Induction of an immune response through the idiotypic network with monoclonal anti-idiotype antibodies in the carcinoembryonic antigen system.

Anti-idiotype antibodies can mimic the conformational epitopes of the original antigen and act as antigen substitutes for vaccination and/or serological purposes. To investigate this possibility concerning the tumor marker carcinoembryonic antigen (CEA), BALB/c mice were immunized with the previously described anti-CEA monoclonal antibody (MAb) 5.D11 (AB1). After cell fusion, 15 stable cloned cell lines secreting anti-Ids (AB2) were obtained. Selected MAbs gave various degrees of inhibition (up to 100%) of the binding of 125I-labeled CEA to MAb 5.D11. Absence of reactivity of anti-Id MAbs with normal mouse IgG was first demonstrated by the fact that anti-Id MAbs were not absorbed by passage through a mouse IgG column, and second because they bound specifically to non-reduced MAb 5.D11 on Western blots. Anti-5.D11 MAbs did not inhibit binding to CEA of MAb 10.B9, another anti-CEA antibody obtained in the same fusion as 5.D11, or that of several anti-CEA MAbs reported in an international workshop, with the exception of two other anti-CEA MAbs, both directed against the GOLD IV epitope. When applied to an Id-anti-Id competitive radioimmunoassay, a sensitivity of 2 ng/ml of CEA was obtained, which is sufficient for monitoring circulating CEA in carcinoma patients. To verify that the anti-Id MAbs have the potential to be used as CEA vaccines, syngeneic BALB/c mice were immunized with these MAbs (AB2). Sera from immunized mice were demonstrated to contain AB3 antibodies recognizing the original antigen, CEA, both in enzyme immunoassay and by immunoperoxidase staining of human colon carcinoma. These results open the perspective of vaccination against colorectal carcinoma through the use of anti-idiotype antibodies as antigen substitutes.

Expression on human thymocytes of the idiotypic structures (Ti) from two leukemia T cell lines Jurkat and HPB-ALL.

The expression on a significant number of thymocytes of idiotypic structures (Ti) restricted to HPB-ALL or Jurkat cells is demonstrated. As many as 2-4% of thymocytes were stained with anti-Ti HPB-ALL or anti-Ti Jurkat monoclonal antibodies, when analyzed by flow microfluorometry. Immunohistochemical localization studies performed on frozen thymus specimens of either fetal or pediatric origin indicated a scattered distribution of Ti-positive cells in both the cortex and the medulla. From lysates of 125I-labeled pediatric thymocytes, anti-Ti HPB-ALL and anti-Ti Jurkat monoclonal antibodies precipitated disulfide-linked heterodimers comparable to those precipitated from 125I-labeled HPB-ALL or Jurkat cells as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis.

Monoclonal antibodies against idiotypic determinant(s) of the T cell receptor from HPB-ALL cells induce IL2 production in Jurkat cells without apparent evidence of binding.

Two monoclonal antibodies (mAb) directed against idiotypic determinants of the T cell receptor (anti-Ti) from HPB-ALL cells induce interleukin 2 (IL2) production in Jurkat T cells without evidence of binding to these cells as judged by fluorescence-activated cell sorter (FACS) analysis, indirect antibody-binding radioimmunoassay and direct binding studies with 125I-labeled mAb. The IL2 response induced by these mAb observed both in the presence and absence of phorbol myristate acetate was in the range of that obtained when Jurkat cells were stimulated with phytohemagglutinin or anti-T3 mAb (Leu 4). The idiotypic specificity of the two anti-HPB-ALL Ti mAb was demonstrated by several criteria. Both mAb bound specifically to HPB-ALL cells as determined by radioimmunoassay or FACS analysis but not with 8 other T cell lines. The anti-HPB-ALL Ti mAb precipitated a disulfide-linked heterodimer of 85 kDa only from 125I-labeled HPB-ALL cells and not from other cell lines tested. Incubation of HPB-ALL cells with anti-T3 abrogated the expression of T3 and induced co-modulation of the idiotypic structures detected by the two anti-HPB-ALL Ti mAb. Conversely, incubation of HPB-ALL cells with either one of the anti-Ti mAb abrogated the expression of T3 and of the idiotypic structures. Our results suggest that mAb with an apparent unique specificity for the receptor of the immunizing T cell line HPB-ALL can activate Jurkat cells by a very weak cross-reaction with these cells, which is not detectable by conventional binding tests.

Dimeric IVIG contains natural anti-Siglec-9 autoantibodies and their anti-idiotypes

Intravenous immunoglobulin (IVIG) preparations are increasingly used for the treatment of autoimmune and chronic inflammatory diseases. Naturally occurring autoantibodies against Siglec-9 and Fas are thought to contribute to the anti-inflammatory effects of IVIG via cell death regulation of leukocytes and tissue cells. Dimeric IVIG fractions are suspected to contain idiotypic (Id)-anti-idiotypic complexes of antibodies, which might also include anti-Siglec-9 and anti-Fas autoantibodies.

Metal Ions Bound To The Human Milk Immunoglobulin A: Metalloproteomic Approach.

The presence of calcium, iron, and zinc bound to human milk secretory IgA (sIgA) was investigated. The sIgA components were first separated by two-dimensional polyacrylamide gel electrophoresis and then identified by electrospray ionization-tandem mass spectrometry (ESI MS MS). The metal ions were detected by flame atomic absorption spectrometry after acid mineralization of the spots. The results showed eight protein spots corresponding to the IgA heavy chain constant region. Another spot was identified as the transmembrane secretory component. Calcium was bound to both the transmembrane component and the heavy chain constant region, while zinc was bound to the heavy chain constant region and iron was not bound with the identified proteins. The association of a metal ion with a protein is important for a number of reasons, and therefore, the findings of the present study may lead to a better understanding of the mechanisms of action and of additional roles that sIgA and its components play in human milk.

Effect of a kickboxing match on salivary cortisol and immunoglobulin A

The hypothesis that salivary cortisol would increase and salivary immunoglobulin A (IgA) decrease after a kickboxing match was tested among 20 male athletes. Saliva samples collected before and after the match were analyzed. Salivary cortisol and salivary IgA concentrations (absolute concentration, salivary IgAabs) and the secretion rate of IgA (salivary IgArate) were measured by enzyme-linked immunosorbent assay. A Wilcoxon test for paired samples showed significant increases in salivary cortisol from pre- to postmatch. No significant changes were observed in salivary IgAabs or secretory IgArate and saliva flow rate. This study indicates that a kickboxing match might increase salivary concentration and thereafter it could be considered a significant source of exercise-related stress. On the other hand, the effect of a kickboxing match on mucosal immunity seems not to be relevant.

Salivary immunoglobulin a response to a match in top-level Brazilian soccer players

Moreira, A, Arsati, F, Cury, PR, Franciscon, C, Oliveira, PR, and Araujo, VC. Salivary immunoglobulin a response to a match in top-level brazilian soccer players. J Strength Cond Res 23(7): 1968-1973, 2009-It has been suggested that several parameters of mucosal immunity, including salivary immunoglobulin A (s-IgA), are affected by heavy exercise either in field sports or in the laboratory environment. Few observations have been made during a true sporting environment, particularly in professional soccer. We tested the hypothesis that salivary IgA levels will be decreased after a 70-minute regulation in a top-level professional soccer friendly match. Saliva samples from 24 male professional soccer players collected before and after the match were analyzed. Salivary immunoglobulin A concentration was measured by enzyme-linked immunosorbent assay and expressed as the absolute concentration (s-IgAabs), s-IgA relative to total protein concentration (IgA-Pro), and the secretion rate of IgA (s-IgArate). Rate of perceived exertion (RPE) was used to monitor the exercise intensity. The paired t-test showed no significant changes in s-IgAabs and s-IgArate (p > 0.05) from PRE to POST match. However, a significant (p < 0.05) increase in total protein concentration (1.46 +/- 0.4 to 2.00 +/- 07) and a decrease in IgA-Pro were observed. The best and most significant correlation was obtained with the RPE and changes in IgA-Pro (rs = -0.43) and could indicate that this expression may be an interesting marker of intensity in a soccer match. However, further investigation regarding exercise intensity, protein concentration, and immune suppression, particularly in team sports, is warranted. From a practical application, the variability of the responses among the players leads us to suggest that there is a need to individually analyze the results with team sports. Some athletes showed a decrease in s-IgA expressions, suggesting the need for taking protective actions to minimize contact with cold viruses or even reducing the training load.

Salivary immunoglobulin a responses in professional top-level futsal players

Moreira, A, Arsati, F, de Oliveira Lima-Arsati, YB, de Freitas, CG, and de Araujo, VC. Salivary immunoglobulin a responses in professional top-level futsal players. J Strength Cond Res 25(7): 1932-1936, 2011-The purpose of this study was to investigate the responses of salivary immunoglobulin A (SIgA) in 10 professional top-level Brazilian futsal players after 2 highly competitive games separated by 7 days. Unstimulated saliva was collected over a 5-minute period at PRE- and POST-match. The SIgA was measured by an enzyme-linked immunosorbent assay and expressed as the absolute concentration (SIgAabs) and secretion rate of IgA (SIgArate). Rate of perceived exertion and heart rate were used to monitor the exercise intensity. A 2-way analysis of variance with repeated measures showed nonsignificant differences between matches to SIgAabs, SIgArate, and saliva flow rate (p > 0.05). However, significant time differences were observed for all these parameters. In summary, we showed that a competitive training match induced a decrease in SIgA levels in top-level futsal players, which suggests an increment of the vulnerability to infections meditated by the training stimulus. This decrease suggests that the athletes were at an increased risk of developing an upper respiratory tract infection, and therefore, it could be necessary to take protective actions to minimize contact with cold viruses or even reduce the training load for athletes.

Comparison of carbohydrate-deficient transferrin, immunoglobulin A antibodies reactive with acetaldehyde-modified protein and acetaldehyde-modified albumin with conventional markers of alcohol consumption

Carbohydrate-deficient transferrin (CDT) has emerged as the best new marker for alcohol abuse. Recently plasma immunoglobulin A (IgA) reactivity with acetaldehyde (AcH)-modified proteins, or the modified proteins per se, have been proposed as a markers for high levels of alcohol consumption. In this study, we have compared CDT, IgA reactivity with AcH adducts (IgA ASR), and AcH-modified albumin with conventional markers of high alcohol intake in groups with well-defined drinking histories, The plasma activity of ALT, AST, and gamma-glutamyltransferase increased steadily with increasing alcohol consumption, CDT and AcH-modified albumin showed a similar pattern, whereas IgA ASR appeared only to be elevated after a threshold level of consumption had been reached, Neither CDT IgA ASR or AcH-modified albumin correlated strongly with any of the conventional markers or each other. This study shows that CDT, IgA ASR, AcH-modified albumin, and the conventional markers are not related, but suggests that the concurrent use of CDT and IgA ASR may lead to better identification of high alcohol intake.

Adult-onset Still`s Disease with Pulmonary and Cardiac Involvement and Response to Intravenous Immunoglobulin

Cardiopulmonary manifestations of adult-onset Still`s disease (AOSD) include pericarditis, pleural effusion, transient pulmonary infiltrates, pulmonary interstitial disease and myocarditis. Serositis are common but pneumonitis and myocarditis are not and bring elevated risk of mortality. They may manifest on disease onset or flares. Previously reported cases were treated with high-dose glucocorticoids and immunosupressants and, when refractory, intravenous immunoglobulin (IVIG). We report an AOSD patient whose flare presented with severe pleupneumonitis and myopericarditis and, following nonresponse to a methylprednisolone pulse, high dose of prednisone and cyclosporine A, recovered after a 2-day 1g/kg/day IVIG infusion.

Leptospirosis pulmonary haemorrhage syndrome is associated with linear deposition of immunoglobulin and complement on the alveolar surface

Leptospirosis is a zoonotic infection associated with severe diseases such as leptospirosis pulmonary haemorrhage syndrome (LPHS). The cause of pulmonary haemorrhage is unclear. Understanding which mechanisms and processes are involved in LPHS will be important in treatment regimens under development for this life-threatening syndrome. In the present study, we evaluated 30 lung specimens from LPHS patients and seven controls using histology and immunohistochemistry (detection of IgM, IgG, IgA and C3) in order to describe the pathological features associated with this syndrome. Immunoglobulin deposits were detected on the alveolar surface in 18/30 LPHS patients. Three staining patterns were observed for the immunoglobulins and C3 in the lung tissues of LPHS patients: AS, delicate linear staining adjacent to the alveolar surface, which was indicative of a membrane covering the luminal surface of type I and II pneumocyte cells; S, heterogeneous staining which was sporadically distributed along the alveolar septum; and IA, weak, focal intra-alveolar granular staining. Human LPHS is associated with individual and unique histological patterns that differ from those of other causes of pulmonary haemorrhage. In the present study, it was found that the linear deposition of immunoglobulins (IgA, IgG and IgM) and complement on the alveolar surface may play a role in the pathogenesis of pulmonary haemorrhage in human leptospirosis.

Increased Immunoglobulin G Anti-Paracoccidioides brasiliensis Serum Antibody Avidity as a Predictor of Favorable Posttherapeutic Evolution in Paracoccidioidomycosis

Paracoccidioidomycosis is endemic in Latin America, and ca. 80% of all cases occur in Brazil. Little is known about antibody avidity or the evolution of such avidity in the posttherapeutic period for the different clinical presentations of the disease. In the present study, we evaluated 53 patients with paracoccidioidomycosis and calculated the avidity index. Medium-and high-avidity antibodies were found in 79.5% of patients with chronic presentation (n = 39). Among patients with the acute form (n = 14), 57.1% of the antibodies presented low avidity. In the posttherapeutic period, there was a significant increase in antibody avidity in patients presenting with the chronic multifocal form. In our preliminary study, which needs to be confirmed using a larger number of samples, the optimized method for studying antibody avidity detected differences among the clinical presentations of the mycosis and indicated the value of the avidity index as a marker of posttherapeutic evolution of patients with a multifocal chronic form of the disease.

Detection of Clonal Immunoglobulin and T-Cell Receptor Gene Rearrangements in Childhood Acute Lymphoblastic Leukemia Using a Low-Cost PCR Strategy

Background Imunoglobulin (Ig) and T cell receptor (TCR) gene rearrangements function as specific markers for minimal residual disease (MRD) which is one of the best predictors of outcome in childhood acute lymphoblastic leukemia (ALL) We recently reported on the prognostic value of MRD during the induction of remission through a simplified PCR method Here we report on gene rearrangement frequencies and offer guidelines for the application of the technique Procedure Two hundred thirty three children had DNA extracted from bone marrow Ig and TCR gene rearrangements were amplified using consensus primers and conventional PCR PCR products were submitted to homo/heteroduplex analysis A computer program was designed to define combinations of targets for clonal detection using a minimum set of primers and reactions Results At least one clonal marker could be detected in 98% of the patients and two markers in approximately 80% The most commonly rear ringed genes in precursor B cell ALL were IgH (75%) TCRD (59%) IgK (55%), and TCRG (54%) The most commonly rearranged genes for TALL were TCRG (100%) and TCRD (24%) The sensitivity of primers was limited to the detection of 1 leukemic cell among 100 normal cells Conclusions We propose that eight PCR reactions per ALL subtype would allow for the detection of two markers in most cases In addition these reactions ire suitable for MRD monitoring especially when aiming the selection of patients with high MRD levels (>= 10(-2)) at the end of induction therapy Such an approach would be very useful in centers with limited financial resources Pediatr Blood Cancer 2010 55 1278-1286 (C) 2010 Wiley Liss Inc

The effect of moderate aerobic exercise and relaxation on secretory immunoglobulin A

A deficiency in secretory immunoglobulin A (sIgA) is associated with recurrent upper respiratory tract infections both in the general community and in elite athletes. The aim of this paper was to investigate the effect of aerobic exercise and relaxation on various indices of sIgA in 12 male and 8 female adults who varied in levels of recreational activity. Salivary samples were obtained before, immediately after and 30 minutes after an incremental cycle ergometer test to fatigue. after 30 minutes of cycling at 30% or 60 % of maximum heart rate, and after 30 minutes of relaxation with guided imagery. Each session was run on a separate day. When expressed in relation to changes in salivary flow rate, sIgA did not change after exercise. However, both the absolute concentration and secretion rate of sIgA increased during relaxation (167 +/- 179 mug ml(-1), p < 0.001: and 37 +/- 71 g(.)min(-1), p < 0.05 respectively). Nonspecific protein increased more than sIgA during incremental exercise to fatigue (decrease in the sIgA/protein ratio 92 +/- 181 g(.)mg protein(-1), p(0.05), but sIgA relative to protein did not change during relaxation. Our findings suggest that sIgA secretion rate is a more appropriate measure of sIgA than sIgA relative to protein, both for exercise and relaxation. These data suggest the possibility of using relaxation to counteract the negative effects of intense exercise on sIgA levels.

The effects of stress management on symptoms of upper respiratory tract infection, secretory immunoglobulin A, and mood in young adults

Objective: To investigate the efficacy of a stress management programme on symptoms of colds and influenza in 27 university students before and after the examination period. Method: The incidence of symptoms, levels of negative affect, and secretion rate of secretory immunoglobulin A (sIgA) were recorded for 5 weeks before treatment, for the 4 weeks of treatment, and for 8 weeks after treatment in treated subjects and in 25 others who did not participate in stress management. Results: Symptoms decreased in treated subjects but not in controls during and after the examination period. Although sIgA secretion rate increased significantly after individual sessions of relaxation, resting secretion rate of sIgA did not increase over the course of the study. Negative affect decreased after examinations in both groups, but was not affected by treatment. Conclusion: Stress management reduced days of illness independently of negative affect and sIgA secretion rate. Although the component of treatment responsible for this effect has yet to be identified, psychological interventions may have a role in reducing symptoms of upper respiratory tract infection. (C) 2001 Elsevier Science Inc. All rights reserved.