Histamine-releasing Factor Activity Research Articles

Secretion of Translationally Controlled Tumor Protein/Histamine Releasing Factor and Its Relevance to Parasitic Infections

Allergic diseases, including atopic dermatitis, allergic rhinitis, and asthma, develop when the immune system is hypersensitized by specific allergens. Translationally controlled tumor protein (TCTP), also known histaminereleasing factor (HRF) because of its cytokine-like activity, mediates late phase reaction leading to allergy and chronic inflammatory diseases in the human. TCTP exhibits HRF activity, when it is released from the cells and becomes dimerized under the inflammatory conditions. It has been shown that TCTP is secreted from cells by a TSAP6-mediated, exosomal route, and exported by H,K-ATPase-mediated process. TCTP is released from various parasitic organisms during parasitic infections. Secreted TCTP is implicated in allergic immune responses to parasites, in the pathogenesis of parasitic infections and also in the evasion of host’s immune activity for parasite survival. This review, briefly collates the current information on the secretion of TCTP/HRF by parasitic species and the biological and clinical implications of such release in parasitic diseases such as malaria.

Cellular Function and Regulation of the Translationally Controlled Tumour Protein TCTP

The ‘translationally controlled tumour protein’ TCTP was originally discovered 30 years ago by researchers interested in proteins regulated at the translational level. Cloning and sequencing confirmed the conservation of this protein among all eukaryotic kingdoms, but did not reveal any functional clue, and TCTP was listed in the databases as a ‘family’ of its own. The functional characterisation of this protein extended over more than a decade, leading to a plethora of individual functions and interactions that have been ascribed to this protein. A major addition to the functional characterisation of TCTP was the identification in 1995 of its histamine releasing factor (HRF) activity in allergic conditions, which for the first time described an extracellular activity for TCTP in human disease. This triggered a host of additional publications aimed at characterising this HRF activity, which are discussed in other articles of this issue. Another milestone in the elucidation of TCTP's function was the demonstration of its anti-apoptotic activity in 2001. Evidence is also accumulating for a role of TCTP in the cell cycle and in early development. This article provides an overview of the main cellular activities of TCTP. The second part will summarise our current knowledge on the mechanisms involved in regulating intracellular TCTP levels.

Studies on the association between immunoglobulin E autoreactivity and immunoglobulin E-dependent histamine-releasing factors.

It has been reported that serum immunoglobulin E (IgE) from certain atopic patients can sensitize basophils to release histamine in response to IgE-dependent histamine-releasing factors (HRFs). It has also been shown that patients suffering from severe forms of atopy may contain IgE autoantibodies. It was investigated whether HRF-responsive sera contained IgE autoantibodies and if there was an association between IgE autoreactivity and IgE-dependent responsiveness to HRF. The presence of HRF-responsive IgE (IgE+) in serum of patients with respiratory atopy was determined by stimulating stripped human basophils sensitized by serum with peripheral blood mononuclear cell (PBMC)-derived HRF, and measuring the release of histamine. In parallel, these sera were screened for the presence of IgE autoantibodies to nitrocellulose-blotted human cellular extracts. The capacity of IgE autoantigen-containing preparations to induce histamine release was tested in the stripped basophil assay. Eleven out of 52 sera contained IgE autoantibodies to blotted cellular extracts of human PBMCs or of the human epithelial cell line A431. No significant association was found between IgE autoreactivity and IgE-dependent responsiveness to HRF: 7/26 IgE+ sera contained IgE to human cellular extracts, and 4/26 of the sera without IgE+ did also. IgE autoantigen-containing extracts did not induce histamine release of appropriately sensitized basophils. By size-exclusion chromatography it was shown that a 32 000 MW autoantigen eluted in the >55 000 MW fraction, which indicates that this protein forms polymers or complexes with other macromolecules. This might explain the discrepancy between binding and histamine-releasing activity. A 20 000 MW IgE-defined autoantigen cross-reacted with a shrimp allergen. Our results indicate that IgE-reactivity to immunoblotted human protein and IgE-dependent HRF activity are distinct entities that may co-occur in atopic patients.

Characterization of histamine releasing factors in diisocyanate-induced occupational asthma

Immunologic mechanisms contributing to diisocyanate-induced occupational asthma (OA) are poorly defined. There is a relatively low incidence of diisocyanate-specific IgE antibody responses. The frequent occurrence of delayed onset asthmatic responses in workers with diisocyanate asthma suggests a role for cellular immune mechanisms. We have shown in vitro production of antigen-specific mononuclear cell-derived histamine releasing factors (HRF) by peripheral blood mononuclear cells (PBMCs) of workers with OA. Monocyte chemoattractant protein-1 (MCP-1) and RANTES (acronym for “regulated on activation normal T expressed and secreted”) are chemokines found in PBMC supernatants that express HRF activity. Diisocyanate-exposed workers were tested for diisocyanate antigen-stimulated enhancement of HRF, MCP-1, and RANTES production in supernatants of PBMCs and for serum specific IgE and IgG antibody levels to diisocyanate antigens bound to human serum albumin (HSA). PBMCs of workers with diisocyanate OA showed significantly increased production of antigen-specific HRF activity and MCP-1 ( >300 ng/ml) compared to diisocyanate-exposed asymptomatic workers ( P < 0.05). Antigen-stimulated enhancement of MCP-1 mRNA was demonstrated by reverse-transcription PCR. RANTES mRNA and chemokine secretion (< 1 ng/ ml) was also demonstrated in PBMCs, but did not show antigen enhancement in OA workers. Hapten specificity for the diisocyanate chemical to which a patient had been exposed was demonstrated for HRF enhancement and for IgG antibody reactions, but not for IgE reactions. HRF production was demonstrated in PBMC subpopulations, including lymphocytes and purified T cells. OA subjects showed increased CD8 + cells by immunofluorescence (mean CD4 +:CD8 + = 1.2 ± 0.2). The results suggest that diisocyanate antigen enhancement of HRF and MCP-1 production are stimulated by hapten-specific T cell reactions. Since a weak association has been found between IgE antibody synthesis and induction of diisocyanate OA, the role of T cell cytokines and chemokines in the pathogenesis of OA requires further investigation.

Effects of systemic corticosteroids on cutaneous histamine secretion and histamine‐releasing factor in patients with chronic idiopathic urticaria

Enhanced skin mast cell releasability of histamine, increased production of histamine releasing factor (HRF), and cutaneous inflammatory process are the hallmarks of chronic idiopathic urticaria (CU). Although H1-antihistamines are known to alleviate the symptoms effectively in most cases, systemic corticosteroids (CS) are given in more resistant patients. Their mode of action remains a matter of controversy. In the present study, the effects of a 7-day course of CS or placebo on histamine content and HRF production in non-lesional skin of 19 CU patients were examined. Using the skin chamber technique, HRF production and histamine content were assessed in normal-appearing skin of patients with CU over a 2-h observation period. Those two parameters were measured before and after treatment, in a double-blind fashion. No significant changes occurred in any parameters after placebo treatment. In contrast with this, significant decrease of HRF activity was observed after 1 week of oral methylprednisolone while no change was documented for histamine secretion. These data suggest that CS therapy improves symptoms of CU in association with a decreased production of HRF in uninvolved skin.

HRF of 30 kDa: evidence for active synthesis.

A variety of mediators are involved in the pathophysiology of a number of clinical conditions. Among them histamine-releasing factors (HRF) act as secretagogue for basophils and mast cells to cause cell degranulation and histamine release. Studies on the kinetics of HRF production by activated mononuclear cells (MNC) have suggested an active synthesis of this cytokine. Using a [35S]-metabolic cell labeling method, we first analyzed the capacity of MNC from patients with allergic rhinitis to ragweed to actively synthesize HRF. We then found that stimulation of MNC with allergen promoted a synthesis of large quantities of proteins, including a protein of about 30 kDa. Secondly, we developed a method for the enrichment of HRF molecules from crude supernatants of ragweed-stimulated MNC using ion exchange and gel filtration chromatography. We observed that the yield of HRF activity is seen just before the chymotrypsinogen marker. The estimated molecular weight was 30-35 kDa.

Characterization of histamine-releasing activity: Role of cytokines and IgE heterogeneity

Histamine-releasing factors (HRFs) are a group of cytokines that cause histamine release (HR) from basophils and mast cells. The concept of the priming effect of cytokines and the heterogeneity of IgE involved in the HRF-induced HR have been emphasized in recent years. In this study, we performed a series of experiments to elucidate the above-mentioned hypotheses. The stock HRF were obtained by stimulating mononuclear cells (MNC) with phytohemagglutinin (PHA). Maximal activity was observed 36 hr after culture. By gel filtration, HRF was eluted with a peak activity ranging from 12 to 18 KD. A large portion (75%) of HRF activity could be neutralized by a combination of antibodies against interleukin 1 (IL-1), IL-3, IL-8, granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha). The stimulation of basophils with 100 ng/ml each of IL-3, IL-6, IL-7, GM-CSF, or TNF-alpha alone caused 10% HR; however, when the cells were pretreated with 10 ng/ml of either IL-3, IL-6, IL-7, IL-8, TNF-alpha, or GM-CSF and then stimulated with anti-IgE, a marked increase in HR was regularly observed. The combination of 100 ng/ml each of IL-1, IL-3, IL-8, GM-CSF, and TNF-alpha could induce only about 20% HR; furthermore, such combinations did not have an additive or synergistic priming effect on anti-IgE-induced HR compared to the effect of single cytokines. Stripping of surface-bound IgE with lactic acid markedly reduced the capacity of basophils to release histamine in response to MNC-HRF and anti-IgE. Passive sensitization of IgE-stripped basophils with high-HRF responders' serum could restore their responsiveness to both MNC-HRF and anti-IgE, but passive sensitization with low-HRF responders' serum could restore responsiveness to anti-IgE only. Moreover, passage of MNC-HRF through high-, but not low-HRF, responders' IgE-Sepharose columns significantly reduced the HR activity of MNC-HRF. Finally, although the eluant could induce only 10% HR, the majority of its HR activity could be restored by the addition of effluent but not by the mixture of IL-1, IL-3, IL-8, GM-CSF, and TNF-alpha, suggesting the presence of a complex interaction among those cytokines. In summary, MNC-HRF contained at least two types of HRF activity; one was IgE dependent and the other was IgE independent.(ABSTRACT TRUNCATED AT 400 WORDS)

Measurement of histamine-releasing factor activity in individual nasal washings: Relationship with atopy, basdphil response, and membrane-bound IgE

We collected individual pools of nasal washings (NWs) from 15 allergic and 15 nonallergic subjects to determine histamine-releasing factor (HRF) activity and to ascertain the relationship of these cytokines with atopic status, basophil releasability, and cell membrane-bound IgE. NWs were concentrated, dialyzed, and assayed with basophils from a single donor. Samples from 12 of 15 allergic subjects and from all the nonallergic subjects revealed ⪰15% histamine release (HR), 33.5% ± 21.3% (mean ± SD) and 38.6% ± 19.6%, respectively ( p > 0.05). When we assayed the same samples with autologous basophils, the allergic group demonstrated higher HR than the nonallergic group (31.9% ± 19.7% versus 4.8% ± 4.3%; p < 0.001). A standard lot of mononuclear cell-derived HRFs was also screened with basophils from both groups. Means for HR from basophils of allergic and nonallergic subjects were 51.9% ± 16.7% versus 26.3% ± 8.2%, respectively ( p < 0.001). Pretreatment of basophils with lactic acid led to abrogation of sensitivity to HRF. Acid-stripped cells incubated with sera from patients with asthma regained their capacity to release histamine. We found that HRF activity can be detected in NWs of most donors, and there is no difference among allergic and nonallergic subjects. Our results suggest that the capacity of these cytokines to induce HR depends on several factors: atopic status, basophil releasability, and membrane-bound IgE.

Monocyte chemotactic and activating factor is a potent histamine-releasing factor for basophils.

Monocyte chemotactic and activating factor (MCAF) is a recently cloned cytokine that causes chemotaxis of basophils. In our pursuit of cytokines affecting basophil function, we studied the effect of MCAF on histamine secretion from basophils. Leukocytes from 20 donors, 10 allergic and 10 normal subjects, were studied. MCAF caused dose-dependent release of histamine at concentrations of 10(-8) and 10(-7) M, and the mean release was 31.25 +/- 2.9% at the highest concentration. In the same experiments the mean histamine release by anti-IgE and histamine releasing factor (HRF) was 27.05 +/- 4% and 32.70 +/- 2.7%, respectively. All 20 subjects responded to MCAF with significant histamine release. Allergic subjects released significantly more histamine than normals in response to anti-IgE (P less than 0.01) but not to MCAF (P = 0.2) and HRF (P = 0.1). The histamine release was significantly correlated between MCAF and HRF (P less than 0.01), but not between MCAF and anti-IgE (P greater than 0.05). The histamine release by MCAF was complete within the first 3 min. MCAF-induced degranulation was a calcium-dependent process. Leukocytes depleted of monocytes responded equally well to MCAF. Using an anti-MCAF affinity column we determined that greater than 50% of HRF activity of crude PBMC supernatant could be attributed to MCAF. Thus, we established that MCAF is a potent secretagogue for basophils.

Respiratory Viruses Induce Production of Histamine-Releasing Factor by Mononuclear Leukocytes: A Possible Role in the Mechanism of Virus-Induced Asthma

Histamine-releasing factor (HRF) is a cytokine produced by mononuclear leukocytes when stimulated with antigens or mitogens. HRF is capable of inducing degranulation of basophils and release of histamine. To determine if respiratory viruses can induce HRF production, mononuclear leukocytes from healthy adult donors were exposed to influenza or respiratory syncytial virus in vitro. HRF activity was tested by culturing the supernatants with fresh peripheral blood leukocytes and measuring the percentage of histamine released. Significant enhancement in histamine release was found in both virus groups compared with that of media controls. Thus, mononuclear leukocytes from normal individuals produce HRF in response to exposure to respiratory viruses, suggesting that this cytokine, which causes basophil degranulation, may play a role in the mechanism of virus-induced bronchospasm.

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.

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.

Altered production of histamine-releasing factor (HRF) activity and responsiveness to HRF after immunotherapy in children with asthma

To delineate the working mechanisms of immunotherapy (IT) (hyposensitization), the production of, and responsiveness to, histamine-releasing factor (HRF) was studied in four groups. These groups consisted of 32 newly diagnosed children with asthma, 40 good responders and 18 poor responders to IT (older than 2 years), and 15 healthy subjects. The results demonstrated (1) peripheral blood mononuclear cells of new patients produced a much greater HRF activity, either spontaneously or after stimulation, than did those of normal subjects, (2) the spontaneous HRF activity decreased significantly in good responders, whereas that of poor responders increased, (3) both the allergen (mite)- and mitogen (phytohemagglutinin [PHA])-stimulated HRF activity was decreased, although decrease was not significant, in good responders, but the activity was not changed in poor responders, (4) the granulocytes of new patients responded to HRF much more vigorously than did granulocytes of normal subjects. The responsiveness diminished significantly in both good and poor responders, although the magnitude of decrease was slightly greater in the former, (5) there was a positive correlation between PHA- and mite-stimulated HRF activity, mite-stimulated HRF activity and responsiveness to HRF, and plasma histamine level and responsiveness to HRF in the new patients, and (6) there was an inverse correlation between PHA-stimulated HRF production and responsiveness to HRF in good responders, but the correlation was positive in poor responders. Thus, IT is able to suppress the HRF activity, particularly the type of spontaneous synthesis, and responsiveness to HRF in clinically benefitted children with asthma, and this effect may be used to explain, partly, the efficacy of IT in a proportion of allergic patients.

Mast cells and pulmonary fibrosis. Identification of a histamine releasing factor in bronchoalveolar lavage fluid.

As elevated bronchoalveolar lavage (BAL) fluid histamine levels are noted in patients with pulmonary fibrosis (PF), we assayed BAL fluid from 16 patients with PF for the presence of a histamine releasing factor (HRF). HRF activity was assayed by measuring release of the preformed mast cell-derived mediators, histamine, or beta-hexosaminidase (beta-hex) from a purified population of IL-3 dependent mouse bone marrow derived mast cells (MBMMC) or human blood basophils. Mean BAL cell free histamine levels in the patients with PF was 1226 +/- 1349 pg/ml, whereas BAL histamine levels in a comparison group of six non-PF patients was 118 +/- 60 pg/ml. HRF was significantly elevated in BAL fluid of patients with PF (mean beta-hex release 24.5 +/- 12.9%; range 6.8 to 52.4%) compared to the non-PF group of patients (mean beta-hex release 7.9 +/- 7.7%; range 1.8 to 20.7%). The PF HRF not only degranulated MBMMC, but also induced the generation of the arachidonic acid metabolite leukotriene C4 from MBMMC (24.6 +/- 4.2 ng leukotriene C4/10(6) MBMMC). The PF HRF did not appear to be a cytokine previously identified in BAL fluid of patients with PF (i.e., platelet derived growth factor or insulin growth factor-1) or a human cytokine able to degranulate human basophils (i.e., IL-1, or granulocyte-macrophage-CSF) as these recombinant human cytokines did not induce MBMMC beta-hex release. Physicochemical characterization of the HRF revealed that it was relatively heat stable, pronase sensitive and on Sephadex G-75 and G-200 column chromatography had an apparent molecular mass of 30 to 50 kDa. The ability of PF BAL to induce beta-hex release from MBMMC was not dependent on IgE as unsensitized or lactic acid treated MBMMC release similar amounts of beta-hex compared to MBMMC sensitized with IgE. Thus, BAL fluid of patients with PF contains an HRF that induces beta-hex release from MBMMC via an IgE-independent mechanism. The presence of the HRF could explain elevated BAL histamine levels in patients with PF.

Detection of Histamine Release Inhibitory Factor- and Histamine Releasing Factor-like Activities in Bronchoalveolar Lavage Fluids

Histamine releasing factors (HRF) are a group of cytokines that cause degranulation of basophils and mast cells. Recently we have described a histamine release inhibitory factor (HRIF) that inhibits HRF-induced histamine release from basophils and mast cells. The objective of this study was to investigate the presence of these cytokines in bronchoalveolar lavage (BAL) fluid from normal subjects. We found that BAL fluids from 12 to 17 volunteers contained a dialyzable (molecular weight cutoff 3500) factor that inhibited basophil histamine release by HRF, anti-IgE, concanavalin A, and N-formyl-methionyl-leucyl-phenylalanine (FMLP). In addition, BAL fluids from 83% of the tested donors contained a nondialyzable inhibitor that blocked HRF-induced histamine release from basophils. The molecular weight of this inhibitor was estimated to be 20 to 30 and 8 to 10 kD by Sephadex G-50 chromatography and TSK 2000 size-exclusion HPLC. None of the unconcentrated BAL fluids showed any HRF activity on initial screening using basophils from allergic subjects. However, when the BAL fluids were concentrated, all BAL samples that were tested (N = 10) demonstrated significant HRF activity. The molecular weight of BAL HRF has been estimated to be in the range of 15 to 25 kD by size-exclusion HPLC, similar to the HRF synthesized by mononuclear cells. Thus we have demonstrated the presence of both HRF and HRIF in the BAL fluids. We speculate that these cytokines may be involved in the local regulation of basophil and mast cell activation.

Activation of guinea pig lymphocytes and mast cells by grain-dust extract

Previous studies have demonstrated that grain dust can stimulate lymphocyte proliferation and the production of interleukin-1 by macrophages. This study was undertaken to investigate whether grain dust could stimulate the production of histamine-releasing factor (HRF) by guinea pig spleen cells. We also studied the direct action of grain dust on guinea pig lung mast cells and basophils. Plastic nonadherent cells from immunized animals were cultured for 24 hours in the presence of grain dust or lipopolysaccharide, and the cell-free supernatants were assayed for HRF activity in the mast cell and basophil histamine release test. Lung mast cells were isolated by enzymatic digestion and discontinuous Percoll gradient centrifugation. It has been demonstrated that grain dust stimulated the production of HRF by spleen cells from the immunized animals but not from the control animals. Stimulation of spleen cells with lipopolysaccharide did not enhance the HRF production. Gel chromatography of grain dust-stimulated supernatant revealed that HRF has a molecular weight in the range of 50 to 70 kd and 5 to 8 kd. We also found that grain dust directly released significant amount of histamine from both mast cells and basophils. The results of this study suggest that grain dust contains some potent active substances that can activate lymphocytes, mast cells, and basophils.

The magnitude of the spontaneous production of histamine-releasing factor (HRF) by lymphocytes in vitro correlates with the state of bronchial hyperreactivity in patients with asthma

In our previous studies we reported that lymphocytes from patients with asthma spontaneously produce histamine-releasing factor (HRF) in vitro. In an effort to examine whether spontaneous HRF production (SpHRF) by lymphocytes from patients with asthma is related to the state of bronchial hyperreactivity (BHR), 20 patients with mild to severe asthma were studied. Peripheral blood lymphocytes were cultured alone in a serum-free medium for 24 hours, and culture supernatant was assayed for HRF activity in two separate histamine-release tests with autologous basophils and normal basophils from known healthy donors. BHR was measured as bronchial reactivity to inhaled histamine and was expressed as a provocation concentration of histamine required to induce a 20% fall in FEV1 (PC20). The result of this study demonstrated that lymphocyte supernatant from all patients with asthma released significant amount of histamine from both autologous and normal basophils. Very high histamine release was usually induced by lymphocyte supernatant from severely ill patients who had PC20 less than 2 mg/ml. Statistical analysis demonstrated that the magnitude of the SpHRF significantly correlated (r = -0.86; p less than 0.001) with PC20. Since mast cell- and basophil-derived mediators have been implicated in the pathogenesis of BHR, high correlation between PC20 and SpHRF by lymphocytes suggests that the latter may contribute to the development of BHR. Further studies are required to disclose the exact relationship between SpHRF and BHR.