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.

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.

[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.

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.

[Immunologic characteristics of women recurrent spontaneous abortion of unknown etiology. II. Characterization of humoral immunology]. / Immunologiczne odrebnosci kobiet z poronieniami nawykowymi o niewyjasnionej etiologii. II. Charakterystyka stanu odpornosci humoralnej.

The purpose of this study was to analyze the humoral immune response of non-pregnant women with unexplained recurrent spontaneous abortion (RSA). The study group was consisted of 117 nonpregnant women with RSA and 44 healthy, non-pregnant multigravidas (as a control). The following immunological parameters were analyzed in peripheral blood of two groups of women: the effect of autologous sera on proliferation of lymphocytes stimulated with phytohemagglutinin (PHA) or paternal lymphocytes (in mixed lymphocyte reaction--MLR), incidence of MLR-blocking antibodies (MLR-BAbs) incidence of antinuclear and anticardiolipin antibodies. The results show that women with RSA of unknown etiology differ in some parameters of immune response--higher incidence of antinuclear and anticardiolipin antibodies, absence of serum factors suppressing lymphocyte proliferation in response to PHA and paternal alloantigens--in comparison with nonpregnant normal multigravidas.

[Efficiency of lymphocyte immunization by the partner in prevention of unexplained recurrent spontaneous abortion. II. Immunologic prognostic factors]. / Skutecznosc immunizacji limfocytami partnera w zapobieganiu poronieniom nawykowym o nie wyjasnionej etiologii. II. Czynniki immunologiczne o znaczeniu prognostycznym.

The prognostic value of some immunological parameters on the course of subsequent pregnancy in 117 women with recurrent spontaneous abortion of unknown etiology subjected to paternal lymphocytes immunization, were estimated. We conclude that neither antinuclear (ANA) nor low titers of anticardiolipin antibodies (ACA), nor mixed lymphocyte reaction--blocking antibodies (MLR-BAbs) have any significant influence on alloimmunization efficiency. However medium or high ACA titers significantly diminish changes for successful alloimmunization and are connected with most complications of subsequent pregnancy.

Tumor necrosis factor alpha but not lymphotoxin is overproduced by blood mononuclear cells in multiple sclerosis.

The cytotoxic cytokines tumor necrosis factor alpha (TNF-alpha) and lymphotoxin (LT) possess toxic activity against myelin and/or oligodendrocytes in vitro. Multiple sclerosis (MS) plaques within the central nervous system (CNS) are infiltrated by peripheral blood mononuclear cells (PBMC). In this study the production of TNF-alpha and LT by PBMC in active MS were measured. PBMC were isolated from the blood of MS patients in relapse and also patients with other neurological diseases (OND) and healthy controls (HC). Isolated cells were cultured unstimulated or stimulated with phytohemagglutinin A (PHA), lipopolysaccharide (LPS) and myelin basic protein (MBP)--a hypothetical autoantigen for MS. Cytokine production was assessed using ELISA method. In the MS group, PBMC without stimulation as well as after stimulation with MBP displayed a significantly increased production of TNF-alpha. LT production was similar in MS and control groups. These results suggest that TNF-alpha but not LT is overproduced by PBMC during MS relapse.

Immunological status of competitive cyclists before and after the training season.

The total number of leucocytes, T lymphocyte subsets, mitogen induced proliferation of lymphocytes, Il-2 generation, adherence capacity and chemiluminescence of granulocytes were measured and a leukergy test performed in fifteen young cyclists. The investigations were carried out at rest at the beginning of a training season and after six months of intensive training and a racing season, cycling approximately 500 km a week. Baseline values of the tested immune parameters were within the range observed in 16 non-trained healthy controls except significantly increased non stimulated neutrophil chemiluminescence. The second cyclo-ergometer test in August showed a marked improvement in the performance capacity of the cyclists. Significant decrease in absolute numbers of CD3+ and CD4+ cells, diminished IL-2 generation and fMLP and PMA stimulated chemiluminescence of neutrophils were noted. Surprisingly, a marked increase in lymphocyte proliferation induced by PHA and anti-CD3 MoAb and normalisation in non stimulated neutrophil chemiluminescence were also observed at rest after the training season. We conclude that long-lasting intensive training may result in significant alterations in lymphocyte number and composition and in neutrophil oxidative burst capacity, but their actual significance for immunity is seen controversially.

The effect of chronic exposure to formaldehyde, phenol and organic chlorohydrocarbons on peripheral blood cells and the immune system in humans.

The aim of this study was to evaluate immunological, hematological and biochemical parameters in subjects chronically exposed to inhaled formaldehyde (F), phenol (Ph) and isomers of organic chlorohydrocarbons (Chc) released from Ksylamit. Twenty-two office workers had been exposed for 6 months to vapors of Ksylamit, used for protection of felt plates inside the office building. The concentration of Ksylamit vapor was measured at the end of the 6-month period and the level of Ph and F in the air inside the building was 1.3 mg/m3 and 0.8 mg/m3, respectively. Most of the workers had ailments due to the irritant effect of inhaled Ksylamit, but no remarkable increase in morbidity was found during the 6 months of exposure and in a 3-year follow-up. Morphological parameters of blood and the number of natural killer (NK) cells in the group of exposed subjects were within the range observed in healthy subjects matched for age and sex. The number of T-lymphocytes and NK cell cytotoxicity were significantly decreased. Phytohemagglutinin (PHA)- and alloantigen-induced lymphocyte proliferation was diminished. Some biochemical parameters suggested liver damage, although these parameters did not correlate with the levels of Ph and methanol excreted in the urine. Eight subjects with the highest levels of Ph in the urine had decreased erythrocyte and T-helper lymphocyte numbers, and increased numbers of eosinophils and monocytes. The results obtained prove that the functions of both the immune and hematopoietic systems could be affected by chronic exposure to these toxic substances.

The effect of short-term myocardial ischemia on the expression of adhesion molecules and the oxidative burst of coronary sinus blood neutrophils.

We examined the influence of transient myocardial ischemia on the number and function of neutrophils in patients with effort angina (EA). We tested fluorometrically the expression of neutrophil membrane molecules (CD11b, CD11c, CD18) and neutrophil oxidative burst using a chemiluminescence (CL) generation system. The estimations were conducted before, 1 min after and 20 min after percutaneous transluminal coronary angioplasty (PTCA) in 15 patients qualified for the treatment because of single-vessel disease. Eight EA patients subjected to coronary arteriography (CA) comprised a control group. We did not observe any marked changes in leucocytosis or lymphocyte number in peripheral blood (PB) or in coronary sinus blood (CSB) after the procedure. The percentage of granulocytes in coronary blood decreased significantly 20 min after reperfusion. No significant changes in white blood cell count were noted in peripheral blood of PTCA patients or in control CA subjects. Oxidative burst of nonstimulated and fMLP, PMA and zymosan stimulated sinus blood neutrophils was significantly depressed 1 min after inflation, and enhanced 20 min after reperfusion. We found a significant increase in the percentage of the CD11c+ neutrophils from 56.7 +/- 7.4% to 64 +/- 6.5% 20 min after inflation and postischemic decrease in the CD11c molecule expression on CSB neutrophils. Significant positive linear correlation (Rval = 0.71) between inflation time and the CD11c molecule expression on CSB immediately after reperfusion was also noted. The results may reflect local activation of neutrophils in ischemic myocardium as a response to ischemia induced increase of activating stimuli.

Effect of Tolpa Peat Preparation on some immune parameters in healthy volunteers. Preliminary data.

Investigations of some immune parameters were carried out in healthy humans receiving TPP in doses of 50, 100, 300 and 600 mg daily for 14 days. The percent and composition of T lymphocyte subsets were studied along with the ability to generate interleukin-1 (Il-1) and interleukin-2 (Il-2) as well as spontaneous and PHA stimulated expression of receptors for Il-2 (TAC) and transferrin receptor (Tr) on lymphocytes. Lymphocytes proliferation induced with mitogens and alloantigens, generation of both Tumour Necrosis Factor alpha(TNF alpha) by monocytes and superoxide radicals by granulocytes were estimated. A 50 mg dose of TPP changed neither the T-cell percent among mononuclear cells nor the composition of T-cell subsets. A higher dosage of 100 mg of TPP caused a significant increase of the percent of T-cells on the 14th day but did not influence their subset composition. On the 14th and 28th day of the observation a statistically insignificant decrease of IL-2 generation was observed but with no change of both TAC receptor expression on lymphocytes and the ability to generate IL-1 by monocytes. TPP in the doses of 50 and 100 mg exerted a slight immunomodulatory potential in healthy humans, however the immunological parameters assessed were within the standard values. In other immune parameters examined during the administration of higher TPP doses in human groups consisted of four volunteers considerable differences have been found but individual differences in a limited experimental group and the lack of a placebo group did not allow for definite conclusions but for preliminary data only.

T lymphocyte subsets and NK cell cytotoxicity in chronic hemodialysis patients. The effect of recombinant human erythropoietin (rHu-EPO) treatment.

We investigated subpopulations of T lymphocytes, NK cell number and cytotoxic activity in 14 chronic uremic patients on regular hemodialysis treatment. We observed a significantly decreased absolute lymphocyte number and percentage of CD3 cells. Relative numbers of CD16 cells were significantly elevated, but NK cell cytotoxic activity was within a normal range. Nine patients with chronic renal anemia on maintenance hemodialysis were enrolled in rHu-EPO treatment trial. The treatment was continued till the hematocrit level reached 30%. Each of the patients had corrected anemia and well-being. After 12 weeks of the treatment we observed in these patients decreases in CD3, CD4, CD8 and CD16 cell numbers and elevation of CD4/CD8 ratio. Cytotoxic activity of NK cells did not change significantly. Presented results indicate that chronic hemodialysis patients have significantly diminished lymphocyte number. rHu EPO treatment affects the T lymphocyte subsets inducing a deep decrease of CD8 and CD16 cell percentage leading to normalisation of the CD4/CD8 ratio.

The effect of thymic hormones (TFX) on lymphocyte subpopulations and proliferation after maximal physical effort.

The studies were performed on healthy well-trained cyclists. Maximal physical exercise was performed on a Monark bicycle ergometer according to individual schemes. Heart rate amounting to about 200 bts/min and oxygen consumption stabilization were considered as criteria for maximal physical exercise. In this study we have investigated the effect of short-term stimulation of conditioned sportsmen with thymic hormones and evaluated T cell subsets, DR antigen and transferrin receptor expression as well as mitogen-induced proliferation of lymphocytes before and after maximal physical effort. The results suggest that intensive physical exercise may be responsible for transient decrease of CD4/CD8 ratio and mitogen responsiveness, and increase of mononuclear cells number bearing HLA DR+ and CD71 antigens. These changes were modified by the treatment with thymic hormones.

Effect of thymic extract on allogeneic MLR and mitogen-induced responses in patients with chronic active hepatitis B.

Cell-mediated immune reactions are involved in the development of chronic active hepatitis (CAH-B). The present studies confirm that lymphocyte from patients with CAH-B have a decreased allogeneic mixed lymphocyte reaction (allo-MLR) and mitogen-induced responses. These abnormalities probably play a role in the immunological dysfunctions underlying chronic liver diseases. Twenty one patients with biopsy-proven CAH-B and a significantly lowered CD4:CD8 cell number ratio were treated with thymic extract (Thymomodulin-TFX) for 6 months. The results of immunological, serological and biochemical findings indicate that this preparation has an immunoregulatory action and produces beneficial clinical effect in patients with CAH-B.

The effect of long-term thymosine factor x/TFx/treatment on T lymphocyte subpopulations and peripheral blood mononuclear cell cytotoxicity in chronic active hepatitis B.

Peripheral blood mononuclear cells (PBMC) cytotoxic activity, percentage of natural killer (NK) cells and T lymphocyte subpopulations in PBMC of 24 patients suffering from chronic active hepatitis B (CAH-B) have been studied. In comparison with healthy subjects CAH-B patients had significantly enhanced both lymphocyte T suppressor (Ts) and NK cell number. Thirteen CAH-B patients with higher Ts number and low lymphocyte T helper/T suppressor ratio (Th/Ts) were subjected to immunostimulation by thymosine factor x (TFx) treatment. After 1 year of TFx treatment all the CAH-B patients improved clinically, the Th/Ts ratio was brought back to normal and 80 per cent of HBe Ag carriers seroconverted to anti-HBe. After one year of TFx administration no significant changes were observed in PBMC cytotoxicity and NK cell number. TFx appears to be a very effective therapeutic agent in some forms of CAH-B. The obtained results show that TFx normalises biochemical parameters of liver lesion and Th/Ts ratio without any effect on NK cell activity in CAH-B patients.

Lymphotoxin production in multiple sclerosis patients.

Lymphotoxin (LT) is a cytotoxic and cytostatic lymphokine produced by activated T and B lymphocytes. Recently, the influence of cytokines on the pathomechanism of demyelinization was investigated. The authors studied LT production in the multiple sclerosis patients. The control group were patients with other neurologic diseases. Spontaneous LT production by T lymphocytes in multiple sclerosis patients was statistically increased during relapse in comparison with the LT production in OND patients. Despite of mitogen stimulation T and B lymphocytes from MS patients produced gradually decreased amounts of LT. The authors suggest that LT might be the product of activated lymphocytes that cause demyelinization during multiple sclerosis.

Treatment with thymic extract TFX for chronic active hepatitis B.

Eighteen patients with biopsy-proven chronic active hepatitis B (CAHB) and a significantly lowered CD4+/CD8+ cell number ratio were treated with thymic factor X (TFX): group I (n = 12) - for 12 months, group II (n = 6) for 6 months. As early as 14 days after starting the treatment a lowering of CD8+ cell numbers with a rise in CD4+/CD8+ cell number ratio were found. These changes continued to progress during the next months and were accompanied by decreased in NK cell numbers with enhancement of NK cell activity. Normalization of the biochemical and immunological parameters occurred after 5-6 months of the treatment. In both groups of patients seroconversion in the HBe system was observed after 9-12 months of the treatment. After two years complete clinical remission persists with normal biochemical and immunological findings and without reversion in the HBe system. The results seem to indicate that TFX has an immunostimulatory action and exerts beneficial effects on the course of CAHB.

Modification of some lymphocyte functions in vitro by opioid receptor agonists and antagonist in chronic uremic patients and healthy subjects.

In ten chronic uremic patients on regular hemodialysis treatment in vitro experiments revealed that stimulation of opioid receptors with morphine did not significantly change the mitogen-induced proliferative response of peripheral blood lymphocytes and interleukin-2 (IL-2) receptor expression on PHA-stimulated lymphocytes, while it appreciably decreased surface transferrin (Trf) receptor expression on PHA-stimulated lymphocytes. However, metenkephalin inhibited mitogen-induced proliferation and surface Trf receptor expression on uremic lymphocytes without affecting IL-2 receptor expression on PHA-stimulated cells. In ten healthy subjects opioid receptor agonists did not significantly affect mitogen-induced proliferation of lymphocytes, except for the inhibitory effect of 10(-8) M morphine in relation to lymphocytes stimulated with an optimal pokeweed mitogen (PWM) concentration. At the same time, opioid receptor agonists depressed surface IL-2 and Trf receptor expression on PHA-stimulated normal lymphocytes. In most of our experiments naloxone itself, a non-selective competitive opioid receptor antagonist, decreased mitogen-induced lymphocyte proliferation and IL-2 and Trf receptor expression on PHA-stimulated lymphocytes. Moreover, most frequently naloxone did not reverse inhibitory effects of opioid receptor agonists on lymphocytes. The results seem to indicate that opioid receptor stimulation by high metenkephalin concentrations, which are observed in the uremic blood plasma, may share the responsibility for immunodeficiency in chronic uremic patients. Next, in the presence of opioid receptor agonists directions of changes in the mitogen-induced proliferative response may not follow the alterations of IL-2 and Trf receptor expression on both uremic and normal lymphocytes. Finally the results also suggest that naloxone may possibly exert effects which are independent of its action on opioid receptors on lymphocytes.

[Comprehensive assessment of cell mediated immunity in petrochemical industry workers]. / Kompleksowa ocena stanu odpornosci komórkowej pracowników przemyslu petrochemicznego.

In the group of 29 workers of Acetobenzene Division of the petrochemical plant in Ptock with occupational exposure to toluene, acetone, and ammonia a set of tests were done evaluating the state of cell-mediated immunity which made possible finding out persons showing abnormalities in the studied parameters. It is suggested to follow-up these person continuously with control immunological testing once a year.

The effects of occupational exposure to hydrocarbons on some immune parameters of workers of the phenol division of a petrochemical plant.

The subjects of investigation included 34 workers at the Phenol Division of the Mazovian Refining and Petrochemical Plants and 18 persons working in administrative positions; the purpose was to elucidate some functions of the immune system cells. The lymphocyte subpopulations (CD3, CD4, CD8) and NK cells were evaluated using monoclonal antibodies and the immunofluorescence method. To search for the functions of lymphocytes and monocytes such as the in vitro production of interleukin-1 (Il-1) and interleukin-2 (Il-2), macrophage inhibitory factor generation (MIF), a cytotoxicity test and T cell proliferation in autologous mixed lymphocyte reaction (AMLR) were also examined. We have shown that workers from the Phenol Division exhibited pronounced deviations in the tested parameters as compared to the administration workers. We found a decrease in CD3 lymphocytes and in Il-1 production. After analysis of the individual results we selected three persons from the Phenol Division who showed abnormal values in four or more parameters (decreased CD4:CD8 ratio, abnormal values of lymphocyte subpopulations, impaired lymphocyte functions in functional tests). These cellular-immunity control tests are promising methods for studies of the biological effects of environmental and/or occupational pollution to toxic derivates of petroleum.