Substance P stimulates release of RANKL via COX-2 expression in human dental pulp cells

Objective: Our previous study found that substance P (SP), a sensory neuropeptide, was expressed in the dental pulp of rats during experimental tooth movement. We examined the effects of SP on the production of prostaglandin (PG) E₂ and the receptor activator of nuclear factor- B ligand (RANKL) by human dental pulp fi broblast-like (HDPF) cells. Materials and methods: SP was added to cultured HDPF cells at concentrations ranging from of 10⁻⁴ to 10⁻¹² mol/L. PGE₂ and soluble RANKL (sRANKL) levels were determined using enzyme-linked immunosorbent assay kits. Gene expression was confi rmed by RT-PCR analysis. Pit formation assays using dentin slices were carried out to examine the effect of SP on osteoclastogenesis. Results: The levels of PGE₂ and sRANKL increased in the presence of SP, though the increases were greater in the experimental groups in both a time- and concentration-dependent manner, and the increase of RANKL was partially mediated by PGE₂ . The gene expression of cyclooxygenase (COX)-2 and RANKL was up-regulated, and conditioned medium samples obtained from HDPF cells treated with SP induced bone resorption. Conclusions: SP stimulated the production of PGE₂ and RANKL, and promoted bone resorption. Therefore, SP may be involved in pulpal inflammation and root resorption during orthodontic tooth movement. Received 11 April 2005; returned for revision 27 July 2005; returned for final revision 20 September 2005; accepted by M. Katori 2 November 2005     

Langerhans cells in human lung tumours: an immunohistological study

In an immunocytochemical study of 41 human lung tumours we have shown that Langerhans cells can be reliably identified using the anti-CD1 monoclonal antibody NA1/34. Langerhans cells are present in all the main varieties of human lung tumour although they are infrequent in both small cell carcinoma and carcinoid tumour. There is considerable variation in numbers of Langerhans cells in both adenocarcinomas and squamous cell carcinomas. In this study tumours were divided into those with high numbers of Langerhans cells (greater than 2 per high power field) and those with low numbers (less than 2 per high power field). Analysing these results against patient survival showed a markedly worse survival in those tumours with a high number of Langerhans cells for all the tumours as a single group and for squamous cell carcinoma as a single entity.     

The A427 anchorage-independence assay as a screening tool to identify potential chemopreventive agents

An in vitro model for screening potential chemopreventive agents using inhibition of anchorage-independent growth of a human lung tumor cell line, A427, is described. A427 cells were selected for the model development, as they are known to be tumorigenic in animals, can grow in soft agarose, and their growth can be inhibited by a well-known chemopreventive agent, 13-cis-retinoic acid. Cells are plated on agarose, allowed to develop colonies for 28 days, the stained colonies are enumerated, and the inhibition of spontaneous colony formation measured. A cytotoxicity test is used concurrently with anchorage independent assay for measuring the relative survival of cells to ensure that any observed inhibition of anchorage independent growth is due to the biological activity of the chemopreventive agents, as it uses human cells as substrates rendering the efficacy data feasible for direct extrapolation to humans.     

Passive immunization: a method of enhancing the immune response against antigen mixtures

Antigenic competition is known to be a widespread phenomenon when using crude extracts of antigens (e.g., Escherichia coli cytoplasmic proteins) for immunization. This non-specific form of immune suppression can be partially overcome by passive immunization with antibodies against dominant antigens (which are the suppressive molecules) before injection of the antigenic mixture. Blocking these immunodominant antigens or antigenic determinants by a passively administered antibody permits antibody responses against hitherto weakly or non-immunogenic molecules.     

Transplantation of cultured fetal human adrenal chromaffin cells to rat brain

Adrenal chromaffin cells, obtained from a therapeutically aborted human fetus of about 11 weeks gestation, were cultured for 3 weeks in vitro and then transplanted to the striatum of rats. Transplanted cells became established through strands of tissue growing into the host striatum. No signs of inflammation or rejection were observed up to the time of sacrifice one month post-transplantation. Histofluorescence examination of the implanted areas showed many clusters of cells having an intensely positive catecholamine fluorescence with some of the cells developing conspicuous processes. This study, showing survival of cultured human adrenal chromaffin cells transplanted into rat brain tissue, might indicate the feasibility of using cultured human material for future human neuronal transplantation studies as a therapeutic measure.     

J chain-positive cells in bursectomized chicks

Using embryonic chickens treated with testosterone propionate, the effects of congenital absence of the bursa of Fabricius determined by the frequency of J chain-positive cells was examined in the spleen, thymus and bone marrow at the embryonic and newly hatched stages. J chain-positive cells in the chicks without bursa were reduced in the spleen. No differences in the numbers of the cells were detected in the thymus and bone marrow. These results imply that removal of the bursa of Fabricius cannot entirely prevent the generation of J chain-positive B cells. Furthermore, these results partly suggest the important role of the bone marrow in the proliferation of some J chain-positive cells in chicks without bursa.     

Are primed CD4+ T lymphocytes different from unprimed cells?

Primed and unprimed lymphocytes are usually classified as separate subsets of cells, based on phenotypic and functional distinctions. In the case of CD4⁺ T lymphocytes, primed cells are thought to proliferate more vigorously, quickly and easily, and to release a different profile of cytokines, than their naive equivalent. However, most of these data were obtained from studies in which populations of lymphocytes were compared before and after antigenic stimulation, and therefore did not distinguish between the effects resulting from the clonal expansion of specific precursor cells within such populations and those due to cell differentiation per se. We have investigated the contribution of precursor cell frequency to some of the functional changes observed in populations of CD4⁺ T cells following antigenic stimulation, using approaches in which antigen-specific precursor frequencies are high in both primary and secondary stimulations: mixed leukocyte reaction responses and cells from αβ T cell receptor transgenic mice. Our data suggest that when equivalent numbers of antigen-specific naive and previously primed CD4⁺ responder T cells are compared, there is no difference in their potency to proliferate but only the previously activated subset can generate cytokines such as interferon-γ.     

Influence of Mycobacterium tuberculosis on differential activation of helper T-cells

Host defence against tuberculosis infection involves T-lymphocyte mediated cellular immune responses. In this study we assessed T-cell activation by studying the early signal transduction events and production of cytokines by human CD4+ T-cells. The study constituted of five groups of subjects: (a) untreated acid fast bacilli (AFB)+ve TB patients who have not started anti-tuberculosis therapy (ATT) [New]; (b) patients who have taken ATT for two months [2T]; (c) patients who have taken ATT for six months [6T]; (d) mantoux positive healthy controls [T+ve]; (e) mantoux negative healthy controls [T-ve]. We found that mantoux positive healthy controls produced significantly higher levels of IP3, intracellular Ca2+ and presented increased PKC activity when CD4+ T-cells were stimulated with M. tuberculosis H37Rv cell lysate as compared to mantoux negative controls. Furthermore, decreased expression of CD54 (ICAM-1) and reduced [Ca2+]i were seen in TB patients as compared to T+ve healthy controls. TB patients showed significantly lower levels of IL-2 and IFNgamma and higher levels of IL-4 as compared to normal healthy controls, suggesting a diminished Th1 response. Thus, the reciprocal changes in cytokines, reduced [Ca2+]i levels, and CD54 expression in patients imply phenotype shifting of Th precursors to Th2 type in TB patients.     

Acute optic neuritis: a virological study in relation to multiple sclerosis.

Previous ultrastructural examination of peripheral blood lymphocytes revealed the presence of intranuclear filamentous structures in multiple sclerosis (MS) and in some optic neuritis (ON) patients. The present investigation was undertaken in the attempt to correlate the presence of such structures with the etiology of ON and MS and possibly to demonstrate the viral origin of the filaments. Suitable virological and serological techniques were used to detect and isolate infectious agents from peripheral blood samples and body excretions of 12 monosymptomatic ON patients at their first acute attack. Nevertheless, any efforts to demonstrate the presence of a virus in these patients have been unsuccessful: no evidence of active viral infection was obtained by serological studies of serum and cerebrospinal fluid samples, nor could viral antigens or inclusions be observed by immunofluorescence and cytochemical analysis. Negative results were also obtained from studies performed in parallel on MS patients and various controls. The significance of the failure to isolate infectious agents from either ON and MS patients is discussed.