Status of cathelicidin IL-37, cytokine TNF, and vitamin D in patients with pulmonary tuberculosis.

Development of Mycobacterium tuberculosis (Mtb) infection depends on the ability of the host to elicit the protective immune response to the pathogen. Cathelicidin plays a role in antibacterial innate immunity mechanisms. This peptide contributes to the barrier function of respiratory epithelium and takes part in controlling pulmonary bacterial infections. LL-37 (leucine-leucine-37) is involved in host defense and innate immune response to mycobacterial infections, as well. This study aims to evaluate the serum concentrations of LL-37 in individuals with active pulmonary tuberculosis (TB) and to determine whether any correlations between peptide LL-37, tumor necrosis factor (TNF) and vitamin D serum levels exist. A total of 46 adults with pulmonary TB were recruited for the study. Sixty-one controls were randomly selected as control group. Serum concentrations of cathelicidin LL-37, vitamin D (25(OH)D), as well as TNF, were measured using an enzyme-linked immunosorbent assay (ELISA) kit. The mean (± SEM) level of LL-37 was significantly higher in the TB group (7.45±1.58) compared with healthy controls (1.41±0.22) (p less than 0.001). Mean serum concentration of TNF was significantly higher in the TB group (8.51±1.92) compared with healthy controls (2.69±0.19) (p less than 0.001). There was no significant difference in mean serum levels of vitamin D between healthy (26.10±1.74) and TB subjects (24.18±1.95). No correlations between LL-37, TNF, and vitamin D levels in patients with TB were observed. Our results indicated that serum levels of peptide LL-37 during TB is raised significantly, and this observation is compatible with the general view of the important role of this cathelicidin in defense mechanisms against Mtb infection.

Serum level of cathelicidin LL-37 in patients with active tuberculosis and other infectious diseases.

A growing body of evidence indicates the role of cathelicidin LL-37, a member of the antimicrobial peptide family, in host innate defense mechanisms. The important role of this peptide in infectious diseases is also suggested, however, to date, data relating to LL-37 expression in the course of bacterial infections are far from complete. Therefore, the aim of the present study was to determine LL-37 serum levels in adult patients with pulmonary tuberculosis (TB). For comparison, circulating LL-37 levels in patients with pneumonia induced by Gram-positive or Gram-negative bacteria species and in healthy subjects were evaluated. Fifty patients with pulmonary TB, 31 patients with pneumonia caused by gram-positive bacteria, 68 individuals with pneumonia caused by Gram-negative bacteria, and 61 randomly selected healthy subjects were enrolled in the study. Serum LL-37 concentration was measured using an enzyme-linked immunosorbent assay (ELISA). We established that the mean level of LL-37 was statistically significantly higher in TB patients than that in patients with Gram-positive bacteria-induced pneumonia (p < 0.001), in patients with Gram-negative bacteria-induced pneumonia (p < 0.001), and in healthy controls (p < 0.001). In patients with TB, no statistically significant correlations between serum LL-37 and CRP concentrations (r = -0.2042; p = 0.189) and between serum LL-37 concentration and WBC count (r = -0.1277; p = 0.414) were observed. Our observations clearly documented that cathelicidin LL-37 plays a role in defense mechanisms against infectious agents, and is particularly important when the infection is caused by an intracellular pathogen.

Mast cells generate cysteinyl leukotrienes and interferon-beta as well as evince impaired IgE-dependent degranulation upon TLR7 engagement.

Mast cells are numerous at anatomical sites close to external environment, virtually at the portals of infection. A few data indicated that these cells express cytoplasmic Toll-like receptors (TLRs) recognizing virus-derived molecules. Accordingly, mast cells could participate in anti-viral defense or/and in viral-related diseases. However, data concerning the influence of viruses on mast cell activity are limited. Thus, the aim of our study was to determine mast cell response to TLR7 ligand, i.e. resiquimod (R848), a synthetic mimic of viral ssRNA. Since mast cells play a central role in allergic reactions the effect of TLR7 agonist was also investigated on FcepsilonRI-dependent mast cell response. Experiments were carried out in vitro on freshly isolated fully mature rat peritoneal mast cells. Mast cells exhibit constitutive TLR7 molecule expression and its up-regulation after the agonist challenge. TLR7-mediated mast cell stimulation resulted in cysteinyl leukotriene (cysLT) and interferon (IFN)-beta synthesis, whereas no histamine and CXCL8 secretion was stated. Moreover, mast cell priming with TLR7 ligand caused the reduction in anti-IgE-induced histamine release. The results suggest that ssRNA viruses could directly activate mast cells to alter their phenotype and to release of potent proinflammatory mediators or indirectly modulate IgE-dependent allergic processes.

Hypertrophic obstructive cardiomyopathy-left ventricular outflow tract shapes and their hemodynamic influences applying CMR.

Hypertrophic cardiomyopathy (HCM) is one of the most common genetic cardiac disorders and is characterized by different phenotypes of left ventricular hypertrophy with and without obstruction. The effects of left ventricular outflow tract (LVOT) obstruction based on different anatomies may be hemodynamically relevant and influence therapeutic decision making. Cardiovascular magnetic resonance (CMR) provides anatomical information. We aimed to identify different shapes of LVOT-obstruction using Cardiovascular Magnetic Resonance (CMR). The study consisted of two parts: An in-vivo experiment for shape analysis and in-vitro part for the assessment of its hemodynamic consequences. In-vivo a 3D depiction of the LVOT was created using a 3D multi-slice reconstruction from 2D-slices (full coverage cine stack with 7 slices and a thickness of 5-6 mm with no gap) in 125 consecutive HOCM patients (age = 64.17 +/- 12.655; female n = 42). In-vitro an analysis of the LVOT regarding shape and flow behavior was conducted. For this purpose, 2D and 4D measurements were performed on 3D printed phantoms which were based on the anatomical characteristics of the in-vivo study, retrospectively. The in-vivo study identified three main shapes named K- (28.8%), X- (51.2%) and V-shape (10.4%) and a mixed one (9.6%). By analyzing the in-vitro flow measurements every shape showed an individual flow profile in relation to the maximum velocity in cm/s. Here, the V-shape showed the highest value of velocity (max. 138.87 cm/s). The X-shape was characterized by a similar profile but with lower velocity values (max. 125.39 cm/s), whereas the K-shape had an increase of the velocity without decrease (max. 137.11 cm/s). For the first time three different shapes of LVOT-obstruction could be identified. These variants seem to affect the hemodynamics in HOCM.

Myocardial injury in patients with acute ischemic stroke detected by cardiovascular magnetic resonance imaging.

BACKGROUND: Patients with acute ischemic stroke (AIS) are at high risk of adverse cardiovascular events. Until now, the burden of myocardial injury derived from cardiovascular magnetic resonance imaging (CMR) has not been established in this population. METHODS: Patients with AIS underwent CMR at 3 Tesla within 120 h after the index stroke as part of a prospective, single-center study. Patients with persistent atrial fibrillation were excluded. Morphology and function of both cardiac chambers and atria were assessed applying SSFP cine. Myocardial tissue differentiation was based on native and contrast-enhanced imaging including late gadolinium enhancement (LGE) after 0.15 mmol/kg gadobutrol for focal fibrosis and parametric T2- and T1-mapping for diffuse findings. To detect myocardial deformation global longitudinal (GLS), circumferential (GCS) and radial (GRS) strain was measured applying feature tracking. Cardiac troponin was measured using a high-sensitivity assay (99th percentile upper reference limit 14 ng/L). T2 mapping values were compared with 20 healthy volunteers. RESULTS: CMR with contrast media was successfully performed in 92 of 115 patients (mean age 74 years, 40% female, known myocardial infarction 6%). Focal myocardial fibrosis (LGE) was detected in 31 of 92 patients (34%) of whom 23/31 (74%) showed an ischemic pattern. Patients with LGE were more likely to have diabetes, prior myocardial infarction, prior ischemic stroke, and to have elevated troponin levels compared to those without. Presence of LGE was accompanied by diffuse fibrosis (increased T1 native values) even in remote cardiac areas as well as reduced global radial, circumferential and longitudinal strain values. In 14/31 (45%) of all patients with LGE increased T2-mapping values were detectable. CONCLUSIONS: More than one-third of patients with AIS have evidence of focal myocardial fibrosis on CMR. Nearly half of these changes may have acute or subacute onset. These findings are accompanied by diffuse myocardial changes and reduced myocardial deformation. Further studies, ideally with serial CMR measurements during follow-up, are required to establish the impact of these findings on long-term prognosis after AIS.

Mycobacterium tuberculosis CwsA interacts with CrgA and Wag31, and the CrgA-CwsA complex is involved in peptidoglycan synthesis and cell shape determination.

Bacterial cell division and cell wall synthesis are highly coordinated processes involving multiple proteins. Here, we show that Rv0008c, a novel small membrane protein from Mycobacterium tuberculosis, localizes to the poles and on membranes and shows an overall punctate localization throughout the cell. Furthermore, Rv0008c interacts with two proteins, CrgA and Wag31, implicated in peptidoglycan (PG) synthesis in mycobacteria. Deletion of the Rv0008c homolog in M. smegmatis, MSMEG_0023, caused bulged cell poles, formation of rounded cells, and defects in polar localization of Wag31 and cell wall synthesis, with cell wall synthesis measured by the incorporation of the [(14)C]N-acetylglucosamine cell wall precursor. The M. smegmatis MSMEG_0023 crgA double mutant strain showed severe defects in growth, viability, cell wall synthesis, cell shape, and the localization of the FtsZ, FtsI, and Wag31 proteins. The double mutant strain also exhibited increased autolytic activity in the presence of detergents. Because CrgA and Wag31 proteins interact with FtsI individually, we believe that regulated cell wall synthesis and cell shape maintenance require the concerted actions of the CrgA, Rv0008c, FtsI, and Wag31 proteins. We propose that, together, CrgA and Rv0008c, renamed CwsA for cell wall synthesis and cell shape protein A, play crucial roles in septal and polar PG synthesis and help coordinate these processes with the FtsZ-ring assembly in mycobacteria.

Effect of scaling and root planing on interleukin-1ß, interleukin-8 and MMP-8 levels in gingival crevicular fluid from chronic periodontitis patients.

BACKGROUND AND OBJECTIVE: There are few data concerning the effect of scaling and root planing on the levels of immune and inflammatory mediators in gingival crevicular fluid from patients with chronic periodontitis. Therefore, in this study the influence of scaling and root planing was determined on amounts of interleukin (IL)-1ß, IL-8 and MMP-8 in gingival crevicular fluid from patients with chronic periodontitis, in relation to clinical parameters. MATERIAL AND METHODS: A total of 51 patients were enrolled in this study. The study population consisted of 30 patients with generalized advanced chronic periodontitis, while 21 periodontally healthy subjects were recruited for the control group. The clinical parameters included approximal plaque index, gingival index, pocket depth and clinical attachment loss. The amounts of IL-1ß, IL-8 and MMP-8 in gingival crevicular fluid were measured by ELISA. Periodontal parameters as well as gingival crevicular fluid humoral factor amounts were evaluated in the control group and in chronic periodontitis patients at baseline and at 1 and 4 wk after scaling and root planing treatment. RESULTS: At baseline, there were significant differences between control subjects and chronic periodontitis patients in terms of clinical attachment loss, pocket depth, gingival index (p < 0.001) and approximal plaque index (p < 0.01). The amounts of IL-1ß, MMP-8 (p < 0.001) and IL-8 (p < 0.01) in gingival crevicular fluid were significantly lower in healthy subjects than in chronic periodontitis patients. Scaling and root planing led to improvement in all examined clinical parameters, apart from clinical attachment loss. Periodontal treatment also resulted in a significant decrease in the amounts of IL-1ß, IL-8 and MMP-8 in comparison to baseline, especially 4 wk after scaling and root planing (p < 0.001); however, the amounts of these humoral factors were still higher than those in control group. CONCLUSION: Our observations indicated that short-term nonsurgical therapy resulted in a significant improvement in periodontal indices and in a marked decrease of IL-1ß, IL-8 and MMP-8 gingival crevicular fluid levels. Nevertheless, no significant correlations were found between clinical parameters and amounts of humoral factors after therapy.

Lipopolysaccharide from Porphyromonas gingivalis stimulates rat mast cells to cysteinyl leukotriene generation and upregulates Toll-like receptor -2 and -4 expression.

Mast cells are found in all tissues of the oral cavity and it is suggested that they take part in the development of oral inflammation. As Porhyromonas gingivalis is widely recognized as a major pathogen in the development and progression of gingivitis and periodontitis, the aim of our study is to determine the effect of P. gingivalis lipopolysaccharide (LPS) on mast cell degranulation, cysteinyl leukotriene (cysLT) generation, and migration, as well as Toll-like receptor (TLR)-2 and -4 expression. Experiments were carried out in vitro on rat peritoneal mast cells. LPS-induced mast cell histamine release was estimated by a spectrofluorometric method and cysLT generation by ELISA test. Mast cell migration in response to this antigen was examined according to Boyden's modified method and TLR expression was determined by flow cytometry. We found that P. gingivalis LPS did not induce mast cell degranulation and histamine release. However, activation of mast cells with this bacterial antigen resulted in generation and release of significant amounts of cysLTs. We also documented that LPS from P. gingivalis did not stimulate mast cell migration, even in the presence of laminin, whereas it strongly upregulated TLR2 and TLR4 expression on mast cells. Observations that P. gingivalis LPS activates mast cells to generate and release proinflammatory mediators such as cysLTs and modulates TLR2 and TLR4 expression indicates that these cells might be involved in the emergency of inflammatory processes evolved in response to P gingivalis infection.

Selected Metabolic Markers in Girls with Turner Syndrome: A Pilot Study.

BACKGROUND: Turner syndrome (TS) predisposes an individual to obesity and related metabolic disorders. As the TS population is at a higher risk of cardiovascular diseases and malformations, research into laboratory markers of metabolic complications has been ongoing. Special significance has recently been attributed to matrix metalloproteinases (MMPs), their inhibitors (TIMPs), and neurotrophic factors, such as BDNF and GDNF. OBJECTIVE: To establish whether cardiometabolic risk in patients with TS is reflected in the concentrations of metalloproteinases and neurotrophic factors. METHOD: The concentrations of circulating MMP-1, MMP-2, MMP-9, TIMP-1, BDNF, GDNF, and VEGF were measured in 17 patients with TS. The control group was composed of 11 girls with nonpathologic short stature and normal karyotype. RESULTS: There were no differences in chronological or bone age. No significant differences were observed in mean weight, although the Z-score BMI was higher in the study group. The mean baseline values of MMP-1 and BDNF were significantly lower in the control group than in the study group (p < 0.001, p = 0.001). Regression analysis revealed a positive correlation between MMP-1 concentrations and Z-score BMI (r = 0.36, p = 0.047) and between BDNF and Z-score BMI (r = 0.48, p = 0.013). CONCLUSION: Our pilot study showed that MMP-1 may be a potential indicator of a higher risk of cardiometabolic complications in girls with TS. The elevated concentrations of BDNF in normal-weight girls with TS need to be studied further, taking into consideration the influence of estrogen-androgen imbalance.

Mast cells participate in chronic low-grade inflammation within adipose tissue.

Obesity is reckoned as one of the civilization diseases, posing a considerable global health issue. Evidence points towards a contribution of multitude immune cell populations in obesity pathomechanism and the development of chronic low-grade inflammation in the expanded adipose tissue. Notably, adipose tissue is a reservoir of mast cells which number in individuals with obesity particularly increased. Some of them tend to degranulation what generate secretion of strong pro-inflammatory and regulatory mediators, as well as cytokines/chemokines. Several lines of evidence suggest that mast cells are strictly associated with pro-inflammatory status in adipose tissue by their indirect impact on immune cell attraction and activation. Furthermore, mast cells affect adipose tissue remodelling and fibrosis by adipocyte differentiation, fibroblast proliferation and enhancing extracellular matrix proteins expression. This review will summarize current knowledge on mast cell features and their role in the development of chronic low-grade inflammation within adipose tissue.

The action of tumor necrosis factor-alpha on rat mast cells.

Taking into account that cytokine tumor necrosis factor-alpha (TNF-alpha) and mast cells (MC) both are involved in inflammation, it seems of great importance to recognize their relationships. Therefore, we have studied whether recombinant human TNF-alpha (rHuTNF-alpha) can cause histamine secretion from rat peritoneal MC. We have also examined the effect of this cytokine on MC reactivity. We have established that TNF-alpha stimulates rat MC to histamine release in a concentration-dependent manner. TNF-alpha-induced histamine secretion was evoked by concentrations > 10-16 M and reached the maximum rate at a concentration of 10-10 M (histamine release 17.1% +/- 1.9%, mean +/- SEM). We have also noticed that pretreatment of MC with TNF-alpha (in a concentration of 10-16 M) significantly inhibited concanavalin A (ConA)-stimulated release of histamine, with the percent release decreasing to 51% of the control value. Treatment of mast cells with TNF-alpha resulted in a decrease of compound 48/80-dependent histamine release as well (the percent released histamine fell to 85% of the control value). This altered MC responsiveness was reversible. After 120 min of resting time, the MC reactivity came back to the initial values. We have concluded that TNF-alpha appears to be a direct stimulus for MC to release histamine, and it may regulate MC secretory function.

Tumor necrosis factor alpha (TNF-alpha) modulates rat mast cell reactivity.

Nowadays there is growing evidence that some cytokines regulate biological functions of the mature mast cells. Therefore, we have studied whether TNF-alpha, the cytokine of multifunctional activities, could directly stimulate rat peritoneal mast cells to histamine secretion and whether it could modulate rat mast cell reactivity in anaphylactic (with ConA) and anaphylactoid (with compound 48/80) reactions. We have established that rat recombinant TNF-alpha does not activate rat mast cells to histamine release. However, TNF-alpha-treatment causes the decrease of spontaneous histamine release up to 85% (TNF-alpha concentration: 2 x 10(-9) M). Pretreatment of mast cells with TNF-alpha inhibits ConA-stimulate release of histamine with the percent release decreasing up to 33.7% of the control value (TNF-alpha concentration: 5 x 10(-9) M) and this decrease is statistically significant. Pretreatment of mast cells with TNF-alpha reduces compound 48/80-dependent histamine release as well and the percent release of histamine fell to 64.7% of the control value. We have concluded that TNF-alpha may play a significant role in regulation of mast cell secretory activity.

Tumor necrosis factor alpha (TNF-alpha) activates human adenoidal and cutaneous mast cells to histamine secretion.

It is widely known that mast cells and cytokine TNF-alpha are both involved in inflammatory reactions. Therefore, we have studied whether TNF-alpha can cause histamine secretion from human adenoidal and cutaneous mast cells. The experiments were performed in vitro on mast cells isolated from tissues by enzymatic dispersion technique. The results of our experiments have clearly shown that this cytokine stimulates mast cells to histamine release. TNF-alpha-induced histamine release was concentration- and time-dependent. Moreover, the release of histamine evoked by TNF-alpha was also dependent on reaction temperature and on glycolytic and oxidative cellular metabolism. We have concluded that TNF-alpha is a potent stimulus for mast cells to release histamine and that it induces histamine release via an active, secretory process.

Histamine-releasing activity of lymphocyte supernatants of guinea pig spleen cell cultures.

We demonstrated the production of a histamine releasing factor (HRF) by 24-h cultures of guinea pig spleen cells which were stimulated or not with specific antigen (ovalbumin, OA) or mitogen (phytohemagglutinins or concanavalin A). HRF induced the release of histamine from homologous mesenteric mast cells in a dose-dependent fashion. The HRF-induced histamine release was not high compared to the release induced by calcium ionophore A23187, but higher than that induced by compound 48/80, polymyxin B and con canavalin A. The mast cells from sensitized guinea pigs released histamine when challenged with OA. We found that HRF-induced histamine release was additive to that induced by antigen, when both agents were added simultaneously to sensitized mast cells. The phenomenon was most significant when a suboptimal dose of antigen was used. Moreover, we did not observe any differences in the magnitude of HRF-induced histamine release between the mast cells from nonsensitized and sensitized guinea pigs. The time course of histamine release induced by HRF was significantly slower than that with specific antigen (10 min and 45 sec, respectively). Our results may suggest that HRF acts on mast cells through a different not immunological mechanism.

Effect of cisplatin and cis-platinum (II) phosphonate complex on murine mast cells.

We investigated the effect of two cis-platinum (II) complexes, cis-[PtCl2(NH3)2] (cisplatin) and cis-[PtCl2(4-pmpe)2] (4-pmpe stands for diethyl 4-pyridylmethylphosphonate), which was recently synthetized in our laboratory, on murine mast cells. We noticed that both tested compounds were able to evoke histamine release from murine mast cells. The histamine secretion was dependent on the concentration of compound and on the time and temperature of the reaction. It was also dependent on metabolic energy (the reaction was diminished in a medium without glucose and abolished in the presence of 2-deoxyglucose). The results indicate that cis-platinum (II) complexes activate mast cells to secrete histamine via a non-cytotoxic, active secretory process.

Histamine secretion from human mesenteric and adenoidal mast cells.

Human mast cells were obtained from adenoids and mesentery by enzymatic dispersion of the tissues with the enzyme collagenase. The digestion of the tissues resulted in a cell suspension which contained 1-2% mast cells. 37.3% (adenoids) and 33.4% (mesentery) of total histamine initially present in the tissues was recovered in the dispersed cell suspensions. More than 90% of the cells were viable. The adenoidal mast cells could be sensitized passively in vitro with homologous reaginic serum and released histamine after challenge with specific antigen. Both populations of mast cells were sensitive to the action of anti-human IgE; the reversed anaphylaxis with anti-IgE was higher in mesenteric mast cells. Both examined mast cell populations were sensitive to the challenge with polymyxin B, concanavalin A and ionophore A23187, however, histamine release was only up to 10% and 20% for adenoidal and mesenteric cells, respectively. Only mesenteric mast cells responded to the action of compound 48/80. Histamine release, induced by polymyxin B, was rapid (maximal release within 5 min), maximal in the presence of 3 mM extracellular calcium ions (but also occurred in the absence of the cation).

The lack of competition between two unrelated IgE antibodies in in vivo sensitization of mouse mast cells.

The relation between IgE antibodies of different specificity in in vivo sensitization of mouse mast cells were studied. In the system of active immunization with EA and KLH mixed with Al(OH)3 no inhibition of an IgE and IgGl response to one antigen by simultaneous administration of other antigen was observed. The lack of inhibition was reflected in serum anti-EA and anti-KLH antibody levels as well as in mast cell sensitization expressed in antigen-induced histamine release. Furthermore, we were unable to detect an inhibitory effect of active immunization of mice with EA on in vivo passive sensitization of peritoneal mast cells. These observations indicated that in such a system immunization actively produced IgE antibodies do not saturate mast cell IgE receptors and do not inhibit subsequent sensitization of these cells with IgE antibodies of another specificity.

Sensitivity of human cutaneous mast cells to anaphylactic and nonimmunological stimuli.

Mast cells represent significant cellular element of the skin. It is now postulated that they play an important role in cutaneous homeostasis and are engaged in some pathological processes as well. The aim of our study was to examine sensitivity of human cutaneous mast cells to anaphylactic and anaphylactoid stimuli. The studies were performed on human cutaneous mast cells obtained from healthy skin by enzymatic dispersion technique. The mast cells were activated in vitro with anti-IgE, concanavalin A (Con A), compound 48/80, substance P (SP) or tumor necrosis factor alpha (TNF-alpha). We have noticed that skin mast cells were susceptible to the challenge with anti-IgE and Con A, and histamine release was dose- and time-dependent. In both cases histamine release was high (44.0 +/- 4.1% with anti-IgE at dilution 1:500 and 20.1 +/- 2.4% with Con A in concentration 500 micrograms/ml). Cutaneous mast cells were challenged in a dose- and time-related fashion with compound 48/80, however histamine release was low (9.8 +/- 2.4%, at concentration of compound 48/80-100 micrograms/ml). SP and TNF-alpha also activated mast cells but the magnitude of histamine release was not high (up to 7.1 +/- 0.9%, SP in concentration 10(-4) M and 17.4 +/- 1.1%, TNF-alpha in concentration 10(-6) M) and maximal after 20 min reaction.

The effect of rat mast cell sensitization with mouse IgE antibody on anaphylactic histamine release by anti-mouse IgE and Con A.

The peritoneal mast cells of rat (Wistar X August) F1 were incubated with purified mouse IgE antibodies and then challenged (in the presence of 50% D2O) with antigen, anti-mouse IgE and concanavalin A. It was found that incubation of mast cells with IgE antibodies led to different level of cell sensitization (in mast cells from different rats), expressed in antigen-induced histamine release (0-52%). Moreover, a) significant antigen-induced histamine release was usually accompanied by high Con A- and anti-IgE-induced histamine release from these cells; the magnitude of release was comparable to Con A- and anti-IgE-induced release from control, nonsensitized cells of the same rat; b) low antigen-induced histamine release was accompanied by the decrease of Con A- and anti-IgE-induced release, as compared to the release from control cells. This fall of reactivity to Con A and anti-IgE was statistically significant and was irreversible during 120 min.

The desensitization of rat mast cells sensitized under various conditions.

Rat peritoneal and pleural mast cells in vitro sensitized with an excess of mouse IgE antibodies were tested for desensitization by preincubation with antigen in calcium-free conditions. It was found that 30 min preincubation in these conditions induced complete desensitization of pleural and peritoneal cells to subsequent action of specific antigen in calcium-containing medium. The extent of desensitization to the action of anti-rat IgE (nonspecific desensitization) depended on: the origin of the cells (pleural mast cells were more sensitive to desensitization), time of incubation of the cells with mouse IgE (longer time of sensitization resulted in higher extent of desensitization), anti-IgE dilutions used to rechallenge histamine release (higher extent of desensitization to higher dilutions of anti-IgE). The results suggest, that nonspecific desensitization occurs in mast cells with higher level of sensitization with mouse IgE. Some functional differences between pleural and peritoneal mast cells are also suggested.