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The significance of indeterminate screening antibody test for human immunodeficiency virus (HIV) serology is still difficult to evaluate, especially in low-risk populations. One hundred twenty-seven blood donors with an initially reactive screening test for HIV antibodies were enrolled in this study. The sera of 95 of these blood donors were reactive on repetition of the test, and none had detectable circulating p24 antigen. Western blot (WB) analysis of the repeatedly reactive sera was as follows: 9 positive, 31 indeterminate, and 55 negative. One of the blood donors with indeterminate WB later presented a seroconversion. On subsequent control 3 to 12 months later, the sera from donors with indeterminate or negative WB did not present any parameters that may indicate a seroconversion. DNA was purified from citrated blood collected from the 127 blood donors at the time of the initial antibody screening. Five micrograms of each DNA sample corresponding to 7 x 10(5) nucleated white blood cells was amplified by polymerase chain reaction (PCR) in the presence of oligonucleotides (primers) corresponding to a highly conserved segment of the pol gene. The detection of amplified DNA was achieved by dot blot and Southern blot using appropriate 32P-labeled oligonucleotides. Ten DNA samples were positive, 9 corresponded to blood donors with a positive HIV serology, and 1 to the blood donor who later presented a seroconversion. These results confirm the sensitivity of the PCR for the diagnosis of HIV infection; they also suggest that repetition of the serology at 3- to 12-month intervals is a valuable procedure for the control of HIV infection status in blood donors.

Development of sensitization in rats monoassocialed with Actinomyces viscosus Ny 1 was estimated by titrating serum agglutinins and by measuring the anamnestic, in vitro response of spleen T lymphocytes to two antigen fractions. These parameters of the immune response were compared with the development of alveolar bone loss using germfree rats as controls. At 46 days of age, 19 days after monoassociation, we observed (i) a mean serum antibody litre of (log2) 3.6, (ii) considerable antigen-dependent T-cell proliferation, and (iii) statistically significant amounts of alveolar bone loss. Thereafter, sensitization to A. viscosus and total bone loss increased cuncomitantly until the end of the experiment at 88 days of age. However, the rate of bone loss decreased with time at some sites, suggesting that negative feedback mechanisms by suppressor cells enable bone remodeling. These results demonstrate that, in experimental periodontal disease elicited by A. viscosus. the immune response to dental plaque bacteria and bone loss occur simultaneously.

The T-cell triggered effector systems have been emphasized as the most important agents mediating periodontal disease in recent years. It was therefore tempting to investigate whether T-cell suppression with cyclosporin A (CS-A) had an effect on the establishment and progression of the disease in rats monoassociated with Actinomyces viscosus Ny 1. The experiment was carried out with two treatments (16 rats each) in two isolators. In one of the isolators CS-A was added to the high sucrose diet 2000 aiming at a dosage of 15 mg per day and kg bodyweight. In the second isolator (control) the same diet without CS-A was fed. At day 25, 38, 52 and 65 four animals from each treatment were removed from the isolators. Dental plaque and alveolar bone loss were evaluated. The cellular composition of blood and histological changes of periodontal lesions on a light and electron microscopic level were monitored. In addition antibody titers and the T-cell suppressing effect of individual serum samples of all animals were determined. Despite clearcut treatment differences in antibody titers as well as an interference by the sera of CS-A treated animals with the Con A dependent lymphocyte activation, bone loss was observed in both groups. Haematological and histological investigation revealed only minor differences between the two treatments. It is concluded that periodontal disease in monoinfected rats seems to be the result of a multitude of pathomechanisms and not to be strictly correlated with T-cell dependent hypersensitivity.

We describe a surface determinant shared by human monocytes and cells of the megakaryocytic axis that has been identified using a mouse monoclonal antibody. This monocyte-platelet antigen (MPA) is expressed on all (greater than 99%) of peripheral blood monocytes, platelets, and megakaryocytes. It is also expressed weakly on the monocytic cell line U937 and the promyelocytic line HL60 and is present on cells from 3 of 4 AML patients examined. It is absent from polymorphonuclear leukocytes, T and B lymphocytes, erythrocytes, and a panel of hematopoietic cell lines. MPA is stripped from monocyte membranes with pronase and is reexpressed overnight. The determinant is carried on a noncovalently linked biomolecular complex with molecular weights of 93,000 and 135,000.

Peripheral T lymphocytes from RIC-Sprague-Dawley rats primed in vivo several months earlier with Actinomyces viscosus Ny 1 reacted with a strong proliferative response upon antigenic challenge in vitro. Two different antigen preparations from A. viscosus Ny 1, a broken cell supernatant (BCS) and an extracellular fraction (EXC) gave maximum responses as measured by the uptake of [3H]thymidine at a concentration of 10 microgram at day 4 of culture. Phytohaemagglutinin (PHA) used as a control provoked a maximum proliferation in the same order of magnitude. T lymphocytes from unprimed, germfree and conventional animals showed a similar stimulation to PHA, but only a marginal reaction to BCS, and were not at all activated by EXC. Evidence is thus presented for efficient removal of cells responding non-specifically to B-cell mitogens.

We describe a surface determinant shared by human monocytes and cells of the megakaryocytic axis that has been identified using a mouse monoclonal antibody. This monocyte-platelet antigen (MPA) is expressed on all (greater than 99%) of peripheral blood monocytes, platelets, and megakaryocytes. It is also expressed weakly on the monocytic cell line U937 and the promyelocytic line HL60 and is present on cells from 3 of 4 AML patients examined. It is absent from polymorphonuclear leukocytes, T and B lymphocytes, erythrocytes, and a panel of hematopoietic cell lines. MPA is stripped from monocyte membranes with pronase and is reexpressed overnight. The determinant is carried on a noncovalently linked biomolecular complex with molecular weights of 93,000 and 135,000.

Numerous pathomechanisms have been proposed as relevant in periodontal disease (1). The T-cell triggered effector systems have been especially emphasized as the most important mediators of this particular chronic inflammation (2–4). It was therefore tempting to investigate whether T-cell suppression by a suitable antilymphocytic agent had a major influence on the establishment and progression of the disease.

Partially purified glucosyltransferases (GTF) isolated from Streptococcus mutans OMZ 176 and respective rabbit antisera were used to study enzyme-antibody interactions. A comparison between sensitive serological techniques and a functional inhibition test based on a radioenzyme assay demonstrated that the latter test system was the only one that discriminated between different antisera. Positive reactions in high dilutions in the former test systems were explained by the involvement of non-GTF contaminants and/or antibodies against enzyme regions distant to the catalytic site. The minute cross-reactions between two enzyme fractions and the respective antisera in the functional inhibition test indicated that the two immunogens contained mainly GTF that differed in the structure of their catalytic region. Control rabbit sera, rat oral fluid, and insoluble and soluble glucans considerably activated the GTF eluted with a 0.5 M phosphate buffer from hydroxapatite. It is suggested that these enzymes had additional binding sites for macromolecules inherent to rabbit sera and rat oral fluid, respectively, and that the observed increase in enzyme activity was due to a more stable enzyme conformation. Possibly the stimulation of GTF by the soluble glucan fraction was caused by a primer and/or acceptor function; however, this was not the case of the insoluble glucan. A stable complex was formed in the absence of the enzyme substrate, sucrose, the activity of which was not readily enhanced. It is concluded that the GTF of strain OMZ 176 are composed of multiple, multi-reactive molecules that enable these enzymes to act as cross-linking agents.

Cell-mediated cytotoxicity against syngeneic fetal rat fibroblasts that require in vitro exposure of effector cells to Actinomyces viscosus Ny1 fractions was investigated by measuring the uptake of radioactivity by fibroblasts during a 2-h pulse with [14C]aminoisobutyric acid after 1 to 3 days of coculture with splenic effector cells. By using splenocytes from inbred RIC-Sprague-Dawley rats as effector cells and syngeneic embryonic rat fibroblasts as target cells, strong cell-mediated cytotoxicity dependent on the in vitro exposure to an A. viscosus Ny1 fraction was observed, but only within a small range of effector-to-target cell ratios (3:1 to 10:1). Concanavalin A and lipopolysaccharide from Escherichia coli induced a comparable cytotoxicity, indicating that the effect might be connected with the mitogenic activity of the A. viscosus NY1 fraction. Splenocytes from rats immunized with A. viscosus Ny1 and from control rats induced similar levels of cytotoxicity in 72-h cytotoxicity assays. In shorter assays (24 h), however, splenocytes from immune animals induced low cytotoxicity, which was, however, significantly higher than that induced by splenocytes from control animals. We conclude that both antigen- and mitogen-dependent cell mediated effector mechanisms are operative in this system and that the two normally overlapping effects can be experimentally separated. This new system describes a fibroblast impairment in the presence of splenocytes and bacterial components and may provide a useful model for studying pathogenic mechanisms operative in periodontal disease.

Protein-coated silica, a macrophage toxin, was used to assess the requirement for accessory cells in the induction of an in vitro proliferative response to (i) antigens from Actinomyces viscosus and (ii) the mitogens conconavalin A (Con A) and phytohaemagglutinin (PHA). T cells were obtained from RIC-Sprague-Dawley rats primed in vivo with A. viscosus Nyl by splenectomy and filtering the spleen cell suspensions through Degalan Ig-anti-IgG columns. In the presence of 100 microgram silica/ml during 4 days of culture, the proliferative response of T lymphocytes was not diminished. In contrast, when the T cells were precultured with silica for 24 h, washed, and subsequently cultured with the antigen fractions, antigen-induced proliferation was abolished. This procedure, however, had no influence on mitogen-induced proliferation was abolished. This procedure, however, had no influence on mitogen-induced T-cell activation. It is therefore concluded that the antigen-dependent anamnestic in vitro response (but not activation by mitogens) of rat T lymphocytes needs help from silica-sensitive macrophages.