Vitamin D status in systemic lupus erythematosus patients and its association with selected clinical and laboratory parameters.

OBJECTIVES: To identify relationships between vitamin D serum levels and the presence of autoantibodies directed against vitamin D and levels of interleukin(IL)-17 and IL-23 in patients with systemic lupus erythematosus (SLE). METHODS: The study included 49 patients with SLE. Serum concentrations of 25(OH)D(3) were measured with electrochemiluminescence immunoassay (ECLIA). Enzyme-linked immunosorbent assays (ELISA) were used to determine antibodies directed against 1,25(OH)(2)D(3) and levels of IL-17 and IL-23 in serum of SLE patients. In evaluation of vitamin D status, the control group consisted of 49 age and gender matched healthy individuals, whereas in assessment of anti-vitamin D antibodies the control group comprised 30 sera from blood donors. RESULTS: Serum concentration of 25(OH)D(3) in SLE patients during the warm season was 18.47 ± 9.14 ng/ml, which was significantly decreased as compared with that of the control group - 31.27 ± 12.65 ng/ml (p = 0.0005). During the cold season a trend toward lower concentration of 25(OH)D(3) in SLE patients was revealed; however, it did not reach statistical significance (11.71 ± 7.21 ng/ml vs. 16.01 ± 8.46 ng/ml; p = 0.054). Results within the recommended range for vitamin D (30-80 ng/ml; 70-200 nmol/l) were observed only in three patients. The 25(OH)D(3) concentration was decreased in SLE patients with renal disease or leucopenia as compared with the levels in patients who did not have either problem (p = 0.006 and p = 0.047, respectively). The cold season was found to be a risk factor for vitamin D deficiency (<20 ng/ml) (odds ratio = 9.25; p = 0.005). Autoantibodies directed against 1,25(OH)(2)D(3) were detected in three SLE patients. No significant difference in 25(OH)D(3) serum concentrations was found between SLE patients with and without these autoantibodies. No link was shown between the existence of autoantibodies against 1,25(OH)(2)D(3) and clinical or laboratory findings, including IL-17 and IL-23 levels. However, serum concentrations of IL-23 were lower in patients with vitamin D deficiency (p = 0.037). CONCLUSIONS: SLE patients, especially those with leucopenia or renal involvement, are at high risk of vitamin D deficiency and require vitamin D supplementation. Some SLE patient sera contained 1,25(OH)(2)D(3) antibodies, but these antibodies do not appear to affect vitamin D levels.

Risk factors for atherosclerosis in healthy employees-a multidisciplinary approach.

BACKGROUND: The aim of the study was to investigate the prevalence of early atherosclerosis in healthy workers and the relationship between classical, psychological, and immunological risk factors and atherosclerosis, as well as their predictive value. METHODS: One hundred healthy managers and 50 office workers aged 35-65 were studied. In all subjects, individual, family, and occupational stress/coping risk factors were evaluated, including plasma levels of biochemical (total cholesterol, LDL, HDL, TG, glucose) and inflammatory-immunological (aCL, anti-beta(2) GPI, oxLDL, HSP, HSCRP) parameters. Carotid artery intima-media thickness (IMT) and atherosclerotic plaques in carotid arteries were assessed with computer analysis of B-mode ultrasound images. RESULTS: In 107 persons (71%) no changes were found in ultrasound images and in 43 individuals (29%) the presence of plaque was shown. The mean IMT value was 0.0618+/-0.013 mm. Cross-domain analysis showed that core predictors for IMT were age, LDL level, smoking, and occupation (being a manager) (beta=0.33, 0.30, 0.23, and 0.20, respectively); the core predictors for plaque were age, total cholesterol level, and an occupational stressor home-work balance (Wald=7, 6.7, and 5.6, respectively). Immunological factors were not independent predictors. CONCLUSIONS: In atherosclerosis, not only traditional risk factors (age, lipid disorders, and lifestyle) but also occupational stress factors may play a role. Immunological factors do not seem to play a role in the development of atherosclerosis in a population of healthy workers. The interplay between occupational stress and atherosclerotic changes requires further investigation.

Topoisomerase I is differently phosphorylated in two sublines of L5178Y mouse lymphoma cells.

Two sublines of LY murine lymphoma, differing in sensitivity to CPT, served as source of topoisomerase I in order to compare the enzyme's properties. The activity of topoisomerase I isolated from LY-S cells of reduced sensitivity to CPT increased about 2-times more upon phosphorylation with casein kinase but was inhibited to a lesser extent upon dephosphorylation with alkaline phosphatase than the enzyme from the CPT-sensitive LY-R cells. The in vitro phosphorylation of LY-S enzyme restored its sensitivity to CPT. The in vitro incorporation of 32P into topoisomerase protein was about 1.7-times higher in LY-S than in LY-R enzyme. A reversed incorporation ratio was observed upon metabolic labelling. The level of topoisomerase I protein, determined by Western blot analysis using scleroderma anti-topoisomerase I antibodies, was about 1.5-times higher in LY-S than in LY-R cells. The level of topoisomerase I mRNA was similar in both sublines. These results indicate that the reduced sensitivity of LY-S cells to CPT is based on the lowered phosphorylation of topoisomerase I protein but does not depend on the expression of topoisomerase I gene.

Arginase isoforms in human colorectal cancer.

Arginase (EC 3.5.3.1) activity was determined in 54 colorectal tissues obtained from patients with primary colorectal adenocarcinoma as well as in serum of 45 patients and 65 healthy individuals. In patients, the preoperative values of the mean serum arginase activity and the activity in colorectal tumors were much higher than in serum of healthy subjects and control tissues. Two isoforms of arginase, anionic and cationic, were identified in colorectal tissues (normal and cancerogenous), and only one, the cationic form, in serum. These arginases were different from the main human liver cationic arginase (pI 9.3). The anionic colorectal arginase was identical with the human liver anionic isoform (pI 7.7), and the cationic arginase from colorectal tissues and blood serum with the human kidney cationic enzyme (pI 8.9). The total activity and the level of protein of the cationic arginase in colorectal cancer was higher than in control tissue, and it was also higher in serum of patients with colorectal cancer than in healthy subjects. Thus, it can be concluded that the increased arginase activity in blood serum and colorectal cancer in studied patients was due to the raised level of the cationic arginase and this isoform seems to be a discriminating parameter for assessing the presence of colorectal cancer.

Evaluation of cerebrospinal fluid for the presense of anticardiolipin antibodies (aCL) in NP-SLE patients.

Many neurological or psychiatric manifestations of SLE (NP-SLE) are related to the presence of anticardiolipin antibodies (aCL) in the patient's sera. The aim of this study was to evaluate the presence of aCL in cerebrospinal fluid (CSF) in SLE patients with NP features. Fifteen SLE patients were studied, all with NP features. CSF was evaluated for intrathecal IgG synthesis, oligoclonal IgG, and blood-brain barrier impairment. Sera and CSF were tested by ELISA for the presence of aCL-IgG and aCL-IgM with and without beta2 glycoprotein (beta2 GPI) cofactor. CSF and sera of 50 low back pain patients served as controls. Six patients were aCL(+) and nine aCL(-). In all patients the general CSF examination was normal. In all patients the value of indices of intrathecal IgG synthesis were normal but oligoclonal protein was present in the CSF of three patients. In none of the patients was the blood-brain barrier impaired. Neither aCL-IgG nor aCL-IgM was detected in the CSF of any NP-SLE patient. Mean levels of aCL in patients without cofactor beta2 GPI and with cofactor were as follows: for IgG class 0.005 and 0.057 OD (negative); for IgM class 0.004 and 0.024 OD (negative). We could not detect aCL in the CSF of patients with NP-SLE, even if sera were positive for aCL.

[Immunological disorders in patients with chronic viral hepatitis type C]. / Zaburzenia immunologiczne u chorych z przewleklym wirusowym zapaleniem watroby typu C.

UNLABELLED: Various immunological disorders have been observed frequently in patients with chronic viral hepatitis C. Hepatitis C virus has been implicated in the pathogenesis of: essential mixed cryoglobulinemia, cryoglobulinemic vasculitis glomerulonephritis, thrombocytopenia, porphyria cutanea tarda, autoimmune thyroiditis (among others). AIM OF THE STUDY: Was to determine the prevalence and clinical meaning of immunological disorders in HCV infected patients. METHODS: 93 HCV infected patients were studied with regard to the presence of cryoglobulins, autoantibodies, rheumatoid factor (RF) and circulating immunological complexes (CIC). RESULTS: 35 patients out of 93 (38%) had detectable cryoglobulins. Cryoglobulins in 90 cases were of type III and in 3--of type II. CIC /by immunoelectrophoresis of the PEG sediments/ were found in sera of 87 patients (93.5%). 38 persons (41%) had detectable rheumatoid factor. Antinuclear antibodies were found in sera of 15 patients (16%), anti-smooth muscles antibodies--in 21 persons (23%) and anti-LKM1 antibodies--in 5 (5%). Titers of autoantibodies were usually low. The most frequent clinical manifestations were arthralgia and skin changes. CONCLUSION: Immunological disorders /circulating immune complexes (93.5%), autoantibodies (60%), rheumatoid factor (41%), cryoglobulinemia type III (38%)/ are frequently found in HCV infected patients. Age, gender, histological score and clinical evidence of liver cirrhosis do not influence on rate of these abnormalities. Clinical manifestations of immunological disorders are usually mild.

The role of cytokines in inflammatory response after total knee arthroplasty in patients with rheumatoid arthritis.

Surgery and trauma is accompanied by changes in blood levels of certain cytokines and chemokines including interleukin-6 (IL-6) and interleukin-8 (IL-8). However, there is little data on correlations between local and systemic levels of these mediators during orthopedic surgeries in rheumatoid arthritis (RA) patients who already show increased levels of proinflammatory cytokines due to their disease. We aimed to measure dynamics of blood and drainage fluid levels of IL-6 and IL-8 in postoperative period in rheumatoid arthritis patients undergoing knee replacement surgery and correlate these changes with blood levels of C-reactive protein (CRP), body temperature and pain. We report that blood and drainage fluid levels of IL-6 and IL-8 showed significantly increasing trend during the 36-h period after the surgery. Drainage fluid levels of both cytokines were significantly higher in comparison with blood, indicating their local production in the operated joint. In contrast, levels of CRP were higher in blood than in drainage fluid. Despite the fact that the levels of tested cytokines had already been high in RA patients before surgery, we conclude that after surgery their levels were being much significantly enough high in drainage fluid to reflect dominated local inflammatory reaction to surgical stress and trauma.

Pathogen-specific humoral response in Treponema pallidum-infected humans, rabbits, and guinea pigs.

Molecular analysis of the humoral response to pathogen-specific polypeptides was done using sera from patients at different stages of syphilis and sera from Treponema pallidum-infected rabbits and guinea pigs collected at various times after infection. The sera were examined by ELISA, fluorescent treponemal antibody absorption test (FTA-ABS), and immunoblot before and after sequential adsorption with cyanogen bromide-activated Sepharose 4B coupled individually to five sonicated nonpathogenic treponemes. Guinea pig antisera were also adsorbed with rabbit proteins. After adsorption, sera from all three species reacted neither by ELISA nor by immunoblot with nonpathogenic treponemes, nor did guinea pig sera react with rabbit proteins. Regardless of the species of treponema used for initial adsorption, none by itself could effectively remove all cross-reactivities. Nine pathogen-specific polypeptides (15, 17, 33, 37, 39, 43, 45, 47, 97 kDa), eight (15-47 kDa) known as integral membrane or endoflagellar components, were commonly recognized by the tree species when chancre immunity developed. The substantial reduction in immune reactivity after adsorption suggests that most of these peptides consist of specific and common treponemal epitopes. Throughout the various stages of the human disease, pathogen-specific antibodies were exclusively of the IgG isotype.

Effect of passive immunization with purified specific or cross-reacting immunoglobulin G antibodies against Treponema pallidum on the course of infection in guinea pigs.

Whole immune serum or highly purified immunoglobulin G (IgG) antibodies to Treponema pallidum exhaustively adsorbed with three strains of nonpathogenic treponemes (TPI-IgG) were used for passive immunization of inbred strain 2 guinea pigs before and after intradermal challenge with 3.4 x 10(7) virulent T. pallidum Nichols organisms. Before challenge, control animals received a similarly purified IgG fraction containing either a cocktail of antibodies against three nonpathogenic treponemes (NPTI-IgG) or IgG prepared from normal guinea pig serum (NGPS-IgG). The purified fractions contained both IgG1 and IgG2 isotypes. The antibody levels (detected by fluorescent treponemal antibody test and enzyme-linked immunosorbent assay) and molecular specificities (immunoblot) of sera obtained from recipient animals before infection reflected those of the purified fractions used for immunization. Three protocols of passive immunization were used. Whole immune serum containing specific and cross-reacting antibodies afforded better protection than TPI-IgG even though asymptomatic animals were not fully protected. A single intradermal injection (0.1 ml) of TPI-IgG or NPTI-IgG into one hind leg 22 h before infection at the same site provided relatively higher protection than multiple intravenous injections (total, 15 ml) of the respective individual preparations. Since purified NGPS-IgG injected in the same animals, into the opposite hind leg, failed to protect against the challenging infection, it is reasonable to assume that specific and cross-reacting antitreponemal antibodies of the IgG1 subclass, which in guinea pigs are homocytotropic, play a relevant role in local protection.

Kinetics of pathogen-specific humoral response in Treponema pallidum-infected young and old inbred strain 2 guinea pigs.

The kinetics of the humoral response to pathogen-specific polypeptides was examined in Treponema pallidum-infected young (3-5 months old) and old (10-20 months old) inbred strain-2 guinea pigs. Sera collected before and at various times after infection were pooled and examined by immunoblotting and two serologic tests (ELISA and FTA) before and after sequential adsorption with CNBr-activated sepharose coupled to normal rabbit proteins and antigens from five nonpathogenic treponemal species. Prior to adsorption the kinetics of the humoral response to T. pallidum antigens did not seem to differ significantly between the two groups. After adsorption, however, a delay in the appearance of detectable antibodies and a milder response to various pathogen-specific polypeptides was observed in the older group. After adsorption, a sharp drop in ELISA-TP, ELISA-TR and FTA titres occurred in both groups. Six months post-infection, between 9 and 10 pathogen-specific polypeptides (97, 57, 47, 45, 43, 39, 37, 33, 17 and 15 kD) were recognized by both groups. The effect of age and levels of natural treponemal antibodies on the clinical symptoms of primary lesions and humoral response to pathogen-specific polypeptides is discussed.

Photosynthetic apparatus in chilling-sensitive plants : III. Contribution of loosely bound manganese to the mechanism of reversible inactivation of hill reaction activity following cold and dark storage and illumination of Leaves.

The levels of both tightly and loosely bound Mn in chloroplasts from fresh, cold and dark stored as well as illuminated leaves of Lycopersicon esculentum Mill. were studied in relation to Hill reaction activity. The tightly bound Mn pool represents one third of the total Mn content in chloroplasts isolated from the fresh leaves, and its level does not change following cold storage and illumination of leaves. Upon cold and dark storage of leaves the loss from the chloroplasts of about 40%-50% of the total amount of Mn is accompanied by an almost complete inactivation of the Hill reactions, as studied with water as an electron donor, as well as by the appearance of an EPR signal characteristic of free Mn(2+) ions. Following illumination of such leaves, the restroration of Hill reaction activity is accompanied by an increase in the total Mn content in chloroplasts of up to 70%-80% of the Mn level measured in the fresh leaves and by disappearance of the EPR signal. In contrast, aging in the cold of isolated chloroplasts does not affect their Mn content. The addition of manganese does not result in the restoration of Hill reaction activity in chloroplasts from cold stored leaves but causes a restoration of this activity inhibited by linolenic acid. The data suggest that the loosely bound Mn pool (extractable with Tris) can be differentiated into two fractions: (1) one functionally inactive in electron transport and (2) one responsible for restoration of Hill reaction activity. Mn of the latter fraction (about 45% of the total Mn content) probably originates from the free Mn ions present in the interior of the chloroplasts following the cold and dark storage of leaves and from Mn reincorporated into chloroplasts from the cytoplasm. Incorporation of Mn from both these sources into thylakoid membrane to form a functionally active, loosely bound Mn pool proceeds during the illumination of leaves and results in the restoration of Hill reaction activity inhibited following the storage of leaves in dark and cold.

Photosynthetic apparatus in chilling-sensitive plants : I. Reactivation of hill reaction activity inhibited on the cold and dark storage of detached leaves and intact plants.

Chloroplast isolated from the detached leaves of chilling-sensitive plants-Phaseolus vulgaris L., Cucumis sativus L., and Lycopersicon esculentum Mill.-stored in the cold for 2-4 days in the dark exhibit an almost complete loss of Hill reaction activity, which on illumination of leaves is restored to almost the original level. In contrast, illumination of either chloroplast suspensions or homogenates from leaves stored in the cold in the dark does not cause restoration of electron transport. Cold and dark storage of leaves of chilling-sensitive plants affects the electron transport before the site of electron donation by diphenylcarbazide and results in an increased sensitivity of the Hill reaction of isolated chloroplasts to exogenous linolenic acid. Illumination of leaves reverses these processes. When tomato plants are exposed to 0°C in intermittent light, Hill reaction activity is not affected while dark storage either at 0°C or 25°C results in a significant decrease of Hill reaction activity after 2-3 days followed by the restoration of electron transport to the original level after 1 or 2 days of the prolonged dark storage of plants. When tomato plants are stored either at 0°C in intermittent light, at 0°C in dark, or at 25°C in dark the sensitivity of the Hill reaction to exogenous linolenic acid remains increased despite a significant restoration of this activity. In conclusion, both darkness and the detachment of leaves from the plant are more effective than cold treatment in damaging photosystem II whereas both light and intact structure of the cell are required for restoration of Hill reaction activity in chloroplasts following cold and dark storage of detached leaves.

Experimental neonatal syphilis in a susceptible (C4D) and a resistant (Albany) strain of guinea pig.

Despite similar levels of natural antibodies and treponemicidal activity, 83% of fourth complement component-deficient (C4D) mother guinea pigs developed ulcerative lesions to a challenge of 5 x 10(7) Treponema pallidum, whereas 75% of offspring 1 to 5 days old were temporarily (2-3 months) resistant to development of dermal lesions. In contrast, only 17% of Albany-strain mothers developed small papular lesions, while 68% of 1- to 5-day-old newborns developed large papular or ulcerative lesions within 9-15 days postinfection. These findings, together with the late development of both dermal lesions and antibodies in C4D neonates, preclude the concept of an antibody-associated natural resistance. T. pallidum infection in either C4D or Albany neonates was not associated with depletion of any particular cell population in lymphoid tissue. However, marked age- and strain-dependent histologic differences were noted. Histologic examination of lymph nodes and spleens from 17-day-old and 3- to 4-month-old animals showed that maturation of lymphoid tissues in C4D animals lagged behind the Albany strain at either age. Moreover, 75% of C4D newborns contained significantly higher levels of immunomodulatory alpha 1 fetoprotein than Albany neonates. The possibility that differences in susceptibility to T. pallidum infection between C4D and Albany guinea pigs as neonates and again as adults is the result of genetically associated changes in immunologic recognition is discussed.

Immunogenicity of three recombinant Treponema pallidum antigens examined in guinea pigs.

The immunogenicity of recombinant treponemal antigens TmpA, TmpB and TmpC incorporated in RIBI adjuvant and injected into inbred strain 2 guinea pigs has been examined. The immune status of these animals has been challenged by infection with Treponema pallidum, Nichols. The immune response evaluated by the fluorescent-antibody test, microhemagglutination test and ELISA demonstrated high titers of antibodies to the T. pallidum antigens. The immunoblot analysis proved that the antibodies were directed to the 43-(Tmp A) 34- (Tmp B) and 35-kdalton (Tmp C) polypeptides. Antibodies cross-reacting with Treponema phagedenis biotype Reiter were, however, also detected. In spite of high titers of antibodies the animals were not protected against challenging infection with 10(8) organisms of T. pallidum.