Local Immunoglobulin Production Research Articles

Does de novo immunoglobulin synthesis occur on the epithelial surface of the human lower respiratory tract?

Although synthesis of immunoglobulins by cells in the human lower respiratory tract has never been directly demonstrated, local production of immunoglobulins by cells within lung parenchyma is thought to play an important role in protecting the normal human lung against a variety of pathogenic organisms, and may contribute to the development of some interstitial lung diseases. To determine whether or not the epithelial surface of the human lower respiratory tract is a site of immunoglobulin synthesis, immunoglobulin production by cells recovered by bronchoalveolar lavage and lung parenchymal tissue from normal subjects and patients with idiopathic pulmonary fibrosis (IPF) or sarcoidosis was evaluated using a reverse hemolytic plaque assay and by measuring incorporation of [35S]methionine into immunoglobulins. Although the reverse hemolytic plaque assay demonstrated immunoglobin-releasing cells to be present on the surface of the lower respiratory tract of normal subjects, and present in increased numbers in patients with IPF or sarcoidosis, these plaque-forming cells were not actively synthesizing immunoglobulins, since preincubation of lavage cells in order to remove immunoglobulin bound to the surface of the cells reduced the number of plaque-forming cells, and inhibition of protein synthesis by cycloheximide did not further reduce the number of plaque-forming cells. Furthermore, direct evaluation of immunoglobulin synthesis using radiolabeling, immunoprecipitation and SDS-polyacrylamide gel electrophoresis failed to detect de novo IgG production by cells recovered from the alveolar surface of 15 of 17 subjects, IgM by 17 of 17, and IgA by 16 of 17.(ABSTRACT TRUNCATED AT 250 WORDS)

IgG4 in human colostrum and human milk: continued local production or selective transport from serum.

Colostrum, mature milk, and paired plasma samples were obtained from 10 postpartum women who had not been previously studied. The geometric mean concentration of IgG4 in colostrum (3.3 micrograms/ml) was similar to the mean concentration in mature milk (3.0 micrograms/ml). The arithmetic mean for the percent of IgG = IgG4 was 10.3 +/- 3.3% for colostrum, 10.3 +/- 3.1% for mature milk, 2.6 +/- 0.3% for early plasma, and 1.7 +/- 0.3% for later plasma. Local mammary production of immunoglobulin was determined by subtracting the estimated serum contribution from the measured concentration in colostrum or milk. Evidence for local mammary production of IgG4 was found in 5 of 10 colostrum samples and 8 of 10 mature milk samples. These observations indicate that the previously observed selective enrichment of IgG4 in colostrum is also true for mature milk. These are the first studies suggesting continued local production of any immunoglobulin other than IgA in mature human breast milk.

Comparative study of natural autoantibodies in the serum and cerebrospinal fluid of normal individuals and patients with multiple sclerosis and other neurological diseases

Using a panel of antigens (actin, myosin, tubulin, albumin, transferrin, peroxidase, thyroglobulin, DNA, prolactin, TNP and myelin basic protein (MBP)), we have tested the antibody activity of serum and cerebrospinal fluid (CSF) from healthy individuals, patients with multiple sclerosis (MS) and individuals with other neurological diseases. No differences in the concentrations and specificities of the serum antibodies were observed among the 3 groups. In contrast, we found that MS patients often had elevated CSF antibody levels against many antigens of the panel. The MS patients with local immunoglobulin production in the central nervous system (CNS) had the highest antibody levels. Restricted antibody activity against a given antigen of the panel was not observed. Compared to the two other groups, the MS group had equivalent titres of anti-MBP antibodies in the CSF. These results suggest that, in MS, a general immune dysregulation exists which leads to a local expansion of B lymphocytes producing autoantibodies with reactivities similar to those of serum natural autoantibodies.

Plasma Cells in the Bronchoalveolar Lavage Fluid of a Patient with Eosinophilic Pneumonia: Morphologic Proof of Local Production of Antibodies

We describe a case of eosinophilic pneumonia in which plasma cells appeared in bronchoalveolar lavage fluid (BALF). A 49-year-old man presented with wheezing, lung infiltrates, peripheral eosinophilia, and extremely high IgE levels in serum and BALF. A differential count of BALF revealed 56.6 percent lymphocytes and 1.3 percent plasma cells. The appearance of plasma cells suggests local maturation of B cells and represents a morphologic proof of local production of immunoglobulins. The increased number of lymphocytes suggests their role in B cell differentiation via the lymphokine network.

Local immunoglobulin production is different in gastritis associated with dermatitis herpetiformis and simple gastritis.

The degree of inflammation and atrophy in gastric body mucosal specimens (n = 38) from 28 patients with dermatitis herpetiformis (DH) was graded histologically. Immunoglobulin (Ig) producing cells were enumerated by paired immunofluorescence staining in a 500 microns wide section area from the muscularis mucosae to the lumen (mucosal 'tissue unit'). The number of immunocytes of the three main classes (IgA, IgM, and IgG) was significantly raised with increasing degree of gastritis. All three classes were increased in specimens showing atrophy compared with those without atrophy. IgA cells predominated as in simple gastritis, but a striking difference was a marked increase of IgM cells in specimens with the most pronounced DH-associated gastritis. Relative class distribution of immunocytes within different mucosal zones showed that the percentage of IgA cells was significantly higher in the luminal than in the basal zone, whereas the contrary was true for IgG cells. IgM cells did not show any zonal preference. No relation was seen between small bowel and gastric lesions. The disproportionate increase of gastric IgM producing cells in DH might nevertheless reflect seeding of precursor cells of the secretory immune system generated in the proximal small intestine where the local IgM response is relatively pronounced.

Interferon-gamma enhances expression of secretory component, the epithelial receptor for polymeric immunoglobulins.

Recombinant interferon-gamma (IFN-gamma) increased in a dose-dependent manner the intracellular pool, the membrane expression, and the shedding of secretory component (SC) in human colonic adenocarcinoma cell line (HT-29). A similar dose-response relationship was observed when we examined the binding of polymeric IgA to HT-29 cells treated with IFN-gamma, thus reflecting expression of functional SC. Because IFN-gamma is produced by T cells during immune responses, activated T cells may be able to promote the external transport of dimeric IgA and pentameric IgM and thereby enhance the efferent limb of the secretory immune system. This is, therefore, the first observation indicating how the secretory transport capacity may be adjusted to increased local immunoglobulin production.

Quantification of immunoglobulins in respiratory tract secretions of the horse

Lavage techniques were used to obtain secretions from the nasal cavity, trachea and bronchi of conscious horses. The techniques, which utilised fibreoptic endoscopy for recovery of tracheal and bronchial secretions, were well tolerated by the horses. The recovery rates of the lavaged fluids were acceptable, but were lowest for bronchial secretions, and there was minimal contamination by blood. The fluids were analysed for IgG and IgM by single radial immunodiffusion, and for IgA and albumin by rocket immunoelectrophoresis. Relative to albumin there was significantly more IgA and IgM, and significantly less IgG, in the nasal cavity than the trachea. IgA and IgM levels were greatest in the nasal cavity and decreased progressively to the bronchi, whereas IgG levels showed the reverse trend. The immunoglobulin: albumin ratios of secretions taken from many levels of the tract were significantly higher than those of serum, suggesting local production of immunoglobulin in the respiratory tract.

Appearance of specific antibody-bearing cells in human bronchial mucosa after local immunization with bacterial vaccine.

The immune response to local in vivo inhalation of a lysed bacteria vaccine was assessed in surgical specimens of main-stem bronchi from patients who had undergone pneumectomy for cancer. The patient population included 22 subjects; 11 of these received the aerosol vaccine twice a day for 10 days prior to surgery, while the remaining 11 patients were used as controls and were not immunized. The submucous glands of immunized subjects showed significantly more cells than did those of the controls, i.e., 62 +/- 8 versus 37 +/- 7, respectively (P less than 0.05). The following five antigens were chosen for study by fluorescence assay: Streptococcus pneumoniae types II and III, Haemophilus influenzae, Streptococcus sp. strain D19, and Klebsiella pneumoniae. An immunization-dependent correlation was found between immunoglobulin A, immunoglobulin A-bearing cells, and specific antibody-bearing cells on the one hand and three of the five antigens (S. pneumoniae types II and III and Streptococcus sp. strain D19) on the other hand. This is the first time that a relationship has been established between bacterial immunization of the lower respiratory tract and local immunoglobulin production in humans.

Juvenile periodontitis. I. Demonstration of local immunoglobulin synthesis.

The purpose of this investigation was to determine the presence and quantity of immunoglobulin G (IgG) and albumin (ALB) in specified periodontal tissues and serum from patients diagnosed as having juvenile periodontitis (JP), using an immunoelectrophoresis technique and to determine which portion of the lesion generated the greatest local immunoglobulin production. Serum and tissue samples were obtained from 19 patients (ages 13-21 years) who were diagnosed as having JP; 18 were female; 16 were black. Normal, diseased, and granulomatous gingival/periodontal tissues were collected during full thickness flap surgery, then minced, homogenized, and centrifuged. Supernatants containing the gingival/periodontal protein and the serum were electrophoresed against rabbit antihuman IgG and ALB. The relative IgG/ALB ratios in each specimen were plotted against known concentrations of IgG using a least squares analysis to provide evidence for local synthesis. In comparing mean IgG/ALB ratios for all tissue types, it was noted that normal gingiva did not differ significantly from serum. Diseased and granulomatous tissues, taken together or separately, had significantly higher IgG/ALB ratios than normal gingiva or serum, demonstrating that 73.6% of the IgG present was due to local synthesis. However, the greatest amounts of locally produced IgG were found in the granulomatous tissue from the deepest areas of the defects.

Changes in tear protein composition in Graves' ophthalmopathy

The tear protein composition of normal volunteers and that of patients suffering from hyperthyroidism and various degrees of infiltrative endocrine ophthalmopathy were studied by polyacrylamide-gel electrophoresis, radial immunodiffusion and laser nephelometry. Increased level of IgG, serum albumin and transferrin was demonstrated in hyperthyroidism and severe infiltrative ophthalmopathy. Elevated IgA, IgM and immune complex (IC) levels were detected in cases with mild and severe infiltrative ophthalmopathy, which showed good correlation with the severity of the disease. A decrease in the level of tear specific proteins showed the dysfunction of the lacrimal glands in long-standing cases of Graves' ophthalmopathy. Elevated immunoglobulin levels in cases with irritation are partly the result of increased vascular permeability. High IgA, IgM and 1C levels observed without permeability changes in mild ophthalmopathy suggest the local production of immunoglobulins in infiltrative ophthalmopathy and their possible role in the development of the eye manifestations of Graves' disease. The use of tear protein determinations in the diagnosis and in the evaluation of different therapeutic procedures is discussed.

CRYPTOGENIC FIBROSING ALVEOLITIS AND EPSTEIN-BARR VIRUS: AN ASSOCIATION?

13 patients with cryptogenic fibrosing alveolitis (CFA) and 12 with interstitial lung disease (ILD) of known cause were studied for their humoral response to herpes simplex virus (HSV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV). Serum antibodies to HSV and CMV were within the normal range in all patients. 10 patients with CFA had raised serum antibodies to EBV, and IgA against viral-capsid antigen (VCA) was detectable in all 13. In the other 12 patients EBV serological profiles were normal and IgA against VCA was detectable in only 1 patient. The EBV antibody levels did not correlate with the level of circulating immune complexes, the presence of rheumatoid factors, or the cytological findings of the alveolitis. The presence of IgG against VCA in 5 CFA patients suggests local production of EBV-specific immunoglobulins. Elevated IgG and IgA against EBV in CFA may indicate non-specific depression of cell-mediated immunity or that EBV plays a part in the aetiology of CFA.

Immunoglobulin levels in serum and cerebrospinal fluid in certain viral infections of the central nervous system.

It is seriously debated whether the presence of immunoglobulins in CSF is due to local production, diffusion of proteins through the blood-brain barrier, or both. Tourtellotte et al [1] strongly suggest that in both healthy and diseased individuals, immunoglobulins are synthesized extravascularly and subsequently diffuse into the CSF. Cohen and Bannister [2] demonstrated that lymphocytes from the CSF of patients with multiple sclerosis could produce IgA and IgG in vitro. Although such evidence suggests the likelihood of local production of immunoglobulins, others believe that elevated protein levels in CSF are due to migration of immunoglobulins from serum to CSF because of damage to the blood-brain barrier [3]. Thus, the elevated IgG level in CSF could be an expression of such an impaired blood-brain barrier. A further implication of this hypothesis is that the more the function of this barrier is impaired, the more extensive is the transudation. This includes the migration of large molecules such as IgM during the inflammatory state, a condition which increases not only the number of theoretical filters but also the size of the pores. IgG levels in CSF and serum were elevated in all five infections studied except viral encephalitis, in which this value remained normal. The serum IgA level was elevated in all five lesions, but the IgA level in CSF was elevated only in viral encephalitis, Guillain-Barré syndrome, and meningeal carcinomatosis.(ABSTRACT TRUNCATED AT 250 WORDS)

The local cell response to human lung carcinomas.

Non-malignant inflammatory cells were studied in tissue sections and cell suspensions from 21 human lung carcinomas. The different cell types were identified and quantified by means of histochemical and immunohistochemical methods. All tumours were surrounded and to a lesser degree also infiltrated by lymphocytes, macrophages, plasma cells containing IgG, IgA, or IgM, and a small number of mast cells. No evident signs of cell necrosis were present in association with these mononuclear cell infiltrates. Central tumour necroses were dominated by large numbers of neutrophils and macrophages, a reaction considered to be secondary to ischemic necroses in rapidly expanding tumours. The weakest mononuclear cell infiltrates were found in tumours with extensive necrotic areas. Of the tumour lymphocytes 3-67% (mean 37%) were T-lymphocytes, 4-32% (mean 19%) were B-lymphocytes. The relation between these markers did not always correlate with the findings in peripheral blood: low percentages of T-lymphocytes were found in some tumours even when the percentage of T-lymphocytes in peripheral blood was within the normal range. The demonstration of both T-lymphocytes and macrophages in and around the tumour tissue may reflect a local immune response against the neoplastic cells, while the presence of plasma cells in all tumours supports the theory that the tumour also induces a local production of immunoglobulin which may interfere with the cellular immunity.

Tumour-associated immunoglobulins and host cell infiltration

Using a variety of techniques, we have investigated the possible interrelationship between tumour-associated immunoglobulins and infiltrating host cells. Initial cytological and histological studies revealed that tumour models which normally have a high immunoglobulin content also contained a relatively high proportion of host cells which could bind immunoglobulin by cytophilic processes. Rosette assays on tumour cell suspensions from mice also challenged with SRBC suggested that some of the immunoglobulin present may be non-specifically bound to host cells or synthesized in situ. Evidence that the local production of immunoglobulin could occur was obtained by protein A-PFC analysis of tumour cell suspensions. These studies also revealed that there were many more IgA-secreting cells within tumours than in spleen or lymph nodes. Finally, immunoglobulin analysis on gradient-separated tumour cell suspensions indicated that immunoglobulin was associated with both tumour and host cell-enriched (i.e., Fc-positive) fractions. These observations emphasize that much of the immunoglobulin previously noted in tumour eluates may not be associated with tumour cells as such, but may be bound to or synthesized by infiltrating host lymphoreticular cells.

Characterization of antibody activity in oligoclonal immunoglobulin G synthesized within the central nervous system in a patient with tuberculous meningitis.

Thin-layer polyacrylamide gel isoelectric focusing of cerebrospinal fluid (CSF) and serum obtained from one patient 48 and 65 days after the onset of tuberculous meningitis revealed five oligoclonal immunoglobulin zones in CSF without any counterpart in serum, indicating local immunoglobulin production. Subsequent immunofixation with specific antisera revealed that three of the zones consisted of immunoglobulin G lambda present simultaneously. Immunofixation with Mycobacterium tuberculosis and bacillus Calmette-Guérin (BCG) as antigens and autoradiography revealed zones of specific antibodies in the CSF which, regarding mobility, corresponded to oligoclonal and polyclonal CSF immunoglobulin G zones. No antibody activity was detectable in the corresponding serum, indicating that the antibodies present in CSF were synthesized within the central nervous system. In seven control patients (three with multiple sclerosis, four with chronic inflammatory central nervous system diseases of unknown cause) with oligoclonal CSF immunoglobulin, no evidence for local production of antibodies against M. tuberculosis or BCG was detectable. Immunofixation with M. tuberculosis or BCG as antigens and autoradiography may prove to be a useful diagnostic complement to conventional techniques in patients with suspected tuberculous meningitis.

Immunoglobulin levels of tonsillar cells

Immunoglobulin G, A and M contents of homogenized tonsillar lymphocytes were measured. Higher Ig values were found in adults than in children. Inflammatory processes increased the immunoglobulin levels of tonsillar lymphocytes. The changes in tonsillar IgG and IgA levels suggest the importance of local immunoglobulin production in the defence mechanism of tonsils. Two isolated IgA deficiencies were detected among the 85 patients.

Normal human bronchial immunoglobulins and proteins: effects of cigarette smoking.

Selected proteins were compared in bronchial lavage fluids recovered from 36 healthy nonsmokers and 19 healthy cigarette smokers. Although fluids lavaged from smokers contained 4 times as many cells (more than 80 per cent macrophages) as fluids from nonsmokers, there were no quantitative differences between the two groups in immunoglobulin A, a1-antitrypsin, a2-globulin, transferrin, or albumin content. However, immunoglobulin G content of bronchial fluids from smokers was twice as high as that from nonsmokers. This selective increase in immunoglobulin is compatible either with stimulated local bronchial immunoglobulin production or the selective exudation of plasma immunoglobulin into alveolar spaces in response to inhaled cigarette smoke.

Immunoglobulin Content in the Bronchial Washings of Patients with Benign and Malignant Pulmonary Disease

The bronchial washings of 58 patients with benign and neoplastic conditions involving the lungs were evaluated for immunoglobulin content. Levels in the washings from the diseased lung were compared with those from the normal contralateral lung. In normal patients and those with either bilateral inflammatory diseases or unilateral bronchiectasis or tuberculosis, the mean variation in igA/K and igG/K ratios between diseased and uninvolved lungs was minimal. In unilateral pneumonitis, however, igG/K was markedly elevated (P less than 0.01). Significant changes in the igA/K occurred on the affected side in patients with cancer. In the group with squamous-cell cancer. the mean elevation was 4.6 times that of contralateral lung (P less than 0.005), whereas with adenocarcinoma and undifferentiated carcinoma, the rise was 6.4 and 2.8 times, respecitvely (P less than 0.01 and P less than 0.01). Thus, carcinoma appears to alter local immunoglobulin production in the affected lung as compared to its normal counterpart. (N Engl J Med 295:694-698, 1976).

The pathogenesis of gluten-sensitive enteropathy.

Gluten-sensitive enteropathy is characterized by flattening of intestinal villi and malabsorption caused by the toxic effect of gluten, a wheat protein. Gluten activates an endogenous mechanism of toxicity that may be the local mucosal immune system: local mucosal immunoglobulin and antigluten antibody production occur soon after gluten ingestion. Approximately 80% of patients with this disease possess HL-A8, a second segregant series antigen. This association also occurs in dermatitis herpetiformis, a disease with vesicular skin lesions and gluten-sensitive flattening of intestinal villi. The association suggests that the fundamental abnormality in enteropathy is a binding reaction involving gluten protein and a binding site on a cell surface, determined in part by the histocompatibility gene; this reaction then results in a local mucosal immune response to gluten. Alternatively, the fundamental abnormality may be the presence of an abnormal immune-response gene linked to the HL-A8 gene or acting in concert with it; this immune-response gene results in local mucosal production of antigluten antibody.

Local monoclonal immunoglobulin production in cancer patient.

La technique directe de l'immunofluorescence a permis de mettre en évidence dans un ganglion lymphatique adjacent à la tumeur cancéreuse du rectum la présence de nombreuses cellules sécrétant de IgG monoclonal.