Interaction between gamma delta T cells and B cells regulating IgG production.

Despite profound knowledge about the molecular structure of the gamma delta T-cell receptor (TCR), the physiological function of gamma delta T cells remains enigmatic. Participation of these cells in complex immune reactions, however, is suggested by the appearance of gamma delta T cells in sites of infectious and autoimmune-induced inflammations. Only a few in vitro models of gamma delta T-cell stimulation have been established: besides a reactivity in the presence of microbial ligands, human gamma delta T cells proliferate upon in vitro challenge with cells from an allogeneic B-lymphoblastic cell line (B-LCL).

Stereologic parameters of nuclear ultrastructure as markers of the steroid receptor status in breast cancer epithelium.

The nuclear specific surface density (Sv/Vv), mean nuclear area (A) and numerical density (Nv) of tumor cell nuclei from 30 primary invasive female breast carcinomas with known concentrations of estrogen and progesterone receptors (ER and PR), were morphometrically analyzed at the ultrastructural level. It was found that carcinomas with concordant positive ER and PR status contained significantly higher number of smaller nuclei per volume unit of epithelium (mean Nv = 1.5 x 10(6) mm-3, mean A = 27 microns 2), than carcinomas of negative concordant receptor status (mean Nv = 0.

Individual differences between nuclear parameters of normal and malignant breast epithelium.

This study attempts to estimate the status of estrogen and progesterone receptors (ER and PR) in tissue of invasive breast carcinoma. Identical stereologic parameters of nuclear ultrastructure, of normal and malignant epithelium in the same breast were compared. Samples of normal and malignant epithelium of 30 surgically amputated breasts were analysed morphometrically.

Expression of candidate cell death genes in cell lines during apoptosis.

We have cloned mouse candidate cell death genes RP-2 and nedd2 and used Northern blot analysis to study their expression in a growth-factor-dependent cell line (FDC-P1) that can be induced to undergo apoptosis by growth factor withdrawal and in a thymoma line (ST4) that undergoes apoptosis when irradiated. RP-2 was expressed in FDC-P1 cells even when not undergoing apoptosis, and mRNA levels did not increase when apoptosis was induced by growth factor withdrawal. FDC-P1 cells expressed two genes that are similar to the Caenorhabditis elegans cell death gene ced-3: a 3.

Neuropeptides in the human penis: an immunohistochemical study.

In the present study, the distribution of neuropeptides in the human penis is demonstrated by immunohistochemistry (IHC). IHC screening detected a complex network of nerve fibers containing vasoactive intestinal polypeptide (VIP), peptide histidine-methionine (PHM), prepro-VIP (111-122), neuropeptide Y (NPY), C-flanking peptide of NPY (C-PON), calcitonin gene-related peptide, substance P, and galanin immunoreactivities. Special attention was also given to the recently isolated, VIP-related lizard peptide helospectin, which could also be detected in neuronal elements in the penis.

DNA ploidy in primary fallopian-tube carcinoma using image cytometry.

Determination of DNA ploidy in 61 samples of primary Fallopian-tube carcinoma (FTC) by use of image cytometry is presented. Of these samples, 48 showed an aneuploid DNA content. Patients with euploid DNA content showed a median survival time of 33.

[Autonomic and peptidergic innervation of the human larynx].

Autonomic and peptidergic innervation of the human larynx (vocal cords, ventricular folds, epiglottis, subglottic region and recurrent nerves) was studied by application of single and double immunocytochemistry and radioimmunoassay. In all tissues investigated, immunoreactivities for a variety of regulatory peptides were detected and included vasoactive intestinal polypeptide (VIP), peptide histidine methionine (PHM), helospectin, neuropeptide Y (NPY), C-flanking peptide of NPY (C-PON), calcitonin gene-related peptide (CGRP), substance P and neurokinin A. In the recurrent nerves, only a few peptide-immunoreactive nerve fibers were found.

NPY- and C-PON-immunoreactive substances in the central nervous system of Helix pomatia.

The distribution of neuropeptide-tyrosin (NPY)- and C-flanking peptide of neuropeptide-tyrosine (C-PON)-immunoreactivities in the central nervous system of the pulmonate gastropod, Helix pomatia, was investigated. NPY- and C-PON-like substances were localized in neuronal somata and neuntes, but were not co-localized within the same cells. NPY-immunoreactive substances were also found in endocrine/paracrine like cells located in the epineurium.

Regulatory peptides in the human larynx and recurrent nerves.

We studied the peptide-innervation of the human larynx (vocal cord, ventricular folds, epiglottis, subglottic region and the recurrent nerves) using immunocytochemical and radioimmunological methods. In the tissues of the larynx investigated, the following regulatory peptides were detected: vasoactive intestinal polypeptide (VIP), peptide histidine methionine (PHM), helospectin, neuropeptide Y (NPY), C-flanking peptide of NPY (C-PON), calcitonin gene-related peptide (CGRP), substance P and neurokinin. In the recurrent nerves only small numbers of peptide-immunoreactive nerve fibers were found.

Autonomic and peptidergic innervation of human nasal mucosa.

The localization and distribution of vasoactive intestinal polypeptide (VIP), peptide histidine methionine (PHM), the novel peptide helospectin, neuropeptide tyrosine (NPY) and its C-flanking peptide (C-PON), substance P and calcitonin gene-related peptide (CGRP) were studied in the middle and inferior turbinate of the human nose using sensitive immunocytochemical and radioimmunological methods. For light microscopy, double immunofluorescence and immunogold-silver staining methods were applied. Ultrastructural immunoelectronmicroscopy was performed using a pre-embedding method.

Application of silver acetate autometallography and gold-silver staining methods for in situ DNA hybridization.

In situ hybridization using biotinylated DNA probes has become an important tool in histopathology. It is well known that the sensitivity of the methods used to demonstrate viral DNA in formalin-fixed and paraffin-embedded specimen depends strongly on the detection system used. In the present study, an optimized in situ DNA hybridization protocol was combined with four different approaches of gold-silver staining methods.

V delta 1+ subset of human gamma delta T cells responds to ligands expressed by EBV-infected Burkitt lymphoma cells and transformed B lymphocytes.

Human gamma delta T cells of peripheral blood can be divided in two groups in terms of their TCR as well as their behavior upon in vitro stimulation. The major subset expresses the TCR V-segments V gamma 9 and V delta 2 and proliferates in response to ligands revealed by various microorganisms, and the cell line Daudi in addition. The minor group is less homogenous on the gamma-chain but is almost completely identified by mAb against the V delta 1 segment; there is no ligand known to promote growth of these cells.

Detection of single HPV copies in SiHa cells by in situ polymerase chain reaction (in situ PCR) combined with immunoperoxidase and immunogold-silver staining (IGSS) techniques.

Detection of HPV by means of in situ hybridization techniques may often present problems if the number of HPV copies is too small, especially when using nonradioactive detection systems. A new way of amplifying extremely small amounts of virus DNA copies is the polymerase chain reaction (PCR), which is mostly used in vitro. In this study, we present a method for in situ PCR combined with immunogold-silver staining (IGSS) and immunoperoxidase (IMP) methods which allows the detection of single copies of HPV-DNA in SiHa cells.

[Peptidergic innervation of human salivary glands (parotid gland and submandibular gland)].

Immunocytochemistry and a radioimmunoassay were used to investigate the existence and distributions of various regulatory peptide immunoreactivities (ir) in human submandibular and parotid glands. Numerous nerve fibers containing vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM), or neuropeptide tyrosine (NPY) and C-flanking peptide of NPY (CPON)-ir were found in close proximity to acini, ducts and blood vessels. Only a few calcitonin gene-related peptide (CGRP)- and substance P (SP)-ir nerve fibers could be demonstrated and were mainly localized around blood vessels and ducts.

Opportunist mycobacteria express ligands that stimulate production of human V gamma 9V delta 2 T lymphocytes.

Human gamma delta T cells are known to respond at high frequencies to pathogenic mycobacteria. Here we show that opportunistic strains of mycobacteria share with pathogenic mycobacteria the ability to trigger at high frequencies human V gamma 9V delta 2 T-cell-receptor-positive T lymphocytes. Stimulating ligands were present in part in a low-molecular-weight fraction of lysates from opportunistic mycobacteria, as has been found for pathogenic strains.

Distribution and co-localization of immunoreactive helospectin with vasoactive intestinal polypeptide and peptide histidine methionine in human nasal mucosa, soft palate and larynx.

Regulatory peptide immunoreactivities reported in the upper respiratory system of man include vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM), which are co-localized in a network of fine varicose nerve fibers. The present study was undertaken to examine the possible occurrence and distribution of the recently described VIP-like peptide helospectin. Double immunofluorescence labelling showed that helospectin is co-localized with VIP and PHM.

Neuropeptides in human salivary (submandibular and parotid) glands.

The existence, distribution and density of various neuropeptides in human submandibular and parotid glands were investigated using immunocytochemistry and radioimmunoassay. Numerous nerve fibers containing vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM), or neuropeptide Y (NPY) and C-flanking peptide of NPY (CPON) immunoreactivities (ir) were found in close association to acini, ducts and blood vessels. Only few calcitonin gene-related peptide (CGRP)- and substance P (SP)-ir nerve fibers could be demonstrated, mainly localized around blood vessels and ducts.

Plasmodium falciparum merozoites primarily stimulate the V gamma 9 subset of human gamma/delta T cells.

Peripheral blood-derived T cells from six unprimed caucasian donors were tested for the in vitro reactivity to Plasmodium falciparum merozoites (PFM). Without exception vigorous proliferative responses were observed within the donors tested. The frequency of PFM-reactive T cells ranged from 1/150-1/300.

Paragangliomas: neuroendocrine features and cytometric DNA distribution patterns. A clinico-pathological study of 22 cases.

Paragangliomas from 22 patients with extraadrenal tumours of this type were studied. Neuroendocrine features were examined using immunohistochemical techniques. Twenty-two antisera raised against neuroendocrine "markers", regulatory peptides, serotonin and intermediate filament proteins were studied in this group and cytometric DNA assessments were made by means of image cytometry.

Regulatory peptides and general neuroendocrine markers in human nasal mucosa, soft palate and larynx.

Various peptide immunoreactivities in the respiratory system have been reported, indicating complex physiological mechanisms. There is only little information on the upper respiratory system of man. The present study was carried out to demonstrate regulatory peptides in the nasal mucosa, larynx (vocal cords and ventricular folds) and soft palate of man using highly efficient immunocytochemical methods.

Gastrin- and somatostatin-immunoreactive cells of the antral mucosa in patients with duodenal or gastric ulcers. An immunocytochemical study.

Gastrin- and somatostatin-immunoreactive cells in biopsies taken from the prepyloric portion of the antrum from 15 patients with duodenal ulcer, 16 patients with gastric ulcer, and a control group of 19 patients without histopathological alterations of the antral mucosa were studied using peroxidase anti-peroxidase and immunogold-silver staining methods in combination with morphometry. Numerical densities and sizes (immunoreactive areas) of the cells demonstrated were measured and compared between all three groups. Gastrin- and somatostatin-immunoreactive cells were located most frequently in the lower midzone of the gastric crypts.

[Application of silver acetate autometallography in histopathology: a new detection method for use in immunogold silver staining, lectin histochemistry and in situ hybridization].

Modern histochemical techniques allow the specific detection of tissue constituents in situ. Routinely formalin fixed, paraffin embedded tissues may present problems to the pathologist since destruction of substances can lead to false negative results. Immunogold-silver staining (IGSS) can be a way to overcome some of these problems.