Research on some parameters of cellular immune response in soldiers undergoing basic training--preliminary report.

BACKGROUND: On the basis of available reports it can be stated that physical stress causes changes in distribution and activity of many components of the immune system. It is believed that psychophysical stress in soldiers can influence their immune system depressively and in consequence increase the risk of upper respiratory tract infections. Therefore, it was decided to conduct studies aimed at the estimation of the influence of military training on the some parameters of cellular immune response. MATERIAL AND METHODS: The study group consisted of 40 draft aged from 18 to 23 years. The research was conducted in the first 8 weeks of service, in the period of the most intense draft stress adaptation. The participants were divide into 2 groups, A and B respectively, 20 soldiers each. Group A derived from an assault unit. Their training induced strenuous physical stress. Group B derived from a support unit. Their training required less physical effort then one of group A. Performed examinations involved: lymphocyte percentage count, lymphocyte proliferative response to mitogen, CD69 antigen expression on T lymphocyte surface, delayed hypersensitivity reaction with CMI Multitest. All assessments were done twice at 8 weeks interval. RESULTS: After 8 weeks of training in the A group a statistically significant increase in the percentage of lymphocytes revealing antigens of the II Class Main Histocompatibility Complex (MHC) was found. In addition, in this group a statistically significant decrease in the value of lymphocyte stimulation index, a statistically significant increase in the percentage of cells revealing CD69 antigen expression after PHA stimulation were observed. During investigated period in the B group following statistically significant changes were found: an increase in the percentage of CD3+ and CD4+ cells, a decrease in the percentage of CD16+CD56+ and an increase in the CD4+ to CD8+ ratio. CONCLUSION: The obtained results show that military service conditions influence some parameters of the cellular immune response but do not result in the clinically significant suppression of the immune system.

[Effect of trimetazidine on biological activity of neutrophils in patients with transient myocardial ischemia induced by exercise testing]. / Wplyw trimetazydyny na aktywnosc biologiczna neutrofilów u chorych z przemijajacym niedokrwieniem miesnia sercowego prowokowanym testem wysilkowym.

We estimated the effect of trimetazidine on biological activity of neutrophils in patients with exercise test--induced transient myocardial ischaemia. The study group comprised 16 patients (10 men and 6 women) aged 40-56 years (mean 48.2 years) with stable angina. Exercise test was performed on cycloer-gometer (Medicor, Hungary). Trimetazidine in a dose 3 x 20 mg/day was applied 24 hours prior to the exercise test. The control group consisted of 14 patients (9 men and 5 women) aged 37-59 years (mean 47.9 years), with stable angina, in whom exercise test was performed but the drug was not administered. Blood samples for examination were taken from basilic vein before the exercise test and 10 min afterwards. To evaluate the ability of neutrophils to aggregate the leukergy test of Fleck modified by Berliner and Aronson was applied. Oxidative metabolism of nonstimulated and stimulated by fMLP and PMA was measured using chemiluminescence method. It was shown that in patients with stable angina trimetazidine caused decrease in neutrophil aggregation. The drug did not inhibit neutrophil biological activity af-ter exercise test--induced myocardial ischaemia.

[Effect of maternal labor and mode of delivery on function of neonatal cord blood neutrophils]. / Wplyw porodu zabiegowego na stan czynnosciowy neutrofilów krwi pepowinowej noworodków.

There are data to suggest that the method of delivery modifies the immunological balance of newborn infants. In this study, an effect of maternal labor and mode of delivery on the neutrophil activity was evaluated in healthy full-term neonates. Neutrophils were obtained from cord blood of two group neonates: 14 vaginally delivered and 12 delivered by elective cesarean section without preceding labor. Neutrophil functions were studied by measuring the chemotaxis, chemiluminescence and phagocytosis activity and by investigating the expression of phagocyte receptors CD11b on neutrophils using receptor-specificMoAbs and immunofluorescence flow cytometry. Infants born by elective cesarean section had significantly higher chemiluminescence activity, chemotaxis and higher expression of CD11b receptors than those delivered vaginally. We suggest that physiological changes associated with stress during labor could explain these differences observed in neutrophil functions.

Tumor necrosis factor-alpha binding by peripheral blood lymphocytes and polymorphonuclear neutrophils in patients with multiple sclerosis.

Tumor necrosis factor-alpha (TNF-alpha) in multiple sclerosis (MS) is responsible for peripheral blood leukocyte priming. The aim of this study was to evaluate fluorescein isothiocyanate (FITC)-labelled TNF binding ability by peripheral blood lymphocytes and polymorphonuclear neutrophils (PMNs) of MS patients, measured using flow cytometry (FACScan). Three groups of MS patients (total 34) were examined. Higher serum levels of TNF soluble receptors sp55 and sp75 were found in the MS patients during MS acute exacerbation (n = 10) and in chronic progressive forms (n = 11) as compared to MS remission (n = 13) and other neurological diseases (n = 14). Peripheral blood lymphocytes and PMNs of patients with acute exacerbation of MS bound TNF-FITC more effectively (p <0.01) as compared with the chronic progressive forms of MS, MS remission and other neurological diseases. The obtained results suggest a greater enhancement of TNF activity during MS acute exacerbation.

Expression of aminopeptidase N (APN) on peripheral blood mononuclear cells' surface as a marker of these cells' transendothelial migration properties in the course of multiple sclerosis.

CD13 Ag and CD11a, CD11b, CD18 molecule expression on peripheral blood mononuclear cells (PBMC) were studied as these cells' adherent or transendothelial migration properties in three different multiple sclerosis (MS) patients groups (total 38): with clinically active MS (acute exacerbation of MS and primary chronic progressive MS (CP-MS)) and with MS remission. The control group consisted of patients, suffering from other non-inflammatory neurological diseases (OND). The results of our study suggest that CD11a/CD18 molecules expression on PB lymphocytes, although higher on these cells' surface in the course of MS as compared to OND, does not differentiate clinical forms of MS. CD11a molecule expression on monocytes did not differ significantly in all tested MS patient groups in comparison to OND. Although the expression of CD11b/CD18 molecules on monocytes' surface shows their activation in the course of MS, it does not differentiate them either. However, CD13 Ag of APN expression on PBMC surface may be an immunological marker of MS clinical form. CD13 Ag expression may also be a sensitive marker of these cells' transendothelial migration properties.

[Evaluation of the hormonal replace therapy (HRT) effect on generation of reactive oxygen intermediates (ROI) by neutrophil in peripheral blood of menopausal women]. / Ocena wplywu hormonalnej terapii zastepczej u kobiet w okresie menopauzalnym na zdolnosc generacji reaktywnych form tlenu przez neutrofile krwi obwodowej.

UNLABELLED: About 30% of women population in Poland is already in the perimenopausal age. HRT (hormonal replacement therapy) is the best way of diminishing the unfavorable symptoms of this state. AIM OF STUDY: The effect of HRT on ROI production by neutrophils before and after HRT, to determinate it's antioxidative influence on cells. MATERIAL: We have examined 40 women both from the Dept. of Menopausal Disease in the Research Institute Polish Mother's Memorial Hospital (RIPMMH) and Out Patient Menopausal Clinic of RIPMMH with average age of 49.8; between October 1997 and January 1998. METHODS: We determined ROI generation by luminol-enhanced chemiluminescence, using standard stimuli like fLMP, PMA and OZ. We used Luminometer 1251, manufactured by Pharmacia-LKB. For statistic evaluation Fisher test, Kolmogorow-Smirnow test and T-student test were used. RESULTS: We found that estrogen-progestagen therapy had a suppressive effect on generation of ROI by neutrophils in vitro and in vivo after using receptor dependent and non-dependent stimuli. ROI generation by neutrophils of peripheral blood induced by PMA, a receptor dependent stimuli, is diminished, both, after HRT or as an effect of estrogen-progestagen action in vitro. The results suggest direct inhibition effect on the neutrophil ability to ROI generation.

Increased expression of neutral endopeptidase (NEP) and aminopeptidase N (APN) on peripheral blood mononuclear cells in patients with multiple sclerosis.

We studied CD10 Ag of neutral endopeptidase (NEP) and CD13 Ag of aminopeptidase N (APN) expression on peripheral blood mononuclear cells (PBMC) as cells activation markers and for their transendothelial migration properties in three groups of MS patients (total 58); with acute exacerbation of MS (n = 18), primary progressive MS (n = 17) and with MS remission (n = 23). The control group (OND) consisted of 24 patients, suffering from other noninflammatory neurological diseases. CD10 Ag and CD13 Ag expression on PBMC was higher in clinically active MS (acute exacerbation and progressive MS) compared to MS remission and OND groups. Our study suggests that CD10 Ag and CD13 Ag can be useful mononuclear cell activation markers in the course of MS. CD13 Ag expression on PBMC may be also the sensitive marker of these cells transendothelial migration properties.

[Impact of chronic psychophysical stress on some neutrophil functions]. / Wplyw dlugotrwalego stresu psychofizycznego na niektóre funkcje neutrofilów.

It is known, that psychophysical stress causes some changes in distribution and activity of many components of the immune system. The research was aimed at examination the influence of different military service conditions on some neutrophil functions. In this study granulocyte chemiluminescence both without stimulation and after stimulation with fMLP, PMA and zymosan was evaluated. In addition, the expression of surface adhesion molecules--CD11b on circulating granulocytes was examined. The studied group consisted of 20 soldiers from an assault unit (group A) and 20 soldiers from a support unit (group B), the age ranged from 18 to 23 years. The examinations were conducted at the beginning of the basic training and after the 8 weeks period. In group A the following results were obtained: a significant increase (p < 0.001) in granulocyte chemiluminescence without stimulation and after stimulation with fMLP, PMA and zymosan, as well as, a significant decrease (p < 0.05) in CD11b expression on granulocytes. And in group B: a significant increase (p < 0.05) in granulocyte chemiluminescence without stimulation but a significant decrease (p < 0.05) after cells stimulation with fMLP and PMA were observed. In this group CD11b expression on granulocytes was significantly (p < 0.001) decreased. The results of the present examination indicate, that military service conditions influence some parameters of the innate immunity.

[Increased percentage of "double phenotype" form T lymphocytes in blood of patients with multiple sclerosis]. / Zwiekszona wartosc odsetkowa "podwójnie dodatniej" formy fenotypowej limfocytów T we krwi chorych na SM.

We studied the percentage of double positive (CD4+CD8+) form of T-cells in a group (total 77) multiple sclerosis (MS) patients, measured by means of monoclonal antibodies anti-CD3, CD4/FITC, CD8/RPE and flow cytometry FACScan (Becton Dickinson). In our study we have shown that the percentage of the double positive T cells is higher (p < 0.05) in the peripheral blood of patients with acute exacerbation of MS (n = 21), and in the course of chronic progressive MS (n = 27) comparing to MS remission (n = 29) and other neurological diseases (n = 25) groups. In the study we have shown that the percentage of double positive T-cells in peripheral blood depends mainly on disease activity.

[TNF-alpha binding ability as a sign of peripheral blood neutrophils preactivation in the course of multiple sclerosis]. / Zdolnosc do wiazania czynnika martwicy nowotworu--alpha przez neutrofile krwi obwodowej chorych na stwardnienie rozsiane.

Usually neglected is the role of neutrophils in causing of immunological disturbances in patients with multiple sclerosis (MS). Nevertheless, it has been indicated over the recent years that these cells possess a sufficient potential to affect both immune response and inflammation. This potential may result in MS through the process of priming of these cells by proinflammatory cytokines like TNF. We studied TNF and its soluble receptors sp55 and sp75 serum levels and binding of fluorescein isothiocyanate (FITC)-stained TNF by neutrophils. We studied three different groups of MS patients: 10 patients in relapse of the disease, 13 in its remission, and 11 in its chronic progressive form (CP-MS). The control was provided by 14 neurological patients (OND) with non-inflammatory diseases. The performed studies showed higher TNF sp55 and sp75 soluble TNF receptors serum levels in the patients with relapse, comparing with other MS patients and OND. TNF binding by neutrophils of MS patients during relapse was also higher, than other MS patients and OND. These result suggest the preactivation of neutrophils in the relapse of MS.

Priming effect of met-enkephalin and beta-endorphin on chemiluminescence, chemotaxis and CD11b molecule expression on human neutrophils in vitro.

The opioid peptides are widely distributed throughout the body, and they are generated during stress and inflammatory reaction. Opioids are involved in the communication between the immune and neuroendocrine systems. In the present study we have investigated the ability of both met-enkephalin and beta-endorphin to stimulate and prime the human neutrophils for enhanced chemiluminescence (CL) and chemotaxis induced with fMLP, OZ or PMA. We have also tested the effect of beta-endorphin and met-enkephalin on CD11a, CD11b, CD18 and CD16 molecule expression on PMN in vitro. PMN from ten healthy donors were incubated in vitro with different concentrations of beta-endorphin or met-enkephalin, and the CL response was evaluated with luminometer. To assess the effect of opioid peptides on CD11a, CD11b, CD18 and CD16 molecule expression the whole blood samples were incubated with different concentrations of the opioids, then the white cells were labelled with respective PE-conjugated MoAb and evaluated by flow cytometry. We have shown that: (1) met-enkephalin and beta-endorphin at physiological concentrations relevant to that of in vivo (10(-8) and 10(-6) M) enhanced fMLP, PMA or OZ stimulated chemiluminescence and induced chemotactic response, (2) High concentrations of beta-endorphin (10(-3) M) or met-enkephalin (10(-5) M) decreased the CL response of PMN in vitro, (3) The opioid peptides at lower concentrations resulted in CD11b and CD18 molecule up-regulation on neutrophils. We may conclude that opioid peptides in physiological concentration are involved in neutrophil priming whereas in higher concentration exert immunosuppressive potency. Opioid peptides like inflammatory cytokines may prime the neutrophils inflammatory response.

Pentoxifylline decreases neutrophil respiratory bursts in patients with stable angina.

We estimated the effect of pentoxifylline (PTX) on the respiratory burst (examined by chemiluminescence method) of unprimed and primed neutrophils with tumor necrosis factor-alpha (TNF-alpha) in patients with stable angina pectoris. Chemiluminescence of non-stimulated as well as formyl-methionyl-leucyl-phenylalanine (fMLP) and phorbol myristate acetate (PMA) stimulated neutrophils was measured. We studied 45 patients with stable angina subjected to percutaneous transluminal coronary angioplasty (PTCA) procedure, who were randomly divided into two groups. The study group consisted of 24 patients who were administered pentoxifylline orally, and the control group consisted of 21 patients without pentoxifylline administration. Blood samples for examination were collected from the coronary sinus and peripheral vein just before the PTCA procedure. Pentoxifylline decreased the respiratory burst of non-stimulated and fMLP-stimulated neutrophils without affecting the chemiluminescence of PMA stimulated neutrophils. Moreover, pentoxifylline diminished the chemiluminescence non-stimulated and stimulated by fMLP but not by PMA of TNF-alpha primed neutrophils. We presume that administration of PTX in stable angina patients may have a beneficial effect.

[Increased expression of aminopeptidase N on lymphocytes in multiple sclerosis]. / Zwiekszona ekspresja aminopeptydazy N na limfocytach w przebiegu stwardnienia rozsianego.

Aminopeptidase N is an ectoenzyme. Its expression on lymphocytes is the effect of cell activation. We studied APN expression on peripheral blood lymphocytes of multiple sclerosis (MS) patients and in the control group of patients with other neurological diseases (OND). We observed increased APN expression on lymphocytes of MS patients during acute exacerbation and in the course of chronic progressive MS compared to MS remission and OND groups. No such differences were found in CD11a molecule of LFA-1 integrin expression on lymphocytes. We suppose that APN expression on lymphocyte surface can be a sensitive marker of these cell activation in the course of MS.

[The effect of pentoxifylline on oxidative metabolism in neutrophils primed with tumor necrosis factor alpha in patients with stable angina pectoris]. / Wplyw pentoksyfiliny na metabolizm tlenowy neutrofilów preaktywowanych przez czynnik martwicy nowotworów alfa u chorych ze stabilna dusznica bolesna.

In 14 patients with stable angina pectoris we examined the effect of pentoxifyline (PTX) on oxidative metabolism of TNF-alpha-priming neutrophils. The control group consisted of 21 patients with stable angina pectoris without pentoxifylline administration. Blood samples for examination were taken from basilic vein (peripheral blood) and coronary sinus immediately before PTCA procedure. In PTX-group was found the significant decrease in spontaneous CL of TNF-alpha-priming neutrophils from coronary sinus blood (1231.0 +/- 119.4 mV x min), in comparison to the control group (1374 +/- 124.4 mV x min). In PTX-group was found the significant decrease in fMLP stimulated CL of TNF-alpha-priming neutrophils from peripheral blood (4219.0 +/- 707.2 mV x min) and coronary sinus blood (4322.0 +/- 664.4 mV x min), in comparison to the control group (5248.0 +/- 595.8 mV x min and 4973.0 +/- 536.5 mV x min; respectively). There were no differences between both groups in PMA stimulated CL of TNF-alpha-priming neutrophils.

[TNF-alpha preactivation (priming) of neutrophils--effect on selected neutrophil functions]. / Preaktywacja (priming) neutrofila przez TNF-alpha--wplyw na wybrane funkcje neutrofila.

Human polymorphonuclear cells (neutrophils) are fundamentally protective, but extracellular release of proteolytic enzymes and oxygen radicals may lead to tissue destruction. Granulocyte function can be rapidly amplified by environmental factors through a mechanism termed "priming". Priming enhances the neutrophils' ability to respond to a secondary agonist, with increased adhesion, respiratory burst, and degranulation. This review summarizes the effects of TNF-alpha priming on the neutrophils' function.

The immunoregulatory abilities of polymorphonuclear neutrophils in the course of multiple sclerosis.

The polymorphonuclear neutrophils (PMN) possess sufficient potential to affect both immune response and inflammation, however it has not been yet described in the course of multiple sclerosis (MS). We have studied binding of fluorescein isothiocyanate (FITC)- stained TNF-alpha by PMN, the expression of CD11a, CD11b, and CD18 molecules of beta2-integrines and the expression of CD10 (neutral endopeptidase-NEP) and of CD13 (aminopeptidase N; APN) antigens on PMN in three different groups of MS patients. The control group included neurological patients (OND) with noninflammatory diseases. The obtained results have proved that during MS exacerbation and in the course of chronic progressive MS, PMN reveal several forms of preactivation, including significantly higher stained-TNF-alpha binding, higher expression of CD11b and CD18, as well as CD10 and CD13 antigens, in comparison with MS remission or OND. We suggest that the increased expression of these molecules on PMN of MS patients in exacerbation of the disease and to a lower degree in the course of CP-MS is a result of PMN priming, and directly prove the PMN involvement in the disease pathogenesis.

Complement activates neutrophils during PTCA procedure in patients with unstable angina pectoris.

We estimated adherence, aggregation and chemiluminescence of neutrophils as well as concentrations of C3c, C4 and C5 complement components and complement haemolytic activity (CH50) in 27 patients with unstable angina pectoris subjected to percutaneous transluminal coronary angioplasty (PTCA). The control group consisted of 12 patients with unstable angina pectoris, in whom coronary angiography was performed but PTCA was decided against for various reasons. Blood samples for examination were taken from coronary sinus and peripheral vein just before, 1 min and 20 min after PTCA or coronary angiography. We observed enhancement of neutrophil adherence, aggregation and chemiluminescence, and decrease in concentrations of C3c, C5 and complement haemolytic activity (CH50) after PTCA procedure. In conclusion we think that ischemia resulting from PTCA causes complement activation in an alternative pathway which seems to be connected with neutrophil activation.

Protease inhibitors diminish lymphocyte stimulation in vitro.

Lymphocyte stimulation initiates activation of signal transduction pathways presumably enzymatic in nature. The next step includes gene activation and specific protein molecule production. Proteases of different specificity are crucial in enzyme activation: protein cleavage and biologically active molecule production. To elucidate the role of proteases in peripheral blood mononuclear cell (PBMC) activation, the broad specificity inhibitors of thiol (PHMB) and serine (TLCK) proteases have been used in vitro. Both inhibitors diminished the PHA-induced lymphocyte stimulation when applied at the beginning of culture; inhibitory effect was abrogated when inhibitors were introduced to the culture system 4 h later. Exogenous IL-2 abolished inhibition. PHMB activity was reversible whereas TLCK was irreversible. The inhibition of T lymphocyte-enriched proliferation is more distinctive as compared to that of PBMC. It can be concluded that proteases are involved in early events of lymphocyte proliferation and IL-2 production which is responsible for different stages of cell growth.