[Natural antibodies against α(1,3) galactosyl epitope in the serum of cancer patients]. / Prirozené protilátky proti α(1,3) galaktosylovému epitopu v séru nemocných s maligními nádory.

BACKGROUND: Natural antibodies against saccharide antigens are found in the human serum; most of them are directed against α-galactosyl epitope (Galα1-3Galß1-4GlcNAc-R). Experimental and initial clinical studies show the potential for use of anti-galactosyl antibodies in the immunotherapy of cancer patients with glycolipids containing the α-galactosyl epitope. This therapeutic approach is based on the presence of these antibodies in the serum of cancer patients. Only scarce literature data is available on the incidence of these antibodies in cancer patients. Data is lacking on their amounts and isotype characteristics in different types of cancer. MATERIAL AND METHODS: An ELISA test with a polyacrylamide-conjugated synthetic disaccharide, Galα1-3Galß, has been designed for quantitative detection of anti-galactosyl IgM, IgG, and IgA antibody isotypes. This test was used to screen the sera from 57 patients with breast, colorectal, or panceatic cancer or malignant melanoma and from 145 healthy controls. RESULTS: The serum concentration of anti-galactosyl antibodies (anti-Gal) is gender dependent: anti-Gal IgM antibodies are present in higher titres in healthy women than in healthy men (p < 0.01). Patients with breast, colorectal, or pancreatic cancer or malignant melanoma had comparable serum levels of anti-Gal IgM, IgG, and IgA antibody isotypes to healthy controls. Male patients with colorectal cancer had higher anti-Gal IgA antibodies than healthy men (p< 0.01). CONCLUSION: Comparable concentrations and isotypes of anti-galactosyl antibodies are found in the serum of cancer patients and healthy controls.

[A simple method for the detection of CD154 (CD40L) on peripheral blood lymphocytes]. / Jednoduchá metoda k prukazu CD154 (CD40L) na lymfocytech v periferní krvi.

OBJECTIVE: CD154 (also called CD40L) is a transmembrane glycoprotein predominantly expressed on the surface membrane of activated CD4+ T cells. Its receptor CD40 is present on all B cells, but also on other cells. The interaction CD154-CD40 is necessary for the optimal development of the adaptive immune response and also has consequences for the modulation of the inflammatory response. A defect in the expression of CD154 is pathognomonic of congenital immunodeficiency called X-linked Hyper-IgM syndrome (XHIGM). To detect the abnormality of CD154 is essential for making the diagnosis of XHIGM. MATERIAL AND METHODS: We worked out a microtest for the detection of CD154 expression on in vitro activated CD4+ T cells in whole blood and compared it with that on isolated cells from peripheral blood. Heparinized peripheral blood was activated with phorbol 12-myristate 13-acetate and ionomycin for 4 hours, labeled with monoclonal antibodies and analyzed by flow cytometry. Considering that the CD4 marker on the plasma membrane surface decreases during the activation, CD4+ T cells are mostly recognized as CD5+/CD8- cells. Their activation is monitored based on the expression of CD69. Three-colour immunofluorescence staining was used for simultaneous detection of CD154. RESULTS: Ten blood donors were tested. As little as 0.5 ml of heparinized whole blood is needed to complete the test. Optimal time for activation and detection of CD154 on T lymphocytes is 4 hours. We found that the number of CD4 molecules on the surface of T cells decreases during the activation. The expression of CD154 in our whole blood microtest is fully comparable with that in the test on isolated leukocytes. CONCLUSION: The presented microtest for the detection of CD154 on activated lymphocytes in whole blood is fast and blood saving, since as little as 0.5 ml of blood is needed to complete it. It can be recommended as the initial test for suspected hyper-IgM syndrome in children. We demonstrate that this screening method can help to detect also carriers of XHIGM.

Serum inflammatory biomarkers in Parkinson's disease.

Numerous recent findings indicate the involvement of a neuroinflammatory reaction in the neurodegeneration in idiopathic Parkinson's disease (PD). We examined 29 consecutive patients with PD, ages 54-84 years, most of whom were moderately impaired (median UPDRS 19; Hoehn-Yahr 3; MMSE 28). A series of serum biomarkers were investigated, and their levels were correlated with the degree of the motor and cognitive impairment. There were no abnormalities of IL-6, acute phase proteins (C-reactive protein, serum amyloid A, alpha 1-antitrypsin, orosomucoid, ceruloplasmin, alpha 2-macroglobulin, transferrin, prealbumin) and factors of the complement system (C1q, C1-INH, C3, C4). A decrease in Mannan-binding lectin (MBL) levels was observed in six patients; an elevation of tumor necrosis factor-alpha (TNF-alpha) was found in 12 patients. No statistically significant correlation was found between the patient's clinical state (neuropsychologic and motor, as expressed by UPDRS III, Hoehn-Yahr, and MMSE) and the immunomarker changes. Our results indicate that the inflammatory process may be reflected in the serum; nevertheless, further research is needed to elucidate the possible clinical implications.

[Early feeding in infancy and atopic dermatitis - a prospective observational study]. / Säuglingsernährung und atopische Dermatitis - eine prospektive Beobachtungsstudie.

BACKGROUND: Recommendations for primary prevention of allergic diseases in high-risk children include hypoallergenic infant formulas (HA) if breastfeeding is insufficient. The primary objective of our study was to investigate the atopic dermatitis (AD) preventive effect of breastfeeding and HA-nutrition in the first 2 years of life and to follow the increase in weight. PATIENTS AND METHODS: Altogether 174 newborns with a hereditary risk for atopy were enrolled in the study, 121 children were investigated at the age of 2 months, 111 at the age of 4 and 106 at the age of 6 months. A total of 45 infants were in the first half-year of life exclusively breastfed and 61 infants were mainly fed with HA. RESULTS: The body weight of initially HA-fed children was 7870G (SD 949) significantly higher as the one of breastfed children (7508 G, SD 912, p=0.0571), in addition the weight increase was also significantly higher in HA-fed infants at the age of 6 months (p=0.0042). The frequency of AD as well as SCORAD score at the age of 6 to 24 months was comparable in both groups. Neither the milk-specific IgE antibodies nor the proliferation of peripheral blood mononuclear cells (PBMC) to bovine beta-Lactoglobulin (BLG) at the age of 6 months had a prognostic value for development of atopic dermatitis. CONCLUSION: The likelihood to develop AD in the first 2 years of life was comparable in exclusively breastfed as in HA-fed infants with hereditary risk for atopy. The initially HA-fed children demonstrated at the age of 6 months higher body weight and weight increase as the exclusively breastfed infants. The efficacy of nutritional intervention on the incidence of AD in high-risk children for atopy could not be predicted by milk-specific IgE antibodies or BLG-specific proliferation of PBMC.

[Ilya Ilich Metchnikov and Paul Ehrlich: 1908 Nobel Prize winners for their research on immunity]. / Ilja Iljic Mecnikov a Paul Ehrlich: laureáti Nobelovy ceny v r. 1908 za práce o imunite.

The Nobel Prize in Physiology or Medicine in 1908 was awarded to Ilya I. Mechnikov and Paul Ehrlich for recognition of their work on immunity. Mechnikov have discovered phagocytes and phagocytosis as the basis of natural cellular immunity. His ,,phagocytic theory" is the principle of immunological concept "self and not self" as the prerequisition of physiological inflammation, and selfmaintaining of organism. Ehrlich developed the methods for standardization of antibody activity in immune sera, described neutralizing and complement-depending effect of antibodies and enunciated the ,"ide-chain" theory of the formation of antibodies. Their concept of the key-stone of immunity was different, but they expressed the basic paradigma of immunology: immunity imply the protection of identity and guarantee the integrity of organism. Both are the founders of immunology as the scientific discipline. Discoveries and conceptions of I. Mechnikov and P. Ehrlich exceedingly influenced development of immunology and are also applicable, instructive and suggestive in contemporary immunology and microbiology.

Lack of dehydroepiandrosterone in type I and II hereditary angioedema and role of danazol in steroid hormone conversion.

BACKGROUND: Hereditary angioedema (HAE) is successfully treated with danazol, a therapeutic steroid compound. To investigate hormones of the hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axis in patients with HAE with and without danazol. METHODS: We included 16 patients with type I HAE, nine patients with type II HAE, and 16 healthy subjects. Serum levels of adrenocorticotropic hormone (ACTH), cortisol, androstenedione, dehydroepiandrosterone (DHEA), free testosterone, and 17beta-oestradiol were measured. RESULTS: Serum levels of ACTH were markedly decreased in patients with type II HAE compared to the other groups (P < 0.001). Serum cortisol was similar between groups but danazol treatment decreased cortisol levels, particularly in women (P = 0.019). Serum levels of DHEA were significantly decreased in all patients with type I and II HAE compared to controls (P < 0.05), which was only partly dependent on prior danazol therapy as patients without danazol had also decreased serum levels of DHEA (P < 0.05). Furthermore, free testosterone serum levels were markedly increased in patients under danazol (P < 0.005) and the ratio of 17beta-oestradiol/free testosterone was significantly decreased in these patients (P < 0.005). CONCLUSIONS: This study demonstrated decreased DHEA in patients with type I and II HAE independent of danazol therapy, which was particularly evident in women. It also demonstrates that danazol induced a marked up-regulation of free testosterone in relation to precursors and downstream 17beta-oestradiol. In HAE, there seems to be a primary lack of the adrenal androgen DHEA.

T and B lymphocyte subpopulations and activation/differentiation markers in patients with selective IgA deficiency.

Selective deficiency of immunoglobulin A (IgAD) and common variable immunodeficiency (CVID) are genetically closely related diseases, both of unknown pathogenesis. A plethora of abnormalities in lymphocyte subpopulations and expression of activation markers were repeatedly documented in CVID patients, while almost no data are available about lymphocyte subpopulations in IgAD patients. We determined basic lymphocyte subpopulations and those subpopulations that were reported to be abnormal in CVID patients (CD25, human leucocyte antigen (HLA)-DR CD45RA, CD45RO, CD27, CD28 and CD29 on both CD4(+) and CD8(+) cells, CD57 and CD38 on CD8(+) cells, CD21, CD27, IgM, IgD on B lymphocytes) in 85 patients with IgAD, 47 patients with CVID and in 65 healthy controls. Statistical analysis was performed by the Mann-Whitney U-test; significant P-values were determined by means of Bonferoni's correction. Our results showed an increase in the relative number of CD8(+) cells and a decrease in the absolute number of CD4(+) cells compared to healthy people, but similar abnormalities in CVID patients were much more expressed. IgAD patients had significantly decreased expression of HLA-DR and increased expression of CD25 on CD4(+) lymphocytes, also CD29 expression was decreased on CD8(+) cells, while other activation/differentiation markers on T cells (including the expression of CD45RA and CD45RO antigens) were not changed. There were no statistically significant abnormalities in B lymphocyte developmental stages in IgAD patients compared to healthy controls. Our observation showed that the majority of T and B lymphocyte subpopulation abnormalities described previously in CVID are not present in IgAD patients.

Age dependency and mutual relations in T and B lymphocyte abnormalities in common variable immunodeficiency patients.

Common variable immunodeficiency (CVID) is primary hypogammaglobulinaemia with an unknown aetiopathogenesis. Although various abnormalities of T and B cells have been described, their pathogenetic roles are unclear. We determined T and B lymphocyte subsets known to be abnormal in CVID in order to disclose possible relations between numerical abnormalities in those cells. Markers associated with B cell development (CD21, CD27, IgM, IgD) were determined on B lymphocytes (CD19+); T lymphocyte development (CD45RA, CD45RO, CD62L) and activation markers (CD25, CD27, CD28, CD29, CD38, CD57, HLA-DR) were determined on CD4+ and CD8+ T lymphocytes in 42 CVID patients and in 33 healthy controls. Abnormalities in CD4+ T lymphocyte activation markers (increase in CD29, HLA-DR, CD45RO, decrease in CD27, CD62L, CD45RA) were observed particularly in patients with a decreased number of memory (CD27+) and mature (CD21+) B cells (group Ia according to the Freiburg group's classification), while abnormalities observed in CD8+ cells (increase in CD27 and CD28 and decrease in HLA-DR, CD57 and CD38) did not depend upon grouping patients together according to B lymphocyte developmental subpopulations. We observed correlations between immature B cells (IgM+ CD21-) and expression of CD27, CD62L, CD45RA, CD45RO and HLA-DR on CD4+ T cells in CVID patients but not in the control group. The expression of CD27 and CD45RA on CD4+ T lymphocytes, such as the percentage of IgD+ CD27- and IgD+ CD27+ cells in B lymphocytes, showed age dependency to be more significant than in the control group. Our study demonstrates that T and B lymphocyte abnormalities in CVID are partially related to each other. Some of those abnormalities are not definite, but may evolve with age of the patient.

[Cow milk-specific humoral and cellular immune response in infants with high risk of atopy under feeding a whey hydrolysate infant formula]. / Kuhmilchspezifische humorale und zelluläre Immunantwort bei Säuglingen mit hohem Atopierisiko unter Ernährung mit einer Molke-Hydrolysatnahrung.

BACKGROUND: Hypoallergenic infant formulas (HAF) were developed for atopy prevention in infants with high risk of atopy if these cannot be breastfed. HAF mount an antigen-specific immune response in infants. The aim of the study was to analyse the immune response in infants fed with a new infant formula based on a whey hydrolysate (HAF) and to compare it with that of exclusively breastfed controls. PATIENTS AND METHODS: Plasma concentrations of cow milk-specific IgE were analysed in 94 infants with high risk of atopy, 44 were exclusively breastfed, 50 were fed with HAF. In addition, cow milk-specific IgG antibodies (26 breastfed, 30 fed with HAF) as well as proliferation of periph-eral blood mononuclear cells to bovine beta-lactoglobulin (BLG) (41 breastfed, 47 fed with HAF) were tested. Specific IgE and IgG antibodies were determined using enzymoimmunometric assay (Alastat). Cellular proliferation was measured using tritiated thymidine incorporation assay after 6 day stimulation with BLG. RESULTS: Elevated IgE to cow milk antibodies (> 0.35 kU/L) were detected in two infants from the breastfed group and in one from the HAF-fed group. The plasma concentrations of milk specific IgG antibodies in HAF-fed infants were insignificantly higher than those in breastfed ones. No significant difference was found in bovine BLG-specific cell proliferation between both groups. CONCLUSION: Concerning the properties investigated like antigenicity, allergenicity and immunogenicity, the extensively hydrolysed whey based hypoallergenic formula does not significantly differ from mother milk in 6 month-old infants with an increased atopy risk.

A concurrent occurrence of cutis laxa, Dandy-Walker syndrome and immunodeficiency in a girl.

UNLABELLED: We report on a 17-y-old girl with inherited cutis laxa, immunodeficiency and Dandy-Walker syndrome. Immunodeficiency manifested itself by decreased and fluctuating levels of IgG, IgA and IgM and intermittent leucopenia causing increased susceptibility to respiratory tract infections. Dandy-Walker syndrome (agenesis of the cerebellar vermis with a large posterior fossa cyst communicating with an enlarged 4th ventricle) was shown on a CT scan but with the exception of macrocrania, no typical signs or symptoms were observed at the age of 17. Loose hyperextensible skin with pendulous skinfolds as a manifestation of cutis laxa was observed from birth. Anomalies of the right pulmonary artery, abnormal branching of the left arteria subclavia (arteria lusoria) from the left aortic arch and bicuspidal aortic valve were also present. CONCLUSION: The combination of the rare disorders cutis laxa, Dandy-Walker syndrome and immunodeficiency is reported here for the first time.

Early manifestation and recognition of C2 complement deficiency in the form of pyogenic infection in infancy.

OBJECTIVE: Although frequently asymptomatic, C2 complement component deficiency may lead to severe pyogenic infections or lupus-like illness. In the present report, we describe infectious manifestations in infancy and childhood in our C2-deficient patients. METHOD: A retrospective study of clinical manifestation in three patients was carried out. C2 deficiency was proved both by undetectable serum C2 level and typical homozygous 28 bp deletion of the C2 gene. RESULTS: All patients were hospitalized at least once by the age of 12 months, each had one episode of meningitis in infancy, one also had arthritis with septicaemia. Infections of the respiratory tract were the causes of other hospitalizations. Two patients also suffered from frequent mild respiratory tract infections; in both patients, decreased immunoglobulin IgA and immunoglobulin IgG2 or immunoglobulin IgG3 levels were recorded. CONCLUSION: Our observations point to an early manifestation of C2 deficiency within the first year of life, with meningitis as the most severe complication. The severity of immunodeficiency may be influenced by concomitant deficiencies of immunoglobulin isotypes.

Dimethyl 2-[[2-(methoxycarbonyl)-1-(methoxycarbonylmethyl)pyrrol-4-yl]methylene]propanedioate.

The title compound, C(15)H(17)NO(8), is a pyrrole-ethene derivative with potential biological activity. Although a large part of the molecule is planar, there is no structural evidence for any conjugation push-pull effect across the ethylenic bond, which is usually observed for substituted ethylenes; pi-electron delocalization appears to be restricted to the 2-(methoxycarbonyl)pyrrole moiety.

cis-6-Phenyl-4bH,6H,11H,13H-isoindolo[1,2-c]benz[2,4]oxazepin-13-one.

The title compound, C22H17NO2, contains an isoindolinone moiety joined to a phenyl-substituted benzoxazepine ring. The isoindolinone moiety is essentially planar and the oxazepine ring adopts a distorted chair conformation, with the phenyl substituent equatorial. Owing to the severe puckering of the central oxazepine ring, the molecule as a whole is non-planar; the benzene ring of the benzoxazepine fragment makes an angle of 67.7 (1) degrees with respect to the isoindoline ring.

4-(4-Methoxyphenylamino)-3-phenylazo-3-penten-2-one.

The title compound, C18H19N3O2, was obtained by an azo-coupling reaction with enaminones and is composed of a planar azoenamine skeleton which forms a six-membered ring through a symmetrical intramolecular hydrogen bond. The compound was found to exist as an equilibrium mixture of major hydrazoimino and minor azoenamine tautomers. Quantification of the relative contribution of the tautomeric forms is obscured by the existence of the hydrogen bond. Comparison of the results with those obtained for a similar structure revealed a substantial effect on the tautomeric equilibria of the nature of the substituent bonded to the amine nitrogen.

[Autoantibodies in relatives of IgA-deficient patients]. / Výskyt autoprotilátek u príbuzných nemocných se selektivním deficitem IgA.

Selective IgA deficiency is frequently associated with autoimmune phenomena. We addressed the question, whether autoantibodies are more frequent in relatives of IgA deficient persons. Fifteen first-degree relatives of patients with a familial form of IgA deficiency and 129 first-degree relatives of patients with a sporadic form of IgA deficiency were evaluated. Following autoantibodies were detected: antinuclear, anti-thyreoglobulin, anti-thyroid microsomal, anti-gastric parietal cell, anti-reticulin, anti-smooth muscle, anti-mitochondrial, anti-neutrophil cytoplasmatic, anti-cardiolipin, rheumatoid factor and antibodies against gliadin. We did not find increase in any of the autoantibodies evaluated in any groups of patients. Our results do not support a significant genetic influence in autoantibodies formation in selective IgA deficiency.

IgA deficiency in Czech healthy individuals and selected patient groups.

BACKGROUND: Selective IgA deficiency (IgAD) is the most common immunoglobulin deficiency with a variety of clinical manifestations. The frequency of IgAD differs depending on the ethnic origin and clinical symptoms of investigated persons. METHODS: The prevalence of IgAD (serum IgA level <0.05 g/l) was determined in 5,310 Czech blood donors, 10,326 patients who had undergone immunological investigation, and 246 first-degree relatives of IgAD and common variable immunodeficiency (CVID) patients. RESULTS: IgAD was detected in 13 (1/408; 0.24%) of the blood donors. The prevalence of IgAD was increased both in children (48/3,113; 1.5%) and adults (33/3,824; 0.9%) referred for frequent respiratory tract infections (in both cases p<0.001) compared to the healthy population. The frequency of IgAD was 12/189 (6%) in first-degree relatives of IgAD patients and 9/57 (16%) in relatives of CVID patients, with the highest frequency observed in children of CVID patients. CONCLUSIONS: The prevalence of IgAD in the Czech healthy population is comparable to that in other Caucasians. The frequency is increased in children with recurrent respiratory tract infections and especially in relatives of patients with immunoglobulin deficiencies.

[IgG subclasses and autoantibodies in adult patients with selective IgA deficiency]. / Podtrídy IgG a autoprotilátky u dospelých pacientu se selektivním deficitem IgA.

Selective IgA deficiency is the most common primary immunodeficiency disease, but the etiopathogenesis is unknown. In a portion of patients disturbed IgA production is accompanied by various immunological abnormalities. Serological laboratory results of 30 female and 22 male adult patients with selective IgA deficiency were compared with sex- and age-matched healthy controls. Hypergammaglobulinemia IgG was observed in 31 patients and only in 2 control persons. Serum IgG1 was increased in 12 and/or IgG3 in 18 patients, the increase in IgG2 was less common (6 persons). The number of persons with increased IgE did not differ from the control group. The occurrence of autoantibodies (antinuclear antibodies, rheumatoid factor, antithyroglobulin and anti-thyroid microsomal antibodies, anti-gastric parietal cells, reticulin, smooth muscle, anticardiolipin, and anti-gliadin antibodies) did not differ significantly from the control group. IgG hypergammaglobulinemia, which, according to our results, is the most frequent accompanying serological abnormality in IgA deficiency, may be caused by compensatory increased production, but may also reflect more profound immunological dysregulation in the disease.

Dimethyl (1-methyl-1,3-benzimidazol-5-yl)aminomethylenepropanedioate monohydrate.

In the title compound, C(14)H(15)N(3)O(4).H(2)O, there is a strong conjugation push-pull effect across the central double bond, as reflected in the molecular dimensions and the planarity of the enaminone portion of the molecule. The molecule has an intramolecular hydrogen bond between the NH and CO groups in the Z configuration, adopting the chelated form. The two pi systems of the molecule (1-methylbenzimidazole and enaminone) are deconjugated and tilted with respect to each other by 15.6 (2) degrees. The solvent water molecule is hydrogen bonded to the N(1) atom of the 1-methylbenzimidazolyl group.

Isoprinosine does not protect against frequent respiratory tract infections in childhood.

UNLABELLED: Isoprinosine, an in vitro immuno-enhancing agent principally acting by stimulating T-lymphocytes, is one of a number of agents sometimes used in an attempt to prevent recurrent respiratory infections in children, although there are no formal trials for this particular drug. We performed a placebo-controlled double-blind trial to assess the efficacy of isoprinosine (50 mg/kg per day) for 6 weeks followed by 50 mg/kg per day twice weekly for 6 weeks in the prevention of frequent acute respiratory tract infections in 102 children aged 4-8 years. A total of 43 children treated with isoprinosine and 41 with placebo finished the study. Despite a transient increase in the total number of CD3+, CD4+ and CD8+ T-lymphocytes after 6 weeks of daily isoprinosine treatment, there was no difference in the number and length of duration of acute respiratory infections, number of antibiotic courses and number of days with cough, pharyngitis, rhinitis and increased body temperature (> or = 37.0 degrees C and > or = 38.0 degrees C). There were no changes in markers of T- or B-lymphocyte activation (CD25, HLA-DR, CD45RA/RO, CD23). CONCLUSION: Attempts at immunomodulation using isoprinosine in the dose and for the duration used may increase the total numbers of both CD4 and CD8 T-lymphocytes but is ineffective in prevention of respiratory tract infections in childhood.