Piotr Kuna

Chemokines in seasonal allergic rhinitis

Monocyte chemotactic and activating factor/monocyte chemoattractant protein-1 (MCAF/MCP-1), RANTES, and macrophage inflammatory protein (MIP)-1 alpha are chemokines known to activate basophils (MCAF/RANTES) and eosinophils (RANTES/MIP-1 alpha). IL-8 inhibits MCAF-induced histamine release from basophils. We questioned whether a relationship exists between the levels of these chemokines and various inflammatory mediators released from mast cells, eosinophils, and basophils as assessed in nasal secretions obtained from patients during the allergy season and out of season. Samples were assessed for MCAF/MCP-1, RANTES, MIP-1 alpha, IL-8, histamine, tryptase and eosinophil cationic protein (ECP) in three subject groups: subjects with allergic rhinitis (n = 18), atopic subjects without rhinitis (n = 9), and healthy individuals (n = 6). Statistically significant differences were apparent only in the subjects with symptoms as follows. MCAF/MCP-1 increased during the season from 336 +/- 47 pg/ml to 829 +/- 137 pg/ml (p < 0.001), whereas IL-8 decreased from a baseline of 1932 +/- 335 pg/ml to 1070 +/- 202 pg/ml (p < 0.028). The ratio of IL-8 to MCAF/MCP-1 decreased during the symptomatic season from the baseline of 6.66 +/- 1.06 seen during winter to 1.3 +/- 0.22 during ragweed season (p < 0.001). Histamine increased from 6.3 +/- 1.5 to 89 +/- 15.5 ng/ml (p < 0.001), ECP increased from 20.6 +/- 6.4 to 237.1 +/- 50.2 ng/ml (p < 0.001), and tryptase increased from 2.34 +/- 0.6 to 9.7 +/- 2.3 U/ml (p < 0.001). Most samples did not have detectable quantities of MIP-1 alpha or RANTES. We also found a correlation between the level of MCAF/MCP-1 and IL-8 and the level of histamine or IL-8 and ECP. Our results suggest that the chemokines MCAF/MCP-1 and IL-8 may participate in the pathogenesis of allergic rhinitis, contributing to the attraction of the proinflammatory cells and mediator release, which might be very important during the late phase of the allergic reaction. Furthermore, the ratio of certain chemokines, such as MCAF/MCP-1 and IL-8 may reflect the magnitude of the reaction, as does the presence of histamine and ECP.

Histamine releasing factors and cytokine-dependent activation of basophils and mast cells

Publisher Summary This chapter describes the various types of histamine releasing factors (HRFs), discusses their interaction with basophils or mast cells, and speculates regarding their role in inflammation in general, and in allergic diseases in particular. The release of histamine from human basophils or mast cells is typically initiated by the interaction of antigen with surface-bound immunoglobulin (IgE) antibody. Histamine releasing factors are defined as products of activated cells that interact with basophils and mast cells to cause the release of histamine. Two general types of HRFs is described, one whose function appears to be dependent upon cell surface IgE and others that are IgE independent. A variety of reports confirmed that lymphoid cells produce such a factor, and HRF-like activities are described as products of T lymphocytes, mixed mononuclear cells, alveolar macrophages, platelets, B lymphocytes, and neutrophils. Such activity also appeared to be present in nasal washings of allergic subjects and in antigen-challenged cutaneous late-phase reactions. Histamine releasing factors cause a noncytotoxic granule exocytosis from basophils when assessed by electron microscopy and cause histamine release as well as release of lipid mediators such as leukotriene C4.

Chemokines and the allergic response

Abstract The β subfamily of chemokines contains‐cytokine‐like factors which are chemotactic for human basophils and eosinophils. They also stimulate these cells to secrete pro‐inflammatory substances such as histamine or eosinophil cationic protein. MCAF/MCP‐1, MCP‐2, MCP‐3, RANTES and MIP‐lα all attract and stimulate basophils; MCP‐I and MCP‐3 are the most potent. RANTES, MCP‐3 and to a lesser degree MIP‐Ia are chemotactic factors and activators of eosinophils. Cytokines such as IL3, IL5 and GM CSF can augment the responses of these cells to the various chemokines and function as primers. These substances may have particular importance as mediators of allergic inflammation, particularly the late phase component of the response.

Effect of neurotropin® on the activation of the plasma kallikrein-kinin system

Bradykinin (BK), an important mediator of allergic reactions and pain induction, is released by the activation of the plasma kallikrein-kinin (K-K) cascade. Neurotropin is a biological material obtained from inflamed rabbit skin inoculated with vaccinia virus and is widely used clinically in Japan as an effective agent for these disorders. Since its mechanism of action is not clearly known, we have investigated the effects of Neurotropin on the human plasma K-K system. In dextran sulfate-activated plasma, Neurotropin inhibited the formation of BK, the cleavage of high molecular weight kininogen (HK) and the formation of kallikrein-C1 inhibitor and activated coagulation factor XII (FXIIa)-C1 inhibitor complexes. Experiments using purified enzyme of the K-K cascade indicated that Neurotropin inhibited surface-mediated activation of coagulation factor XII (FXII) and the activation of prekallikrein by FXIIa. Neurotropin also inhibited the binding of FXII and HK to the activating surface. These data suggest that the ameliorating effects of Neurotropin in allergic disorders and pain syndromes may be related to this ability to inhibit activation of the K-K cascade and consequently the formation of BK.

Chemokines of the α, β-subclass inhibit human basophils' responsiveness to monocyte chemotactic and activating factor/monocyte chemoattractant protein-1

Abstract Monocyte chemotactic and activating factor (MCAF) is the most potent cytokine that activates basophils to release histamine. The response of human basophils to either simultaneous or sequential addition of the chemokines RANTES, macrophage inflammatory protein (MIP)-1α, MIP-1β, platelet factor (PF)4, connective tissue activating peptide III (CTAP-III), interleukin (IL)-8, and inflammatory protein (IP)-10 on MCAF-induced histamine release was studied. Simultaneous addition of MCAF and any of the chemokines studied evoked an augmented response as measured by histamine release, whereas preincubation of leukocytes or purified basophils (80%) with these chemokines decreased MCAF-induced histamine release in a dose-dependent manner. Histamine release by anti-IgE remained unchanged. When tested at 5 × 10 -9 mol/L, the decrease in histamine release by RANTES was 69.2% ± 3.5%, by MIP-1α 48.8% ± 3.1%, by MIP-1β 42.9% ± 3.1%, by PF4 56.5% ± 2.9%, by IL-8 41.2% ± 2.2, by CTAP III 27% ± 4.4%, and by IP-10 15.3% ± 2.6%. The peak inhibition of histamine release by the chemokines was reached within 10 minutes of preincubation with basophils and remained unchanged thereafter. Washing basophils after preincubation with chemokines abolished the inhibition, with the exception of desensitization by low concentrations of MCAF. With the exclusion of MCAF and RANTES, none of the chemokines (at the concentration range of 5 × 10 -8 to 5 × 10 -11 ) induced significant (>10% above spontaneous) histamine release from basophils. Preincubation of basophils with C5a (5 × 10 -10 mol/L) did not affect histamine release, whereas preincubation with granulocyte-macrophage colony-stimulating factor (10 ng/ml) or IL-5 (10 ng/ml) enhanced MCAF-induced histamine release by 121.8% ± 10.1% and 108% ± 10.8%, respectively. We have therefore characterized RANTES, MIP-1α, MIP-1β, CTAP III, PF4, IL-8, and IP-10 as inhibitors of MCAF-induced histamine release. Although the results are consistent with receptor blockade, the α and β chemokines appear to interact with separate receptors linked to G proteins; thus, a mechanism of receptor class desensitization is proposed. Interaction of this group of cytokines at the site of allergic inflammation may modulate a function of basophils to initiate, augment, or inhibit histamine release. (J ALLERGY CLIN IMMUNOL 1995;95:574-86.)

Histamine-Releasing Factors

Histamine-releasing factors (HRF) are cell-derived products which cause histamine release from basophils and/or mast cells. We have isolated HRF from human mononuclear cells and platelets and have purified 3 molecular species having molecular weights of 8-10, 15-17 and 35-41 kilodaltons (kDa). We prepared monoclonal antibodies to the 8- to 10-kDa form and have isolated it by affinity chromatography. A broad band was seen upon sodium dodecyl sulfate gel electrophoresis in 15% gels as well as immunoblotting, and the band was divided into an upper and a lower half. Amino acid sequence analysis of the upper half indicated that it is closely homologous to connective-tissue activating peptide III (CTAP III). The lower half also aligned with CTAP III beginning with amino acid 16; thus, proteolysis and occurred removing the N-terminal 15 amino acids. This corresponds to neutrophil-activating peptide 2. Both appear to be active on basophils with a dose-response between 250 ng up to 10 micrograms. Although interleukin-3 and granulocyte/macrophage-colony-stimulating factor have similar histamine-releasing capability at lower effective concentrations, they do not account for HRF activity in mononuclear cell/platelet supernatants, and the 15- to 17 and 40- to 41-kDa moieties appear to be unique gene products unrelated to previously described cytokines.

Further characterization of histamine releasing chemokines present in fractionated supernatants derived from human mononuclear cells

Background Histamine releasing factors (HRF) are members of the β chemokine family of cytokines and have been characterized using recombinant proteins. Mononuelcar cell and/or platelet supernatants have been shown to contain HRF and the initial void peak obtained using Mono Q anion exchange chromatography possesses such activity, as do two later peaks eluted from the column.

Histamine releasing factors

Histamine-releasing factors (HRF) are cell-derived products which cause histamine release from basophils and/or mast cells. We have isolated HRF from human mononuclear cells and platelets and have purified 3 molecular species having molecular weights of 8-10, 15-17 and 35-41 kilodaltons (kDa). We prepared monoclonal antibodies to the 8- to 10-kDa form and have isolated it by affinity chromatography. A broad band was seen upon sodium dodecyl sulfate gel electrophoresis in 15% gels as well as immunoblotting, and the band was divided into an upper and a lower half. Amino acid sequence analysis of the upper half indicated that it is closely homologous to connective-tissue activating peptide III (CTAP III). The lower half also aligned with CTAP III beginning with amino acid 16; thus, proteolysis and occurred removing the N-terminal 15 amino acids. This corresponds to neutrophil-activating peptide 2. Both appear to be active on basophils with a dose-response between 250 ng up to 10 micrograms. Although interleukin-3 and granulocyte/macrophage-colony-stimulating factor have similar histamine-releasing capability at lower effective concentrations, they do not account for HRF activity in mononuclear cell/platelet supernatants, and the 15- to 17 and 40- to 41-kDa moieties appear to be unique gene products unrelated to previously described cytokines.

Relationship of histamine-releasing factors to the human intercrine/chemokine group of cytokine-like molecules

Histamine Releasing Factors have been characterized as a product of human mononuclear cells and platelets. MCAF/MCP-1, a monocyte-derived product is the most potent one described which acts rapidly (w