Association between Expression of Insulin-like Growth Factor-1 (IGF-1), IGF-1 Receptor (IGF-1R), and Hypertension-Mediated Organ Damage (HMOD) Parameters in Leukocytes and Plasma of Children/Adolescents with Primary Hypertension.

A decrease in IGF-1 is often linked to inflammation. Low systemic and local IGF-1 production and downregulation of IGF-1R expression may precede and predict PH development in children/adolescents. Leukocyte mRNA expression of IGF-1 and its receptor (IGF-1R) and plasma IGF-1 were measured in a group of 39 PH children/adolescents (29 boys and 10 girls) and 35 age-matched normotensive children (19 boys and 16 girls) using the RT-PCR and ELISA tests. The expression of the IGF-1R protein was assessed by flow cytometry. Plasma IGF-1 concentration was evaluated with ELISA. The expression of IGF-1 and IGF-1R and plasma concentrations of IGF-1 did not differ between groups. However, the PH children had a decreased percentage in IGF-1R-bearing lymphocytes (p = 0.02) and monocytes (p = 0.0003), as well as a low density of IGF-R in monocytes (p = 0.02). The IGF-1 expression was negatively correlated with pulse-wave velocity (PWV) (r = -0.49), systolic blood pressure (SBP) (-0.44), and carotid intima-media thickness (cIMT) (-0.43). The IGF-1R expression was negatively correlated with PWV (r = -0.42) and SBP (r = -0.41). Our results suggest that early subclinical hypertensive arterial injury is associated with lower activity of IGF-1-IGF-1R expression and loss of protective actions.

Implications of the Matrix Metalloproteinases, Their Tissue Inhibitors and Some Other Inflammatory Mediators Expression Levels in Children Obesity-Related Phenotypes.

OBJECTIVES: Matrix metalloproteinases (MMPs) are calcium-dependent zinc-containing endo-peptidases engaged in many biological processes including adipogenesis, angiogenesis, and tissue remodeling. Fat tissue infiltration by peripheral leukocytes plays an important role in transition of fat tissue residual, non-inflammatory status into the pro-inflammatory one, resulting in fat tissue inflammation and expansion as well as production of many mediators like adipokines and cytokines. The aim of this study was to investigate the expression of MMPs, their endogenous tissue inhibitors (TIMPs), and selected inflammatory mediators in leukocytes and plasma of children with simple obesity to find their associations with obesity-related phenotypes. MATERIAL AND METHODS: Twenty-six overweight/obese children and twenty-three healthy volunteers participated in the study. The leukocyte mRNA expression levels of MMP-2, -9, -12 -14, TIMP-1, -2, and IL-6 were analyzed by the real time quantitative PCR. Plasma MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios as well as the concentrations of MMP-9, TIMP-1, IL-1 beta, IL-6, TNF- alpha, leptin and resistin were tested by ELISA assays. Gelatin zymography was used to assess the activity of the leukocyte MMPs proteins. RESULTS: The obese children showed the following: a) increased expression of leukocyte TIMP-1 and slight elevation (close to statistical significance) of leukocyte MMP-9 (p = 0.054), the decline in MMP-2, b) elevation of plasma MMP-9, leptin, and MMP9/TIMP1 ratio, c) reduced expression of plasma TNF-alpha and MMP-2/TIMP-2 ratio. Several negative correlations were found: TIMP2 vs. ALT (r = -0.536), AST (r = -0.645) and TTG (r = -0.438), IL-6 vs. GGTP (r = -0.815), and MMP12 vs. TTG (r = -0.488), leptin vs. ALT (r = -0.569), MMP-9 vs. total cholesterol (r = -0.556). The only positive correlation was that of plasma leptin level vs. GGTP (r = 0.964). CONCLUSIONS: At the beginning of obesity development (children), possibly compensatory reactions prevail, reflected here by an increase in the expression of leukocyte MMPs inhibitor TIMP-1, decrease in the level of leukocyte MMP-2 and plasma MMP-2, MMP2/TIMP-2 ratio, low plasma TNF-alpha and negative correlations between the expression of TIMP-2 and liver (AST, ALT) or fat (TTG) inflammatory markers.

Expression of Matrix Metalloproteinases in the Circulating Immune Cells in Children with Helicobacter pylori Infection-Correlation with Clinical Factors.

H. pylori gastritis is strongly associated with the upregulation of the expression of several matrix metalloproteinases (MMPs) in the gastric mucosa. However, the role of MMP-2 and MMP-9, and their inhibitors (tissue inhibitors of metalloproteinases -TIMPs) produced by immune cells in infected children have not been clearly defined. Moreover, the effects of H. pylori eradication therapy on MMPs and TIMPs production has not been evaluated. A total of 84 children were studied: 24-with newly diagnosed H. pylori gastritis, 25-after H. pylori eradication therapy (17 of them after successful therapy), 24-with H. pylori-negative gastritis, and 11-controls. Plasma levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 by ELISA; MMPs and TIMPs expression in lymphocytes; neutrophils and monocytes in peripheral blood by multiparameter flow cytometry; and mucosal mRNA expression levels of MMPs and TIMP-1 in gastric biopsies by RT-PCR were evaluated. Children with H. pylori-related gastritis showed the following: (1) increased MMP-2 and TIMP-2 plasma levels, (2) increased intracellular expression of MMP-2 in the circulating lymphocytes and neutrophils, (3) low frequencies of circulating TIMP-1+ and TIMP-2+ leukocytes, and (4) high expression of mRNA for MMP-9 along with low expression of mRNA for MMP-2 in the gastric mucosa. Unsuccessful H. pylori eradication was associated with the following: (1) high plasma levels of MMP-9 and TIMP-1, (2) increased pool of TIMP-1+ lymphocytes as well as high expression of MMP-9 in circulating lymphocytes, and (3) high expression of mRNA for MMP-9 in the gastric mucosa. Our data suggest that MMPs are important contributors to stomach remodelling in children with H. pylori-related gastritis. Unsuccessful H. pylori eradication is associated with increased MMP-9 in plasma, circulating lymphocytes, and gastric mucosa.

Distribution and maturation state of peripheral blood dendritic cells in children with primary hypertension.

Dendritic cells (DCs) play an important role in T cell alterations in primary hypertension (PH). We determined the numbers and maturation markers of peripheral blood total DCs (tDCs), myeloid cells (mDCs), and plasmacytoid cells (pDCs) and their association with hypertension-mediated organ damage (HMOD) markers and selected immune parameters in 30 adolescents with white coat hypertension (WCH), 25 adolescents with PH and a group of 35 age- and sex-matched children with normotension. Using multicolor flow cytometry, expression of maturation markers (CD86 and CD83) in tDCs (Lin1-/HLA-DR+), myeloid DCs (Lin1-/HLA-DR+/CD11c+) (mDCs), and plasmacytoid DCs (Lin1-/HLA-DR+/CD123+) (pDCs) and the distribution of peripheral Th17-bearing and T-reg cells were defined. In subjects with hypertension, carotid intima-media thickness (cIMT), left ventricular mass index (LVMI), and pulse wave velocity (PWV) were assessed. Compared with WCH and subjects with normotension, subjects with hypertension had reduced tDC and pDC numbers, an increased percentage of mDC subsets, an elevated mDC/pDC ratio, an increased population of mature mDC and pDC subsets bearing CD83 of high density, a decreased subset of CD86-bearing pDCs, and increased expression of DC activation markers (HLA-DR, CD86), as well as CD11c (mDCs) and CD123 (pDCs). PWV, LVMI, and cIMT values correlated negatively with tDCs and pDCs and positively with mDC numbers. Expression of DC maturation/activation markers (CD83, CD86, HLA-DR, CD11c, and CD123) correlated positively with PWV. Certain DC characteristics of WCH subjects resembled those of PH subjects (decreased tDC frequency and upregulation of activation marker expression). These changes correlated with HMOD. WCH subjects presented a DC phenotype that was intermediate between the normotensive and hypertensive phenotypes.

Expression of Matrix Metalloproteinases and Their Tissue Inhibitors in Peripheral Blood Leukocytes and Plasma of Children with Nonalcoholic Fatty Liver Disease.

Gene expression profiles of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) were evaluated in peripheral blood leukocytes of children with nonalcoholic fatty liver disease (NAFLD). Gene expression patterns were correlated with their plasma protein counterparts, systemic parameters of liver injury, and selected markers of inflammation. The MMP-2, MMP-9, MMP-12, MMP-14, TIMP-1, TIMP-2, TGF-ß, and IL-6 transcripts levels were tested by the real-time PCR. Plasma concentrations of MMP-9, TIMP-1, MMP-9/TIMP-1 ratio, MMP-2/TIMP-2 ratio, sCD14, leptin, resistin, IL-1 beta, and IL-6 and serum markers of liver injury were estimated by ELISA. The MMP-9, TIMP-2 expression levels, plasma amounts of MMP-9, TIMP-1, and the MMP-9/TIMP-1 ratio were increased in children with NAFLD. Concentrations of AST, ALT, GGT, and leptin were elevated in serum patients with NAFLD, while concentration of other inflammatory or liver injury markers was unchanged. The MMP-2 and MMP-9 levels correlated with serum liver injury parameters (ALT and GGT concentrations, respectively); there were no other correlations between MMP/TIMP gene expression profiles, their plasma counterparts, and serum inflammatory markers. Association of MMP-2 and MMP-9 expression with serum liver injury parameters (ALT, GGT) may suggest leukocyte engagement in the early stages of NAFLD development which possibly precedes subsequent systemic inflammatory responses.

Regulatory T-cell subset distribution in children with primary hypertension is associated with hypertension severity and hypertensive target organ damage.

BACKGROUND: The relationship between circulating regulatory T-cell (Tregs) subset distribution and hypertension severity in children with primary hypertension is not known. We aimed to find out if target organ damage (TOD) in children with primary hypertension is related to defects in Tregs distribution reflected by their phenotype characteristics. METHODS: The study constituted 33 nontreated hypertensive children and 35 sex-matched and age-matched controls. Using multicolor flow cytometry technique, we assessed a distribution of the total Tregs (CD4CD25CD127) and their subsets (CD45RA-naive Tregs, CD45RA memory/activated Tregs, CD45RACD31 recent thymic emigrants Tregs and mature naive CD45RACD31 Tregs) in the whole blood. RESULTS: Hypertensive children showed decreased percentage of the total Tregs, the CD45RA-naive Tregs, the total CD31 Tregs and the recent thymic emigrants Tregs but elevation of the CD45RA memory/activated Treg and mature naive CD45RACD31 Tregs. Decreased frequency of the total Tregs, naive Tregs and CD31-bearing Treg cell subsets (CD31 total Tregs, CD45RACD31 recent thymic emigrants Tregs) negatively correlated to TOD markers, arterial stiffness and blood pressure elevation. In contrast, increased percentage of memory Tregs and CD31 Tregs subsets positively correlated to organ damage markers, arterial stiffness and blood pressure values. These changes were independent of BMI, age, sex and hsCRP. CONCLUSION: Both diagnosis of hypertension, TOD and arterial stiffness in hypertensive children were associated with decreased population of total CD4 Tregs, limited output of recent thymic emigrants Tregs, and increased pool of activated/memory Tregs. Hypertension was an independent predictor of the circulating Treg subsets distribution irrespective of hsCRP.

Loss of CD31 receptor in CD4+ and CD8+ T-cell subsets in children with primary hypertension is associated with hypertension severity and hypertensive target organ damage.

BACKGROUND: Primary hypertension is associated with still poorly known T-cell dependent immunity defects that participate in the disease development. However, the relationship between peripheral T-cell subset distribution and disease severity in humans is not known. The aim of the study was to find out if target organ damage in adolescents with primary hypertension is associated with thymus-dependent lymphocytes renewal reflected by changes in the T-cell subset phenotype characteristics. METHODS: Using seven-color flow cytometry technique, we assessed CD31, CCR7 and CD28 receptors expression in CD45RA and CD45RO bearing peripheral CD4 and CD8 T-cell subsets. The study included 32 hypertensive children/adolescents and 35 sex-matched and age-matched controls. RESULTS: Children with primary hypertension had slightly increased CD4 T-cell pool but decreased population of CD31 expressing CD4 T-cell subsets (recent thymic emigrants). Frequency of the CD4 and CD4/CD45RA+ T cells lacking CD31 correlated positively with the hypertensive organ damage markers (pulse wave velocity, central blood pressure, left ventricular mass index). Left ventricular hypertrophy was associated with decreased CD4/CD45RA:CD4/CD45RO ratio, loss of the CD31 receptor in the CD4 and CD8 T-cell subsets and increased population of effector/memory T cells bearing CD8/CD28 and CD8/CD45RA+/CCR7 phenotype. Regression analysis revealed that these associations were independent of age, sex, and BMI. CONCLUSION: The results suggest that subclinical arterial injury and left ventricular hypertrophy in adolescents with primary hypertension is associated with declined thymic function and increased pool of T cells bearing effector/memory phenotype.

Distinct cellular phenotype linked to defective DNA interstrand crosslink repair and homologous recombination.

Repair of DNA interstrand crosslinks (ICLs) predominantly involves the Fanconi anemia (FA) pathway and homologous recombination (HR). The HR repair system eliminates DNA double strand breaks (DSBs) that emerge during ICLs removal. The current study presents a novel cell line, CL­V8B, representing a new complementation group of Chinese hamster cell mutants hypersensitive to DNA crosslinking factors. CL­V8B exhibits increased sensitivity to various DNA­damaging agents, including compounds leading to DSBs formation (bleomycin and 6­thioguanine), and is extremely sensitive to poly (ADP-ribose) polymerase inhibitor (>400­fold), which is typical for HR­defective cells. In addition, this cell line exhibits a reduced number of spontaneous and induced sister chromatid exchanges, which suggests likely impairment of HR in CL­V8B cells. However, in contrast to other known HR mutants, CL­V8B cells do not show defects in Rad51 foci induction, but only slight alterations in the focus formation kinetics. CL­V8B is additionally characterized by a considerable chromosomal instability, as indicated by a high number of spontaneous and MMC­induced chromosomal aberrations, and a twice as large proportion of cells with abnormal centrosomes than that in the wild type cell line. The molecular defect present in CL­V8B does not affect the efficiency and stabilization of replication forks. However, stalling of the forks in response to replication stress is observed relatively rarely, which suggests an impairment of a signaling mechanism. Exposure of CL­V8B to crosslinking agents results in S­phase arrest (as in the wild type cells), but also in larger proportion of G2/M­phase cells and apoptotic cells. CL­V8B exhibits similarities to HR­ and/or FA­defective Chinese hamster mutants sensitive to DNA crosslinking agents. However, the unique phenotype of this new mutant implies that it carries a defect of a yet unidentified gene involved in the repair of ICLs.

Altered matrix metalloproteinase 9 and tissue inhibitor of metalloproteinases 1 levels in children with primary hypertension.

BACKGROUND: Matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) are involved in cardiovascular remodeling in hypertension. Because metabolic abnormalities typical of metabolic syndrome is the dominant phenotype of primary hypertension in children, we hypothesized that MMP-9 and TIMP-1 plasma concentrations are altered in hypertensive children and correlate with metabolic abnormalities and target organ damage. METHOD: A total of 109 children (15.6, 10-17 years) with untreated primary hypertension were included to the study. The control group consisted of 74 healthy, normotensive children. RESULTS: Plasma MMP-9, TIMP-1 concentrations, and MMP-9/TIMP-1 ratio were significantly elevated in hypertensive boys in comparison with normotensive boys (P = 0.0001, P = 0.04, and P = 0.001, respectively), whereas there were no differences between hypertensive and normotensive girls. The levels of MMP-9 and TIMP-1 as well as MMP-9/TIMP-1 ratio were not associated either with hypertension stage, left ventricular hypertrophy, or carotid intima-media thickness. However, in a subgroup of 30 hypertensive patients in whom arterial stiffness was measured, TIMP-1 concentrations correlated with aortic pulse pressure (P < 0.05; r = 0.367), augmentation pressure (P < 0.05; r = 0.428), and augmentation index (P < 0.05; r = 0.404).Only hypertensive boys presented negative correlations of both MMP-9 and TIMP-1 levels with high-density lipoprotein cholesterol (r = -0.254, P = 0.01 and r = -0.241, P = 0.02, respectively). CONCLUSION: Hypertensive boys but not girls had elevated MMP-9 and TIMP-1 plasma concentrations, which indicates sex-related role of MMP/TIMP system in pediatric hypertension. The correlation between serum TIMP-1 and markers of arterial stiffness indicates on the involvement of TIMPs in arterial remodeling.

Melanoma cell adhesion molecule as an emerging biomarker with prognostic significance in systolic heart failure.

BACKGROUND: Melanoma cell adhesion molecule (MCAM) is a marker of endothelial damage. MCAM diagnostic and prognostic value was assessed in chronic heart failure (CHF). MATERIALS & METHODS: 130 CHF patients and 32 controls were included in the study. Telephone follow-up lasted one year. End points were: death from all causes, and hospitalization with CHF exacerbation. RESULTS: MCAM was higher in patients than in controls (p = 0.01). Receiver operator curve analysis revealed that MCAM may serve as a predictor of death (area under the curve: 0.8404; p < 0.002). Patients with MCAM above 500 ng/ml had worse prognosis (p = 0.03). NT-proBNP and age were independent predictors of death in multivariate analysis. CONCLUSION: The increased MCAM indicates endothelial damage in CHF and may serve as a marker of worse prognosis in these patients.

Phenotype of NK Cells Determined on the Basis of Selected Immunological Parameters in Children Treated due to Acute Lymphoblastic Leukemia.

Acute lymphoblastic leukemia (ALL) is the most frequent pediatric malignancy. The chemotherapy for ALL is associated with a profound secondary immune deficiency.We evaluated the number and phenotype of natural killer (NK) cells at diagnosis, after the intensive chemotherapy and following the completion of the entire treatment for patients with ALL. The fraction, absolute number, and percentage of NK cells expressing interferon-γ were determined in full blood samples. The fraction of NK cells expressing CD158a, CD158b, perforin, A, B, and K granzymes was examined in isolated NK cells.We have shown that patients assessed at ALL diagnosis showed significantly lower values of the fraction of NK cells and percentage of NK cells with the granzyme A expression. Additionally, the absolute number of NK cells, the expression of CD158a, CD158b, perforin, and granzyme A were significantly lower in patients who completed intensive chemotherapy. Also, there was a significantly higher fraction of NK cells expressing granzyme K in patients who completed the therapy.Abnormalities of NK cells were found at all stages of the treatment; however, the most pronounced changes were found at the end of intensive chemotherapy.

Does the liposuction method influence the phenotypic characteristic of human adipose-derived stem cells?

Adipose-derived stem cells (ASCs) possess a high differentiation and proliferation potential. However, the phenotypic characterization of ASCs is still difficult. Until now, there is no extensive analysis of ASCs markers depending on different liposuction methods. Therefore, the aim of the present study was to analyse 242 surface markers and determine the differences in the phenotypic pattern between ASCs obtained during mechanical and ultrasound-assisted liposuction. ASCs were isolated from healthy donors, due to mechanical and ultrasound-assisted liposuction and cultured in standard medium to the second passage. Differentiation potential and markers expression was evaluated to confirm the mesenchymal nature of cells. Then, the BD LyoplateTM Human Cell Surface Marker Screening Panel was used. Results shown that both population of ASCs are characterized by high expression of markers specific for ASCs: cluster of differentiation (CD)9, CD10, CD34, CD44, CD49d, CD54, CD55, CD59, CD71 and low expression of CD11a, CD11c and CD144. Moreover, we have noticed significant differences in antigen expression in 58 markers from the 242 studied. Presented study shows for the first time that different liposuction methods are not a significant factor which can influence the expression of human ASCs surface markers.

Expression of Adiponectin Receptors on Peripheral Blood Leukocytes of Hypertensive Children Is Associated with the Severity of Hypertension.

The aim of the study was to find out whether peripheral blood leukocyte adiponectin receptors 1 and 2 (AdipoR1, AdipoR2) protein expression patterns (flow cytometry) differ between the primary hypertension children (n = 57) and healthy controls (n = 19) and if their expression levels are related to selected clinical parameters. The group of 26 patients [AdipoR(-)] showed lower and the group of 31 patients [AdipoR(+)] showed higher AdipoRs protein expression than the control and each other (P < 0.01 for neutrophils, P < 0.05 for monocytes). The AdipoR(+) leukocytes expressed higher AdipoR1 mRNA levels (RT-PCR) than AdipoR(-) ones and controls (P = 0.022 and P = 0.007, resp.). Despite greater BMI, the AdipoR(-) patients had unchanged serum adiponectin levels. In contrast, AdipoR(+) patients had lower serum adiponectin concentrations than the AdipoR(-) ones and controls (P < 0.001). The AdipoR(+) patients had higher blood pressure (P = 0.042) and greater carotid intima-media thickness (P = 0.017) than the AdipoR(-) ones. The stage of hypertension was associated with increased neutrophil but not monocyte AdipoR1 density (AdipoR1 MFI) (P < 0.05). Severe ambulatory hypertension was presented more often in AdipoR(+) patients than in AdipoR(-) ones (51.6% versus 26.9%, resp.; P < 0.01). In conclusion, neutrophil AdipoRs upregulation was associated with early stages of vascular injury, hypertension severity, and low serum levels of adiponectin.

Lactic acid bacteria strains exert immunostimulatory effect on H. pylori-induced dendritic cells.

The aim of this study was to find out if selected lactic acid bacteria (LAB) strains (antagonistic or nonantagonistic against H. pylori in vitro) would differ in their abilities to modulate the DCs maturation profiles reflected by their phenotype and cytokine expression patterns. Methods. Monocyte-derived DCs maturation was elicited by their direct exposure to the LAB strains of L. rhamnosus 900 or L. paracasei 915 (antagonistic and nonantagonistic to H. pylori, resp.), in the presence or absence of H. pylori strain cagA+. The DCs maturation profile was assessed on the basis of surface markers expression and cytokines production. Results. We observed that the LAB strains and the mixtures of LAB with H. pylori are able to induce mature DCs. At the same time, the L. paracasei 915 leads to high IL-10/IL-12p70 cytokine ratio, in contrast to L. rhamnosus 900. Conclusions. This study showed that the analyzed lactobacilli strains are more potent stimulators of DC maturation than H. pylori. Interestingly from the two chosen LAB strains the antagonistic to H. pylori-L. rhamnosus strain 900 has more proinflammatory and probably antibactericidal properties.

[Lactic acid bacteria and health: are probiotics safe for human?]. / Bakterie kwasu mlekowego i zdrowie: czy probiotyki sa bezpieczne dla czlowieka?

The effect of Lactobacillus and Bifidobacterium on human health has been examined for many years. Numerous in vivo and in vitro studies have confirmed the beneficial activity of some exogenous lactic acid bacteria in the treatment and prevention of rotaviral infection, antibiotic-associated diarrhea, inflammatory bowel disease and other gastrointestinal disorders. Probiotics support the action of the intestinal microflora and exhibit a favorable modulatory effect on the host's immune system. However, it should be remembered that relatively harmless lactobacilli can occasionally induce opportunistic infections. Due to reaching almost 20x10(12) probiotic doses per year which contain live cultures of bacteria, it is essential to monitor the safety aspect of their administration. In recent years, infections caused by Lactobacillus and Bifidobacterium made up 0.05% to 0.4% of cases of endocarditis and bacteremia. In most cases, the infections were caused by endogenous microflora of the host or bacterial strains colonizing the host's oral cavity. According to a review of cases of infections caused by bacteria of the genus Lactobacillus from 2005 (collected by J.P. Cannot'a), 1.7% of infections have been linked directly with intensive dairy probiotic consumption by patients. Additionally, due to the lack of a precise description of most individuals' eating habits, the possible effect of probiotics on infection development definitively should not be ruled out. The present paper describes cases of diseases caused by lactic acid bacteria, a potential mechanism for the adverse action of bacteria, and the possible hazard connected with probiotic supplementation for seriously ill and hospitalized patients.

Collaborating with the enemy: function of macrophages in the development of neoplastic disease.

Due to the profile of released mediators (such as cytokines, chemokines, growth factors, etc.), neoplastic cells modulate the activity of immune system, directly affecting its components both locally and peripherally. This is reflected by the limited antineoplastic activity of the immune system (immunosuppressive effect), induction of tolerance to neoplastic antigens, and the promotion of processes associated with the proliferation of neoplastic tissue. Most of these responses are macrophages dependent, since these cells show proangiogenic properties, attenuate the adaptive response (anergization of naïve T lymphocytes, induction of Treg cell formation, polarization of immune response towards Th2, etc.), and support invasion and metastases formation. Tumor-associated macrophages (TAMs), a predominant component of leukocytic infiltrate, "cooperate" with the neoplastic tissue, leading to the intensified proliferation and the immune escape of the latter. This paper characterizes the function of macrophages in the development of neoplastic disease.

Peripheral blood T lymphocyte subsets in children with congenital asplenia.

The aim of the current study was to examine whether a congenital lack of the spleen changes distribution, state of activation and function of peripheral lymphocyte T subsets. Seven children with congenital asplenia (CA) aged 1.5-17 years and seven age-matched controls were tested. By triple-color flow cytometry we examined: (1) the expression of CD3(+), CD4(+), CD8(+), CD19(+), and CD56(+) on lymphocytes; (2) the distribution of CD45RA(+) and CD45RO(+) in CD4(+) and CD8(+); (3) the expression of CD27(+) in the CD4(+) and CD8(+) T-cell-bearing CD45RA(+), CD45RO(+), or CD45RB(+). Lymphocyte proliferative responses and cytokines production (IFN-gamma, IL-6, TNF-alfa, and IL-10) in anti-CD3-induced peripheral blood mononuclear cells were tested. The results indicate (1) a normal distribution of the basic lymphocyte subsets, (2) low CD3(+)/CD8(+) percentage but expressing CD8(+high) and non-significantly elevated CD4(+)/CD8(+) ratio, (3) CD45RA(+high) and CD27(+high) in the CD4(+) and CD8(+) T cell, and (4) CD45RB(+high) in the CD4(+) and CD45RO(+high) in the CD8(+). The distribution of CD27(+) in the CD45RA(+) and CD45RO(+) CD4(+) T cells remained unchanged. However, the percentage of CD8(+)/CD45RO(+)/CD27(+) T cells tended to be elevated. Altogether, these data indicate that CA is connected with (1) the presence CD4(+) T cells expressing the "naive" phenotype (CD45RA(+high) RB(+high) and CD27(+high)), (2) high numbers of activated CD8(+) T cells shifted toward the memory phenotype (CD45RO(+high)) but still showing high CD27(+) expression, which may indicate failure in T CD8(+) cytotoxic effectors differentiation, and (3) a tendency to the rather pro-inflammatory status of cells, low IL-10 expression, and suboptimal lymphocytes responses to mitogenic stimulation.

Expression of adhesion and activation molecules on circulating monocytes in children with Helicobacter pylori infection.

OBJECTIVES: The aim of this study was to assess the cell surface expression of adhesion (CD11a, CD11b, CD11c, CD18, CD54, and CD58) and activation (CD14, HLA-DR, and CD16) molecules on the circulating monocytes in Helicobacter pylori (H. pylori)-infected and noninfected children with gastritis, with the goal of comparing the results with those obtained from the controls. MATERIALS AND METHODS: Ninety-four children were studied: 47 of them with H. pylori infection (of those 25 children after the failure of eradication therapy) and 26 children with gastritis where H. pylori infection was excluded, as well as 21 controls. H. pylori infection status was assessed based on [¹³C] urea breath test, rapid urease test, and histology. Analysis of the monocyte surface molecule expression was carried out by flow cytometry. RESULTS: H. pylori-infected children and children who experienced a failure of the eradication therapy differed significantly in the expression of adhesion and activation molecule on circulating monocytes. A decrease, both in the proportion of CD11c- and CD14-bearing monocytes, and the expression of CD11c and CD14 molecules on circulating monocytes, was found in children in whom the eradication therapy failed (p < .05). Low expression of CD11b (p = .04) and CD18 (p = .02) integrins on monocytes was also observed. Additionally, the percentage of HLA-DR-bearing monocytes was decreased (p = .04), while the CD16 density receptor was increased (p = .02). Compared with the controls, low percentage of CD16-positive monocytes was noted in noninfected children with gastritis (p = .01). CONCLUSION: H. pylori eradication therapy in children causes inhibition of inflammatory response via a reduction in CD11b, CD11c, and CD18 beta2 integrin monocyte expression.

[Higher incidence of alveolar lymphocytes (AL) apoptosis in smokers depends on BCL-2 expression and specific response to tumor necrosis factor alpha (TNFalpha). Bronchoalveolar lavage (BAL) material analysis from selected interstitial lung diseases (ILD) and healthy controls]. / Czestsza apoptoza limfocytów pecherzykowych (alveolar lymphocytes, AL) u palaczy papierosów zalezy od ekspresji BCL-2 i swoistej odpowiedzi na czynnik martwicy nowotworów alpha (tumor necrosis factor alpha, TNFalpha). Analiza materialu z plukania oskrzelowo-pecherzykowego (bronchoalveolar lavage, BAL) chorych z wybranymi chorobami sródmiazszowymi pluc i osób zdrowych.

BACKGROUND: We have previously described the increased apoptosis rate in smokers alveolar lymphocytes (AL) that was independent from the FASL/ FAS system activation. Consequently, the role of intrinsic apoptosis pathway and other ligand/death receptor pairs as TNFalpha/TNFR1 and TRAIL/DR4 important for apoptosis regulation should be considered in this phenomenon. The purpose of the study was to evaluate the impact of tobacco consumption on expression of selected BCL-2 family members and ligand/receptors pairs in bronchoalveolar lavage (BAL) harvested from patients with pulmonary sarcoidosis (PS), idiopathic pulmonary fibrosis (IPF) and healthy volunteers. The results were analyzed in the context of AL apoptosis rate. METHODS: AL apoptosis from PS (n=36, incl. 22 smokers), IPF (11, incl. 5 smokers) and controls (n=17, incl. 9 smokers) was evaluated by flow cytometry (sub-G1 of cell cycle). AL were stained for BCL-2, BCL-xL, BAK, TNFR1 (CD120A) TNFR2 (CD120B) and DR4. ELISA assay was used to evaluate the BAL supernatant levels of TNFalpha and TRAIL. RESULTS: According to previous observations, AL apoptosis rate was significantly higher in smoker subgroups as compared to nonsmoking counterparts. Decreased AL BCI-2+ relative number was observed in smoking PS (80.5 +/- 6.2 vs 91 +/- 9.8% in nonsmokers) and controls (59 +/- 14.1% vs 75 +/- 16.1%, p<0.05). TNFalpha concentration in BAL supernatant was significantly higher only in healthy smokers (2.32 +/- 0.77 vs 0.42 +/- 0.27 pg/ml, p<0.05), whereas TRAIL levels were remarkably enhanced in IPF smokers (44.8 +/- 12.8 vs 13.5 +/- 5.0 pg/ml, p<0.05) only. However, TUNEL. detected AL apoptosis positively correlated with TNFalpha. in smokers (p<0.05) and negatively with AL CD120B:CD120A expression ratio. Paradoxically, TNFalpha levels were positively correlated with AL BCL-2 expression in nonsmokers (Rs +0.58, p<0.01), but not in smokers. No differences were observed in all subgroups in respect to AL expression of DR4, BCL-xL or BAK. CONCLUSIONS: 1. AL were not sufficiently protected against apoptosis in smokers. 2. The most likely mechanisms involve down-regulation of BCL-2 expression and altered AL susceptibility to TNFalpha, mediated by imbalance between AL membrane expression of TNF receptor type 1 (death receptor) and type 2 (survival mediator). 3. Mechanisms regulating the increased AL apoptosis in smokers seem to be different in each tested group.

Pediatric Helicobacter pylori infection and circulating T-lymphocyte activation and differentiation.

BACKGROUND: In this study, H. pylori-infected and noninfected children with gastritis were compared to a control group with respect to circulating CD4(+) and CD8(+) T lymphocytes expressing activation and differentiation markers. Additionally, the lymphocyte phenotypes of children with gastritis were correlated with the gastric inflammation scores. MATERIALS AND METHODS: H. pylori infection status was assessed based on [¹³C]urea breath test, rapid urease test, and histology. Analysis of the lymphocyte surface molecule expression was carried out by triple-color flow cytometry. RESULTS: The group of H. pylori-infected children showed an elevated proportion of peripheral B cells with CD19(low) , along with a twofold increase in the percentage of memory (CD45RO(+)) CD4(+) and CD8(+) T-cell subsets (p < .05). Moreover, a positive correlation between the age and the percentage of these subsets was seen (r = .38, p = .04 and r = .56, p < .01, respectively). Children with gastritis but without infection had a slightly increased percentage of CD8(+) T cells and CD56(+) NK cells, CD3(high) T cells and CD45RO(high) CD4(+) T-cell subsets (p < .05). Both H. pylori-infected and noninfected children with gastritis were characterized by an increased percentage of memory/effector CD4(+) T cells, the presence of NK cells with CD56(high), memory T-cell subset with CD4(high), and naive, memory, memory/effector, and effector T-cell subsets with CD8(high) (p < .05). Gastric inflammation scores correlated positively with the percentage of CD4(+) T lymphocytes in H. pylori-infected children (r = .42, p = .03). In noninfected children, gastric inflammation scores correlated positively with the percentage of B cells (r = .45, p = .04). CONCLUSION: In H. pylori-negative children, gastritis was associated with an increased percentage of activated NK and T cells, and intermediate-differentiated peripheral blood CD4(+) T cells, which was more pronounced in H. pylori-positive children who also showed an increased B-cell response. However, increased inflammation was only associated with the elevation of CD4(+) T-cell percentage in H. pylori-positive children as well as B-cell percentage in H. pylori-negative children with gastritis.