IL-35, an anti-inflammatory cytokine which expands CD4+CD25+ Treg Cells.

Interleukin 12 (IL 12) p35/p40 is a heterodimeric cytokine which plays a critical role in inflammation, immunity and tissue proliferation, and also plays a relevant function in T helper (Th) cell polarization and Th1 T-cell differentiation. IL-12 family members, IL-12p70, IL-23, IL-27 and IL-35, play an important role in influencing helper T-cell differentiation. EBV-induced gene 3 can be associated with the p35 subunit of IL-12 to form the EBI3/p35 heterodimer, also called IL-35. It has been shown that IL-35 has biological activity and able to expand CD4+CD25+ Treg cells, suppress the proliferation of CD4+CD25- effector cells and inhibit Th17 cell polarization. IL-35 has been shown to be constitutively expressed by regulatory T (Treg) cells CD4(+)CD25(+)Foxp3(+) and suggested to contribute to their suppressive activity. IL-35 is a crucial mediator which provokes CD4+CD25+ T cell proliferation and IL-10 generation, another well-known anti-inflammatory cytokine, along with TGFbeta cytokine. These studies suggest that IL-35, together with other successfully discovered cytokine inhibitors, represents a new potential therapeutic cytokine for chronic inflammation, autoimmunity and other immunological disorders.

Stimulation of CCL2 (MCP-1) and CCL2 mRNA by substance P in LAD2 human mast cells.

Chemokines are cytokines with chemotactic properties on inflammatory cells and other cell types. Chemokine (C-C motif) ligand 2 (CCL2), which is also called monocyte chemotactic protein 1 (MCP-1), is a potent chemotactic molecule that attracts lymphocytes, monocytes, mast cells, and memory T cells, but not neutrophils. CCL2/MCP-1 represents a link between the activation of monocytes, lymphocytes, basophils, mast cells, and eosinophils in inflammatory disorders, such as the late-phase allergic reaction. This C-C chemokine also plays a role in regulating Th-cell cytokine production and leukocyte trafficking. Laboratory of allergic diseases (LAD) cells is the first reported human mast cell line that closely resembles a primary culture of CD34+-derived human mast cells. These cells were cultured in vitro and treated with different concentrations of substance P (SP) for the production of CCL2/MCP-1. We used calcium ionophore as a positive control for stimulating transcription and translation of CCL2/MCP-1. The stimulation of SP on CCL2/MCP-1 was statistically significant (P < 0.05) compared with the control (untreated cells). In this study, we determined the expression and secretion of CCL2/MCP-1 from SP-activated LAD2 human mast cells in vitro. The levels of CCL2/MCP-1 from SP-activated LAD2 human mast cells were higher at 10 microM and at 18 h incubation compared with controls. This effect was also revealed on CCL2/MCP-1 messenger RNA (mRNA) expression, as determined by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. Our data suggest that SP is an important neurotransmitter that can stimulate the chemokine CCL2, which plays a fundamental role in inflammation by recruiting inflammatory cells to specific cites.

Expression of Co-stimulatory molecules on langerhans cells in lesional epidermis of human atopic dermatitis.

Langerhans cells (LC) are immature dendritic cells (DC) present in the skin epithelium. To understand the molecular and cellular mechanisms governing the inflammatory reaction in atopic dermatitis (AD), the expression of the LC specific marker CD1a, a member of major histocompatibility (MHC)-like glycoproteins, and the co-stimulatory molecules CD80 and CD86, expressed on functionally mature dendritic cells, were counted in lesional biopsies and normal epidermis by an immunohistochemical method. CD1a specific staining was observed in both normal and AD lesion specimens. CD80 and CD86 positive cells with morphological characteristics of the LC were found in lesional AD epidermis, suggesting a high level of functional maturity of these cells and their involvement in chronic inflammatory disease.

Role of flavonoids and vitamins in cancer.

Flavonoids are naturally occurring polyphenolic plant compounds that are capable of inhibiting histamine and cytokine release from several cells. Many studies suggest that flavonoids are anticancer agents with an apoptotic effect on tumor cells. Studies with animal tumour models have found vitamin deficiency to enhance susceptibility to chemical carcinogenesis and large doses of anti-oxidant vitamins and flavonoids to inhibit carcinogenesis. In some studies flavonoids and/or vitamins were found to reduce the predisposition to develop tumours in animals and humans. In conclusion, in this review we describe the role of flavonoids and vitamins in cancer.

Inhibitory effect of quercetin on tryptase and MCP-1 chemokine release, and histidine decarboxylase mRNA transcription by human mast cell-1 cell line.

Mast cells are important in reactions of allergic disease and are also involved in a variety of neuroinflammatory diseases. Mast cells can be immunologically activated by IgE through their Fc receptors, as well as by neuropeptides and cytokines to secrete mediators. Here we used a human mast cell-1 (HMC-1) cell line cultured and treated with a physiological activator, anti-IgE, and a nonphysiological activator, calcium ionophore A23187, for tryptase and MCP-1 generation and transcription of histidine decarboxylase. We used quercetin, a potent antioxidant, cytoprotective and anti-inflammatory compound capable of inhibiting histamine and some cytokines released from several cell types, as an inhibitor of immunological and nonimmunological stimulus for mast cells. In this study quercetin inhibits, in a dose-response manner, tryptase and MCP-1. Moreover, using RT-PCR quercetin inhibited the transcription of histidine decarboxylase, the rate-limiting enzyme responsible for the generation of histamine from histidine, and MCP-1. Our data suggest that quercetin is an important and good candidate for reducing the release of pro-inflammatory mast cell mediators activated by physiological and nonphysiological stimulators.

Relevance of plant lectins in human cell biology and immunology.

Protein-carbohydrate interactions are used for intercellular communication. Mammalian cells are known to bear a variety of glycoconjugates. Lectins, first discovered in plants, are proteins which can specifically bind carbohydrates. Given the high affinity of plant lectins for carbohydrates, they have always been important as molecular tools in the identification, purification and stimulation of specific glycoproteins on human cells. Lectins have provided important clues to the repertoire of carbohydrate structures in animal cells. The discovery of plant lectins gave a great impulse to modern glycobiology. They represent important biochemical reagents for numerous applications in the biomedical field and in research. Sequence determinations and structural characterization helped to understand the mechanism of action in many biologic systems. Plant lectins have been fundamental in human immunological studies because some of them are mitogenic/activating to lymphocytes. Understanding the molecular basis of lectin-carbohydrate interactions and of the intracellular signalling evoked holds promise for the design of novel drugs for the treatment of infectious, inflammatory and malignant diseases. It may also be of help for the structural and functional investigation of glycoconjugates and their changes during physiological and pathological processes.

Inhibitory effects of cadmium on peripheral blood mononuclear cell proliferation and cytokine release are reversed by zinc and selenium salts.

Zinc (Zn) and selenium (Se) exert regulatory activities on immune functions, while cadmium (Cd) is an immunotoxic agent. The object of this study was to detect effects of 10(-4), 10(-5), and 10(-6) M Cd sulphate, Zn sulphate, and sodium selenite, and their combinations on human peripheral blood mononuclear cell (PBMC) proliferation and IFN-gamma and TNF-alpha production. Only 10(-5) M Zn sulphate significantly enhanced spontaneous PBMC proliferation, which was unaffected by the other salts. At 10(-4) and 10(-5) M, Cd sulphate exerted a dose-response inhibitory action on phytohemagglutinin- (PHA-) stimulated PBMC proliferation and cytokine release, while 10(-4) M and 10(-5) M Zn sulphate and 10(-5) M sodium selenite induced a stimulatory effect on both proliferation and cytokine release; 10(-4) M sodium selenite enhanced only the PBMC proliferation; at 10(-6) M, none of the salts changed the PHA-stimulated immune activity. Moreover, 10(-4) and 10(-5) M Zn and 10(-5) M Se strongly upregulated IFN-gamma (a Th1 cytokine) release, even in presence of 10(-5) M Cd, and reduced the inhibitory effects of Cd on PBMC proliferation and TNF-alpha release. This study confirms that Zn and Se both strongly enhance cytokine release induced by mitogenic stimulation, showing also that Zn acts with a broader range of concentrations than Se. This suggests that dietary excess of Se may not have beneficial effects.

Different effects of platinum, palladium, and rhodium salts on lymphocyte proliferation and cytokine release.

The effects of graded concentrations of Pt, Pd, and Rh salts on spontaneous and PHA-stimulated peripheral blood mononuclear cell (PBMC) proliferation and IFN-gamma, TNF-alpha, and IL-5 release were the focus of this study. Spontaneous PBMC proliferation was inhibited by all 10(-4) M salts (with the exception of PtCl2), while it was enhanced by 10(-5) M PtCl2 as well as by 10(-5) and 10(-6) M (NH4)2[RhCl6] and RhCl3 (but not by 10(-7) M salts). Pt, Pd, and Rh compounds showed similar effects on PHA-stimulated PBMC proliferation and cytokine release; however, the effects on IFN-gamma release were stronger. Thus, 10(-4) and 10(-5) M (NH4)2[PtCl6] and 10(-4) M (NH4)2[PtCl4] inhibited the PHA-stimulated immune activity; 10(-4) M PtCl2 did not exert activity, while 10(-6) M (NH4)2[PtCl6] and 10(-5) and 10(-6) M (NH4)2[PtCl4] and PtCl2 enhanced PBMC proliferation and/or cytokine release. (NH4)2[PdCl6] showed stronger dose-related inhibitory effects (present also at 10(-7) M concentration) on PHA-stimulated proliferation and cytokine release than (NH4)2[PdCl4], PdCl2, or Rh salts; the inhibitory activity of (NH4)2[RhCl6] was slightly higher than that of RhCl3. In conclusion, this study shows that: (a) the immune capacity of Pt, Pd, and Rh depends on speciation; (b) low concentrations of Pt salts stimulate spontaneous and PHA-stimulated immune responses; (c) the in vitro activity of Pd compounds (which are only inhibitory) is higher than that of Pt and Rh salts. These findings are consistent with the observations that sensitization and allergic contact dermatitis in response to Pd are increased in the general population, although the roles of cross-sensitization to Pd and Ni are difficult to determine.

Differential production of RANTES and MCP-1 in synovial fluid from the inflamed human knee.

Synovial production of chemokines may play an important role in the recruitment of phagocytic leukocytes during inflammation. MCP-1, as well as RANTES mediate many different inflammatory diseases and are important in the recruitment of diverse leukocytes. We set out to study the different production of MCP-1 and RANTES in three different inflammatory conditions of the knee: arthrosynovitis, mechanical trauma, and hyperuricemia. In this study we evaluated if in each pathological condition mentioned above, there was a prevalence in production of one chemokine over the other. ELISA method was used to determine base production of the chemokines in the synovial fluid, serum and in supernatants from activated inflammatory cells. RANTES and MCP-1 messenger RNA (mRNA) was measured by semi-quantitative RT-PCR. Protein expression was detected by Western blot analysis. The synovial fluid cells from the knee of patients affected with arthrosynovitis, trauma, and hyperuricemia, expressed RANTES and MCP-1 and RANTES was produced in higher quantities than MCP-1 in all three pathological conditions. In patients treated with non-steroidal antiinflammatory drugs (NSAD) and dexamethasone, the levels of the two chemokines was reduced in serum and in synovial fluid. In addition, the synovial fluid cells from these patients released less RANTES and MCP-1 when compared to untreated patients. We conclude that in arthrosynovitis, trauma and hyperuricemia, RANTES and MCP-1 are both expressed and RANTES is produced in higher quantities. The fact that these chemokines are found in the three inflammatory diseases suggests that RANTES and MCP-1 are not specific to these inflammatory diseases, however they play a key role in inflammation by recruiting mononuclear leukocytes in the inflamed knee joint.