Relationship of blood protein levels to outdoor air pollutant concentrations in a semicohort of school-age children living in urban areas differing by quality of air

Variations in some humoral immune responses to polluted air were studied in two semicohorts of children, initial age 10 years, from two urban communities differing from each other by the degree of ambient air pollution. The material for analysis (blood, saliva) was collected every autumn and spring in 3 successive years, giving a total of 6 sets of specimens for each examinee. All blood specimens were examined for the serum level of immunoglobulins (IgG, IgA, IgM), lysozyme (LYS), total serum protein (TP) and the level of the acute reactants alpha 2 macroglobulin (A2M), alpha 1 antitrypsin (A1AT), transferrin (TRF) and ceruloplasmin (CPL). The saliva specimens were examined for the level of lysozyme (sLYS) and secretory IgA (sIgA). The mean protein concentrations for each of the 6 sampling series were correlated with the mean of 24-h emission concentrations measured in the last 3 months preceding the autumn or spring sampling series. In the community area characterized by a low-degree non-industrial pollution of air the correlations of immunoglobulins to SO2 and floating particles (FP) in air were as a rule inversed while the response from TP, LYS and acute reactants was direct. In the community contaminated by industrial pollutants, correlations between proteins and SO2 were markedly weaker, but there was a significant positive correlation between H2S and levels of IgA and A2M in blood and sIgA and sLYS in the saliva. A high degree of positive correlation was also observed between H2S and levels of IgM and LYS. Inverse correlations were only between levels of LYS and FP, SO2 and H2S. Significant correlations were also between contaminant concentrations and FP. The associations found between the contaminant concentrations in air and levels of blood and saliva proteins supports the hypothesis that quality of air may have considerable impacts on defense mechanisms. Seasonal variations in the quality of air may increase the rates of childhood morbidity for acute upper respiratory tract infections.     

Immuno-biochemical findings in groups of individuals occupationally and non-occupationally exposed to emissions containing nickel and cobalt

Immune reactions elicited in the sera of individuals exposed to nickel and cobalt were assessed by changes in the concentration of serum immunoglobulins IgG, IgA and IgM and serum proteins alpha 2 macroglobulin (A2M), transferrin (TRF), alpha 1-antitrypsin (A1AT), ceruloplasmin (CPL) and lysozyme (LYS). Examinations were carried out in workers occupationally exposed to Ni (38 individuals) or Co (35 individuals) and in groups of non-occupationally exposed children living in areas with a different degree of air pollution from a nearby source of Ni and Co emissions (one group was made up of 54 exposed children, the other one of 64 "less exposed" children of the same age). Groups of non-exposed controls were represented by a group of 42 male adults matched by age and by a group of 48 children from a non-polluted area. Significantly increased average values were obtained for IgG, IgA and IgM in group of workers exposed to Ni, for IgA in workers exposed to Co and for A1AT, A2M, CPL and LYS in both groups of occupationally exposed adults (p less than 0.001 - p less than 0.005). Among non-occupationally exposed children the group of the most exposed had significantly elevated average values for A2M and A1AT which were higher than those recorded in groups of "less exposed" and control children (p less than 0.02 and p less than 0.05, respectively). The biomedical importance of these findings is discussed in detail.     

Indicators of humoral immunity in smokers and nonsmokers working underground

A group of 215 underground shale miners (148 smokers and 67 nonsmokers) was examined for levels of immunoglobulins in sera (IgG, IgA, IgM) and saliva (sIgA), transferrin (TRF), alpha 2-macroglobulin (A 2M) and lysozyme levels in sera (LYS) and saliva (sLYS). The control group consisting of 95 healthy men was selected to match the age of the exposed miners. The mine environment was characterized by the presence of ionizing radiation, due to a geological configuration of the rock. The study revealed that the decline of IgG, IgA and sLYS levels in smokers was significantly deeper than in nonsmokers and controls. The levels of sIgA and serum LYS levels were increased both in smokers and nonsmokers. The change in the serum LYS and A 2M levels in miners did not depend on the smoking habits, except of A 2M levels in miners employed for more than 21 years. The differences were, as a rule, less pronounced in younger miners and tended to increase with the increasing length of work underground. The distribution analysis revealed some inter-miner differences in the susceptibility to these changes and the formation of smaller subgroups of miners with more or less subnormal test values, which supports the hypothesis on the heterogeneity of this population group. The tests showed that the statistical analysis of abnormal test values might play an important role in the assessment of the health risk in individuals. A simultaneous detection of subnormal values in some of pairs of tests may provide decisive data allowing a reliable differentiation between the afflicted miners and the healthy control subjects.     

Seasonal changes in the relationship of blood calcium levels to immunoglobulins and some of the blood proteins in schoolchildren

Blood and saliva were collected in the autumn and spring from a group of schoolchildren (39 girls, 35 boys) with a mean age of 11.4 years. Serum immunoglobulin IgG, IgA, IgM and IgE, alpha 1-antitrypsin (A 1-AT), alpha 2 macroglobulin (A 2M), transferrin (TRF), ceruloplasmin (CPL), lysozyme (LYS) and pertussis (PE) antibody levels were determined. Calcium (Ca2+) and total serum protein levels were also determined. Secretory IgA (sIgA) and secretory lysozyme (sLYS) levels were assessed in the saliva. A highly significant drop in Ca2+ levels was found in the spring in boys, while in girls there was only a greater scatter of the values. Mean IgG, IgA and IgM values fell significantly in the spring in both sexes, but IgE levels fell significantly only in boys. PE levels rose significantly in the spring in girls. Among the other proteins, all the values rose in boys, except for TRF, whose levels fell. In girls, LYS and TRF levels rose, but all the other values fell. The coefficients of correlation between Ca2+ and the tested proteins showed a significant relationship only for A 2M and PE in girls and only for the total protein level in boys; in boys, the determination coefficient for sIgA and IgM was over 10%. The results do not testify to the existence of a close relationship between blood Ca2+ levels and Ig and other blood protein levels.     

Immunological profiles in workers of a power plant burning coal rich in arsenic content

A group of 47 male adults working in a thermal power plant burning coal containing 900 to 1,500 g of arsenic per ton dry weight was examined on the blood serum immunoglobulins IgG, IgA and IgM content and levels of acute reactants alpha-1-antitrypsin (A1AT), alpha-2-macroglobulin (A2M), transferrin (TRF), orosomucoid (ORO) ceruloplasmin (CPL), and lysozyme (LYS). Investigations in the control group comprising 27 workers from another power plant in the same district where the coal content of arsenic was more than 10 times lower were analogous. The inter-group differences in means were evaluated by t-test, differences in the association of values by F-test, and the correlations with age and the length of exposure were assessed using the regression analysis method. The differences in mean IgG, IgA, IgM, LYS and A2M levels between the exposed and control groups of workers were insignificant or of borderline significance only. In contrast, differences in TRF, ORO and particularly CPL levels were statistically highly significant, in all instances P less than 0.001. In the control group, persons with abnormal values in at least two immunobiochemical tests used accounted for 3.7%, in the group of the exposed for 51% (P less than 0.002). All these findings, especially the rise in CPL concentration levels in the exposed group are discussed on the background of the rise in cancer mortality rates found previously in this group of power plant workers.     

A sensitive solid phase radioimmunoassay for secretory IgA: its application for the determination of urinary levels of secretory IgA in children and adults

A sensitive solid phase radioimmunoassay method was established for the specific quantitative determination of secretory IgA (sIgA) by taking advantage of the dual antigenicities of sIgA, one specific for alpha-chain and the other for secretory component (SC). The sIgA and IgA in the sample were first bound by anti-IgA antibodies coated on the polystyrene tube, then the amount of bound sIgA was quantified by the use of 125I-labeled anti-SC antibodies. This method is quite sensitive and allows us to distinguish sIgA from IgA and free SC which usually coexist in exocrine secretions. Linear relationship was observed between the bound radioactivity of radioiodinated anti-SC and the amount of sIgA in the range of 5 to 60 ng of sIgA. With this method, the urinary sIgA levels in normal children and adults were measured. Urinary sIgA was detected in half of infants within 7 days after birth, while it could be demonstrated in almost all of the infants by 14th day. Then, it gradually increased and reached about a half of the adult level by the age of puberty. This method will be useful for the sensitive and specific measurement of sIgA in various exocrine secretions.     

Study of the antibody response against Mycobacterium tuberculosis antigens in Warao Amerindian children in Venezuela

This study was aimed at investigating alternate methods for serodiagnosis of tuberculosis (TB), which are needed because bacteriologic diagnosis of childhood TB is difficult. A selection of 80 serum and saliva samples were tested from Warao indigenous children under 15 years of age; 34 high TB suspects (28 positive and 6 negative for the tuberculin skin test, TST) and 46 healthy contact children (32 positive and 14 negative for the TST). Several enzyme-linked immunosorbent assay (ELISA) serological tests were developed to test for Mycobacterium tuberculosis-specific antibodies, including serum IgA, IgG, IgE, and secretory IgA (sIgA) in saliva against 3 specific antigens (PPD, HSP60, 38 kDa). Of these, 2 antigens, PPD and 38 kDa, showed significantly higher reactivity. The sensitivity and specificity of these tests for diagnosis remained limited, between 26.5% and 38.2%, and 77.4% and 97%, respectively. Of all the samples studied and combinations realized between all isotypes and antigens combined with 3 isotypes (anti-PPD IgG, IgE, and anti-38kDa sIgA) managed to detect the largest number of patients, showing an improved sensitivity level of 64.7%, although specificity levels dropped to 81.8%. These results were compared with the Omega diagnostics commercial kit results. The commercial kits showed significantly lower reactivity (sensitivity of 20% and 13.33% to Myco G and Complex Plus, respectively) and a specificity of 100%. This study shows that in indigenous populations of Venezuela, where invasive procedures cannot be used to select samples but evaluation with a chest X-ray for radiological studies is available, the combination of 3 specific isotypes may be a useful tool to increase diagnostic accuracy with pulmonary TB in this population, when used together with clinical and epidemiological criteria.     

Mesenteric lymph nodes: cells with surface and sytoplasmic immunoglobulins

The distribution and morphology of cells with surface and cytoplasmic immunoglobulins were investigated in mesenteric lymph nodes (MLN) from rats, using both frozen and (fixed) paraffin sections, with a two-step immunoperoxidase technique. Anti-IgA, -IgE, -IgG and -IgM sera were used. Surface Ig-cells (sIg) of all four isotypes studied were found in MLN, mainly localized in the interfollicular area and within the follicle corona. The percentages of sIgA, IgE, IgG and IgM were about 15, 5, 45 and 35%, respectively. In addition, sIgM- and sIgG-cells were found around high endothelial venules. The percentages of cells containing IgA, IgG or IgM (cIg-cells) were about 60, 25 and 15%, respectively; only a few cIgE-cells were found. cIg-cells were not only present in the interfollicular areas and the medulla but also within the germinal centers of the follicles. These results are discussed with regard to the interaction between Peyer's patches (PP) and MLN.     

Plant expression of chicken secretory antibodies derived from combinatorial libraries

Delivery of secretory IgA antibodies (sIgA) to mucosal surfaces is a promising strategy to passively prevent infectious diseases. Plants have been proposed as biofactories for such complex immunoglobulin molecules. Recently, the molecular characterization of all four monomers of chicken sIgA (IgA immunoglobulin heavy and light chains, J-chain and secretory component) has been completed, allowing recombinant, up scaled production of chicken sIgA and extension of passive immune strategies to poultry. To test the suitability of the plant cell factory for bulk production of chicken sIgA, we studied the expression of chicken IgA, dIgA and sIgA in planta. To that end, new cassettes were designed that allowed the grafting of immunoglobulin variable regions derived from combinatorial libraries into full-size chicken IgA frames ready for plant expression. Using this system, 10 individual phage display clones, which had previously been selected against Eimeria acervulina antigens, were transferred "from phage to plant". Plant-made chicken antibodies showed strong differences in expression levels, which seemed governed mainly by the stability of their respective light chains. Finally, with the co-expression of chicken IgA heavy and light chains, J-chain and secretory component in N. benthamiana leaves we showed that plant cells are suitable biofactories for the production of assembled chicken sIgA complexes.     

Factors of humoral resistance in workers exposed to vinyl chloride with a view to smoking habits

Serum samples were assayed in 110 workers (59 smokers and 51 non-smokers) at PVC manufacturing factory, the results being compared with those obtained in a group of age-matched healthy controls. Non-smokers had significantly higher levels of immunoglobulins (IgG, IgA, IgM), while in smokers there was an increase in IgM only. Lysozyme levels (LYS) were elevated in all exposed subjects, but there was a highly significant decrease in the total protein (TP) content. Alfa-2-macroglobulin (A2M) and orosomucoid (ORO) were also elevated in exposed workers. A significant increase was found in ceruloplasmin (CPL), with smokers having higher levels than non-smokers. No difference was detected between the examined groups in transferrin (TRF) and alfa-1-antitrypsin (A1AT). Exposure duration did not correlate with any of the assayed parameters. The positive correlation of IgA levels and inverse correlation of CPL with age in the control group could not be confirmed in the exposed subjects where a significant inverse correlation between age and AlAT levels, which was found to be positive in exposed non-smokers, proved to be inverse in the subgroup of smokers. The authors discuss possible explanations of the reported findings.     

Air pollution effects on structure,proteins and flavonoids in pollen grains of Thuja orientalis L. (Cupressaceae)

Increase in the levels of air pollution due to the increase in industrial and agricultural technology has prompted investigation of mechanisms that contribute to air pollution tolerance in plants. Pollen grains of Thuja orientalis were collected from controlled (less polluted) and polluted areas (mainly SO₂, NO₂, CO, HC and APM). Thuja pollen is considered inaperturate and granulate. The exine is shed during rehydration, leaving the male gametophyte naked. The pollen grains collected from polluted areas are smaller and more fragile compared to control ones. The exine splitting is faster and higher in polluted pollen grains. SDS–PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) pattern do not show significant differences in polluted pollen than those in the control group. HPLC (high pressure liquid chromatography) analysis demonstrates that air pollution induces flavonoids accumulation to significantly higher levels in polluted pollen than in controlled ones. These observations suggest that plants try to respond suitably by adjusting their metabolism so that minimum damage is done due to air pollutants. Their protective responses may include an increase in antioxidant enzymes and metabolites and induction of protection-related secondary metabolite genes especially flavonoids.     

合生元益生菌冲剂对提高儿童消化道黏膜免疫的作用

目的探讨健康儿童口服法国合生元益生菌冲剂后的唾液中sIgA(分泌型IgA)含量的变化,从而判定其对提高消化道黏膜免疫的作用。方法选择正常健康儿童28例,根据年龄分为4组,其中从每组随机抽出1或者2例作为对照组,对照组共为7例,而试验组则为21例。收集口服合生元益生菌冲剂前后正常健康儿童唾液标本经放射免疫分析法检测sIgA的含量。结果对照组儿童之间唾液sIgA分泌量差异较大,个体的唾液sIgA水平会随着身体健康状况和饮食条件等呈不稳定情况,而试验组儿童的唾液sIgA分泌量在连续口服益生菌冲剂6天后有了一个新的提高,并在继续服用益生菌冲剂的7天内获得比较高而稳定的水平。结论口服法国合生元益生菌冲剂可有效诱导消化道黏膜免疫应答,提高并维持儿童的免疫力。     

丹参对毛细支气管炎患儿体液免疫调节作用的观察

目的:探讨丹参佐治毛细支气管炎的疗效及其可能的机理。方法:73例毛细支气管患儿随机分为常规组和丹参组,均用常规方法治疗,但后者加用复方丹参静脉滴注。在治疗前、后用全自动散射比浊法测定并比较两组患儿血清IgE、IgG、IgA、补体C3和C4水平。结果:丹参组和常规组总有效率依次为94.44%和78.38%,差异具有统计学意义(P<0.05)。治疗后,丹参组患者血清IgE、C3和C4水平下降,IgG、IgA水平上升,与治疗前相比均有一定程度差异(P<0.01-0.05)。治疗后,常规组患者仅IgE水平下降(P<0.01),但其余指标与治疗前差异无统计学意义(P>0.05)。结论:丹参佐治毛细支气管炎,临床效果较好,且可调节机体的体液免疫应答,增强机体的抗感染能力。     

Novel probiotic Enterococcus faecium IS-27526 supplementation increased total salivary sIgA level and bodyweight of pre-school children: A pilot study

Enterococcus faecium IS-27526 is a novel probiotic isolated from dadih, an Indonesian traditional fermented buffalo milk. A 90 days randomized double-blind placebo-controlled study ofpre-post trial was conducted in pre-school children with two groups, placebo and probiotic group. Ultra High Temperature low fat milk was used as a carrier in each group. The aims of this study were to investigate the effect of E. faecium IS-27526 in milk on humoral immune response and on bodyweight of pre-school children. Total serum IgA and total salivary sIgA were measured by sandwich ELISA. The bodyweight of young children was measured. The results showed that total serum IgA did not significantly increase in the probiotic group compared with the placebo group. Total salivary sIgA level and the bodyweight significantly increased (p < 0.05) in probiotic groups compared to the placebo. Changes of total salivary sIgA level were significantly higher in underweight children supplemented with probiotic. Weight gain was observed significantly in children with normal bodyweight supplemented with probiotic. Neither mortality nor weight loss was recorded throughout the study. Taken together, novel probiotic E. faecium IS-27526 has significant positive effects on humoral immune response, salivary sIgA, in underweight pre-school children, and on weight gain of pre-school children.     

Immunopathological responses in women with chronic inflammatory diseases of the uterus and appendages and their therapeutic correction

A total of 145 women with chronic inflammatory diseases of uterus and appendages (IDUA) were examined. Bacterioscopy of smears and culture of vaginal, cervical canal and uterine cavity contents were performed in all patients. Direct immunofluorescence and PCR were used for detection of chlamydiae in scrapes and smears and in biopsy specimens taken from endometrium. The number of T lymphocytes and their subpopulations, B lymphocytes as well as the level of plasmatic cells synthesizing IgA, IgM and IgG and secretory IgA (sIgA), immune complexes and C3 component of the complement were determined in biopsy specimens of the endometrium. Pathomorphological examination revealed structural changes characteristic of the immunocomplex disease (secondary deficiency of sIgA, pronounced suppression of IgA production by plasmocytes along with an essential increase in the number of IgG synthesizing cells and immune complexes fixed to the venule endothelium) were detected. Lectin-histochemical study revealed considerable changes in secretory activity of endometrial epitheliocytes manifested by severe suppression of bactericidal activity of the uterine mucus. The detected disturbances of local protective reactions in the endometrium of women IDUA gave grounds for including immunomodulators into the traditional treatment scheme. The latter made it possible to increase essentially the therapeutic effect.     

Relationship between canine mucosal and serum immunoglobulin A (IgA) concentrations: serum IgA does not assess duodenal secretory IgA

A double-sandwich enzyme immunoassay method was developed for determination of serum immunoglobulin A (S-IgA) and mucosal secretory immunoglobulin A (sIgA) in duodenal brush samples obtained via endoscopy and the relationship between enteric mucosal sIgA, salivary sIgA and S-IgA in dogs was examined. Twenty healthy dogs underwent routine endoscopy. A brush sample from the duodenal mucosa was obtained and washed in PBS, with a serum sample being taken concurrently. A saliva sample was collected from twelve of these dogs. S-IgA and sIgA with total protein concentrations in the duodenal washings and saliva samples were determined. A significant negative correlation (r = -0.64, P = 0.0059) was found between duodenal sIgA/protein ratios and S-IgA concentrations. Saliva sIgA/protein ratios did not correlate with sIgA/protein ratios of duodenal samples. The method described here allows for direct assessment of duodenal IgA; therefore indirect measures based on serum IgA or salivary IgA can be avoided. In addition, these indirect measures appear to be poor indicators of duodenal sIgA competence in dogs.     

Observation of antibody activity against antibiotics in patients with atopic disease

Antibodies to antibiotics were studied in patients hypersensitive to these preparations and in patients with other types of allergic disease. In addition to examining these antibodies, a whole series of other immunological tests were carried out (IgG, IgA, IgM, IgE, C'3 and OPI). Antibodies to antibiotics were found in a high percentage of subjects, not only in those hypersensitive to antibiotics but also in patients with asthma bronchiale and rhinitis pollinosa. The considerable differences in IgG and IgE observed in patients with high titres of haemagglutinating antibodies to antibiotics point out the blocking character of these antibodies.     

Using the Pollen Viability and Morphology for Fluoride Pollution Biomonitoring

The methods using plants for biomonitoring of air and soil quality are simple, cheap, and fast and can supplement the classical physicochemical methods. In this study, biological pollen characterization of some collected legume species from an aluminum smelter area in Iran (IRALCO) was carried out to determine the actual value of pollen as a bioindicator of the effects of soil and atmospheric pollution. Young buds and flowers of six legumes (Cercis siliquastrum L., Medicago sativa L., Robinia pseudoacacia L., Melilotus officinalis (L.) lam, Trifolium repens L., and Sophora alopecuroides L.) in polluted and control plants were removed and compared. Studies of light and electron microscopic preparation showed some abnormalities during pollen development in affect of fluoride pollution. The viability of pollen grains estimated by staining with acetocarmine shows sharp differences in smearing advanced pollen grains from abnormal ones. Except M. officinalis, the pollen grains of C. siliquastrum, M. sativa, R. pseudoacacia, T. repens, and S. alopecuroides in polluted areas showed light, partial, or no staining with acetocarmine, whereas almost all of the control ones clearly stained. Observation of the pollen grains by light microscopy and scanning electron microscopy showed the significant effect of fluoride on shapes and sizes of pollen grains. The stimulation and inhibition of these pollen characteristics depend on the pollen species as well as on the pollutant and its concentration. Therefore, pollen grains provide essential information on biological impact of pollutants and they are good candidates for biomonitoring the atmospheric and edaphic pollutions.     

Specific antibodies bound to secretory IgA in the sera of patients with intestinal infections]

Specific IgA and sIgA antibodies were studied in the sera of patients suffering from various intestinal diseases (dysentery, salmonellosis, typhoid fever, chronic typhoid carrier state) and in the sera of healthy persons immunized by parenteral route with typhoid alcohol vaccine. The nature of antibodies was identified in Coombs' test, using monospecific antisera to alpha-chain and to the secretory component. IgA and sIgA antibodies were revealed most frequently in the sera of dysentery patients and of chronic typhoid carriers. No sIgA antibodies were found in the sera of subcutaneously immunized persons. The presence of specific sIgA antibodies in the serum reflects the participation of local immune mechanisms in the formation of systemic immunity in the intestinal infections.     

Resistance of normal serum IgA and secretory IgA to bacterial IgA proteases: evidence for the presence of enzyme-neutralizing antibodies in both serum and secretory IgA, and also in serum IgG

Normal serum IgA and secretory IgA (sIgA) of subclass IgA1 were isolated from pooled human serum and milk, respectively. They were tested for their susceptibility to bacterial IgA proteases from Haemophilus influenzae, Streptococcus pneumoniae, Neisseria gonorrhoeae, and Neisseria meningitidis that cleave IgA of only the IgA1 subclass. They were also tested for susceptibility to a novel IgA-protease from Clostridium ramosum that cleaves IgA of the IgA1 as well as the IgA2 subclass of the A2m(1) allotype. Both normal serum IgA1 and sIgA1 exhibited resistance to most IgA proteases. The one exception was the IgA protease from C. ramosum which readily cleaved both the serum IgA1 and sIgA1 into Fab and Fc fragments. Secretory component (SC) had nothing to do with the resistance of these IgAs. The resistance of these IgAs to most of the IgA proteases was found to be due to their enzyme-neutralizing antibody activity, since the Fab but not the Fc fragment of sIgA1 showed enzyme-inhibitory activity against these IgA proteases. Similar enzyme-neutralizing antibody activity was found in the pepsin-digested normal serum IgG-(Fab')2 fragment. These results indicate that the induction of the enzyme-neutralizing antibodies against the bacterial IgA proteases took place not only in mucosal sIgA but also in serum IgA and IgG. No enzyme-neutralizing antibody activity against the novel IgA-protease of C. ramosum was detected in any immunoglobulin preparations used in the present study or in the serum of a patient who carries the IgA protease-producing strain of C. ramosum in his feces.