to TsF-H-mediated suppression, responses by bm12 spleen cells are nonsuppressible under the same conditions. Responses of both C57BL/6 and bm12 spleen cells are suppressed by monoclonal GAT-specific suppressor factors. The inability of TsF-H to suppress responses by the bm12 spleen cells presumably reflects the effects of the mutation in the beta-chain of the I-A antigen in this strain on the required I-A restriction between TsF-H and target cell for manifestation of suppressive activity. The data are discussed in terms of involvement of I-A or recognition of I-A in mediating suppression.

Absorption↗