Intracellular cytokine production by Th1/Th2 lymphocytes and monocytes of children with symptomatic transient hypogammaglobulinaemia of infancy (THI) and selective IgA deficiency (SIgAD)

Intracellular expression of several cytokines was assessed in lymphocytes and monocytes of children with transient hypogammaglobulinaemia of infancy (THI) and selective IgA deficiency (SIgAD). THI was characterized by an increased frequency of CD3+/CD4+ lymphocytes expressing tumour necrosis factor alpha (TNF-alpha), TNF-beta and interleukin 10 (IL-10), while in SIgAD elevated numbers of these cells containing TNF-alpha and interferon gamma (IFN-gamma) were observed. No changes in the number of CD4+ T cells expressing IL-4 in both diseases were noted. The proportion of CD33+ monocytes containing TNF-alpha both in THI and SIgAD was unchanged. The secretion of IL-12 by peripheral blood mononuclear cells (PBMCs) of patients with THI and SIgAD was significantly elevated and associated with an increased frequency of IL-12 expressing monocytes in THI but not in SIgAD. IL-18 secretion was slightly, but not significantly, elevated in both diseases. Intracellular Th1 and Th2 type cytokines within CD3+/CD4+ lymphocytes were also determined in the normal blood donors that showed high or low production of IgG and IgA in vitro. In low producers of IgG an increased proportion of CD3+/CD4+ cells expressing TNF-alpha and IFN-gamma was found, while in low IgA responders only elevated TNF-alpha positive CD3+/CD4+ cells were observed. These results suggest that THI and SIgAD may represent diseases with an excessive Th1 type response that is associated with an up-regulation of IL-12 secretion and, at least in THI, elevated numbers of monocytes expressing intracellular IL-12. Up-regulation of IL-12 may be the essential factor in the patomechanism(s) of these diseases as already described in common variable immunodeficiency (CVID).     

Different familial association patterns of autoimmune diseases between juvenile-onset systemic lupus erythematosus and juvenile rheumatoid arthritis.

The aim of this study was to determine if the prevalence of autoimmune disorders in the relatives of patients with systemic lupus erythematosus (SLE) is greater than that of relatives of patients with juvenile rheumatoid arthritis (JRA). Interviews were used to obtain histories of the following autoimmune disorders among living or deceased first-, second-, and third-degree relatives of 91 SLE and 110 JRA families: ankylosing spondylitis, SLE, rheumatoid arthritis (RA), JRA, multiple sclerosis, juvenile dermatomyositis, Sj?gren's syndrome, myasthenia gravis, psoriasis, and thyroid diseases. There were statistically significant differences between the SLE and JRA probands in mean age and gender ratio (19.1 +/- 4.8 vs 14.0 +/- 5.5 years; M (male)/F (female): 17/74 vs 62/48, p<0.005). The prevalence rate of autoimmune diseases in relatives of SLE families (20.9%) was greater than in JRA families (11.8%), but not statistically significantly so. The mean age (18.0 +/- 5.3 vs 14.0 +/- 4.3 years), mean age at diagnosis (13.4 +/- 4.3 vs 7.9 +/- 3.9 years) and gender ratio (F/M, 16/3 vs 5/8) of the patients with affected relatives between these 2 groups all had statistically significant differences. A higher prevalence of SLE in relatives was found in SLE families than in JRA cases. Furthermore, this study revealed a higher incidence of autoimmune disorders among second- and third-degree relatives of SLE or JRA probands versus first-degree ones, especially sisters (including 1 pair of twins) and the maternal aunt in SLE families. These data demonstrate that the prevalence of autoimmune disorders in the relatives of patients with SLE is greater than those of relatives of patients with JRA. This suggests that clinically different autoimmune phenotypes may share common susceptibility genes, which may act as risk factors for autoimmunity.     

Serum IgE levels in patients with selective IgA deficiency.

In this study serum IgE levels were measured by a double-antibody radioimmunoassay in 31 patients with serum IgA concentration less than 0.01 mg/ml who were followed in the arthritis and allergy clinics. On a group basis there was no significant difference in mean serum IgE levels between the IgA deficient patients and normal subjects of the same age. However, in the absence of atopic disease, IgA deficient patients had significantly lower serum IgE levels. When atopy was associated with IgA deficiency IgE levels were the same as in the normal subjects but significantly lower than those of atopic non-IgA deficient patients. IgE levels in those with recurrent respiratory tract infection were not different. Adults with anti-IgA antibodies had significantly lower IgE values. IgE levels in patients with RA, JRA or SLE were not significantly different. Selective IgA deficient patients may have a relative deficiency of serum IgE depending on the comparison group.     

Autoantibodies to a regulatory T cell subset in human ageing.

Anti-T cell autoantibodies were detected in some aged humans. Non-immunoglobulin-bearing (Ig-) cells were isolated from the peripheral blood of normal human donors by negative selection through Ficoll, using sheep erythrocytes coated with rabbit anti-human Ig. The Ig- cells were then reacted with sera from 83 individuals ranging in age from 60 to 99 years; 36% of the serum samples were noticeably reactive with the Ig- cells (average reactivity 28%). The peripheral blood lymphocytes from some of the aged individuals were also tested for levels of Ig-secreting cells in a reverse haemolytic plaque assay; there was a six- to eight-fold increase in the number of plaque-forming cells (PFC) from those individuals whose sera contained appreciable amounts of anti-T cell antibody, as compared with those whose sera contained little or no anti-T cell antibody. Isolated Ig- cells from these individuals were also examined for the presence of regulatory T cell subsets, using sera from juvenile rheumatoid arthritis (JRA) patients. The Ig- cells from the subjects who had no detectable anti-T cell antibodies in their sera and near normal PFC levels were reactive with the JRA sera, whereas the Ig- cells from individuals with increased numbers of PFC and with serum anti-T cell antibodies were only slightly reactive with the JRA sera. These data suggest that a majority of the regulatory JRA+ subset of T cells had been lost in the latter group. When sera from aged individuals containing anti-T cell autoantibodies were reacted with JRA-, Ig- cells isolated from a normal human donor, little positive reactivity was seen, indicating that the autoantibodies in sera from aged humans and from some JRA patients are directed against similar T cell subsets.     

Antibodies to basement membrane proteins nidogen and laminin in sera from streptococcal-related diseases and juvenile rheumatoid arthritis patients.

Using the ELISA technique, antibodies against two different basement proteins, laminin and nidogen (ALNA), were determined in 226 children suffering from one of 37 different inflammatory or infectious diseases. These included 80 patients with streptococcal infection and 40 with juvenile rheumatoid arthritis. Forty-eight percent of the streptococcus-infected patients (or 75% of those in the acute phase) and 60% of juvenile rheumatoid arthritis patients had significantly elevated ALNA levels compared with healthy controls. Interestingly 10 adult rheumatoid arthritis patients displayed normal ALNA levels, suggesting a particular immune process occurring in children affected by juvenile rheumatoid arthritis. By means of periodate oxidation and glycosidase treatments we have shown that ALNA positive sera recognized terminal alpha-galactose as the reactive epitope.     

Comparison of the frequency of atopic diseases in children with severe and partial IgA deficiency

Similar frequencies of atopic diseases and of elevated total serum IgE levels were observed in 40 children with serum IgA levels below 5 mg/dl and absence of salivary IgA (severe selective IgA deficiency, SIgAD) and in 40 children with serum IgA levels above 5 mg/dl but below -2 SD of age-normal mean values and presence of salivary IgA (partial SIgAD). These findings suggest that the absence of secretory IgA, which has been postulated to play a protective role by excluding allergens at the mucosal level, does not appear to play a crucial role in the pathogenesis of atopic diseases.     

Immunoregulatory aberrations in patients with polyarticular juvenile rheumatoid arthritis

The presence of hypergammaglobulinemia and various circulating autoantibodies in children with polyarticular juvenile rheumatoid arthritis (JRA) implies an immunoregulatory disorder. We report here experiments planned to elucidate the underlying cellular aberrations in this disease. Twelve children with polyarticular JRA were studied. Percentages of Leu-1, Leu-2, and Leu 3 T cells were comparable to those of normal individuals. Immunofluorescent double staining studies demonstrated elevated numbers of activated (DR+) T cells of both Leu-2 and Leu-3 phenotype. B cells characterized both phenotypically (Leu-12) and functionally (as spontaneous plaque-forming cells, PFC) were elevated. In vitro PFC responses to pokeweed mitogen (PWM) and Epstein-Barr virus (EBV) were diminished. The levels of concanavalin A-induced suppressor cells of the PWM-stimulated PFC responses were comparable to control values. In contrast, the EBV-associated suppressor T cells were significantly impaired in both EBV-seropositive and EBV-seronegative patients. These studies indicate that peripheral blood B-cell activity is abnormal in polyarticular JRA. Defective T-cell responses in vitro suggest that this may be due to disruption of normal regulatory circuits between B and T cells and may contribute to the pathogenesis of this disease.     

Expression of the major rheumatoid factor cross-reactive idiotype in pediatric patients with systemic lupus erythematosus

Rheumatoid factor cross-reactive idiotype (RF-CRI) is expressed in high concentrations in the sera of some patients with rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). To determine if RF-CRI is specifically expressed in rheumatic disease or if it is secondary to polyclonal B-cell activation, we examined sera of 23 children with SLE, 16 adolescents with infectious mononucleosis (IM), and age-matched pediatric controls for RF-CRI expression. Concentrations of RF-CRI in serum, determined by an inhibition ELISA, were 24 +/- 17 micrograms/ml (mean +/- SD) in 25 normal children, 31 +/- 17 in 16 young adults with IM, and were significantly increased, 70 +/- 80 micrograms/ml, in the 23 children with SLE (p less than 0.036). Eleven of 23 SLE patients had serum RF-CRI greater than the mean +/- 2 SD for normal children. Ten of 23 SLE sera contained IgM rheumatoid factor (RF) activity. One patient with IM had a borderline elevated RF-CRI level, and 5 IM patients had RF in their sera. The serum IgM concentrations in sera were: SLE (192 +/- 93 mg/dl) and IM (234 +/- 77 mg/dl) sera. These levels were significantly elevated compared to controls (132 +/- 44 mg/dl), p less than 0.031 for SLE and p less than 0.001 for IM, suggesting that polyclonal activation of B cells was present in SLE and IM patient groups. Increased expression of RF-CRI in the SLE patients correlated directly with high titer anti-DNA antibody values (r = 0.3965, p less than 0.05) and RF activity when human IgG (r = 0.5026, p less than 0.05) was used as the RF binding substrate and inversely with serum C3 levels (r = 0.3925, p less than 0.05). RF-CRI expression did not correlate with RF that bound rabbit (r = 0.3123, p greater than 0.05). Increased serum RF-CRI expression is not a result of polyclonal B-cell activation. RF-CRI may be selectively up-regulated in patients with SLE.     

The effect of pregnancy on ankylosing spondylitis, psoriatic arthritis, and juvenile rheumatoid arthritis.

It has long been established that rheumatoid arthritis improves during pregnancy. The gestational course of other inflammatory arthritides like ankylosing spondylitis (AS), psoriatic arthritis (PsA), and juvenile rheumatoid arthritis (JRA) has been less well studied. The present review summarizes the results of our retrospective and prospective studies on the interaction between these diseases and pregnancy. The results showed clear differences for their gestational course. Patients with PsA improved or even remitted in 80% of the pregnancies, whereas 80% of the AS patients had unaltered or aggravated disease symptoms. The 20% of AS patients who markedly improved while pregnant all had AS with accompanying diseases like psoriasis, ulcerative arthritis, or small joint arthritis. Quiescent JRA was not reactivated by pregnancy, and active disease at conception ameliorated in about 60%. Fetal outcome was not adversely affected by AS, PsA, or JRA nor did there occur serious intercurrent diseases during pregnancy. In AS and PsA patients delivery was mainly uncomplicated. Sequelae of JRA were a frequent cause for cesarean section in JRA patients. A postpartum flare during the first 3 months after delivery occurred in about 90% of the AS pregnancies, 70% of the PsA pregnancies, and about 50% of the JRA pregnancies.     

Distribution of Borrelia burgdorferi specific antibody among patients with juvenile rheumatoid arthritis in Korea.

Lyme disease, a multi-systemic infection occurring worldwide, has yet to be reported in Korea, although the spirochete B. burgdorferi, known as the causative organism of the disease, has recently been isolated from the vector tick Ixodes persulcatus in the region. To contribute to revealing whether Lyme disease exists in Korea or not, B. burgdorferi specific antibodies (IgG, IgM, and/or IgA) were measured by three individual enzyme-linked immunosorbent assays (ELISA) utilizing different antigens in 38 patients with juvenile rheumatoid arthritis (JRA) which shares a number of clinical features with Lyme arthritis. The antibody prevalence rates in patients with JRA were various depending on the antigens (21% of IgG and IgM antibodies to purified organisms, 0% for IgG antibody to purified native flagella, and 5% for IgG, IgM, and IgA antibodies to recombinant p39) and were not different compared to 39 controls (21%, 0%, and 0% respectively). The antibody prevalence rates compared in various subgroups of patients with JRA according to types of JRA, length of illness, age, and sex were not different. Comparing the three different antigens, the greatest number of positive responders were yielded by purified organisms followed by p39 and purified flagellin, however the possibility of nonspecificity with purified organisms remained. The data indicate that serologic tests using ELISA fail to illustrate Lyme disease among 38 patients with JRA in Korea.     

Prevalence of anti-Fab antibodies in patients with autoimmune and infectious diseases.

Sensitive ELISA were devised to examine the specificity of circulating IgM and IgA autoantibodies for whole human IgG, Fc and Fab fragments of human IgG. Sera from patients with autoimmune and infectious conditions such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), tuberculosis (TB), infectious mononucleosis (IM) and cystic fibrosis (CF) were studied. Results of the ELISA assays using whole human IgG as antigen revealed that a proportion of patients in each of the groups studied had circulating IgM and IgA rheumatoid factors (RF). Fifteen normal individuals studied were negative. In the latex positive RA group, IgM RF and IgA RF had primarily anti-Fc reactivity (100% and 93% respectively), although 3/15 patients also showed IgM anti-Fab reactivity and one patient had high IgA anti-Fab activity. Patients with SLE and TB who had detectable RF levels also revealed predominantly anti-Fc specificity. In contrast, examination of 25 patients with IM showed positivity for IgM RF activity in 8% of patients using whole IgG as antigen, 24% positivity using purified Fc fragments as antigen and 45% positivity when plates were coated with Fab fragments. Similarly, a large number of CF patients (54%) also showed predominantly IgM anti-Fab activity. Of interest, 69% of the CF patients who were all studied at the time of bacterial infection had detectable IgA RF levels, with 46% of these patients showing both IgA anti-Fc and anti-Fab activity. These findings suggest that autoantibody specificities in autoimmune and infectious diseases are different.     

Quantification of cross-reactive idiotype-positive rheumatoid factor produced in autoimmune rheumatic diseases. An indicator of clonality and B cell proliferative mechanisms.

The aetiology of sustained autoantibody production in human autoimmune diseases is unknown. Evidence for structural similarities and common clonal origin among autoantibodies have been demonstrated through the expression of cross-reactive idiotype (CRI). In the present study we use four monoclonal antibodies (MoAbs) with specificity for non-overlapping CRI on human rheumatoid factor (RF) autoantibodies to define the structural features of polyclonal RF characteristic of patients with autoimmune rheumatic diseases. The pattern of CRI expression in the serum of 12 patients with rheumatoid arthritis (RA), eight with systemic lupus erythematosus (SLE) and 20 with primary Sjögren''s syndrome and 34 normal individuals were determined in parallel with the level of IgM RF, IgA RF and autoantibodies to the cellular antigens SS-A, SS-B, Sm, nRNP and dsDNA and cryoglobulins. The results demonstrate significant elevation in the level of IgM and IgA expressing VHI (G6 and G8) and VHIII (B6 and D12) associated CRI in the serum of patients with autoimmune rheumatic diseases compared with normal individuals. These increases paralleled, but did not equal the increase in the level of immunoglobulins and RF. However, when expressed as proportion of immunoglobulin, only the VHI-associated CRI were significantly elevated in patients compared with normal individuals. The proportion of IgM RF expressing the VHI-associated CRI was higher in patients with Sjögren''s syndrome compared with SLE and RA. Furthermore, the proportion of IgA RF expressing the G6 CRI was higher than G6+ IgM RF. These findings imply that different mechanisms contribute to RF production in autoimmune diseases. It is suggested that polyconal B cell activation is likely to be a contributing mechanism. However, such polyclonal activation is unlikely to be random since a selective elevation in the level of specific autoantibodies and VHI-associated CRI is observed. Furthermore, the data demonstrate that a proportion of autoantibodies in autoimmune diseases are immunoglobulin germline gene encoded. This is more evident in some patients with primary Sjögren''s syndrome, where RF is likely to be oligoclonal or monoclonal in individuals with lymphoproliferation.     

Immunity to ocular and collagen antigens in childhood arthritis and uveitis

Humoral and cellular immunity to ocular antigens (S antigen and alpha, beta heavy, beta light, and gamma crystallins) and connective tissue antigens (type I, II, III, and IV collagens) were studied in children with juvenile rheumatoid arthritis (JRA) with or without uveitis and in controls. There was no association between the presence of uveitis and antibody to any collagen type or of lymphocyte transformation to type II collagen. Antibodies to all crystallins were more common in children with JRA than in controls, and antibodies to BH crystallins were more frequent in children with JRA and uveitis than in those with JRA alone. Such reactivity did not appear to represent antibody cross-reactive with collagen antigens. Lymphocyte proliferation to alpha crystallin was increased in JRA with or without uveitis, whereas lymphocyte response to S antigen was associated with the presence of uveitis.     

The level of IgA antibodies to human umbilical vein endothelial cells can be enhanced by TNF-alpha treatment in children with Henoch-Schönlein purpura

Anti‐endothelial cell antibodies (AECA) have been found to play an important role in many vascular disorders. In order to determine the presence of AECA in children with Henoch–Schönlein purpura (HSP), and to elucidate the pathogenic and clinical value of their measurement in this disease, AECA were detected by immunofluorescence staining and a human umbilical vein endothelial cell (HUVEC)‐based enzyme‐linked immunosorbent assay (ELISA) in 20 children with HSP, 10 children with juvenile rheumatoid arthritis (JRA) without vasculitis and 10 normal healthy children. Antibodies against another endothelial cells, human dermal microvascular endothelial cells (HMVEC‐d) were also detected by cell‐based ELISA. In some experiments, we compared the binding activity of antibodies to HUVEC with and without tumour necrosis factor‐α (TNF‐α) or interleukin‐1 (IL‐1) pretreatment. Patients with acute onset of HSP had higher serum levels of IgA antibodies, both against HUVEC and against HMVEC‐d, than healthy controls (P = 0·001, P = 0·008, respectively). Forty‐five per cent of patients had positive IgA AECA to HUVEC, and 35% had positive IgA AECA to HMVEC‐d. The titres of IgA antibodies to HUVEC paralleled the disease activity. After TNF‐α treatment, the values of IgA AECA to HUVEC in HSP patients were significantly increased (P = 0·02). For IgG and IgM AECA, there was no difference between HSP patients and controls (P = 0·51, P = 0·91). Ten JRA children without vasculitis had no detectable IgG, IgM or IgA AECA activity. The results of this study showed that children with HSP had IgA AECA, which were enhanced by TNF‐α treatment. Although the role of these antibodies is not clear, IgA AECA provide another immunological clue for the understanding of HSP.     

The clinical associations of antinuclear antibodies in juvenile rheumatoid arthritis

To clarify further the clinical correlates of antinuclear antibodies (ANA) in children with juvenile rheumatoid arthritis (JRA) this study compared the features of 60 ANA positive and 25 ANA negative children with JRA. ANA was more likely to be present in those with pauciarticular JRA than polyarticular JRA particularly if the ANA was of high titer. ANA positive subjects were more likely to have extraarticular manifestations, especially iridocyclitis. No significant differences were observed in onset ages, sex distribution, season of disease onset, family histories, or prognosis. There was no correlation between ANA titer and disease activity. Thus, while certain clinical features do correlate with ANA positivity in JRA, most clinical manifestations do not occur with distinctively different frequencies in the ANA negative and ANA positive groups.     

Juvenile rheumatoid arthritis: cellular hypersensitivity and selective IgA deficiency

Although humoral immune mechanisms are currently thought to be of pathogenetic significance in juvenile rheumatoid arthritis (JRA), little is known about the role of cellular hypersensitivity in this disease. A possible association between abnormalities of humoral and cellular immunity exists in patients with ataxia-telangiectasia, who may have absent IgA, abnormal delayed hypersensitivity, or both. As IgA deficiency has been noted in 2–3% of patients with JRA, we have studied selected aspects of humoral and cellular hypersensitivity in patients with JRA and IgA deficiency and in patients with JRA and normal IgA levels. All patients had normal serum levels of complement, IgG, IgM, and IgD.

Cellular hypersensitivity was evaluated by cutaneous delayed-type hypersensitivity, in vitro migration inhibitory factor production, and antigen induced 3H-thymidine incorporation by lymphocytes using Candida and Streptokinase–Streptodornase antigens. Two of four IgA deficient patients had positive in vitro but negative in vivo responses to antigens. Seven of fourteen JRA patients with normal immunoglobulin levels exhibited a similar dissociation of in vivo and in vitro manifestations of delayed hypersensitivity. This pattern of cellular immune response was associated with activity and chronicity of disease; it was independent of IgA deficiency.

     

Detection of antibodies to streptococcal mucopeptide in patients with rheumatic disorders and normal controls

Bacterial mucopeptide is an integral part of bacterial cell walls and is therefore ubiquitous in our environment. An enhanced degree of humoral immunity has ben detected not only in patients with acute rheumatic fever (ARF), with a known recent response to streptococci, but also in patients with adult and juvenile rheumatoid arthritis (RA and JRA). Our studies confirmed this association with ARF and JRA using a precipitin system as well as a radioimmunoassay to detect IgG anti-mucopeptide antibodies. In those with adult RA, either IgM or IgA rheumatoid factors or IgM or IgA antibodies specific for mucopeptide were responsible for the increased incidence of precipitins to mucopeptide in the RA patients detected in this and other studies. No differences in the specificities of the anti-mucopeptide antibodies were noted between the various patient populations as there were no lines of partial identity or nonidentity when examined by Ouchterlony double diffusion analyses. Additionally, no differences of anti-mucopeptide antibody were observed when the sera from these same patient populations were examined employing inhibition studies utilizing N-acetylglucosamine and rhamnose.     

Complement activation by antibodies to DNA in systemic lupus erythematosus measured by enzyme immunoassay

An enzyme immunoassay to detect complement-fixing antibodies to DNA (CF-antiDNA) was developed. Of SLE sera, 64% had these antibodies as did 6% of 50 rheumatoid arthritis and 3.2% of 93 normal human sera. The mean CF-antiDNA level was higher in the sera of SLE patients with renal disease than those SLE patients who had no renal disease (P less than 0.0001), and higher in those SLE patients with active rather than inactive renal disease (P = 0.006). CF-antiDNA was more closely associated with renal activity than total IgG-antiDNA or CH50. These observations suggest that both the quality and quantity of anti-DNA antibodies play a role in the pathogenesis of renal disease, and that modern enzyme immunoassays help distinguish the relative importance of complement-fixing antibodies to anti-DNA from that of total anti-DNA.