The centrosome moves out of a nuclear indentation in human lymphocytes upon activation.

Using a beta-tubulin specific antibody, centrosomes were labeled in paraformaldehyde fixed human lymphocytes. Cells were kept in suspension to preserve the three-dimensional (3D) morphology as much as possible. The centrosome was generally identified as the focus of the microtubule array.

The nuclear position of pericentromeric DNA of chromosome 11 appears to be random in G0 and non-random in G1 human lymphocytes.

The nuclear topography of pericentromeric DNA of chromosome 11 was analyzed in G0 (nonstimulated) and G1 [phytohemagglutinin (PHA) stimulated] human lymphocytes by confocal microscopy. In addition to the nuclear center, the centrosome was used as a second point of reference in the three-dimensional (3D) analysis. Pericentromeric DNA of chromosome 11 and the centrosome were labeled using a combination of fluorescent in situ hybridization (FISH) and immunofluorescence.

Plasmid rescue of an oncogenic sequence containing a mouse mammary tumor virus gene.

Plasmid rescue was performed on an oncogenically transformed cell line established by transfection of NIH/3T3 cells with normal mouse DNA and plasmids containing a murine leukemia virus long terminal repeat, and a selectable marker. One of the rescued plasmids contained newly acquired DNA 3,500 basepairs in length. This sequence was present in several extra copies in the parental cell line.

The mitogen-induced generation of interferon-gamma producing cells in cultures of rhesus monkey peripheral blood mononuclear cells is age-dependent.

An enzyme-linked immunoabsorbent spot (ELISPOT) assay has been developed for measuring the frequency of interferon-gamma (IFN-gamma) producing cells in rhesus monkeys. Aged monkeys revealed, upon mitogenic stimulation, a significantly higher percentage of IFN-gamma secreting peripheral blood mononuclear cells (PBMC) compared to young animals. No correlation was found between the frequency of IFN-gamma producing PBMC and the mitogen-driven proliferation, indicating that in rhesus monkeys no direct correlation exists between these two activation parameters.

Occurrence of a soluble nonspecific suppressor factor in the serum early after birth.

A nonspecific suppressor factor has been identified in serum of newborn rats and calves. This factor, designated SUF-s, was shown to interfere--across species barriers--with lymphocyte responses in vitro and in vivo. SUF-s interferes in vitro with T- and B-cell proliferation induced by different mitogens and IL-2.

Resistance to collagen-induced arthritis in rats and rhesus monkeys after immunization with attenuated type II collagen.

Immunization of susceptible rodent or primate species with type II collagen (b-CII) from bovine origin induces type II collagen-induced arthritis (CIA). The disease is characterized as a systemic polyarthritis associated with humoral and cellular autoimmunity to CII and shares similarity with human arthritic diseases. The objective of this study was to develop a procedure for induction of resistance to CIA in animals, which possess a certain major histocompatibility complex phenotype that makes them prone to develop CIA (susceptible).

Major histocompatibility complex class II DQ diversity in rhesus macaques.

By the use of restriction fragment length polymorphism analysis 10 Taq I fragments could be identified for the MhcMamu-DQA1 region. A strong correlation exists between the occurrence of Mamu-DQA1/Taq I fragments and Mamu-DQA1 allelic sequence variation. Most restriction fragments correspond with a unique Mamu-DQA1 allele, with one exception being the Taq I 4.

In vivo transfer and expression of the lacZ gene in the mouse lung.

In vivo gene transfer by lipofection was studied in the mouse lung to develop a gene therapy protocol for disorders in which the lung is affected, such as cystic fibrosis. The bacterial lacZ gene encoding beta-galactosidase was used as a reporter, and the X-GAL staining procedure was optimized for cryostat sections of mouse lung. Three to five days after intratracheal instillation of a lacZ DNA-liposome mixture, lacZ expression was shown in a high percentage of airway epithelium cells.

Mhc-DQB repertoire variation in hominoid and Old World primate species.

Comparison of 87 distinct Mhc-DQB sequences, obtained from 13 primate species, demonstrates that five out of eight trans-species Mhc-DQB allele lineages are at least 30 million years old and predate divergence of hominoid and Old World primate species. One lineage may be much older because its members are not only traced back in higher primates, but also are present in a New World primate species. Comparing Mhc-DQB repertoire variation in distinct species, allows one to pinpoint when certain polymorphisms were lost or gained in primate evolution.

Endocrine profile during acquisition of free-choice alcohol drinking in rhesus monkeys; treatment with desglycinamide-(Arg8)-vasopressin.

This study concerns the effect of spontaneous acquisition of alcohol drinking in rhesus monkeys on plasma levels of beta-endorphin, ACTH, prolactin, cortisol and testosterone. Twelve monkeys had free-choice access to water and two ethanol/water solutions (1%, 2%, v/v) for 4 weeks. During the first 2 weeks, six monkeys were injected (i.

Resistance to collagen-induced arthritis in a nonhuman primate species maps to the major histocompatibility complex class I region.

Type II collagen-induced arthritis (CIA) is an experimentally inducible autoimmune disorder that is, just like several forms of human arthritis, influenced by a genetic background. Immunization of young rhesus monkeys (Macaca mulatta) with type II collagen (CII) induced CIA in about 70% of the animals. One major histocompatibility complex (MHC) class I allele was present only in young animals resistant to CIA and absent in arthritic animals.

Low doses of morphine reduce voluntary alcohol consumption in rhesus monkeys.

Experimental opioid modulation has been found to influence the consumption of alcohol in animals. Whereas it has generally been agreed upon that opiate antagonists reduce alcohol consumption, the results with opiate agonists are less consistent. The present study reports on the effect of low doses of morphine in 8 adult male rhesus monkeys that had a free choice in drinking water, a 16% and a 32% ethanol/water solution, (a) during continuous ad libitum access (Experiment I), and (b) after 2 days of alcohol abstinence (Experiment II).

Assessment of the inhibitory effect of immunosuppressive agents on rat T cell interferon-gamma production using an ELISPOT assay.

The immunospot (ELISPOT) assay has proven to be an efficient and sensitive method for the enumeration of single cells secreting antibodies or cytokines. Here we show that the generation of interferon-gamma producing cells (IFN-gamma pc) in rat spleen cell cultures stimulated with concanavalin A (ConA) is dose-dependently inhibited by a wide variety of immunosuppressants such as cyclosporin A, FK506, hydrocortisone, dexamethasone, azathioprine and ART-18, a monoclonal antibody (mAb) with established immunosuppressive activity in organ transplantation and autoimmunity. The minimal inhibitory concentration (m.

Acquired resistance to type II collagen-induced arthritis in rhesus monkeys is reflected by a T cell low-responsiveness to the antigen.

Ten out of 14 rhesus monkeys developed arthritis after a single immunization with bovine type II collagen (B-CII). In contrast to primary resistant monkeys, arthritic animals showed a B-CII specific T cell proliferation during the induction phase of the disease. All surviving animals showed a full remission of the disease.

Remission induction of adjuvant arthritis in rats by total body irradiation and autologous bone marrow transplantation.

Following the demonstration that adjuvant arthritis in rats can be cured with total body irradiation (TBI) and allogeneic or syngeneic bone marrow, the efficacy of autologous bone marrow was investigated in the experiments reported here. Bone marrow from arthritic rats, harvested at the same time that the recipients were irradiated, and real autologous bone marrow were found to be similarly effective as bone marrow grafts from naive syngeneic donors. Sublethal TBI with lower doses was less effective, but the highest tolerated doses of 8 Gy approached the effect of 9 Gy and bone marrow rescue.

Immunomodulation by quinones. A model for the use of quinones in the treatment of inflammation.

The therapeutic application of quinones in areas other than oncology, such as in chronic inflammation, has been proposed. However, because of the adverse side-effects on the function and vitality of almost all investigated cell types the therapeutical margin is small. The thiol-conjugating capacity of quinones may, however, be applied to reduce the tissue-damaging effects of stimulated neutrophils.

Development and characterisation of a cyclophosphamide resistant variant of the BNML rat model for acute myelocytic leukaemia.

A cyclophosphamide resistant subline (BNML/CPR) was developed in vivo in the BN rat acute myelocytic leukaemia (BNML) model. Full resistance was achieved after in vivo exposure of leukaemic animals to cyclophosphamide with, in total, 15 intraperitoneal injections of 100 mg/kg. The CPR line was cross-resistant to ifosfamide, but less so to mafosfamide.

The importance of non-human primates for preclinical testing of immunosuppressive monoclonal antibodies.

Monoclonal antibodies (MAb) specific for lymphocyte markers can be considered as very specific immunomodulating drugs for treatment of allograft rejection and autoimmune diseases. Although the selection of potentially useful specificities of MAb can be made in rodents, human specific MAb can only be evaluated in man or a closely related species in which these human specific MAb are equally reactive. Because of the restricted reactivity of human specific MAb, non-human primates are the only available species for efficacy and safety studies.

Antiarthritic activity of the newly developed neutrophil oxidative burst antagonist apocynin.

The plant-phenol 4-hydroxy-3-methoxyacetophenone (trivial name apocynin) is a strong inhibitor of neutrophil superoxide anion (O2-) release in vitro. In vitro the inhibitory effect of apocynin is restricted to cells with the capacity to release peroxidase and reactive oxygen species (ROS). Peroxidase deficient cells are insensitive to apocynin.