Preparation and Characterization of Antibodies Specific for the 2,4,6-Trinitrophenyl Group<sup>*</sup>

Little,; Hn, Eisen

doi:10.1021/bi00875a001

Cited by 161 publications

(77 citation statements)

References 16 publications

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“…Guinea pig albumin (GPA) and ovalbumin (OVA) were obtained from Miles Laboratories Inc., Miles Research Products, Elkhart, Ind. 2,4,6-trinitrophenyl (TNP)-GL, TNP-GPA, and TNP-OVA were prepared according to the method of Little and Eisen (9).…”

Section: Methodsmentioning

confidence: 99%

The histocompatibility restrictions on macrophage T-helper cell interaction determine the histocompatibility restrictions on T-helper cell B-cell interaction.

Yamashita¹,

Shevach²

1978

The Journal of Experimental Medicine

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The role of the major histocompatibility complex (MHC) 1 in the regulation of immunocompetent cell interactions has been intensively studied in several experimental systems including the interaction of antigen-pulsed macrophages and the primed T cell which proliferates on exposure to antigen in vitro (1, 2) and the interaction of the primed T-helper cell and the B cell in the generation of an antibody response (3). Both of these interactions appear to be controlled by the products of genes which map in the I-region of the MHC (2, 4, 5). The results of these experiments have suggested that the primed T cell is activated by carrier determinants of the nominal antigen in association with I-region associated (Ia) antigens on macrophages and the helper T cell, in turn, activates B cells which bear the same Ia antigens and determinants of the nominal antigen bound to immunoglobulin receptors on their surface. Although a large body of experimental evidence is consistent with this hypothesis, it has proven to be difficult to directly test experimentally because the majority of studies on macrophage T-cell interaction have used T-cell proliferation as a measure of T-cell activation, whereas the role of the macrophage as an antigenpresenting cell in the in vitro generation of a secondary antibody-forming cell response has proven to be difficult to define (6). In an attempt to directly correlate the results of studies on macrophage T-cell interaction with those on T-and B-cell interaction, we have examined the helper cell activity of primed guinea pig T cells which have been cultured in vitro for 7 days with antigen-pulsed macrophages under conditions which have been previously shown (7, 8) to be highly efficient in the antigen-specific selection of T cells which proliferate on re-exposure to antigen. We will demonstrate that these selected T cells function efficiently as T-helper cells only when mixed with syngeneic, but not allogeneic, hapten-primed B cells. Furthermore, when F1 T cells are selected with antigen-pulsed parental macrophages, they will only cooperate with B cells of the same parental strain as the macrophages used in the selection culture. In addition, we will demonstrate in the well characterized poly-L-lysine system in ZAbbreviations used in the paper: CFA, complete Freund's adjuvant; GL, a copolymer of e-glutamic acid and e

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Section: Methodsmentioning

confidence: 99%

The histocompatibility restrictions on macrophage T-helper cell interaction determine the histocompatibility restrictions on T-helper cell B-cell interaction.

Yamashita¹,

Shevach²

1978

The Journal of Experimental Medicine

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“…Mouse Ig was prepared from CBA/J mouse sera and conjugated with TNP hapten (11,12). A single preparation with a level of substitution of 40 TNP molecules per Ig molecule (TNP 40 -Ig) was used throughout.…”

Section: Reagentsmentioning

confidence: 99%

Epicutaneous Immunization with Protein Antigen in the Presence of TLR4 Ligand Induces TCRαβ+CD4+ T Contrasuppressor Cells That Reverse Skin-Induced Suppression of Th1-Mediated Contact Sensitivity

Ptak¹,

Majewska

Bryniarski³

et al. 2009

The Journal of Immunology

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Our previous work showed that epicutaneous (EC) immunization of mice with different protein Ags applied on the skin in the form of a patch induces a state of subsequent Ag-nonspecific unresponsiveness due to suppressor CD4+8+ T cells (Ts) that inhibit Th1-mediated contact sensitivity (CS) reactions via released TGF-β. In the present work we show that EC immunization with Ag together with the TLR4 ligand LPS induced cells that could prevent suppression by the Ag-nonspecific Ts. These up-regulatory cells, called contrasuppressor T cells (Tcs), belong to a population of Ag-specific TCRαβ CD4+ lymphocytes and are different from Th1 CD4+ cells that mediate the CS reaction. Experiments using knockout mice showed that EC induced contrasuppression is MyD88, INF-γ, and IL-12 dependent, whereas IL-6 is not involved in this phenomenon. Additional experiments with anti-IFN-γ mAb showed that IFN-γ is required for induction of Tcs cells but does not play a crucial role in the effector phase of contrasuppression. Additionally, treatment of CS effector cells with rIL-12 makes them resistant to EC induced suppression without affecting Ts cells, whereas IL-12 neutralization in vitro abrogates contrasuppression. These data show that IL-12 is indeed involved in the effector phase of EC induced contrasuppression and that this cytokine does not act directly on Ts cells. The mechanism of action of Tcs protects Th1 effector cells mediating CS from the nonspecific Ts, leaving suppression to other Ags intact. Ts and Tcs cells do not influence each other and can be induced simultaneously in the same animal.

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“…Trinitrophenyl-BSA (TNP-BSA) and TNP-OVA were prepared as described elsewhere (26,27, respectively). mrIL-4 and mrIFN-γ were purchased from Genzyme (Cambridge, MA, USA) and Genentech Inc. (South San Francisco, CA, USA), respectively.…”

Section: Reagentsmentioning

confidence: 99%

Aging reduces the primary humoral response and the in vitro cytokine production in mice

Simioni

Costa

Tamashiro

2007

Braz J Med Biol Res

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Aging is accompanied by a decrease in several physiological functions that make older individuals less responsive to environmental challenges. In the present study, we analyzed the immune response of female BALB/c mice (N = 6) of different ages (from 2 to 96 weeks) and identified significant age-related alterations. Immunization with hapten-protein (trinitrophenyl-bovine serum albumin) conjugates resulted in lower antibody levels in the primary and secondary responses of old mice (72 weeks old). Moreover, young mice (2, 16, and 32 weeks old) maintained specific antibodies in their sera for longer periods after primary immunization than did old mice. However, a secondary challenge efficiently induced memory in old mice, as shown by the increased antibody levels in their sera. The number of CD4 + and CD8 + T cells in the spleen increased until 8 weeks of age but there was no change in the CD4 + /CD8 + ratio with aging. Splenic T cells from old mice that had or had not been immunized were less responsive to concanavalin-A and showed reduced cytokine production compared to young mice (IL-2: 57-127 vs 367-1104 pg/mL, IFN-γ: 2344-12,836 vs 752-23,106 pg/mL and IL-10: 393-2172 vs 105-2869 pg/mL in old and young mice, respectively). These data suggest that there are significant changes in the organization of the immune system throughout life. However, the relevance of these alterations for the functioning of the immune system is unknown.

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Preparation and Characterization of Antibodies Specific for the 2,4,6-Trinitrophenyl Group^*

Cited by 161 publications

References 16 publications

The histocompatibility restrictions on macrophage T-helper cell interaction determine the histocompatibility restrictions on T-helper cell B-cell interaction.

The histocompatibility restrictions on macrophage T-helper cell interaction determine the histocompatibility restrictions on T-helper cell B-cell interaction.

Epicutaneous Immunization with Protein Antigen in the Presence of TLR4 Ligand Induces TCRαβ+CD4+ T Contrasuppressor Cells That Reverse Skin-Induced Suppression of Th1-Mediated Contact Sensitivity

Aging reduces the primary humoral response and the in vitro cytokine production in mice

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Preparation and Characterization of Antibodies Specific for the 2,4,6-Trinitrophenyl Group*

Cited by 161 publications

References 16 publications

The histocompatibility restrictions on macrophage T-helper cell interaction determine the histocompatibility restrictions on T-helper cell B-cell interaction.

The histocompatibility restrictions on macrophage T-helper cell interaction determine the histocompatibility restrictions on T-helper cell B-cell interaction.

Epicutaneous Immunization with Protein Antigen in the Presence of TLR4 Ligand Induces TCRαβ+CD4+ T Contrasuppressor Cells That Reverse Skin-Induced Suppression of Th1-Mediated Contact Sensitivity

Aging reduces the primary humoral response and the in vitro cytokine production in mice

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About

Preparation and Characterization of Antibodies Specific for the 2,4,6-Trinitrophenyl Group^*