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Phospholipase A2 receptor 1 (PLA2R1) exists important role in membranous nephropathy. In this study, we evaluate a PLA2R1 in a middle-aged rat model of renal function repair to further investigate the molecular mechanisms of membranous nephropathy. We analyzed the PLA2R1 knockout (KO) model and PLA2R1 knock in (KI) model in rats, extending the time to 85 weeks of age. Urinary biochemical indicators were detected using a fully automated biochemical analyzer. The complement C3, IgG, and Nephrin were detected using the immunofluorescence method. Western blot was used to detect the expression levels of complement C3, IgA and PLA2R1 in middle-aged models. The KO model continues to display glomerular proteinuria, complement C3 aggregation, and IgA and IgG deposition. Comparing with the KO model, the deposition of complement C3 and IgA in the glomerulus of the KI chimeric model still exists and IgG expression weakened. Inserting humanized PLA2R1 into rats can continuously repair partial renal function and reduce proteinuria, which will help investigate the pathogenesis of membranous nephropathy and complement activation signaling pathways., (© 2024 John Wiley & Sons Ltd.)

In recent years, there has been increasing attention towards understanding the relationship between age-related alterations in the oral microbiota and age-associated diseases, with reports emphasizing the significance of maintaining a balanced oral microbiota for host health. However, the precise mechanisms underlying age-related changes in the oral microbiota remain elusive. We recently reported that cellular senescence of ileal germinal center (GC) B cells, triggered by the persistent presence of commensal bacteria, results in diminished IgA production with aging and subsequent alterations in the gut microbiota. Consequently, we hypothesize that a similar phenomenon may occur in the oral cavity, potentially contributing to age-related changes in the oral microbiota. Examination of p16-luc mice, wherein the expression of the senescent cell marker p16 INK4a can be visualized, raised under specific pathogen-free (SPF) or germ-free (GF) conditions, indicated that, unlike ileal GC B cells, the accumulation of senescent cells in GC B cells of cervical lymph nodes increases with age regardless of the presence of commensal bacteria. Furthermore, longitudinal studies utilizing the same individual mice throughout their lifespan revealed concurrent age-related alterations in the composition of the oral microbiota and a decline in salivary IgA secretion. Further investigation involving Rag1 -/- mice transplanted with B cells from wild-type or p16 INK4a and p21 Waf1/Cip1 -double knockout mice unveiled that B cell senescence leads to reduced IgA secretion and alteration of the oral microbiota. These findings advance our understanding of the mechanism of age-associated changes in the oral microbiota and open up possibilities of their control., (© 2024 The Author(s). Aging Cell published by Anatomical Society and John Wiley & Sons Ltd.)

Galactose-deficient IgA1 (Gd-IgA1) plays a crucial role in the development of Immunoglobulin A nephropathy (IgAN), however, the underlying pathogenic mechanisms driving Gd-IgA1 production in B cells are not well understood. In this study, RNA-seq analysis identified 337 down-regulated and 405 up-regulated genes in B cells from 17 patients with IgAN and 6 healthy controls. Among them, ST6Gal1, which was associated with IgAN in a previous genome-wide association study (GWAS), was up-regulated in IgAN and significantly positive correlated with elevated Gd-IgA1. In addition, we identified increased plasma ST6Gal1 levels in 100 patients with IgAN, which were associated with higher levels of proteinuria, plasma IgA, Gd-IgA1 levels, greater degrees of systemic complement activation including C3a, Bb, C4d, MAC and a lower proportion classified as C2 grade (crescent proportion ≥25%). Interesting, in vitro, recombinant ST6Gal1 (rST6Gal1) exposure reduced the production of Gd-IgA1 in cultured peripheral blood mononuclear cells from IgAN patients. rST6Gal1 stimuli also increased expression of C1GALT1, which were well-known proportional to the decrease in galactose deficiency of IgA1. In conclusions, we identified increased plasma ST6Gal1 levels and the association of ST6Gal1 with disease severity of IgAN. Additionally, rST6Gal1 administration in vitro increased expression of C1GALT1 and reduced the production of Gd-IgA1., (© 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)

Problem: Chlamydia infections in women can ascend to the upper genital tract, and repeated infections are common, placing women at risk for sequelae. The protective role of anti-chlamydia antibodies to surface exposed antigens in ascending and incident infection is unclear., Method of Study: A whole-bacterial ELISA was used to quantify chlamydia-specific IgG and IgA in serum and cervical secretions of 151 high-risk women followed longitudinally. Correlations were determined between antibody and cervical burden, and causal mediation analysis investigated the effect of antibody on ascension. We examined the relationship of antibody to incident infection using the marginal Cox model., Results: Serum and cervical anti-chlamydia IgG and cervical IgA levels correlated inversely with cervical burden. While lower burden was associated with reduced ascension, causal mediation analysis revealed that the indirect effects of antibody mediated through reductions in bacterial burden were insufficient to prevent ascension. Analysis of women uninfected at enrollment revealed that serum and cervical anti-chlamydia IgG were associated with increased risk of incident infection; hazard ratio increased 3.6-fold (95% CI, 1.3-10.3), and 22.6-fold (95% CI, 3.1-165.2) with each unit of serum and cervical IgG, respectively., Conclusion: Although anti-chlamydia IgG and IgA correlated with reduced cervical chlamydia burden, they failed to prevent ascension and increased levels of anti-chlamydia IgG were associated with increased risk for incident infection., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Imbalances of T-cell subsets are hallmarks of disease-specific inflammation in psoriasis. However, the relevance of B cells for psoriasis remains poorly investigated., Objective: To analyse the role of B cells and immunoglobulins for the disease-specific immunology of psoriasis., Methods: We characterized B-cell subsets and immunoglobulin levels in untreated psoriasis patients (n = 37) and compared them to healthy controls (n = 20) as well as to psoriasis patients under disease-controlling systemic treatment (n = 28). B-cell subsets were analysed following the flow cytometric gating strategy based on the surface markers CD24, CD38 and CD138. Moreover, immunofluorescence stainings were used to detect IgA in psoriatic skin., Results: We found significantly increased levels of IgA in the serum of treatment-naïve psoriasis patients correlating with disease score. However, IgA was only observed in dermal vessels of skin sections. Concerning B-cell subsets, we only found a moderately positive correlation of CD138 + plasma cells with IgA levels and disease score in treatment-naïve psoriasis patients. Confirming our hypothesis that psoriasis can develop in the absence of functional humoral immunity, we investigated a patient who suffered concomitantly from both psoriasis and a hereditary common variable immune defect (CVID) characterized by a lack of B cells and immunoglobulins. We detected variants in three of the 13 described genes of CVID and a so far undescribed variant in the ligand of the TNFRSF13B receptor leading to disturbed B-cell maturation and antibody production. However, this patient showed typical psoriasis regarding clinical presentation, histology or T-cell infiltrate. Finally, in a group of psoriasis patients under systemic treatment, neither did IgA levels drop nor did plasma cells correlate with IgA levels and disease score., Conclusion: B-cell alterations might rather be an epiphenomenal finding in psoriasis with a clear dominance of T cells over shifts in B-cell subsets., (© 2018 European Academy of Dermatology and Venereology.)

Bronchiolitis obliterans is a small-airway obstructive lung disease for which immunologically mediated pathogenesis is supposed. Frequent association of bronchiolitis obliterans with paraneoplastic pemphigus is well known, but its association with other autoimmune bullous diseases has not been reported except for a case of anti-laminin-332-type mucous membrane pemphigoid in a patient with chronic graft-versus-host disease. We report a case of non-paraneoplastic autoimmune subepidermal bullous disease associated with fatal bronchiolitis obliterans in a patient without transplantation. Although the patient's serum contained immunoglobulin (Ig)A antibodies to the 180-kDa bullous pemphigoid antigen/type XVII collagen and IgG antibodies to laminin-332, diagnosis of either linear IgA bullous dermatosis or mucous membrane pemphigoid could not be made because of the failure to detect linear IgA deposition at the basement membrane zone by direct immunofluorescence and the lack of mucous membrane lesions. Physicians should be aware that autoimmune bullous diseases other than paraneoplastic pemphigus can also associate with this rare but potentially fatal lung disease., (© 2016 Japanese Dermatological Association.)

Background: Leukocytoclastic vasculitis (LCV) in children is a complex group of conditions., Objectives: This study presents the demographics, clinical features, direct immunofluorescence (DIF) results and suspected aetiologies of 56 biopsy-confirmed cases of leukocytoclastic vasculitis in children., Methods: Retrospective review of 56 children seen at Mayo Clinic in Rochester, Minnesota, from 1993 to 2013 with clinical features and cutaneous biopsy consistent with LCV., Results: Twenty-seven (48%) cases were found to be due to IgA vasculitis (Henoch-Schonlein purpura). The remaining cases were found to be due to cutaneous small-vessel vasculitis (n = 19, 34%), urticarial vasculitis (n = 5, 9%), ANCA-associated vasculitis (n = 4, 7%) and acute haemorrhagic oedema of infancy (n = 1, 2%). IgA vasculitis was found to be associated with abdominal pain (P = 0.008), whereas the non-IgA vasculitis group was associated with headache (P = 0.052). Children with IgA vasculitis had palpable purpura (P = <0.001), petechia (P = 0.057), vesicles (P = 0.009) and involvement of the buttock (P = 0.004) more frequently than the non-IgA vasculitis group. On DIF, perivascular IgA was positive in IgA vasculitis compared to non-IgA vasculitis cases (P = <0.001), the other conjugates were similar between the two groups., Conclusion: The most common subtype of biopsy-confirmed LCV in children is IgA vasculitis. Clinical features, exam characteristics and DIF results can be helpful in determining the subtype of cutaneous vasculitis in children., (© 2016 European Academy of Dermatology and Venereology.)

Problem: The pIgR mediates transport of IgA into the lumen of mucosal tissues preventing pathogenic infection. Despite this, the expression of pIgR during chlamydial infections of the male and female reproductive tracts remains poorly understood., Method of Study: The expression of pIgR in response to hormone cycling or over the course of chlamydial infection was determined in vitro and in vivo by Western blot or immunohistochemistry., Results: PIgR was upregulated in response to Chlamydia spp. infection of human epithelia, in both male and female mouse reproductive tracts. PIgR expression was found to be highest during estrus in the cervicovaginal and uterine epithelia and lowest during diestrus or following hormonal synchronization with Depo-Provera. Chlamydial infection of mice mediates upregulation of pIgR and transcytosis of IgA into the lumen., Conclusions: Our results suggest that chlamydial infection enhances IgA secretion and pIgR expression by epithelia in the lower reproductive tracts of females and males, and hormone synchronization downregulates pIgR expression and transcytosis of IgA prior to challenge., (© 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

A 63-year-old Japanese man with non-Hodgkin B-cell lymphoma presented with erythematous skin lesions on his entire body, with oral, ocular and anal mucosal lesions. The patient was diagnosed with paraneoplastic pemphigus. Immunofluorescence showed both immunoglobulin (Ig)G and IgA antibodies to keratinocyte cell surfaces. Various immunoblot and enzyme-linked immunosorbent assays showed both IgG and IgA antibodies to various autoantigens, including desmogleins, desmocollins, envoplakin, periplakin and bullous pemphigoid antigens. This was a unique case with a very rare autoantibody profile in paraneoplastic pemphigus., (© 2016 Japanese Dermatological Association.)

Intestinal ischaemia/reperfusion (I/R) severely disrupts gut barriers and leads to high mortality in the critical care setting. Transforming growth factor (TGF)-β1 plays a pivotal role in intestinal cellular and immune regulation. However, the effects of TGF-β1 on intestinal I/R injury remain unclear. Thus, we aimed to investigate the effects of TGF-β1 on gut barriers after intestinal I/R and the molecular mechanisms. Intestinal I/R model was produced in mice by clamping the superior mesenteric artery for 1 hr followed by reperfusion. Recombinant TGF-β1 was intravenously infused at 15 min. before ischaemia. The results showed that within 2 hrs after reperfusion, intestinal I/R disturbed intestinal immunoglobulin A class switch recombination (IgA CSR), the key process of mucosal IgA synthesis, and resulted in IgA dysfunction, as evidenced by decreased production and bacteria-binding capacity of IgA. Meanwhile, the disruptions of intestinal microflora and mucosal structure were exhibited. Transforming growth factor-β1 activated IgA CSR as evidenced by the increased activation molecules and IgA precursors. Strikingly, TGF-β1 improved intestinal mucosal IgA dysfunction, dysbiosis and epithelial damage at the early stage after reperfusion. In addition, SB-431542, a specific inhibitor of activating mothers against decapentaplegic homologue (SMAD) 2/3, totally blocked the inductive effect of TGF-β1 on IgA CSR and almost abrogated the above protective effects on intestinal barriers. Taken together, our study demonstrates that TGF-β1 protects intestinal mucosal IgA immunity, microbiota and epithelial integrity against I/R injury mainly through TGF-β receptor 1/SMAD 2/3 pathway. Induction of IgA CSR may be involved in the protection conferred by TGF-β1., (© 2016 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

We report a case involving a 62-year-old woman with in vivo-bound immunoglobulin (Ig)G and IgA antibodies in both the intercellular space (ICS) and basement membrane zone (BMZ). Her clinical and histopathological features were identical with those of pemphigus vulgaris, while the immunopathological findings suggested IgG/IgA pemphigus. Direct immunofluorescence (IF) showed in vivo-bound IgG and IgA antibodies in the ICS and BMZ, whereas indirect IF showed circulating IgG but not IgA antibodies in the ICS and BMZ. The anti-ICS IgG bound to desmoglein-3, while the anti-BMZ antibodies bound to the epidermal side of 1 mol/L NaCl-split skin. To the best of our knowledge, only two similar cases have been reported so far. Furthermore, we also examined IgG subclass distribution of the in vivo-bound and circulating anti-ICS and BMZ antibodies, and found that IgG1, IgG2 and IgG4 bound to ICS of the lesional skins, while IgG1 and IgG3 bound to the BMZ. The circulating anti-ICS antibodies belonged to IgG1 and IgG4, while the circulating anti-BMZ antibodies to IgG1, IgG2 and IgG4. We report a case involving a 62-year-old woman with in vivo-bound immunoglobulin (Ig)G and IgA antibodies in both the intercellular space (ICS) and basement membrane zone (BMZ). Her clinical and histopathological features were identical with those of pemphigus vulgaris, while the immunopathological findings suggested IgG/IgA pemphigus. Direct immunofluorescence (IF) showed in vivo-bound IgG and IgA antibodies in the ICS and BMZ, whereas indirect IF showed circulating IgG but not IgA antibodies in the ICS and BMZ. The anti-ICS IgG bound to desmoglein-3, while the anti-BMZ antibodies bound to the epidermal side of 1 mol/L NaCl-split skin. To the best of our knowledge, only two similar cases have been reported so far. Furthermore, we also examined IgG subclass distribution of the in vivo-bound and circulating anti-ICS and BMZ antibodies, and found that IgG1, IgG2 and IgG4 bound to ICS of the lesional skins, while IgG1 and IgG3 bound to the BMZ. The circulating anti-ICS antibodies belonged to IgG1 and IgG4, while the circulating anti-BMZ antibodies to IgG1, IgG2 and IgG4., (© 2015 Japanese Dermatological Association.)

Aim: The rotavirus has been identified as the most common cause of severe gastroenteritis in young children. This study compared the levels of rotavirus-specific antibodies in breast milk and serum samples in Swedish and Bangladeshi mothers to identify any seasonal variations., Methods: Breast milk and serum samples were collected from 69 Swedish and 52 Bangladeshi mothers during all months of the year. Sera were analysed for Immunoglobulin A (IgA) and Immunoglobulin G (IgG) titres and breast milk for secretory IgA (SIgA) antibody levels against rotavirus., Results: Significantly, higher SIgA antibody levels against rotavirus were found in breast milk from Bangladeshi than Swedish mothers (p < 0.0001). Seasonality of rotavirus antibody levels was only detected in breast milk from Swedish mothers, with samples collected during the spring having significantly higher rotavirus titres than samples collected during the autumn (p = 0.0203)., Conclusion: This study found significantly higher SIgA antibody levels against rotavirus in breast milk from Bangladeshi than Swedish mothers, together with a seasonal variation among Swedish mothers only. These results suggest that Swedish children who are being breastfed should ideally be vaccinated during seasons when the rotavirus antibody levels in breast milk are low., (©2014 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.)

Objective: To evaluate the generation of rheumatoid arthritis (RA)-related autoantibodies in the lung., Methods: Simultaneous collection of serum and induced sputum was performed in 21 healthy controls, 49 at-risk subjects without inflammatory arthritis but at risk of RA due to family history or seropositivity for anti-citrullinated protein antibodies, and 14 subjects with early RA. Samples were tested for anti-cyclic citrullinated peptide 2 (anti-CCP2), anti-CCP3, anti-CCP3.1, rheumatoid factor isotypes IgM, IgG, and IgA, and total IgM, IgG, and IgA., Results: One or more autoantibodies were present in sputum of 39% of at-risk seronegative subjects, 65% of at-risk seropositive subjects, and 86% of subjects with early RA. In at-risk seronegative subjects, the rate of anti-CCP3.1 positivity and the median number of autoantibodies were elevated in sputum versus serum. In subjects with early RA, the rate of positivity for several individual autoantibodies and the median number of autoantibodies were higher in serum than in sputum. Results in at-risk seropositive subjects were intermediate between these groups. In at-risk subjects with autoantibody positivity in sputum, the ratios of autoantibody to total Ig were higher in sputum than in serum, suggesting that these autoantibodies are generated or sequestered in the lung., Conclusion: RA-related autoantibodies are detectable in sputum in subjects at risk of RA and in subjects with early RA. In a subset of at-risk subjects, the presence of sputum autoantibodies in the absence of seropositivity, and the increased autoantibody-to-total Ig ratios in sputum, suggest that the lung may be a site of autoantibody generation in the early development of RA. These findings suggest an important role of the lung in the pathogenesis of RA., (Copyright © 2013 by the American College of Rheumatology.)

Dermatitis herpetiformis (DH) is common in some Caucasian populations but extremely rare in Japanese, probably because of different immunogenetic backgrounds. We report two Japanese DH cases with typical clinical, histological and direct immunofluorescence features. However, no symptom of gluten-sensitive enteropathy was shown. The diagnosis was confirmed by eliminating other autoimmune blistering diseases by indirect immunofluorescence, enzyme-linked immunosorbent assays and immunoblotting. However, circulating immunoglobulin (Ig)A anti-endomysium, reticulin and gliadin antibodies were not detected. IgA antibodies to tissue and epidermal transglutaminases were also negative. One case was associated with lung cancer and the other one with autoimmune pancreatitis. On review of 17 cases of DH reported in Japan over the previous 10 years, including our cases, one case was associated with gluten-sensitive enteropathy, four with malignant neoplasms, two with autoimmune systemic disorders and one with psoriasis. Although our cases were typical of DH in clinical, histopathological and IgA deposit features, they showed different human leukocyte antigen haplotypes, no gluten-sensitive enteropathy and no DH-specific IgA antibodies, including those to epidermal and tissue transglutaminases. These results suggest that studies of unique characteristics in Japanese DH patients should facilitate further understanding of pathogenesis in DH., (© 2012 Japanese Dermatological Association.)

Objective: Many different genes or mediators have been implicated in promoting the development of vasculitis, although little is known regarding the mechanisms that normally act to suppress lesion formation. Endothelial nitric oxide synthase (eNOS) has been shown to inhibit vascular inflammation in many different model systems, but its roles in the pathogenesis of vasculitis have not been elucidated. This study was undertaken to determine the functions of eNOS in the initiation and progression of vasculitic lesion formation., Methods: MRL/MpJ-Fas(lpr) mice lacking the gene for eNOS (Nos3(-/-) ) were generated and comprehensively evaluated and compared to controls with regard to the development of autoimmune disease, including vasculitic lesion formation and glomerulonephritis., Results: Nos3(-/-) MRL/MpJ-Fas(lpr) mice exhibited accelerated onset and increased incidence of renal vasculitis compared to Nos3(+/+) controls. In contrast, no significant differences in severity of glomerulonephritis were observed between groups. Vasculitis was also observed in other organs of eNOS-deficient mice, including in the lungs of several of these animals. Ultrastructural analyses of renal lesions revealed the presence of electron-dense deposits in affected arteries, and IgG, IgA, and C3 deposition was observed in some vessels in the kidneys of Nos3(-/-) mice. In addition, Nos3(-/-) MRL/MpJ-Fas(lp) mice showed increased levels of circulating IgG-IgA immune complexes at 20 weeks of age, compared to Nos3(+/+) MRL/MpJ-Fas(lpr) and Nos3(-/-) C57BL/6 mice., Conclusion: These findings strongly indicate that eNOS serves as a negative regulator of vasculitis in MRL/MpJ-Fas(lpr) mice and further suggest that NO produced by this enzyme may be critical for inhibiting lesion formation and vascular damage in human vasculitic diseases., (Copyright © 2012 by the American College of Rheumatology.)

Aims: This study evaluated the salivary biochemical and immunological status of children with cancer undergoing to antineoplasic treatment in an attempt to identify alternatives for a less invasive and less painful monitoring of these patients., Materials and Methods: Unstimulated whole saliva samples were obtained from 115 children without cancer (control) and 32 children with cancer (CA). Children with cancer were also evaluated after antineoplasic treatment (CAT, n = 17). The salivary concentrations of glucose, triglycerides, total cholesterol, alkaline phosphatase, gamma-glutamyltransferase (GGT), urea, insulin, thyroid-stimulating hormone (TSH), triiodothyronine (T3), levothyroxine (T4), and immunoglobulin A (IgA) were determined., Results: Acute lymphocytic leukemia, acute myeloid leukemia, and Hodgkin's lymphoma were the most frequent cancers, although cases of non-Hodgkin's lymphoma, medulloblastoma, ependymoma, osteosarcoma, nephroblastoma, Ewing's sarcoma, and endodermal sinus tumor were also observed. The salivary concentration of cholesterol, triglycerides, or GGT did not differ between groups. Instead, the concentrations of alkaline phosphatase and T4 were higher in patients with cancer, irrespective of treatment. TSH levels were higher in the CA group and urea concentration was lower in the CAT group. T3 was undetectable in all groups. Antineoplasic treatment increased the glucose level and decreased the insulin concentration. Salivary concentration of total IgA was lower in children with cancer, irrespective of treatment., Conclusions: Cancer and antineoplasic treatment affected biochemical and immunological parameters in the saliva of children, shedding new light on the potential usefulness of saliva for monitoring children with cancer, especially to patients undergoing immunosuppressive therapy., (© 2012 John Wiley & Sons A/S.)

Vogt-Koyanagi-Harada disease is characterized by marked bilateral uveitis associated with symmetric vitiligo, alopecia, poliosis and dysacousia. Linear immunoglobulin (Ig)A bullous dermatosis (LABD) is characterized by small, tense, subepidermal bullae caused by IgA type autoantibody targeting the basal lamina. LABD patients sometimes show coexistence of IgG type autoantibody, termed linear IgA/IgG bullous dermatosis (LAGBD). We reported a 35-year-old Japanese male case of combined LAGBD and Vogt-Koyanagi-Harada disease. His human leukocyte antigen typing was -A24, B52, C*1202, DR*1502, DQ*0601. Immunoblot revealed that patient sera reacted to both 180- and 230-kDa proteins at the IgA and IgG level. Because Vogt-Koyanagi-Harada disease and LABD are reported to be associated with other autoimmune diseases, it is probable that Vogt-Koyanagi-Harada disease and LAGBD in our case may be associated with each other in the pathomechanism. However, we cannot exclude the possibility of this being mere coincidence., (© 2011 Japanese Dermatological Association.)

Background: An association between microscopic colitis and coeliac disease (CD) has been suggested in literature; however, population-based data are limited., Aims: To estimate the degree of association between these two diseases and to identify possible risk factors for disease concomitance., Methods: A population-based review of all patients diagnosed with CD and microscopic colitis in a large Canadian centre over a 5-year period. Endoscopy and pathology databases were searched to identify all cases of CD and microscopic colitis diagnosed within the Calgary Health Region between 2004 and 2008. Incidence rates were age-standardised and gender-standardised to 2006 Canadian Census data. standardised incidence ratios (SIR) were used to assess disease concomitance., Results: Over 5 years, 763 patients were diagnosed with CD, and 1106 were diagnosed with microscopic colitis. The standardised incidence of CD ranged from 10.4 to 15.7 per 100,000 population. The standardised incidence of microscopic colitis ranged from 16.9 to 26.2 per 100,000 population. Forty patients were diagnosed with both CD and microscopic colitis, 21 of whom were middle aged (40-60 years) females. Within the CD cohort, microscopic colitis occurred at an annual rate of 11.4 per 1000 cases of CD with an overall SIR of 52.7., Conclusions: There exists a strong association between microscopic colitis and CD with disease concomitance being approximately 50-times that expected in the general population. The concomitant diagnosis of CD and microscopic colitis largely occurs in middle-aged women. Therefore, middle-aged women with CD and diarrhoea as a presenting or persistent symptom should undergo lower endoscopy with biopsies to rule out microscopic colitis., (© 2011 Blackwell Publishing Ltd.)

Aim: This article describes the changes and relationships between biochemical and immunological parameters in the colostrum and serum of diabetic women., Methods: Colostrum and blood samples were collected from 30 diabetic and 15 normoglycaemic women. Glucose, total protein, antibody, complement proteins (C3 and C4), fat and calorie content, amylase, lipase and superoxide dismutase (SOD) were determined., Results: Glucose was higher in both the colostrum and serum of diabetic mothers compared to that of their normoglycaemic counterparts. In both groups, total protein was higher in colostrum than in serum. IgA and IgG were lower in the colostrum of hyperglycaemic mothers, whereas IgM did not vary between the groups. Colostral C3 protein was significantly lower in diabetic mothers, but colostral C4 protein was similar between normoglycaemic and hyperglycaemic mothers. Fat content was lower in the colostrum of the diabetic mothers, although calorie content did not vary between the groups. Amylase was lower in colostrum than in serum in both groups. Lipase was higher in the colostrum and serum of diabetic mothers. Colostral SOD was similar between the groups., Conclusions: Our results support the hypothesis that the colostrum of diabetic mothers suffers biochemical and immunological alterations that affect the levels of its components., (© 2010 The Author(s)/Acta Paediatrica © 2010 Foundation Acta Paediatrica.)

Problem: Chlamydia trachomatis is the most common sexually transmitted infection worldwide. While infection in females requires a Th1 response for clearance, such a response in males may disrupt the immune privileged nature of the male reproductive tract, potentially contributing to infertility., Method of Study: We investigated the role of IgA in protection against an intrapenile Chlamydia muridarum infection of C57BL/6 and pIgR-/- mice., Results: Here, we show that the poly immunoglobulin receptor is the main pathway for IgA transport into the male reproductive tract. The high levels of IgA seen in prostatic fluid of wild-type mice correlate with reduction in chlamydial infection both in vitro and in vivo., Conclusion: These findings indicate that a Chlamydia vaccine that induces neutralizing IgA in the prostate will aid in the protection against infection in males.

Objective: In patients with systemic lupus erythematosus (SLE) and lupus nephritis, the presence of antiphospholipid antibodies (aPL) is considered to be an indication of increased risk of thrombotic microangiopathy, a serious complication of SLE. Previous studies have demonstrated a critical role for activation of the classical pathway of complement that leads to thrombotic injury in the presence of aPL. This study was undertaken to investigate whether C4d deposition in lupus nephritis is related to circulating aPL and the presence of renal microthrombi., Methods: Deposition patterns of C4d in 44 renal biopsy samples obtained from 38 patients with biopsy-proven lupus nephritis were determined by staining with a polyclonal anti-C4d antibody. A phosphotungstic acid-hematoxylin stain was used to identify fibrin microthrombi. Clinical data (serum creatinine levels and presence or absence of aPL) were obtained and correlated with findings in the renal biopsy specimens. Patients were categorized as having aPL (n = 20) or not having aPL (n = 18)., Results: A strong relationship between the intensity of glomerular C4d staining and the presence of microthrombi was found (P < 0.002). Intense glomerular C4d deposition was present in 7 of 8 biopsy samples showing renal microthrombi. Neither C4d deposition nor the presence of microthrombi was correlated with aPL status., Conclusion: Our findings suggest that activation of the classical pathway of complement plays a pathogenic role in the development of renal tissue injury leading to thrombosis, irrespective of the type of circulating antibodies present. Immunodetection of glomerular C4d deposition in renal biopsy samples could be a convenient method of identifying patients at risk of thrombotic microangiopathy.

Background: Human salivary proteins: peroxidase, lysozyme, lactoferrin, and IgA, participate in the protection of oral tissues, as well as upper digestive and respiratory tracts, against a number of microbial pathogens. In the current study, we investigated the effect of acute consumption of a large dose of ethanol on representative human salivary proteins of the innate and adaptive immune systems., Methods: Eight healthy male volunteers drank an average of 2.0 g (1.4 to 2.5 g/kg) body weight of ethanol, in the form of vodka, in the 6-hour period. Samples of resting whole saliva were collected 12 hours before, then 36 and 108 hours after, the alcohol consumption. The levels of total protein, immunoglobulin A, lysozyme and lactoferrin as well as peroxidase activity were determined in saliva., Results: At 36 hours after alcohol consumption, salivary protein and lysozyme concentrations as well as peroxidase activity were significantly decreased (p = 0.002, p = 0.043, and p = 0.003, respectively), in comparison to the values obtained at 12 hours before drinking. Between 36 and 108 hours after alcohol consumption, the salivary protein and lysozyme concentrations, as well as peroxidase activity showed a tendency to increase, although at 108 hours after the drinking session, the concentration of protein and peroxidase activity were still significantly lower than before drinking. There was no significant change in the level of lactoferrin, after the drinking session. The salivary concentration of IgA tended to increase at 36 hours after alcohol consumption, and at 108 hours it was significantly higher (p = 0.028), when compared to IgA concentration in the saliva collected before drinking (from 8% to 26% and 32% of total protein content, respectively)., Conclusion: Our report is the first to show that acute ingestion of relatively large, yet tolerable dose of alcohol, significantly disturbs salivary antimicrobial defense system. Reduced lysozyme level and decreased peroxidase activity may contribute to increased susceptibility to infections, when acute alcohol intake coincides with exposure to pathogens.

Background: Environmental factors such as nutrition or exposure to infections play a substantial role in the pathogenesis of type 1 diabetes (T1D). We have previously shown that gluten-free, non-purified diet largely prevented diabetes in non-obese diabetic (NOD) mice. In this study we tested hypothesis that early introduction of gluten-enriched (gluten+) diet may increase diabetes incidence in NOD mice., Methods: Standard, gluten-free, gluten+ modified Altromin diets and hydrolysed-casein-based Pregestimil diet were fed to NOD females and diabetes incidence was followed for 310 days. Insulitis score and numbers of gut mucosal lymphocytes were determined in non-diabetic animals., Results: A significantly lower diabetes incidence (p < 0.0001) was observed in NOD mice fed gluten-free diet (5.9%, n = 34) and Pregestimil diet (10%, n = 30) compared to mice on the standard Altromin diet (60.6%, n = 33). Surprisingly, gluten+ diet also prevented diabetes incidence, even at the level found with the gluten-free diet (p < 0.0001, 5.9%, n = 34). The minority of mice, which developed diabetes on all the three diabetes-protective (gluten+, gluten-free, Pregestimil) diets, did that slightly later compared to those on the standard diet. Lower insulitis score compared to control mice was found in non-diabetic NOD mice on the gluten-free, and to a lesser extent also gluten+ and Pregestimil diets. No substantial differences in the number of CD3(+), TCR-gammadelta(+), and IgA(+) cells in the small intestine were documented., Conclusions: Gluten+ diet prevents diabetes in NOD mice at the level found with the non-purified gluten-free diet. Possible mechanisms of the enigmatic, dual effect of dietary gluten on the development of T1D are discussed., (2007 John Wiley & Sons, Ltd)

Problem: Chlamydial infections are often associated with various fertility-related disorders. Serological prediction of these has limitations, as they do not differentiate between past and current infections. Thus, we looked for local markers that could predict more precisely women at higher risk of developing severe complications., Method of Study: A total of 320 Chlamydia trachomatis positive women with or without fertility disorders were tested for the prevalence of immunoglobulin A antibodies to synthetic peptides of chlamydial heat-shock protein 60 (cHSP60) and cHSP10 along with cervical interferon-gamma (IFN-gamma) and serum C-reactive protein (CRP) levels., Results: Positive IFN-gamma level was the single best predictor for fertility disorder [odds ratio (OR) 15.4]. The predictive value of IFN-gamma could be significantly improved only by the addition of CRP test (OR 37.9)., Conclusion: Positive IFN-gamma levels in cervical washes along with elevated CRP levels could be used to predict women who are at higher risk of developing fertility disorders.

Background: Currently, no biochemical assay involving gingival crevicular fluid is utilized routinely as a screening test for periodontal disease., Objective: The objective of the present study was to evaluate the potential of gingival crevicular fluid assay as a screening methodology., Methods: The subject population was comprised of 27 volunteers. Nine participants were classified as 'subject with periodontal destruction' (SPD) exhibiting at least one site with pocket depth and attachment loss>3.5 mm, whereas the remaining individuals were categorized as 'subject with minimal periodontal destruction' (SMD). Gingival crevicular fluid was collected from fixed sites via a standardized method. Biochemical assays of 12 substances (hemoglobin, albumin, transferrin, alpha(1)-antitrypsin, fibronectin, IgA, IgG, IgM, lactoferrin, myeloperoxidase and neutrophil elastase) were conducted at a commercial laboratory. Power transformation of total quantities in gingival crevicular fluid was performed for statistical analysis., Results: Relationships between total quantity of each substance and periodontal disease status were unclear. Logistic regression analysis yielded six predictive models, which consisted of substance pairs: neutrophil elastase/IgA, neutrophil elastase/hemoglobin, neutrophil elastase/alpha(1)-antitrypsin and neutrophil elastase/IgG, and IgA/albumin and IgA/transferrin (p<0.05). Regression lines for SPD and SMD on a scattergram of IgA and neutrophil elastase were nearly parallel within the range of amounts in gingival crevicular fluid. The predictive model derived from both substances afforded sensitivity and specificity of 88% and 94%, respectively., Conclusions: These results indicated that the combination of IgA and neutrophil elastase in gingival crevicular fluid may be crucial for prediction of periodontal disease status. Furthermore, these data suggested that biochemical assays employing both substances in gingival crevicular fluid may provide a satisfactory screening test for periodontal disease.

Whether the response to chemotherapy is a prognosticator in multiple myeloma (MM) is still not known. Therefore, the relationship between survival and the rate of monoclonal protein (M-protein) decrement during the first cycles of therapy was prospectively assessed in 262 patients with newly diagnosed MM that were included in a phase III trial (HOVON-16). M-proteins were collected monthly during melphalan-prednisone therapy (MP: melphalan 0.25 mg/kg, prednisone 1.0 mg/kg orally for 5 d every 4 weeks). Patients with light chain disease (n = 18), immunoglobulin M (IgM)-MM (n = 1) and no immunotyping (n = 1) were excluded. Of the 242 patients studied, 75% had IgG M-protein and 25% IgA; MM stages: I: 1%, II: 35% and III: 64%. The median M-protein decrease after the first cycle of MP was 21% for IgG and 27% for IgA, and declined to < 5% after four cycles. An obvious survival advantage was seen for patients who had an M-protein decrease of at least 30% after the first MP cycle, which became significant when an M-protein decrease of 40% or more was reached. As established prognostic parameters (Salmon & Durie stage, serum creatinine, and haemoglobin) also remained prognostically significant, we concluded that early response to MP predicts for survival in MM.

FcalphaRI is the predominant receptor for IgA in the serum. Nevertheless, the interaction between the molecules that finally leads to an immune response is poorly understood. To investigate the structural requirements for IgA binding, the extracellular region of FcalphaRI was cloned and overexpressed in Escherichia coli. The resulting inclusion-body protein was refolded and purified. Despite its deglycosylated state, this recombinant FcalphaRI retained its ability to bind human IgA. The protein crystallized spontaneously as microcrystalline needles. Recrystallization yielded crystals belonging to a primitive monoclinic space group. A complete 2.8 A resolution X-ray diffraction data set was collected using synchrotron radiation.

Background: Chronic urticaria has been described in patients with Helicobacter pylori infection. We studied the titer of IgG and IgA type antibodies against H. pylori in patients with and without urticaria of unknown etiology. We also investigated the prevalence of antibodies against H. pylori-associated lipoprotein 20 (lpp20) in patients with and without chronic urticaria., Methods: The concentration of anti-H. pylori antibodies (IgG and IgA) was determined by the RIDA test. The level of anti-lpp20 antibodies was determined by Western blot using various H. pylori antigens (from 19 to 120 kDa)., Results: Patients with chronic urticaria and H. pylori infection (subgroup 1, n = 33) had high IgG and IgA titers whereas all patients with chronic urticaria and without H. pylori infection (subgroup 2, n = 23) were seronegative (P = 0.0128 for IgG and P = 0.003 for IgA). Titers in subgroup 1 did not differ significantly from a control group (n = 33) with severe H. pylori-associated gastritis without urticaria. The prevalence of the anti-lpp20 antibodies was significantly higher in subgroup 1 compared to the control group (93.9 vs 21.2%, P < 0.0001 for IgG, and 46.1 vs 6.3%, P < 0.0029 for IgA)., Conclusions: We suggest that IgG and IgA antibodies to H. pylori-associated lpp20 may play role in the pathogenesis of chronic urticaria.

Urinary cross-linked N-telopeptide of type I collagen (NTx) has been reported to be a sensitive and specific marker of bone resorption in multiple myeloma (MM). In this study, we measured the levels of NTx in 30 newly diagnosed MM patients and 25 controls. We examined its association with the overall score of skeletal involvement measured by Tc-99m-MIBI scintigraphy and other biochemical markers of bone disease (tumour necrosis factor a (TNF-a), serum calcium and creatinine). We further studied the correlation of NTx with the stage of disease (according to Durie-Salmon criteria) and bone marrow infiltration by plasma cells. High levels of NTx, bone marrow infiltration, TNF-alpha, calcium and creatinine were noted at advanced stages of disease (p < 0.05). NTx and TNF-a were found at significantly higher concentrations in patients with a high overall score (3 and 4) in Tc-99m-sestaMIBI in comparison to a low score (0, 1 and 2; p < 0.05). Positive correlations were found between NTx and TNF-a, as well as between bone infiltration and TNF-a or calcium. In conclusion, NTx is a useful marker for the monitoring of bone resorption in MM and correlates with imaging findings on Tc-99m-sestaMIBI and other biochemical markers of disease activity., (Copyright 2002 John Wiley & Sons, Ltd.)

Salivary fluid secretion is dependent upon reflex stimuli mediated by autonomic nerves. In order to determine if immunoglobulin A (IgA) and salivary proteins are secreted in the absence of nerve stimulation, small volumes (< 2 microl) of saliva were consecutively collected from the submandibular duct of anaesthetised rats following rest pauses in order to sample the protein contents of the ductal system. Within the first 5 microl of such saliva collected by parasympathetic nerve stimulation, IgA and other salivary proteins reached peak concentrations that were over 20-fold greater than levels in parasympathetically stimulated saliva subsequently collected during a 5 min period of stimulation. Confocal microscopy of TRITC-labelled IgA added to live, acutely isolated submandibular acini indicated that it did not enter the lumina by paracellular leakage. IgG is thought to enter saliva by paracellular leakage but did not accumulate in luminal saliva in the present study. Electrophoresis suggested that the major proteins secreted in the absence of stimulation were the same as those present in subsequently stimulated saliva. It can be concluded that IgA and other major submandibular proteins are secreted into glandular lumina in the absence of nerve stimulation. The functional significance of such unstimulated protein secretion is at present unclear.

Background: Apart form bacterial virulence factors of Helicobacter pylori, certain host factors influence the pathogenesis of H. pylori gastritis. In particular, antigastric autoantibodies that are detectable in the sera of a substantial proportion of H. pylori were shown to correlate with the development of gastric atrophy. The aim of this study was to analyze the possible antigastric autoimmune response in H. pylori gastritis at the site where the action is, i.e. , in the gastric mucosa., Material and Methods: Gastric biopsy specimens from antrum and corpus mucosa of 24 H. pylori-infected and of 33 noninfected patients were cultured for 3 days, and tissue culture supernatants were analyzed for the amount of locally produced IgA and IgG. Antigastric autoantibodies were screened in the sera and in the supernatants by means of immunohistochemistry., Results: The infected patients had significantly higher concentrations of locally produced IgA, whereas the IgG concentrations were virtually the same in infected and noninfected patients. IgG or IgA antigastric autoantibodies, or both, were detectable only in the sera (38%) and supernatants (17%) of infected patients. Interestingly, the patient with the strongest local autoimmune response showed body-predominant H. pylori gastritis, with destruction of gastric glands and atrophy of the body mucosa., Conclusions: These results demonstrate that antigastric autoimmune reactions are detectable at the site of the disease and might be relevant for the pathogenesis of gastric mucosa atrophy in H. pylori gastritis.

Secretion of fluid and proteins by salivary cells is under the control of parasympathetic and sympathetic autonomic nerves. In a recent study we have shown that, in the rat submandibular gland, autonomic nerves can also increase the secretion of IgA, a product of plasma cells secreted into saliva as SIgA (IgA bound to Secretory Component, the cleaved poly-immunoglobulin receptor). The present study aimed to determine if parotid secretion of SIgA is increased by autonomic nerves and to compare SIgA secretion with other parotid proteins stored and secreted by acinar and ductal cells. Assay of IgA in saliva evoked by parasympathetic nerve stimulation immediately following an extended rest period under anaesthesia indicated that it had been secreted into intraductal saliva in the absence of stimulation during the rest period. The mean rate of unstimulated IgA secretion (2.77+/-0.28 microg min(-1) g(-1)) and the 2.5-fold increase in IgA secretion evoked by parasympathetic stimulation were similar to results found previously in the rat submandibular gland. Sympathetic nerve stimulation increased SIgA secretion 2.7-fold, much less than in the submandibular gland. SDS-PAGE and Western blot analysis with anti-IgA and anti-Secretory Component antibodies confirmed that SIgA was the predominant form of IgA in saliva. Acinar-derived amylase and ductal-derived tissue kallikrein were more profoundly increased by parasympathetic and particularly sympathetic stimulation than SIgA. Overall, the results of the present study indicate that SIgA forms a prominent component of unstimulated parotid salivary protein secretion and that its secretion is similarly increased by stimulation of either autonomic nerve supply. The secretion of other parotid salivary proteins that are synthesized and stored by acinar or ductal cells is upregulated to a much greater extent by parasympathetic and particularly sympathetic stimulation.

Salivary secretion of proteins from rat submandibular glands was studied using graded stimulation of the parasympathetic nerve in isolation, and then at a fixed rate in combination with graded sympathetic nerve stimulation. Increasing the frequency of parasympathetic nerve stimulation per se caused a gradual increase in the secretion of peroxidase (from acini) but only small increases in proteinase (from ductal cells) and IgA outputs. Dual stimulations, with an increasing frequency of sympathetic nerve stimulation on a background of low frequency parasympathetic nerve stimulation, showed that maximal acinar secretion of peroxidase required only a low frequency of additional sympathetic stimulation, whereas ductal secretion of kallikrein was greatest with the highest frequency of additional sympathetic stimulation (20 Hz in bursts). IgA secretion also required high frequency additional sympathetic stimulation in bursts for greatest output. Although a synergism occurred with parasympathetic plus sympathetic nerve stimulation for the secretion of both peroxidase and kallikrein it was not evident for the secretion of IgA. This presumably reflects a difference for exocytosis of proteins stored in granules (e.g. peroxidase and kallikrein) compared to those proteins continuously transported across the plasma membrane in vesicles by transcytosis. This work confirms that vesicular movement of secretory IgA can be increased by both parasympathetic and sympathetic nerve stimulation, but the frequency parameters differ for each nerve.

Treponema denticola has been shown to be associated with periodontitis in man and animals. The organism ferments amino acids and thrives on the proteins in the periodontal pocket. Accordingly, T. denticola possesses various proteolytic enzymes, including a chymotrypsin-like protease, capable of hydrolyzing a whole range of proteins, including immunoglobulins. Yet, it is not clear whether the intact cells of T. denticola can degrade immunoglobulins and albumin. The purpose of this study was to clarify this point. Three strains of T. denticola were cultured in liquid medium, and cells were harvested by centrifugation. Protein degradation in cell suspensions was assayed by capillary electrophoresis and immunonephelometry. None of the T. denticola strains appeared to be able to degrade IgA, IgG, or albumin, while a strain of P. gingivalis completely hydrolyzed these proteins. The findings suggest that, in the periodontal pocket, T. denticola depends on proteinases from other bacteria for utilization of the available serum proteins. This is in accordance with clinical data showing a close relationship between T. denticola and strongly proteolytic bacteria, such as Porphyromonas gingivalis and Bacteroides forsythus.

Background: Helicobacter pylori is a major cause of chronic antral gastritis and peptic ulcer diseases. Many researchers have examined the possibility of immunologically-mediated prevention of H. pylori infection using an oral vaccine. The purpose of this study is to investigate whether mucosal and systemic immune responses are induced by oral immunization with H. pylori lysate-loaded poly(D, L-lactide-coglycolide)[PLG] nanoparticles, and if so, how the distribution of serum IgG subclasses are produced., Methods: PLG nanoparticles (H. pylori-PLG) with encapsulated H. pylori lysates were prepared by the solvent evaporation method, and the physical properties of the nanoparticles were investigated. Following the oral immunization of the H. pylori-PLG nanoparticles into mice, antibody induction was assayed in serum and gut washings, and the pattern of serum IgG subclasses was determined by ELISA., Results: The prepared H. pylori-PLG nanoparticles were spherical, nonporous particles with a mean diameter of less than 1 microm. The multiple oral immunization with H. pylori-PLG nanoparticles induced significantly H. pylori-specific mucosal IgA response as well as serum IgG responses. The serum antibody subclasses elicited were predominantly IgG1 and IgG2b., Conclusion: Our results suggested that oral immunization of H. pylori-PLG nanoparticles induced the H. pylori-specific mucosal and systemic responses in mice and enhanced Th2-type responses.

Problem: The transport of various proteins across the human placenta was investigated by comparing maternal and fetal concentrations of tetanus antigen (TT-AG), anti-tetanus (TT)-immunoglobulin G (IgG) (following maternal vaccination), IgA, human chorionic gonadotropin (hCG), human placental lactogen (hPL), and alpha-fetoprotein (AFP) at term., Method of Study: The concentrations of the six proteins were determined using enzyme-linked immunosorbent assay in serum of maternal venous and umbilical (fetal) vein samples obtained at delivery from uncomplicated term pregnancies (n = 16)., Results: The ratios (mean +/- standard deviation) of fetal (umbilical) to maternal level were 1.41 +/- 0.33 (anti-TT-IgG), 0.91 +/- 0.37 (TT-AG), 0.002 +/- 0.001 (IgA), 0.003 +/- 0.001 (hCG), and 0.008 +/- 0.004 (hPL), while the maternal:fetal concentration ratio of AFP was 0.002 +/- 0.002. IgA, hCG, hPL, and AFP showed a close correlation between maternal and fetal levels varying between r2 = 0.47 to 0.73 (P < 0.004-0.0001). Because AFP is produced by the fetus while IgA originates in the mother, the appearance of small amounts of these two proteins in the maternal or fetal compartment, respectively, suggests a slow rate of diffusion following a high concentration gradient. The detection of hCG and hPL in fetal serum is also interpreted as diffusion from the maternal into the fetal blood. Anti-TT-IgG has a significantly higher concentration in the fetal as compared with the maternal serum, which is in line with the well-documented active transfer of IgG. Fetal TT-antigen levels were similar to maternal concentrations, showing a close correlation (r2 = 0.74, P < 0.0001) between the two proteins., Conclusions: The correlation between maternal and fetal concentrations of various proteins like IgA (150,000 Da), hCG (42,000 Da), and hPL (21,000 Da) suggests passive diffusion of these macromolecules across the placenta from the maternal to the fetal side, albeit at a slow rate. A similar process is postulated for AFP (70,000 Da) diffusing in the opposite direction from the fetus to the mother. There was no significant difference between the transplacental fetomaternal gradient of IgA and hCG and the maternal-fetal gradient of AFP. In view of the substantially larger volume of circulating maternal as compared with fetal blood, a significantly higher rate of crossing of AFP as compared with the other proteins must be assumed. It is uncertain whether a difference in the rate of transplacental transfer in the two directions or an additional source of AFP production in the maternal compartment explains the high maternal level. Anti-TT-IgG concentration is significantly higher in fetal than in maternal serum suggesting active transfer from the mother to the fetus. Furthermore, there is considerable transfer of TT-AG and a close correlation of fetal:maternal ratios of anti-TT-IgG (150,000 Da) and TT-AG (150,000 Da) could be an indication for a specific transfer of the antigen antibody complex.

The time of appearance and pattern of expression of several alpha-granule proteins, von Willebrand factor (VWF), fibrinogen and immunoglobulins (Ig) were examined and compared in human bone marrow megakaryocytes (MK) using an immunocytochemical approach. VWF is synthesized by immature MK, whereas it has been shown that fibrinogen is incorporated from the plasma into alpha-granules. The present study was undertaken in order to determine whether there are chronological and morphological differences in the expression of VWF and fibrinogen in vivo in human MK. Seven paraffin-embedded biopsies of normal human bone marrow were labelled with specific antibodies for VWF and for fibrinogen, detected by the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. and analysed by immunomorphometry. We found a clear, statistically significant. difference in the labelling pattern of VWF and fibrinogen. The expression of other endocytosed alpha-granule proteins, immunoglobulins G and A, was therefore studied in bone marrow MK from two patients with multiple myeloma, one with monoclonal IgG and one with monoclonal IgA. The immunostaining pattern was similar to that of fibrinogen and different from VWF, and characteristic of endocytosed alpha-granule proteins. This study demonstrates that: (i) immunohistochemical staining of MK alpha-granules proteins distinguishes the peripheral cockade distribution pattern of endocytosed protein from the perinuclear pattern of endogenously synthesized proteins; (ii) VWF is present in human bone marrow MK when fibrinogen is not yet detectable: (iii) VWF synthesis ceases while fibrinogen is still being incorporated: (iv) immunoglobulins can be detected in MK cytoplasm, with a staining pattern resembling that of fibrinogen.

Socioeconomic development has led to a progressive increase of alcohol consumption in Taiwan, with an accompanying increase in alcohol-related psychiatric problems, traffic accidents, and liver disease. The prevalent rates of alcohol dependence for Han Chinese and Fomosan aborigines were 0.1% and 1%, respectively in 1950. The rate of alcohol dependence increased to 23% for aborigines in 1995. The number of cases of death and serious injuries due to alcohol-related traffic accident has decreased, and the number of fatalities resulting from these accidents has decreased from third to eighth since the inception of a program of random traffic stops with alcohol breath test in 1997. Alcohol liver disease (ALD) was defined as daily alcohol consumption of 60 g, for a duration of longer than 5 years. We classified ALD patients into two groups: (1) those whose average daily consumption of alcohol exceeded 120 g for a duration longer than 15 years (group A); and (2) all other patients (group B). The case records of 33 cases of biopsy-confirmed ALD were obtained for study. The average of daily alcohol consumption in these cases was 160 g. All but one of these patients were male, age ranged from 26 to 69 years, with an average of 43.1. Clinically, ill-defined gastrointestinal symptoms were the most common presentation (61%), and hepatomegaly was the main physical sign (73%). The average mean corpuscular volume values of ALD and non-ALD patients were 102.3 +/- 10.94 and 94.5 +/- 8.1, respectively (p < 0.01). The mean corpuscular volume values of group A and group B were 102.9 +/- 9.7 vs. 96.5 +/- 9.11 (p < 0.05). Result from serum SGOT/SGPT and gamma-glutamyltransferase/alkaline phosphatase for ALD and non-ALD revealed statistically significant differences between these groups. Using the avidin-biotin complex technique, tissue IgA deposition for ALD patients was found to be different from that of non-ALD patients. Ten of 13 ALD patients vs. 2 of 13 non-ALD patients had continuous-form IgA deposition. Histologically, 45.5% of ALD patients had alcoholic cirrhosis, whereas alcoholic hepatitis was present in only 9.1% of patients. Overall, 88% of cases showed various severity of fatty metamorphosis.

Problem: Placental transport of various proteins present in human serum, such as immunoglobulins (IgG, IgA), specific anti-tetanus IgG (anti-TT-IgG), and tetanus toxoid-antigen (TT-AG), was investigated. In addition, the transport of IgG modified with biotin (IgG-BT) and 14C-bovine serum albumin (14C-BSA, a permeability marker for macromolecules), was assessed., Method of Study: During the perfusion of an isolated cotyledon from human term placenta the perfusate was recirculated on both maternal and fetal sides. After an initial stabilisation phase of 2 hr (control phase), media on both sides were exchanged and perfusion was continued comparing two different conditions (experimental phase). In the first group (control experiments [A, n = 3]), no test proteins were added during the experimental phase (4-6 hr). In the second group (B, n = 5), during the experimental phase (6 hr) the maternal perfusion medium contained IgG (Sandoglobuline, 6-10 g/L), anti-TT-IgG (21-25 mg/L), TT-AG (0.19-0.24 mg/L), and IgA (0.13-0.19 g/L). IgG-BT (2 g/L) and 14C-BSA (30-40 nCi/ml) were added to the medium on the maternal side. IgGs and TT-AG were determined by specific enzyme-linked immunosorbent assay., Results: Both groups showed stable metabolic conditions with constant rates of glucose consumption, lactate production, and hormone (human chorionic gonadotropin, human placental lactogen) release observed throughout the experiment. Washout levels of endogenous IgG and IgA observed in the maternal circuit at the end of the control period were 5 and 1000 times higher than in the fetal circuit. In the experimental phase these levels remained constant at 50-80% of control levels with no change in the last 4 hr of perfusion (group A). In group B, with addition of extra proteins, trace amounts of IgG-BT, IgA, and 14C-BSA were detectable in the fetal circuit within 1 hr, with no significant further increase in circulating levels in the following 4 hr of the perfusion. In contrast, the detection of IgGs in the fetal circuit was delayed by 2 hr; thereafter, a continuous linear increase was observed for all IgGs. TT-AG in fetal perfusate was below the detection limit. TT-AG was found on the fetal side only after ultrafiltration of samples obtained at the end of the experiment. For permeability comparison, the ratio between concentrations on the fetal and maternal side multiplied by 100 ([F:M] x 100), as detected after 6 hr of perfusion, was assessed (n = 5, mean +/- SD). Labelling of IgG with biotin (IgG-BT) reduced its placental transfer by a factor 10 (0.04 +/- 0.01) when compared with the natural IgG (0.49 +/- 0.08) or the specific antibody (anti-TT-IgG). The relative fetal-to-maternal ratio found for TT-AG (0.48 +/- 0.12) was similar to anti-TT-IgG (0.46 +/- 0.11), and approximately 4 and 50 times that of 14C-BSA (0.12 +/- 0.03) and IgA (0.01 +/- 0.01), respectively. Considering that the molecular weights of TT-AG and anti-TT-IgG were at least twice that of BSA and similar to IgA, the difference in transfer suggests a specific mechanism of transport., Conclusions: Compared with other proteins there is a significantly increased transfer of IgGs across the in vitro perfused human placenta from the maternal to the fetal side, indicating a specific transport mechanism. The similarity in transfer of anti-TT-IgG and tetanus antigen may suggest the transport as antibody-antigen complex.

Thrombocytopenia frequently occurs immediately after orthotopic liver transplantation. We have investigated the cause of this phenomenon in a cohort of 45 consecutive liver transplant recipients. The median preoperative platelet count (range) of 129 x 10(9)/l (14-719) fell to 56 x 10(9)/l (23-334) by the fourth postoperative day. The median preoperative reticulated platelet percentage (range) of 6.7% (2.2-23.9) increased to 16.4% (4.6-40.8) on day 7. There was a significant rise in prothrombin fragment F1.2 by the first postoperative day which was followed by rises in fibrinogen and fibrin degradation products. There was no increase in platelet-associated immunoglobulin or markers of endothelial activation. We conclude that there is an increased rate of platelet consumption associated with thrombin generation that reflects the magnitude of liver transplant surgery.

Objective: A qualitative and quantitative analysis of the functional, antigen-specific B cell receptor repertoire of patients with rheumatoid arthritis (RA) in synovial and peripheral compartments., Methods: B cells were activated to grow and differentiate at high efficiency in vitro under limiting-dilution conditions. Isotype and specificity of the secreted Ig were tested by enzyme-linked immunosorbent assay., Results: In contrast to peripheral B cells, most synovial B cells had already switched to IgG/IgA in vivo. The frequencies of B cells specifically recognizing foreign antigens were decreased within the synovial population, whereas the frequencies of B cells specific for type II collagen, mycobacterial heat-shock protein 60 (hsp60), or IgG Fc fragments were significantly increased, revealing a negative correlation in terms of frequencies., Conclusion: B cells specific for human type II collagen, hsp60, and IgG Fc fragments are produced and/or expanded locally within the affected joints of RA patients. Thus, the specific immune system is definitely involved in the local inflammatory and destructive processes.

The production of IgA protease in twelve strains of Streptococcus sanguis isolated from patients with Behcet's disease (BD) was examined. Protease activity was detected in 10 out of 12 strains. The protease was purified from one representative strain, S. sanguis 113-20, by employing Rotofor and DEAE-Sephacel chromatography. The molecular mass of the purified protease was approximately 100 kDa, and it cleaved the proline-threonine site of the IgA. Both IgG and IgA titers against the cells (113-20) and the purified IgA protease in the sera of BD patients and healthy controls, 36 each, were assayed. The IgG titers against the cells and protease were not significant in the BD patients or controls, but the IgA titers against the cells and protease in the BD patients were significantly higher than those of the controls. These data indicate that the BD patients are infected with IgA protease-producing S. sanguis strains, which cause an increase of IgA titer against these organisms and IgA protease antigen. Since the organisms can proliferate in BD patients for a long period of time (years), it seems that IgA antibodies cannot effectively eliminate the organisms.

Ten calves were used to elucidate the ultrastructure of enterocytes before and 24 h after colostral intake. Tissue samples were obtained from duodenum, jejunum (5 locations) and ileum. Protein A-gold technique was applied to immunoelectron-microscopically demonstrate colostral IgA. The prominent feature of the precolostral enterocytes are intracytoplasmic vacuoles. The frequency of vacuoles increases from cranial jejunum to ileum and from the villi bases to the tips. The appearance of absorptive vacuoles after colostral administration correlates with the incidence of precolostral empty vacuoles. Bovine IgA was detected in absorptive vacuoles and within the intestinal lumen of postcolostral calves. In addition to a diffuse IgA labelling of most vacuoles, a few corresponding enterocytic vacuoles labelled inhomogenously or negatively. This study demonstrates morphologically that the main site of colostral absorption is the middle-to-caudal region of the small intestine. Immunoelectron microscopy of IgA labelling provides indications of a selective IgA absorption in addition to pinocytosis.