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G Stingl

Publications and source records attributed to G Stingl.

At least 19 recordsLinked to original sources
PubMed1995

The high affinity IgE receptor (Fc epsilon RI) mediates IgE-dependent allergen presentation.

The discovery that the high affinity IgE receptor (Fc epsilon RI) is expressed on APCs of patients with atopic diseases raised the possibility that the functional importance of Fc epsilon RI in the pathogenesis of atopy may extend beyond its role in type I allergic reactions. Here we show that, following removal of in vivo-bound IgE by lactic acid treatment, targeting of allergens to monocytes by Ag-specific IgE critically depends on Fc epsilon RI expression. Even more importantly, lactic acid-treated, monocyte-enriched PBMCs present allergen to T cells 100- to 1000-fold more effectively if the allergen has been targeted to Fc epsilon RI on these cells via allergen-specific IgE. This mechanism may critically lower the atopic individual's threshold to mount allergen-specific T cell responses capable of promoting IgE production and delayed-type hypersensitivity reactions.

Allergens↗
PubMed1995

Priming of tumor-specific T cells in the draining lymph nodes after immunization with interleukin 2-secreting tumor cells: three consecutive stages may be required for successful tumor vaccination.

Although both CD4+ and CD8+ T cells are clearly required to generate long-lasting anti-tumor immunity induced by s.c. vaccination with interleukin 2 (IL-2)-transfected, irradiated M-3 clone murine melanoma cells, some controversy continues about the site and mode of T-cell activation in this system. Macrophages, granulocytes, and natural killer cells infiltrate the vaccination site early after injection into either syngeneic euthymic DBA/2 mice or athymic nude mice and eliminate the inoculum within 48 hr. We could not find T cells at the vaccination site, which argues against the concept that T-cell priming by the IL-2-secreting cancer cells occurs directly at that location. However, reverse transcription-PCR revealed transcripts indicative of T-cell activation and expansion in the draining lymph nodes of mice immunized with the IL-2-secreting vaccine but not in mice vaccinated with untransfected, irradiated M-3 cells. We therefore propose that the antigen-presenting cells, which invade the vaccination site, process tumor-derived antigens and, subsequently, initiate priming of tumor-specific T lymphocytes in lymphoid organs. These findings suggest a three-stage process for the generation of effector T cells after vaccination with IL-2-secreting tumor cells: (i) tumor-antigen uptake and processing at the site of injection by antigen-presenting cells, (ii) migration of antigen-presenting cells into the regional draining lymph nodes, where T-cell priming occurs, and (iii) circulation of activated T cells that either perform or initiate effector mechanisms leading to tumor cell destruction.

Animals↗
PubMed1995

Impaired survival of T cell receptor V gamma 3+ cells in interleukin-4 transgenic mice.

The mouse epidermis contains a network of Thy-1+ dendritic T cells. Most of these cells express a homogeneous T cell receptor (TCR) configuration (V gamma 3/V delta 1) with only negligible junctional diversity. Because fetal thymocytes are precursors of these dendritic epidermal T cells (DETC) and the addition of interleukin (IL)-4 to fetal thymic organ cultures causes an early arrest in thymopoiesis, we examined DETC development in transgenic (tg) mice expressing IL-4 under the control of major histocompatibility complex class I regulatory sequences. Immunohistologic examination of epidermal sheets and polymerase chain reaction analysis of total skin RNA from IL-4 tg mice failed to reveal TCR V gamma 3+ DETC and V gamma 3 mRNA, respectively. In contrast, the sizes of TCR gamma delta subpopulations in lymphoid organs were unchanged in these mice. Although the numbers and staining intensities of TCR V gamma 3+ thymocytes in early fetal (days 14-17) IL-4 tg mice were similar to those of littermate controls, we observed a preferential death of these cells in thymic organ cultures from IL-4 tg mice. We observed further that epidermal sheets prepared from 9-day-old mice whose mothers had been treated with an IL-4-neutralizing antibody from day 12 to day 18 of pregnancy contained DETC numbers similar to those of controls. However, upon termination of the anti-IL-4 treatment, DETC ceased to expand. We conclude that IL-4 impairs the survival of TCR V gamma 3+ cells.

Animals↗
PubMed1995

Phenotypic changes that TCR V gamma 3+ fetal thymocytes undergo during their maturation into dendritic epidermal T cells.

Murine Thy-1+, TCR V gamma 3/V delta 1+ dendritic epidermal T cells (DETC) express CD2 antigens, but differ from most other T-cell subsets in their absence of CD4, CD5, and CD8 antigens. To determine whether negativity for those antigens is an intrinsic feature of a given T-cell population or whether such triple-negative T cells go through a maturational stage during which they express these antigens, we determined the phenotype of TCR V gamma 3+ fetal thymocytes, which are the precursor cells of DETC. We found that TCR V gamma 3+ fetal thymocytes at day 17 of gestation are CD2+, CD5+, mostly CD8+, and partly CD4+. The expression of CD5 is highest on early TCR V gamma 3+ thymocytes; these cells express intermediate levels of CD5 when they leave the thymus and lose CD5 expression until or shortly after arrival in the epidermis. A similar loss of CD5 expression by TCR V gamma 3+ cells was observed in vitro under various culture conditions. To determine whether expression of CD5 is important for the maturation of DETC, we searched for these cells in the epidermis of CD5-deficient mice. There was no alteration in the number of Thy-1+/TCR V gamma 3+ dendritic cells in the epidermis of CD5-/- mice. Even though the latter finding speaks against a pivotal role of CD5 during the maturation of DETC, the described cell system may serve as a useful tool in further experiments aimed to clarify the function of the CD5 glycoprotein as well as the mechanism(s) regulating its expression.

Animals↗
PubMed1995

Immunologic host defense in melanoma: delineation of effector mechanisms involved and of strategies for the augmentation of their efficacy.

There exists substantial evidence that the immune system plays an important role in the prevention and control of cancer. This evidence includes 1) the occasional clinical observation of spontaneous tumor regression, 2) the correlation of this phenomenon with the presence of tumor-infiltrating lymphocytes, and 3) the in vitro demonstration of the specificity of tumor-infiltrating lymphocytes for the autologous tumor. Because of the only weak immunogenicity of and the occurrence of active immunosuppression by the cancer, this response often does not suffice to combat the neoplasm successfully. One strategy for amplifying the anti-tumor immune response is vaccination of patients or experimental animals with cancer cells, the immunogenicity of which has been enhanced by the introduction of genes encoding immunostimulatory molecules. Several investigators have shown that transfection of certain types of cancer cells with the interleukin-2 gene reduces their tumorigenicity and that immunization with interleukin-2-transduced cancer cells protects animals from challenge with a tumorigenic dose of wild-type cancer cells. We have recently established a murine melanoma model (M-3) and have used it to elucidate the mechanism by which interleukin-2-transfected cancer cells can induce protective immunity. We will demonstrate the following: 1) that the mechanisms leading to the loss of tumorigenicity of interleukin-2-expressing cancer cells are somewhat different from those leading to the rejection of wild-type cancer cells in immunized animals, 2) that immunity resides within both CD4- and CD8-positive T cells, and 3) that host antigen-presenting cells are probably important in the induction of this protective anti-tumor immunity.

Animals↗
PubMed1995

Immunoglobulin E-binding structures on antigen-presenting cells present in skin and blood.

In atopic individuals, cutaneous antigen-presenting cells (APC), i.e., Langerhans cells and dermal dendritic cells, frequently display anti-IgE reactivity. Although earlier observations suggested that this phenomenon results from the binding of (complexed) IgE to the low-affinity IgE receptor (Fc epsilon RII/CD23), we and others demonstrated recently that Langerhans cells, dermal dendritic cells, and peripheral blood monocytes from atopic individuals can bind monomeric IgE via the high-affinity receptor for IgE (Fc epsilon RI). These new observations re-stimulated investigations aiming to unravel the nature and functionality of the relevant in vivo IgE-binding moiety(-ies) on APC. New data demonstrate that Fc epsilon RI, both quantitatively and qualitatively, is the pivotal serum IgE-binding structure on APC of atopics and, even more important, that Fc epsilon RI on APC functions as an allergen-focusing molecule. Thus, it is likely that allergens may be more efficiently taken up, processed, and presented to T cells after targeting to APC via Fc epsilon RI as compared with allergen binding to APC in the conventional manner. In vivo, Fc epsilon RI-IgE-dependent allergen presentation may critically lower atopic individuals' threshold to mount allergen-specific T-cell responses. This would result in the perpetuation of allergen-specific IgE production (type I reactions) and perhaps even the occurrence of T-cell-mediated, delayed-type hypersensitivity reactions in allergen-exposed tissues.

Antigen-Presenting Cells↗
PubMed1994

Fetal skin-derived MHC class I+, MHC class II- dendritic cells stimulate MHC class I-restricted responses of unprimed CD8+ T cells.

Dendritic cells are very potent, if not the most effective, stimulator cells for the induction of primary T cell immune responses. We have established, from murine fetal skin, growth factor-dependent cell lines with a pronounced dendritic shape and a phenotype similar to that of fetal Langerhans cells (i.e., MHC class I+/II). Functionally, these lines induce a vigorous proliferation of allogeneic, but not syngeneic, CD8+ lymphocytes. T cell blasts thus generated are capable of lysing various target cells in an MHC class I-restricted fashion. Our contention that this skin cell-induced MHC class I-restricted activation of CD8+ lymphocytes occurs in the absence of CD4+ T cells is based on 1) the lack of FACS-detectable CD4+ T cells in the purified CD8+ T cell population, 2) the lack of reactivity of purified CD4+ T cells to MHC class I-disparate fetal skin cell lines, and 3) the inhibition of the fetal skin cell-induced MLR by anti-CD8/MHC class I, but not anti-CD4/MHC class II, mAb. Skin cell-induced activation of unprimed CD8+ T cells was found to be critically dependent on physical contact between stimulator and responder cells and the expression of the costimulatory molecule B7 on fetal skin cell lines. Lines described in this study may represent a powerful tool for studying the molecular events occurring in the induction of MHC class I-restricted primary immune responses, understanding their pathophysiologic role, and perhaps may prove useful for vaccination purposes against selected pathogens.

Animals↗
PubMed1994

T-cell receptor repertoire of lymphocytes infiltrating cutaneous melanoma is predominated by V alpha specificities present in T-cells of normal human skin.

Lymphocytes infiltrating solid tumors can be propagated in vitro with interleukin 2 and are then capable, as CD8+ cytotoxic T-cells, of specific lysis of autologous tumor targets in a class I-restricted manner. Since the specificity of these cells is determined by their T-cell receptor (TCR) configuration, the aim of this study was to delineate the TCR V alpha repertoire of tumor-infiltrating lymphocytes within 24 melanoma specimens and to compare these data with the TCR expression pattern of unaffected peritumoral and normal human skin. While lymphocytes within all skin specimens tested expressed a substantial, albeit limited, heterogeneity of V alpha specificities with an average number of 9.0 different V alpha gene segments, the V alpha repertoire within cutaneous melanoma lesions was significantly more restricted (mean of V alpha expression, 3.86; P < 0.001) with a predominance of only 3 V alpha families (V alpha 13, V alpha 15, and V alpha 16), all of which were also found to be expressed within normal skin. Collectively, the fact that the TCR V alpha repertoire in melanoma is skewed toward the predominance of only a few V alpha regions may be indicative of a limited number of melanoma-associated antigenic determinants being involved in the anti-tumor immune response.

Adult↗
PubMed1994

Expression of functional high affinity immunoglobulin E receptors (Fc epsilon RI) on monocytes of atopic individuals.

Suggestive evidence indicates that immunoglobulin E (IgE)-dependent activation of mononuclear phagocytes plays an important pathogenic role in allergic tissue inflammation. Prevailing opinion holds that low affinity IgE receptors are the relevant IgE-binding structures on monocytes/macrophages and that functional events occurring after cross-linking of membrane-bound IgE on these cells are mediated by these receptors. Here we demonstrate that peripheral blood monocytes can bind monomeric IgE via the high affinity IgE receptor (Fc epsilon RI) and that Fc epsilon RI expression on these cells is upregulated in atopic persons. Further, we demonstrate that, upon monocyte adherence to substrate, bridging of monocyte Fc epsilon RI is followed by cell activation. We propose that direct interaction of multivalent allergen with Fc epsilon RI(+)-bound IgE on mononuclear phagocytes results in cell signaling via Fc epsilon RI and that the biological consequences of this event may critically influence the outcome of allergic reactions.

Animals↗
PubMed1994

CD5- dendritic epidermal T cells are derived from CD5+ precursor cells.

Murine Thy-1+, TcR V gamma 3/V delta 1+ dendritic epidermal T cells (DETC) differ from most other T cell subsets by the absence of CD4 and CD8 antigens as well as the lack of CD5 expression. To see whether negativity for those antigens is an intrinsic feature of a given T cell population or if such triple-negative T cells go through a maturational stage where they express these antigens, we determined the phenotype of TcR V gamma 3+ fetal thymocytes which are the precursor cells of DETC. We found that TcR V gamma 3+ fetal thymocytes phenotypically differ from mature DETC in that they are CD5+, mostly CD8+ and partly CD4+. The injection of fetal thymic suspensions containing TcR V gamma 3+/CD5+ (but not TCR V gamma 3+/CD5-) thymocytes into Thy-1-disparate athymic nude mice resulted in the appearance of donor-type TcR V gamma 3+/CD5- dendritic cells in the recipients' epidermis, indicating that TcR V gamma 3+ thymocytes are indeed the precursors of CD5- DETC. Tracing CD5 expression on DETC precursors during their intrathymic maturation and their migration to the fetal skin, we found that (i) the earliest DETC precursor cells as defined by TcR V gamma 3 expression express high levels of CD5 antigen (day 15 of gestation), (ii) after day 16 of gestation 70% of TcR V gamma 3+ thymocytes express high and 30% express intermediate levels of CD5, (iii) TcR V gamma 3+ cells in the fetal blood express low levels of CD5, (iv) the first TcR V gamma 3+ cells entering the epidermis express very low levels of this antigen and (v) TcR V gamma 3+ epidermal cells later than day 19 of gestation are CD5-. A similar down-regulation of CD5 expression on DETC precursors was also noted when TcR V gamma 3+ cells were cultured in vitro. Even the addition of PMA and ionomycin, which up-regulates CD5 expression on TcR alpha/beta-bearing thymocytes and lymph node T cells, could not prevent CD5 down-regulation on DETC precursors. The described cell system may serve as a useful tool in further experiments aimed to clarify the function of the CD5 glycoprotein as well as the mechanism(s) regulating its expression.

Animals↗
PubMed1994

Immunohistological analysis of anti-melanoma host responses.

Various clinical and experimental observations point to the existence of an immunological host defense in cutaneous malignant melanoma. To identify the major effector mechanisms mediating the specific anti-tumor immune response, we examined 23 benign and neoplastic melanocytic lesions (3 nevi, 14 primary melanomas, and 3 cutaneous and 3 systemic metastases) by quantitative immunohistology, and correlated these results with the histopathological and clinical subtypes of malignant melanoma. Our analyses indicate that CD3+ T-cell receptor alpha/beta-expressing lymphocytes are the prevailing leukocyte subset in primary as well as secondary malignant melanoma. We further observed that in early lesions (< 0.75 mm) of superficial spreading melanoma the vast majority of tumor-infiltrating lymphocytes (TIL) belong to the CD4+ subset and frequently express CD45RA antigens. In more advanced tumors, the contribution of CD8+ TIL gradually increases, indicating that the quality of the anti-tumor immune response changes during the course of the disease. Finally, we found that a varying percentage of cutaneous TIL express the cutaneous leukocyte antigen which is defined by the monoclonal antibody HECA 452 and preferentially expressed by skin-seeking memory T cells. In contrast, extracutaneous melanoma metastases (liver, brain, ovary) were completely devoid of HECA 452-reactive lymphocytes. These findings suggest that lymphocytes infiltrating cutaneous melanomas belong to a memory/effector T-cell subset functionally associated with the skin.

Antibodies, Monoclonal↗
PubMed1994

T-cell receptor diversity in dendritic epidermal T cells in the rat.

The rat epidermis contains a population of dendritic CD3+ cells. For a better characterization of these cells and to investigate their relationship to epidermal lymphocytes of other species, we stained rat epidermal sheets using a variety of monoclonal antibodies against rat leukocyte differentiation antigens in an indirect immunofluorescence procedure. Additionally, we attempted to define their T-cell receptor (TCR) isotype at both the nucleic acid and protein level. Results obtained showed that the majority of the CD3+ dendritic epidermal cells are CD45+, CD2+, TCR alpha beta-, major histocompatibility complex class II-, Thy-1-, asialo GM1-, CD4-, CD5-, and CD8- lymphocytes. We further observed that, in contrast to the mouse system, the rat epidermis additionally harbors a small but distinctive portion of dendritic CD3+ cells that exhibit reactivity with an anti-pan TCR alpha beta monoclonal antibody. Our further finding that rat epidermal cells enriched for CD3+ lymphocytes express full-length C delta mRNA suggests that the vast majority of rat epidermal T cells carry surface-bound TCR gamma delta moieties. On the basis of these findings, one may speculate that the indigenous T-cell population of the epidermis is not necessarily programmed to uniformly express monomorphic TCR gamma delta molecules but, to effectively fulfill its role in host defense, is capable of adaptation to the specific challenges encountered by a given species.

Animals↗
PubMed1994

Immunomorphologic characterization of Fc epsilon RI-bearing cells within the human dermis.

Recently we reported that the high-affinity receptor for IgE, Fc epsilon RI, is constitutively expressed on normal epidermal Langerhans cells (LC) and on certain cells within the dermis. To study the nature of these cells we performed immunofluorescence double-labeling experiments using an anti-Fc epsilon RI reagent (MoAb 15-1) as well as monoclonal antibodies (MoAb) against leukocyte differentiation antigens expressed on LC, interdigitating cells and macrophages. Avidin-fluorescein isothiocyanate was used to distinguish mast cells. We found that dermal Fc epsilon RI+ cells are bone marrow derived (CD45+). Further, we found that a subset of 15-1+ dermal cells coexpresses antigens present on certain members of the LC/DC family: the majority of Fc epsilon RI+ cells reacted with MoAb anti-HLA-DR and RFD1, the latter recognizes an antigenic moiety on interdigitating cells, and a small subpopulation coexpressed CD1a. In reverse fashion, virtually all CD1a+ cells and most RFD1+ cells reacted with the anti-Fc epsilon RI reagent. Approximately one third of 15-1+ cells represented avidin-FITC+ mast cells whereas Fc epsilon RI expression was not detected on FXIIIa+ dermal dendrocytes or CD3+ lymphocytes. By immunoelectronmicroscopy, we found that perivascularly located 15-1-reactive cells exhibited pronounced dendrites, an indented nucleus, numerous mitochondria, and abundant endo-/lysosomal structures. However, Birbeck granules or granules specific for basophils or eosinophils were never detected in these cells. Collectively, our data suggest that the pool of dermal Fc epsilon RI+ cells consists mainly of cells of the LC (CD1a+)/DC(RFD1+) lineage and mast cells but does not include FXIIIa+ dermal macrophages.

Antibodies, Monoclonal↗
PubMed1994

Expression of monoclonal antibody HECA-452-defined E-selectin ligands on Langerhans cells in normal and diseased skin.

The cutaneous lymphocyte-associated antigen recognized by the monoclonal antibody HECA-452 has been thought to play a major role in the homing of memory T-cell subsets to the skin by virtue of its ability to bind to E-selectin of dermal microvascular endothelial cells. Considering that the homing of different leukocyte populations to the skin may involve similar mechanisms, we studied the expression of HECA-452-reactive molecules on CD1a+ epidermal Langerhans cells. Immunofluorescence double-labeling of cryostat sections and epidermal sheets of normal skin revealed HECA-452 immunoreactivity on a subpopulation of dermal and epidermal CD1a+ cells, whereas upon flow-cytometric analysis of epidermal single cell suspensions virtually all CD1a+ cells bound HECA-452 antibodies. We observed a marked upregulation of HECA-452-antigen expression on CD1a+ epidermal cells and a pronounced increase in the number of HECA-452+/CD1a+ dermal cells in lesional skin from inflammatory and neoplastic lymphocytic skin diseases, compared to normal skin. The molecule detected by the HECA-452 antibody on Langerhans cells is neuraminidase sensitive and contains a CD15 (LewisX) carbohydrate backbone. Because Langerhans cells react with the sialyl-LewisX-specific antibody CSLEX1, it is very likely that the HECA-452-reactive structure is or contains sialyl-LewisX. Our data are compatible with the view that i) resident epidermal Langerhans cells upregulate HECA-452-antigen expression due to the cytokine profile generated in the disease process or ii) that Langerhans cell precursors express HECA-452-antigens and show an enhanced immigration into lesional skin. The characterization of HECA-452+ cells in peripheral blood may not only clarify this issue but may also help to identify the still elusive Langerhans cell-precursor.

Antibodies, Monoclonal↗
PubMed1994

Epidermal langerhans cells of AIDS patients express HIV-1 regulatory and structural genes.

In this study we searched for the presence of human immunodeficiency virus (HIV-1)-spliced mRNA in epidermal cells from AIDS patients. Using reverse polymerase chain reaction and Southern hybridization we detected mRNA for HIV-1 regulatory (tat, rev, nef) and structural genes (env) in epidermal cells highly enriched for Langerhans cells, but not in Langerhans cell-depleted epidermal cells in two of three HIV-1-infected patients tested. In contrast to the expression of HIV-1 mRNA, T-cell-specific mRNA was readily detectable in all preparations of Langerhans cell-depleted but in only one preparation of Langerhans cell-enriched epidermal cells. This indicates that contaminating T cells are a very unlikely source of virus-specific mRNA in our epidermal cell preparations. Our data suggest that Langerhans cells are the main, if not the only, cell type infected with HIV-1 within the epidermis of AIDS patients and demonstrate that HIV-1 regulatory as well as structural genes are transcribed in these cells.

Acquired Immunodeficiency Syndrome↗
PubMed1994

A review on Fc epsilon RI on human epidermal Langerhans cells.

In order to show the surface expression of Fc epsilon RI on human epidermal Langerhans cells (LC), we examined monomeric IgE binding to LC. We demonstrated that the majority of epidermal LC were able to bind monomeric IgE. IgE binding to LC could neither be prevented by preincubation of the tissue with monoclonal antibodies (mAb) against either Fc epsilon RII/CD23 or Fc gamma RII/CD32, nor by the addition of lactose, but could be entirely abrogated by preincubation with the anti-Fc epsilon RI mAb 15-1. These data show that epidermal LC express Fc epsilon RI molecules.

Humans↗
PubMed1994

Genetic modification of cells by receptor-mediated adenovirus-augmented gene delivery: a new approach for immunotherapy of cancer.

Most concepts of gene therapy of cancer are based on the generation of an enhanced immune response against the cancer by means of vaccination with gene-modified cancer cells. We have investigated the applicability of a new gene transfer technique which uses the receptor-mediated endocytosis pathway and the endosome disruption activity of adenovirus for the generation of a cancer vaccine consisting of interleukin-2 (IL-2)-transfected, irradiated murine melanoma cells (clone M-3). This technique resulted in very high IL-2 expression (in the range of 30,000 Units IL-2/10(6) cells/24 hrs) in the transfected cells without the need to selection of stably expressing cell clones. We found that this high IL-2 expression of the melanoma cells correlates with high efficacy of the vaccine. Immunization of animals with this vaccine elicits a systemic T-cell-mediated immune response which protects from tumor development after implantation of highly tumorigenic doses of wild-type melanoma cells.

Adenoviridae↗