Characterization of a galectin-like activity from the parasitic nematode, Haemonchus contortus, which modulates ovine eosinophil migration in vitro.

The development of eosinophilia is a characteristic feature of helminth infection, although the exact nature of the interaction between eosinophils and parasites remains to be fully defined. Previously, it has been reported that Haemonchus contortus and other nematodes produce eosinophil-specific chemoattractants. This paper describes studies aimed at isolating and identifying the factor(s) responsible. Initial studies showed that soluble extracts of infective larvae (L3) of H. contortus provoked a chemokinetic, rather than chemotactic, response in ovine bone marrow eosinophils in vitro. This activity was inhibited by lactose to a markedly greater extent than sucrose suggesting a galectin-like identity. Lactose affinity chromatography of soluble H. contortus extracts resulted in the isolation a specific bound fraction which retained biological activity. SDS-PAGE gel electrophoresis indicated a single Coomassie-stained band at between 31 and 41kDa. Subsequent, mass spectrometric analysis confirmed that the bound fraction contained a mixture of nematode galectins. The results confirm that H. contortus larvae produce several galectin-like proteins, at least one of which demonstrates eosinophil chemokinetic activity in vitro. The possibility of the parasite-derived factor mimicking the mammalian galectin-9, a known eosinophil chemokine, is discussed.

Nasal lavage fluid concentrations of eotaxin-1 (CCL11) in naturally occurring allergic rhinitis: relationship to disease activity, nasal luminal eosinophil influx, and plasma protein exudation.

BACKGROUND: Eotaxin-1 (CCL11) is a CC chemokine whose nasal eosinophilic chemotactic activity in vivo and in vitro has been demonstrated primarily using nasal allergen challenge models. The extension of these challenge findings to the in vivo setting has been limited. OBJECTIVE: To obtain nasal lavage fluid from volunteers with perennial and seasonal (in- and out-of-season) allergic rhinitis (AR) and non-atopic non-rhinitic controls for the measurement of eotaxin-1 concentrations and to relate these findings to the symptomatic disease severity, the percentage of lavage eosinophils, and to alpha(2)-macroglobulin (alpha(2)-MG) lavage concentrations, as a marker of vascular permeability and an index of airway inflammation. METHODS: Thirty-seven volunteers with AR (16 seasonal and 21 perennial) and 20 non-atopic non-rhinitic volunteers were recruited and phenotyped. Nasal lavage fluid was obtained by standardized protocol. The nasal lavage fluid concentrations of eotaxin and alpha(2)-MG were measured by ELISA, and differential cell counts performed on cytospins. RESULTS: Eotaxin-1 nasal lavage fluid concentrations were significantly higher in both the perennial and seasonal (in-season) AR groups compared with the controls, and significantly related to the severity of symptom expression and to the percentage of lavage eosinophils. The lavage eosinophil counts were significantly higher in both the symptomatic rhinitis groups compared with the control groups and correlated with the lavage concentrations of alpha(2)-MG. alpha(2)-MG levels were significantly increased in seasonal (in-season) rhinitics compared with both non-atopic controls and seasonal (out-of-season) rhinitics. A significant correlation was observed between the levels of alpha(2)-MG and levels of eotaxin in the symptomatic allergic rhinitic groups. CONCLUSIONS: This study clearly demonstrates the relevance of eotaxin-1 to the pathogenesis of naturally occurring AR.

Increased eosinophil-lineage committed progenitors in the lung of allergen-challenged mice.

BACKGROUND: There is increasing evidence that hemopoietic progenitor cells may traffic from bone marrow to sites of allergen exposure in asthma and undergo in situ differentiation, contributing to ongoing airway inflammation. However, the isolation and detailed phenotyping of true CD34 + progenitors from lung tissue during an allergen-induced airway eosinophilia has not been performed. OBJECTIVE: We attempted to isolate and investigate the in vivo kinetics of hemopoietic progenitor cells and production of eosinophilopoietic mediators in the lung. METHODS: In a mouse model of allergic airway inflammation, cells were extracted from lung tissue by enzymatic digestion. Total (CD34 + 45 + ) and eosinophil lineage committed (CD34 + 45 + IL-5Ralpha + ) progenitors were enumerated by flow cytometry. Outcome measurements were made 2, 6, 12, 24, 48, and 72 hours and 7 and 14 days after allergen challenge. RESULTS: Compared with saline control, CD34 + 45 + progenitors were elevated between 6 and 48 hours ( P < .05), attenuated by 72 hours and subsequently increased by 14 days ( P > .05). CD34 + 45 + IL-5Ralpha + progenitors were transiently elevated at 6 hours ( P < .05) before a return to preallergen levels by 12 hours and a subsequent increase at 14 days ( P < .05). Bronchoalveolar lavage eosinophils were increased at 2 hours, peaking at 72 hours ( P < .00625) and declining by 14 days. Both IL-5 and eotaxin levels were increased by 2 hours, peaking at 12 hours ( P < .05) and 24 hours ( P < .05), respectively. CONCLUSION: We propose that the increase in CD34 + 45 + IL-5Ralpha + cells and the eosinophilopoietic mediators IL-5 and eotaxin in the lung after allergen exposure may promote in situ differentiation of eosinophils that contribute to ongoing allergic airway inflammation.

Lung eosinophilic inflammation and airway hyperreactivity are enhanced by murine anaphylactic, but not nonanaphylactic, IgG1 antibodies.

BACKGROUND: Chronic airway inflammation is a fundamental feature of bronchial asthma, which is characterized by the accumulation and activation of inflammatory cells, such as mast cells and eosinophils, that are tightly regulated by TH2 cytokines and chemokines. Recently, we demonstrated, in a murine model of asthma with immunosuppressed mice reconstituted with antigen-specific IgE or IgG1 antibodies, that IgE, but not IgG1, participates in potentiation of airway inflammation and induction of airway hyperreactivity (AHR). The IgG1 antibody, however, did not elicit passive cutaneous anaphylactic reactions, which was in contrast to IgE. OBJECTIVES: Because 2 types of murine IgG1 have been demonstrated with regard to anaphylactic activity, the present experiments were undertaken to determine the role of anaphylactic and nonanaphylactic IgG1 antibodies in the development of antigen-induced eosinophilia and AHR in this model. METHODS: Dinitrophenyl-conjugated, heat-coagulated hen's egg white was implanted in immunosuppressed mice reconstituted with anaphylactic or nonanaphylactic IgG1. Intratracheal challenge with aggregated dinitrophenyl-ovalbumin was performed on day 14, and lung inflammatory and mechanical parameters were evaluated after 48 hours. RESULTS: Our results demonstrated that reconstitution of immunosuppressed mice with anaphylactic IgG1 antibodies in contrast to nonanaphylactic IgG1 antibodies potentiates their ability to have pulmonary eosinophilic inflammation and AHR. IL-5 and eotaxin levels in bronchoalveolar lavage fluid from anaphylactic IgG1-reconstituted mice were also higher than those in nonanaphylactic IgG1-reconstituted mice. CONCLUSIONS: These results indicate that the anaphylactic property of murine IgG1 molecules is essential for their capacity to enhance lung eosinophilic inflammation and to induce AHR.

Eotaxin expression in oral squamous cell carcinomas with and without tumour associated tissue eosinophilia.

AIM: Eotaxin is a powerful and selective eosinophil chemoattractant. The purpose of this study was to compare the expression of eotaxin in oral squamous cell carcinomas with and without tumour associated tissue eosinophilia (TATE). The mechanisms that control the recruitment of eosinophils to these tumours are not clearly established. METHODS: A total of 60 patients with oral squamous cell carcinomas (OSCC) with TNM stages II and III, located in the tongue, oral floor, retromolar area and inferior gingiva were divided in two groups: 1--OSCC with intense eosinophilic inflammatory infiltrate and 2--OSCC with absent/low eosinophilic inflammatory infiltrate. The eotaxin expression was analyzed by immunohistochemistry using standard streptavidin-biotin-peroxidase complex technique with monoclonal (mouse anti-human eotaxin) and polyclonal (rabbit anti-human eotaxin) antibodies. RESULTS: The eotaxin expression was identified in normal oral mucosa as well as in both OSCC groups including malignant epithelial cells, eosinophils, neutrophils, plasma cells and fibroblasts. The eosinophils showed intense immunopositivity for eotaxin. CONCLUSION: These results suggest that the eotaxin expressed in oral squamous cell carcinomas, mainly derived from eosinophils, is probably involved in the mechanisms of eosinophils chemotaxis to the tumour and in the maintenance of TATE in these malignant tumours.

Pleural fluid levels of vascular cell adhesion molecule-1 are elevated in eosinophilic pleural effusions.

STUDY OBJECTIVES: The mechanisms responsible for the accumulation of eosinophils in pleural fluid are not fully understood. The objective of the present study was to examine the relationship between pleural fluid eosinophilia and the levels of vascular cell adhesion molecule (VCAM)-1, eotaxin, RANTES (regulated upon activation, normal T-cell expressed and secreted), and interleukin (IL)-4 in pleural effusions. PATIENTS AND METHODS: Thirty-one patients with eosinophilic pleural effusion (EPE) [eosinophil percentage > 10% of the pleural fluid nucleated cells] and 10 patients without EPE were evaluated. VCAM-1, eotaxin, RANTES, and IL-4 in all pleural fluids were measured using enzyme-linked immunosorbent assay kits. IL-5 levels of the same fluids were measured in a previous study. RESULTS: VCAM-1, eotaxin, and RANTES but not IL-4 were detectable in the pleural fluids. The mean level of VCAM-1 in EPE (336 +/- 85 ng/mL) was significantly higher (p = 0.011) than that in the noneosinophilic effusions (260 +/- 34 ng/mL) [mean +/- SD]. VCAM-1 levels were significantly correlated with the eosinophil count and percentage in all pleural fluids (r = 0.43, p = 0.005, and r = 0.37, p = 0.019, respectively). Multiple linear regression analysis disclosed that both IL-5 (beta, 0.63; p < 0.001) and VCAM-1 (beta, 0.27, p = 0.025) are independent predictors of the number of eosinophils in all pleural fluids. RANTES and eotaxin did not differ significantly between EPEs and non-EPEs, and were not correlated with the number of pleural fluid eosinophils. CONCLUSION: The levels of VCAM-1 are increased in EPE, suggesting that VCAM-1 is important in the pathogenesis of EPE. Neither eotaxin nor RANTES is associated with pleural fluid eosinophilia.

Up-regulation of cysteinyl leukotriene 1 receptor by IL-13 enables human lung fibroblasts to respond to leukotriene C4 and produce eotaxin.

Cysteinyl leukotrienes (CysLTs) play an important role in eosinophilic airway inflammation. In addition to their direct chemotactic effects on eosinophils, indirect effects have been reported. Eotaxin is a potent eosinophil-specific chemotactic factor produced mainly by fibroblasts. We investigated whether CysLTs augment eosinophilic inflammation via eotaxin production by fibroblasts. Leukotriene (LT)C(4) alone had no effect on eotaxin production by human fetal lung fibroblasts (HFL-1). However, LTC(4) stimulated eotaxin production by IL-13-treated fibroblasts, thereby indirectly inducing eosinophil sequestration. Unstimulated fibroblasts did not respond to LTC(4), but coincubation or preincubation of fibroblasts with IL-13 altered the response to LTC(4). To examine the mechanism(s) involved, the expression of CysLT1R in HFL-1 was investigated by quantitative real-time PCR and flow cytometry. Only low levels of CysLT1R mRNA and no CysLT1R protein were expressed in unstimulated HFL-1. In contrast, stimulation with IL-13 at a concentration of 10 ng/ml for 24 h significantly up-regulated both CysLT1R mRNA and protein expression in HFL-1. The synergistic effect of LTC(4) and IL-13 on eotaxin production was abolished by CysLT1R antagonists pranlukast and montelukast. These findings suggest that IL-13 up-regulates CysLT1R expression, which may contribute to the synergistic effect of LTC(4) and IL-13 on eotaxin production by lung fibroblasts. In the Th2 cytokine-rich milieu, such as that in bronchial asthma, CysLT1R expression on fibroblasts might be up-regulated, thereby allowing CysLTs to act effectively and increase eosinophilic inflammation.

Eotaxin level in induced sputum is increased in patients with bronchial asthma and in smokers.

BACKGROUND: Airway eosinophilia is one of the hallmarks of asthma. Eotaxin may play an important role in eosinophil recruitment. OBJECTIVES: To examine the relationship between eotaxin levels in the sputum and eosinophilic inflammation. METHODS: The sputum was obtained from 11 non-smokers, 14 smokers and 13 asthmatic patients using a sputum induction method. Eotaxin and interleukin (IL)-5 levels in the sputum were determined by ELISA and immunocytochemical analysis. RESULTS: Asthmatic patients had eosinophilia and smokers showed neutrophilia in their sputum. The eotaxin level in the sputum was significantly higher in smokers (median 412.5, range 91.1-872.2 pg/ml) and asthmatic patients (351.0, 185.0-928.0 pg/ml) compared with non-smokers (123.2, 0-369.0 pg/ml; both p < 0.05). IL-5 was detected in the sputum of 1 non-smoker, none of the smokers and 4 asthmatic patients. The percentage of eotaxin-positive cells was higher in smokers and asthmatic patients than in non-smokers, but the percentage of IL-5-positive cells was significantly higher only in asthmatic patients (p < 0.05). CONCLUSIONS: These findings suggest that the elevated eotaxin level in the sputum does not always accompany the increase in eosinophils, and cooperation with another cytokine such as IL-5 may be required for the recruitment of eosinophils.

The kinetics of allergen-induced eotaxin level in nasal lavage fluid: its key role in eosinophil recruitment in nasal mucosa.

Eotaxin (CCL11) is a potent eosinophil chemoattractant belonging to the C-C chemokine. To evaluate the role of eotaxin in eosinophilic inflammation in nasal mucosa, we investigated the levels of eosinophil chemoattractants in nasal lavage fluids obtained after antigen challenge, compared with eosinophil counts and eosinophil protein X (EPX) levels. In subjects with allergic rhinitis, allergen challenge led to parallel increases in eosinophil counts, levels of EPX, and eotaxin concentrations in nasal lavage fluid. The levels of eotaxin in lavage samples showed strong correlation with lavage levels of eosinophil counts and EPX. Normal subjects had few, if any, eosinophils and EPX as well as the measured parameters in their nasal lavage fluids before and after antigen challenge. In our experiments of eosinophil endothelial transmigration (TEM) assay using the nasal microvascular endothelial cells, eotaxin showed the most potent effect among various eosinophil chemoattractants. In addition, treatment of eosinophils with anti-CCR-3 mAb significantly blocked eosinophil TEM induced by homogenate of nasal mucosa. These results indicate that eotaxin has an important role in eosinophil-dependent inflammation in nasal mucosa and suggest that blocking eotaxin or CCR-3 might be useful for new therapeutic tools of allergic rhinitis.

Characterization of airway inflammation after repeated exposures to occupational agents.

BACKGROUND: Little is known about the comparative kinetics of eosinophil recruitment after exposure to low- and high-molecular-weight sensitizers in subjects with occupational asthma (OA). OBJECTIVES: The aims of the study were to investigate the kinetics of changes in inflammatory mediators associated with eosinophil infiltration (IL-5 and eotaxin) and to examine the nature of the airway inflammation induced in response to different types of occupational agents. METHODS: We investigated 15 subjects with OA caused by high- and low-molecular-weight agents. The subjects were exposed to increasing doses of the relevant occupational agent over 3 to 4 days until a 20% fall in FEV(1) occurred. Methacholine challenge and sputum induction were performed at the end of each day of exposure. Sputum samples were assessed for differential cell counts, including eosinophils, IL-5, and eotaxin messenger RNA. RESULTS: There was an increase in sputum eosinophils, eotaxin, and IL-5 on the day preceding the occurrence of asthmatic reaction, although there was no change in functional parameters (FEV(1) and PC(20)). Increase in sputum eosinophils was more prominent in subjects exposed to low-molecular-weight agents than to high-molecular-weight agents. CONCLUSION: Changes in eosinophils, IL-5, and eotaxin precede functional changes after exposure to occupational agents in subjects with OA. Eosinophil inflammation is a feature of exposure to both high- and low-molecular-weight agents. Induced sputum may be a useful tool in the early diagnosis of OA.

Increased coexpression of eotaxin and interleukin 5 in bullous pemphigoid.

While the presence of eosinophils in the skin lesions of bullous pemphigoid is well documented, the chemotactic factors responsible for eosinophil recruitment into the tissue still remain to be defined. In this study, eotaxin and interleukin-5 (IL-5) concentrations were determined in the blister fluid and sera of patients with bullous pemphigoid (acute and remission phase, n=6) in comparison with normal healthy controls (n=6) using the enzyme-linked immunosorbent assay (ELISA) technique. Eotaxin and IL-5 levels were increased in the blister fluid compared with the acute and remission phase sera, as well as compared with the sera of normal controls. In addition, immunoreactivity for eotaxin was predominantly found in the inflammatory cell infiltrate of lesional bullous pemphigoid biopsy specimens. In conclusion, the data provide evidence that co-operation of eotaxin and IL-5 may play an essential role in activating and recruiting eosinophils, which ultimately contribute to the tissue damage in bullous pemphigoid.

Quantitative analysis of eotaxin and RANTES messenger RNA in nasal polyps: association of tissue and nasal eosinophils.

OBJECTIVES/HYPOTHESIS: Nasal polyps develop in the ethmoidal and middle turbinate area, often in relation to inflammatory conditions. Their exact etiology and pathogenesis are still under debate. Histologically, the polyps are infiltrated by a number of inflammatory cells, with eosinophil predominating in most specimens. This finding suggests that the nasal polyp is an inflammatory growth that is controlled by the local environment. The chemokines eotaxin and RANTES (regulated on activation normal T cell expressed and secreted) have been postulated to be involved in the recruitment and activation of eosinophils to certain inflamed tissues. The purpose of this study was to investigate eotaxin and RANTES mRNA expression in nasal polyps and its effect on tissue and nasal eosinophils. METHODS: Nasal polyps (917 allergic and 30 nonallergic cases) were obtained from endoscopic sinus surgery, and 15 normal inferior turbinates also were taken. Immunohistochemical staining for eosinophils and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) tests for eotaxin and RANTES mRNA expression were performed, and the concentration of nasal eosinophil cationic protein (ECP) was measured. RESULTS: The amounts of eotaxin mRNA in the allergic nasal polyps were 11.4 times higher and the levels in the nonallergic polyps were 6.4 times higher than in the normal inferior turbinate. However, the RANTES mRNA expression did not show any differences among the three groups. Tissue eosinophilia and nasal ECP levels were significantly correlated with eotaxin mRNA level but not with RANTES mRNA expression. CONCLUSION: Nasal polyp eosinophilic infiltration and activation correlate mainly with increased eotaxin gene expression rather than with RANTES expression.

Eotaxin levels in pleural effusions: comparison with monocyte chemoattractant protein-1 and IL-8.

OBJECTIVE: In order to investigate the role of eotaxin in pleural diseases, we measured eotaxin in pleural effusions and studied the relationship between eotaxin levels and recruitment of inflammatory cells, particularly eosinophils. Interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) levels were also measured for comparison. METHODS: We evaluated 47 pleural effusion samples, 7 transudates and 40 exudates. The exudates consisted of 19 malignant, 11 tuberculous, and 5 parapneumonic effusions, and 5 effusions of other etiologies. Chemokine levels were measured by specific sandwich enzyme-linked immunosorbent assays. RESULTS: Eotaxin was detected in all samples examined, but the levels did not differ significantly among the exudates. There was no significant correlation between the levels of eotaxin and MCP-1 or IL-8. The level of eotaxin but not the others was significantly higher in eosinophilic effusions (>10% eosinophils among white blood cells in the fluid) than in non-eosinophilic fluids. The number of eosinophils in pleural effusions was significantly correlated with the eotaxin levels, but not with the levels of other chemokines. The number of neutrophils was significantly correlated with IL-8 but not with the others. CONCLUSIONS: Results suggest that eotaxin contributes to the migration of eosinophils in pleural inflammation. Taken together with the correlation between IL-8 and neutrophils, it appears that the predominant type of pleural inflammatory infiltrate is controlled, at least in part, by the subgroup of chemokines expressed in the pleural space.

Dermal fibroblasts represent a potent major source of human eotaxin: In vitro production and cytokine-mediated regulation.

Accumulating evidence indicates that eotaxin plays an integral role in tissue recruitment of eosinophils in humans as well as in animals. To clarify which types of cells are actually important as sources of human eotaxin, we used a specific enzyme-linked immunosorbent assay (ELISA) to compare various types of hemopoietic and nonhemopoietic cells for the ability to produce eotaxin protein. Regardless of various conditioning, we failed to determine any significant eotaxin generation by peripheral leukocytes and vein endothelial cells (less than 20 pg/ml). A small amount of immunoreactive eotaxin was detected in cultures of A549 bronchial epithelial cell line cells. In contrast, dermal fibroblasts were capable of generating extremely high, and potentially biologically relevant, amounts of eotaxin protein (on the order of ng/ml). The eotaxin generation was induced by tumour necrosis factor alpha (TNF-alpha) or IL-4, and the production was drastically increased by combined use of these cytokines. Because fibroblasts are ideally situated within the interstium at the sites of allergic responses, our finding that these cells represent an important cellular source of eotaxin suggests that fibroblast-derived eotaxin may act to regulate eosinophil recruitment in a paracrine fashion.

Eotaxin expression in Onchocerca volvulus-induced dermatitis after topical application of diethylcarbamazine.

In persons with onchocerciasis, topical application of the anthelminthic diethylcarbamazine (DEC) induces clinical and histologic responses similar to acute papular onchodermatitis, including recruitment of eosinophils to the skin. To determine whether the eosinophil chemokine eotaxin is likely to be associated with eosinophil recruitment in onchodermatitis, DEC was applied to a 5-cm2 area on the skin of infected persons, and biopsies were taken from lesions 24 h later. Histologic analysis showed elevated dermal and epidermal eosinophils compared with tissue from an adjacent (untreated) site. Reverse transcription-polymerase chain reaction showed that eotaxin gene expression in DEC-treated skin was elevated 2- to 17-fold compared with control tissue. Eotaxin immunoreactivity was noted in mononuclear cells and eosinophils in the perivascular region of the dermis and in lymphatic and vascular endothelial cells. Together, these observations are consistent with a role for eotaxin in recruitment of eosinophils to the dermis in early stage onchocercal skin disease.

Eotaxin contributes to renal interstitial eosinophilia.

BACKGROUND: A potent eosinophil chemotactic cytokine, human eotaxin, is directly chemotactic for eosinophils. Therefore, the specific expression of eotaxin in tissue might play a crucial role in tissue eosinophilia. However, the precise molecular mechanism of the recruitment and activation of eosinophils in human renal diseases remains to be investigated. We evaluated the role of eotaxin in the pathogenesis of human diffuse interstitial nephritis with marked infiltration of eosinophils. METHODS: In this study, we examined 20 healthy volunteers. 56 patients with primary or secondary glomerular diseases and two hypereosinophilic syndrome patients without renal involvement. Urinary and serum eotaxin levels were determined by an enzyme-linked immunosorbent assay. We also detected the presence of eotaxin protein immunohistochemically. RESULTS: On the one hand, urinary levels of eotaxin were significantly higher before the initiation of glucocorticoid administration in the patient with interstitial nephritis with marked infiltration of eosinophils. On the other hand, urinary eotaxin levels were not detected in any patients with nephrotic syndrome, interstitial nephritis without eosinophils, hypereosinophilic syndrome without renal involvement or other renal diseases. Serum eotaxin levels were not detected in any of the patients. Therefore, the detection of eotaxin in the urine was specific for renal interstitial eosinophilia. Moreover, endothelial cells, infiltrating mononuclear cells and renal epithelial cells in the tubulointerstitial lesions were immunostained with specific anti-eotaxin antibodies. Furthermore, the elevated urinary levels of eotaxin decreased dramatically during glucocorticoid-induced convalescence. HYPOTHESIS: We hypothesize that in situ expression of eotaxin may provide a new mechanism to explain the renal interstitial eosinophil infiltration.

Eosinophil chemotactic factors from cysticercoids of Hymenolepis nana.

A comparative study of eosinophil chemotactic factors was carried out using cysticercoids and oncospheres of Hymenolepis nana. Cysticercoids showed twice the chemotactic activity for eosinophils than the oncospheres. Eosinophilia induced by oncospheres and cysticercoids observed in secondary and primary infections, respectively, were discussed from the view point of the immunobiology of this parasite.

[Lymphocyte chemotactic factor in tuberculous pleural effusion].

To evaluate whether lymphocyte chemotactic factor is involved in the accumulation of lymphocytes in tuberculous pleurisy, we measured lymphocyte chemotactic activity in tuberculous pleural effusions, and compared with that in malignant pleural effusions and transudate. The lymphocyte chemotactic activity was measured in vitro with chemotactic chamber. The cells suspended in the culture medium was added to the upper well and the effusions, normal human serum (NHS), or culture medium were placed below nitrocellulose filter. The lymphocyte migration was quantified by counting the number of cells migrating beyond a distance of 70 microns from top of the filter in 5 selected fields. The chemotactic activity of the effusions was expressed as a percentage of the control migration in the culture medium. When we used the freshly isolated peripheral blood lymphocytes as responding cells, the chemotactic activity was 176.0 +/- 41.3% in tuberculous effusions, 115.1 +/- 53.8% in malignant effusions, 87.1 +/- 16.1% in transudate, and 113.3 +/- 24.2% in NHS, respectively. The activity of tuberculous effusions was significantly higher than that of transudate and NHS. When we used PHA-activated lymphocytes, the activity was 284.4 +/- 159.3% in tuberculous effusions, 123.1 +/- 77.6% in malignant effusions, 75.8 +/- 10.3% in transudate, and 52.6 +/- 10.1% in NHS, respectively. The activity of tuberculous effusions was significantly higher than that of malignant effusions, transudate and NHS. The chemotactic activity of tuberculous effusions to PHA-activated lymphocytes was significantly higher than that to freshly isolated lymphocytes. The activity was specific for T lymphocytes, and showed both chemotaxis and chemokinesis by checkerboard analysis. Gel filtration performed with Sephacryl S-200 revealed that the chemotactic activities in a tuberculous fluid had three peaks located in the regions between blue dextran and immunoglobulin G, near to human albumin and cytochrome c marker. The most potent activity was found at the region near human albumin. Lymphocyte chemotactic factor in tuberculous effusion may stimulate the migration of T lymphocytes, especially the activated T lymphocytes to the pleural spaces.

Cell infiltration, ICAM-1 expression, and eosinophil chemotactic activity in asthmatic sputum.

We have applied the technique of sputum induction by hypertonic saline in asthmatics and nonatopic control subjects to study an array of indices of airway inflammation believed to be relevant to asthma pathogenesis. Compatible with a central role for eosinophils and mast cells in asthma, sputum of asthmatic subjects contained increased numbers of eosinophils and levels of eosinophil cationic protein (ECP) and mast cell tryptase. Eosinophil numbers, and ECP and histamine levels correlated with the degree of methacholine airways responsiveness, and ECP, tryptase, and histamine correlated with raised concentrations of albumin. Using the micro-Boyden chamber technique eosinophil chemotactic activity was identified only in the sputum from asthmatics. The correlation between the raised levels of total IgA, IL-8/IgA complexes, and tryptase and the degree of sputum eosinophilia and ECP levels, suggests possible mechanisms for eosinophil chemotaxis and activation in asthma. Row cytometric analysis of sputum lymphocytes showed an increase in CD4+ T cells and T cells expressing intercellular adhesion molecule-1 (ICAM-1) in asthma which, together with the finding of raised levels of soluble ICAM-1 in the sputum, indicates upregulation of this adhesion molecule. Finally, the proportion of CD16+ natural killer (NK) cells was reduced in the sputum of asthmatics. These observations highlight the importance of the airway inflammation in causing asthma and further confirm the usefulness of sputum induction as a tool in asthma research.