Immunological Profile in Children with Minimal Change Nephrotic Syndrome

ABSTRACT. Peripheral blood from patients with active stage of minimal change nephrotic syndrome (MCNS) was examined for concanavalin A (ConA)-inducible suppressor T cell activity, proliferative response to phytohemagglutinin (PHA) and in the autologous (AMLR) and allogeneic (MLR) mixed lymphocyte reaction, proportions of T cells with receptors for IgM (Tu) or IgG (Tγ) and the levels of serum immunoglobulin M, G and A. Six of 9 patients with MCNS studies showed deficiency of ConA-induced suppressor cell activity. In the AMLR, only one of 9 patients with MCNS demonstrated depressed proliferative response (p<0.05). In the allogeneic MLR, T cells from 5 of 9 patients with MCNS demonstrated poor proliferative response when stimulated with normal control non-T cells. Five of 9 patients with MCNS had depressed proliferative response to PHA. The proportion of total T cells, Tu cells and Ty cells in the patient group were comparable to healthy control group. Serum IgG was significantly decreased in 7 of 11 patients. This study demonstrates multiple immunological abnormalities in patients with MCNS that might play a role in its pathogenesis.     

Immunological profile in children with minimal change nephrotic syndrome

Peripheral blood from patients with active stage of minimal change nephrotic syndrome (MCNS) was examined for concanavalin A (ConA)-inducible suppressor T cell activity, proliferative response to phytohemagglutinin (PHA) and in the autologous (AMLR) and allogeneic (MLR) mixed lymphocyte reaction, proportions of T cells with receptors for IgM (Tu) or IgG (T gamma) and the levels of serum immunoglobulin M, G and A. Six of 9 patients with MCNS studies showed deficiency of ConA-induced suppressor cell activity. In the AMLR, only one of 9 patients with MCNS demonstrated depressed proliferative response (p less than 0.05). In the allogeneic MLR, T cells from 5 of 9 patients with MCNS demonstrated poor proliferative response when stimulated with normal control non-T cells. Five of 9 patients with MCNS had depressed proliferative response to PHA. The proportion of total T cells, Tu cells and T gamma cells in the patient group were comparable to healthy control group. Serum IgG was significantly decreased in 7 of 11 patients. This study demonstrates multiple immunological abnormalities in patients with MCNS that might play a role in its pathogenesis.     

T cell subsets in glomerulonephritis

Abnormal lymphocyte function has been postulated to have a pathogenetic role in nephrotic syndrome. In an attempt to investigate the pathogenetic role of lymphocyte subsets in human glomerular disease, we studied 110 children suffering from nephritis during the acute nephrotic phase or nephritis without steroid treatment, 4 weeks later after steroid treatment, in remission and relapse. These patients included minimal change nephrotic syndrome (MCNS) 15 cases, focal segmental glomerular sclerosis (FGS) 6 cases, mesangial cell proliferative nephropathy (MesPGN) 42 cases, membranoproliferative glomerulonephritis (MPGN) 2 cases, hepatitis B surface antigenemia associated with membranous nephropathy (HBVMN) 10 cases, IgA mesangial nephropathy (IgAN) without nephrotic syndrome 7 cases, poststreptococcal glomerulonephritis (PSGN) 24 cases and chronic glomerulonephritis (CGN) 4 cases. There was no significant difference in the total lymphocyte count of each different pathological group of nephritis except that lymphopenia was noted in the CGN patients. When the lymphocyte phenotypic profile was examined, OKT8 cells were significantly increased in the MesPGN patients and both OKT4 and OKT8 cells were significantly increased in HBVMN. Comparison of MCNS and MesPGN during the acute nephrotic phase showed the OKT4/OKT8 ratio decreased significantly in MesPGN. Four weeks after steroid treatment, OKT4 cells decreased both in MCNS and MesPGN being pronounced in MCNS. In the remission stage with steroid treatment the OKT4/OKT8 ratio decreased in MCNS and was mildly elevated in MesPGN. In relapse, the OKT4/OKT8 ratio was the same as it was during the onset of nephrotic phase. MCNS cases were steroid responsive whereas in MesPGN there were frequent relapses or partial steroid response.(ABSTRACT TRUNCATED AT 250 WORDS)     

Selective enhancement of human IgE production in vitro by synergy of pokeweed mitogen and mercuric chloride

Selective stimulation of IgE production by pokeweed mitogen (PWM) plus mercuric chloride (HgCl₂) was further investigated in peripheral blood mononuclear cells (MNC) from non-atopic donors, who are high and low responders to PWM stimulation with respect to IgG, IgM and IgA production, and from various patients with elevated serum IgE levels. Non-atopic high responders showed a strong selective increase in IgE plaque forming cells (PFC) without changes in IgG, IgM and IgA PFC, whereas low responders showed a slight increase in IgE PFC only when B cells were co-cultured with mitomycin C (MMC)-treated T cells. In patients with elevated serum IgE levels, PWM plus HgCl₂ caused a variable selective increase in IgE PFC, however, there was no correlation between IgE PFC production and serum IgE levels.     

T Cell Subset and Steroid Sensitivity in Idiopathic Nephrotic Syndrome in Children

The relationship between T cell subset of peripheral blood lymphocytes and steroid sensitivity was studied in children with idiopathic nephrotic syndrome (INS). The subjects were 28 INS children, aged 2 to 16 years. T cells bearing receptors for IgG (Tγ) were identified by double rosette assay with sheep erythrocytes and IgG-sensitized ox erythrocytes. T cells bearing receptors for IgM (Tμ) were identified by IgM-sensitized ox erythrocytes rosette assay in purified T cells after incubating overnight. In steroid sensitive INS children, decreased numbers of Tγ cells and increased numbers of Tμ cells in onset or relapse were found and they were normalized with corticosteroid administration. Tγ cells of steroid nonsensitive INS children remained decreased even after steroid therapy. Tγ cells have been suggested to be, at least in part, suppressor T cells. Alteration of peripheral blood Tμ cells in this observation therefore may suggest the abnormality of suppressor T cells in INS children.     

Transient remission after intercurrent measles infection in a patient with hyperimmunoglobulin E syndrome

A 5-yr-old patient with hyper IgE syndrome contracted measles. This was accompanied by a temporary disappearance of his skin lesions. The patient had a long history of recurrent infections, chronic eczematoid pruritic dermatitis, and elevation of serum IgE level since infancy. Immunologic studies revealed decreased suppressor T cells (OKT8 cells) with increased IKT4/OKT8 ratio, defect in suppressor T cell function, and decreased chemotactic index. In February 1985, when he developed an interrcurrent measles infection at age of 5, the eczematoid pruritic dermatitis disappeared completely and immunologic defects improved transiently, with normalization of OKT4/OKT8 ratio, decrease in in vitro IgE synthesis, in vivo serum IgE level, and interleukin-2 production, decreases in IgG Fc receptor-bearing cells and autologous mixed lymphocyte reaction, and normalization of chemotactic index. One month later, the eczematoid skin lesion relapsed and immunologic defects reappeared. These studies suggested that the pathogenesis of hyper IgE syndrome involved a hypofunction of suppressor T cell. The transient remission associated with measles infection is probably related to the effect of the virus on the helper T cells, resulting in a normalization of OKT4/OKT8 ratio and IgE production.     

Lymphocyte subpopulations in primary immunodeficiency disorders.

Venous blood mononuclear cells from 42 children with primary immunodeficiency disorders and from controls matched for age were studied for lymphocyte subpopulations by E rosetting, surface immunoglobulin, and a panel of anti T cell monoclonal antibodies (OKT series). In 3 cases of severe combined immunodeficiency (SCID) due to adenosine deaminase deficiency, very few circulating T or B cells were found. The other 7 cases of SCID all had normal or, in 3 cases, very high numbers of circulating B cells, but in 6 of these very few cells showed T cell markers. One child had very high numbers of B cells and T cells with an immature pattern of reactivity similar to that found on common thymocytes. In T cell deficient children no consistent pattern was found, but in those with cartilage hair hypoplasia with immunodeficiency there was a low helper (OKT4) to suppressor (OKT8) ratio and high numbers of circulating OKT10 positive cells. In cases of X-linked agammaglobulinaemia circulating B cells were not found but the pattern of T cell markers was normal. In cases of common variable hypogammaglobulinaemia there was a wide scatter of helper (OKT4) to suppressor (OKT8) cell ratios. Five children were studied before and after treatment with the synthetic thymic hormone preparation TP5. There were appreciable alterations in the pattern of staining with anti T cell monoclonal antibodies in 4 of these cases, but in 1 case only was this accompanied by improvements in clinical and immune function.     

Selective IgA deficiency

Of 140 patients referred to the Pediatric Immunology Clinic during of 12-month period with the symptoms of recurrent infections or allergic respiratory illness, 21 (75%) were found to have selective IgA deficiency defined as serum concentration ≤ 5 mg. with normal levels of IgG and IgM. T lymphocyte number was reduced in the patients where as B cells with surface membrane IgA were increased. Autoantibodies and circulatory immune complexes were found more often in IgA-deficient subjects than in controls. Follow-up beyond the age of 9 years showed a spontaneous increase in serum IgA in 6, whereas 15 continued to have IgA deficiency. The latter group of children were characterized by more frequent infections, a higher prevalence of atopic disease, lower T cell count and serum IgG concentration, higher serum IgE level and a higher prevalence of food antibodies and immune complexes. These observations highlight the natural history and immunologic features of selective IgA deficiency.     

Serum anti-streptococcal IgA,IgG and IgM antibodies in IgA-associated diseases

Serum anti-streptolysin-O antibody (ASO) and anti-streptococcal polysaccharide antibody (ASP) of IgA, IgG and IgM classes were measured using an enzyme-linked immunosorbent assay in 41 children with IgA nephropathy (Group A), 15 children with uncomplicated anaphylactoid purpura (Group B) and 13 children with purpura nephritis (Group C). The serum concentrations of the IgA, IgG and IgM classes were measured by single radial immunodiffusion. When compared with sex- and age-matched controls, the concentrations of serum IgA (but not of IgG or IgM) were significantly increased in the three groups studied. The titers of ASO of the IgA and IgM classes, and those of ASP of the IgA and IgG classes, were significantly increased in Group A. In Group B, only the ASP titers of the IgA class were significantly increased. No significant difference was noted in the titers of either ASO or ASP of any class in Group C. Thus, increased antibody response in IgA nephropathy is not restricted to IgA. Anaphylactoid purpura with or without renal disease appears to be different in its humoral anti-streptococcal response from IgA nephropathy.     

T and B lymphocytes before, during, and after cytostatic therapy of acute lymphoblastic leukemia (ALL) in children (author's transl)]

In 56 children with acute lymphoblastic leukemia (ALL) T and B lymphocytes and immunoglobulins (IgG, IgM, and IgA) were studied before, during, and after therapy. At the time of diagnosis T lymphocytes were normal, the number of B lymphocytes was increased, and immunoglobulins usually were normal. Only two of 30 children had abnormally low immunoglobulin levels. Under therapy a progressive lymphopenia was observed. B cells were depressed most extensively. IgG and IgA levels showed a nadir during early treatment, i.e. after remission induction for IgG and 4-8 months after diagnosis for IgA. IgM was markedly elevated after meningosis prophylaxis. After cessation of therapy lymphocytes increased with an intense rebound of B cells. The recovery was not fully completed after one year free of therapy.     

Long-term study on T lymphocyte subsets in newly diagnosed type 1 diabetes mellitus

T lymphocyte subsets in peripheral blood from 16 newly diagnosed type 1 diabetic children were studied prospectively at four time intervals: as soon as possible after diagnosis and 1, 4 and 12 months later. T lymphocyte subsets were analysed using monoclonal antibodies and counted by cytofluorimetry. The percentage of T lymphocytes (OKT3+ cells) did not change at the four study times. The percentage of helper/inducer T cells (OKT4+ cells) was high at the diagnosis (43.1 +/- 2.1%), but decreased after 1 and 4 months with no difference in the control values. The percentage of suppressor/cytotoxic T lymphocytes (OKT8+ cells) was low at the diagnosis, but increased after 1 and 4 months. The OKT4/OKT8 ratio was 2.31 +/- 0.22 at the diagnosis study, decreasing to 1.83 after 1 month, compared with 16 sex- and age-matched control children. The high percentage of helper/inducer T lymphocytes and low number of suppressor/cytotoxic T cells at onset of diabetes favour immune reactions that lead to beta-cell damage.     

Parvovirus B19 infection as a cause of anemia in pediatric acute lymphoblastic leukemia patients during maintenance chemotherapy

PURPOSE: Persistent parvovirus B19 tends to occur in immunocom-promised patients and manifests as pure red cell aplasia and chronic anemia. This study aimed to detect the contribution of parvovirus B19 infection to anemia in children with acute lymphoblastic leukemia (ALL) receiving chemotherapy. PATIENTS AND METHODS: Two groups of ALL patients were studied during maintenance chemotherapy: 50 patients with persistent anemia (ie, extending for >2 weeks) and 34 patients without anemia (controls). Serum parvovirus B19 IgG and IgM were investigated by an enzyme-linked immunosorbent assay, and the virus DNA was sought in bone marrow cells by a nested polymerase chain reaction assay. RESULTS: Parvovirus B19 DNA was detected in 11 of the 50 (22%) ALL children with anemia, 4 of whom were also IgM positive. In addition, IgM positivity was observed in nine (18%) other children who were negative for parvovirus B19 DNA. The children without anemia were found to be significantly different than those with anemia in terms of parvovirus B19 DNA positivity and DNA + IgM positivity (P = 0.03 and 0.01, respectively). IgG was found to be positive in a total of 19 (38%) cases, with B19 DNA present in 6 of them. CONCLUSIONS: These findings indicate the high frequency of parvovirus B19 in anemia in children with ALL and the importance of testing for its DNA in the bone marrow cells together with IgG and IgM antibodies in the serum of immunocompromised patients. It is important to consider parvovirus B19 infections as a cause of anemia and suppressed erythropoiesis in children with ALL who are receiving ongoing treatment.     

Restoration of suppressor T-cell functions in children with AIDS following intravenous gamma globulin treatment

Suppressor T-cell function was analyzed in seven children with acquired autoimmunodeficiency syndrome (AIDS) or AIDS-related complex (ARC). Four of the patients had markedly increased serum IgG levels. All patients had elevated percentages and absolute numbers of peripheral blood T8 cells. In vitro concanavalin A generation of suppressor cells for T-cell mitogenic responses and suppression of pokeweed mitogen-driven immunoglobulin secretion were diminished in all patients. After intravenous treatment with gamma globulin, four patients regained in vitro suppression of pokeweed mitogen-driven gamma globulin secretion. Treatment with intravenous gamma globulin also modified in vitro suppressor T-cell functions in children with AIDS or ARC.     

Increased Serum Levels of Anti-Mumps and Rubella Viruses Antibodies in Children with Kawasaki Disease

Serum IgM and IgG antibodies against mumps virus and rubella virus were quantitated serially by enzyme-linked immunosorbent assays in 15 children with Kawasaki disease. IgM antibody against mumps virus and IgG antibody against rubella virus were increased significantly in children with Kawasaki disease who had no previous history of mumps virus or rubella virus infection. Booster-like effect in IgG antibody levels against mumps virus was observed in children with Kawasaki disease who had a previous history of exposure to mumps virus. Since none of the patients manifested clinical symptoms of mumps or rubella, the increased antibody levels against mumps and rubella may represent a part of the nonspecific increase in serum immunoglobulins in children with Kawasaki disease.     

Serum immunoglobulin G, M and IgG:IgM ratio as predictors for outcome of childhood nephrotic syndrome

Background  Nephrotic syndrome is an immune mediated disorder of the kidney associated with T cell dysfunction and secondary disturbance of B cell with changes in levels of immunoglobulin and IgG:IgM ratio. These changes in immunoglobulin levels can be used as a proxy marker to understand the clinical variety and prognosis of nephrotic syndrome. Methods  We studied 43 children with nephrotic syndrome during January 2003 to January 2005 in the Pediatric Nephrology Unit, Department of Pediatrics, Bangabandhu Sheikh Mujib Medical University, Dhaka, Bangladesh. Blood samples were collected from the 43 patients, and serum levels of IgG, IgM and IgG:IgM were measured by liquid phase immunoprecipitation assay. Another 20 healthy children attending the laboratory for blood grouping and hepatitis B screening test were enrolled as controls. Results  In the 43 children with nephrotic syndrome, 24 had steroid sensitive nephrotic syndrome (SSNS) and 19 steroid resistant nephrotic syndrome (SRNS). Compared with healthy children, the IgG level was low, IgM level was high, and IgG:IgM ratio was low (P<0.05). The serum IgG level and IgG:IgM ratio were significantly lower in children with SRNS and in children with frequent relapse (FRNS) combined with steroid dependent nephrotic syndrome (SDNS) than in those with infrequent relapse nephrotic syndrome (IFRNS) (P<0.05, respectively). Conclusions  Management of different nephrotic syndromes is based on the levels of immunoglobulins along with clinical and biochemical parameters. The decrease of IgG level as a predictive marker for unfavorable prognosis of nephrotic syndrome in children needs further evaluation in larger scale studies.     

Immune deficiencies following cancer treatment in children

The aim of this study was to determine serum immunoglobulins, IgG subclasses, lymphocyte subsets, and serum protective antitoxin levels of tetanus and diphtheria, and to investigate specific antibody response to tetanus and diphtheria vaccines in children with cancer who have been treated for leukemias and solid tumors. Forty patients with different types of childhood malignancies were enrolled in this study and their lymphocyte subsets, serum Ig A, M, G and IgG subclass concentrations were determined at completion of chemotherapy and 6 months later. We measured serum diphtheria (D) and tetanus (T) antitoxin levels and investigated specific antibody responses against DT vaccines at 6 months. Only the leukemic children had low CD19+ cells at completion of chemotherapy and 6 months later. The patients with solid tumors had reduced CD4+ cells, but increased natural killer cells at completion of chemotherapy. Serum IgA and IgM levels were decreased in leukemic patients after chemotherapy. There were no IgG subclass deficiency. Forty-two per cent of the patients did not have protective serum T antitoxins. All patients produced high levels of DT antibodies by vaccination. Immune system changes recover by 6 months after cancer therapy in children. Children with solid tumors, as well as leukemias, should be followed-up in terms of immune deficiencies. A repeat dose of tetanus toxoid should be recommended at 6 months.     

The immunological profile of children with Down's syndrome

In 1965 Benda demonstrated that bioptic and autoptic material of children with Down's syndrome showed hypoplasia of the thymus with poor histological differentiation between the cortex and medulla and impairment of Hassal's corpuscles which were also fewer than normal. In the seventies studies revealed the increased susceptibility to infections and higher incidence of leucosis in Down's syndrome patients as well as changes of immunologic defences (in particular cell mediated immunity). This study examines 35 children (16 boys and 19 girls) aged 6 months-20 years. Subjects were divided into a group of 18 cases in poor health with a history of recurrent infections and a group of 17 children in good health. Skin tests were performed by inoculating 0.1 ml of a solution formed by 1 ml physiological solution and 0.1 ml tetanus toxoid. Skin reaction was evaluated 48 hours later. Lymphocyte typing tests were performed with the rosette method and with monoclonal antibodies for T lymphocytes and with the determination of surface immunoglobulins for B lymphocytes. OKT4 (T helper), OKT8 (T suppressor) subsets assayed and the OKT4/OKT8 ratio was determined. Skin tests were negative in 3 cases (8.6%). The number of B lymphocytes was normal in all children. Total number of lymphocytes was decreased in 51.4% of cases. Two subjects had a reduction of OKT4 and 14 had an increased of OKT8 and 16 a significantly lower OKT4/OKT8 ratio. It is clear that skin tests were normal also in those children with low total lymphocyte values. The most closely related parameter to mobility was the OKT4/OKT8 ratio and the most distantly related on was the skin test. Only 3 cases had modifications of all 3 parameters together. Apart from the constant and complete immunological deficit described by many authors and which we cannot confirm the results of this study are in agreement with those of other authors.     

Clinicl Significance of Mesangial IgM Deposits in Minimal Change Nephrotic Syndrome

The clinicopathologic features and the response to corticosteroid therapy were compared in 9 patients with minimal change nephrotic syndrome (MCNS) and diffuse mesangial IgM deposits (Group I) and in 32 of those without IgM deposits (Group II). However, serum IgM levels in Group I in both relapse and remission were significantly higher than those of Group II and controls (p<0.001). In Group I mesangial IgM deposits were diffuse in 9 (100%), mesangial C_1q was present in 4, IgA and fibrinogen were each observed in 1, respectively. Electron dense deposits in the mesangium were also present in 2 to 5 patients in Group I. No significant differences were found between the two groups in age of onset, sex ratio, laboratory data except for serum IgM level, duration before biopsy, follow-up periods, outcome, and response to steroid therapy. Our data suggest that a more severe degree of either impairment of mesangial clearance of IgM or overproduction of IgM may be involved in patients with MCNS and mesangial IgM deposits but that these patients could not be considered a distinct group of patients.     

T and B lymphocyte subpopulations and leukocyte terminal deoxynucleotidyl-transferase in energy-protein undernutrition.

Children with energy-protein undernutrition showed a reduction in the number of circulating T lymphocytes identified on the basis of their ability to form rosettes with sheep red blood cells. T cells with a receptor for IgM (Tmu) were decreased whereas T cells with a receptor for IgG (T gamma) were increased. Surface immunoglobulin bearing B cells were comparable in well nourished and malnourished subjects but the proportion of B alpha was increased in the latter. "Null" cells without the conventional markers of T or B cells were proportionately increased. Leukocyte terminal deoxynucleotidyl-transferase activity was elevated in the majority of undernourished children and correlated with the proportion of "null" cells. The significance of these observations is discussed and it is suggested that "null" cells represent immature undifferentiated T lymphocytes.     

Enteroviral meningoencephalitis as a complication of X-linked hyper IgM syndrome

We describe 5 children from 2 families with mutations in the CD40 ligand (CD40L) gene leading to absent expression of CD40L on activated CD4 cells. All subjects presented with interstitial pneumonia with low serum IgG and normal serum IgM. One child had normal and one child had elevated serum IgA. Four had confirmed Pneumocystis carinii pneumonia. In spite of intravenous immunoglobulin treatment yielding therapeutic serum immunoglobulin levels, 3 children had enteroviral encephalitis. When assessed by flow cytometry, the 3 surviving affected male children had absent CD40L expression on activated CD4(+) T cells. The affected children from both families were shown to have the same single nucleotide insertion (codon 131) resulting in frameshift and early termination within exon 4 (extracellular domain). This observation demonstrates that persistent enteroviral infection is not only observed in X-linked agammaglobulinemia but may also occur in patients with X-linked hyper IgM syndrome.