The quantity and quality of α-gal-specific antibodies differ in individuals with and without delayed red meat allergy.

BACKGROUND: IgG to galactose-α-1,3-galactose (α-gal) are highly abundant natural antibodies (Ab) in humans. α-Gal-specific IgE Ab cause a special form of meat allergy characterized by severe systemic reactions 3-7 h after consumption of red meat. We investigated 20 patients who experienced such reactions and characterized their α-gal-specific IgE and IgG responses in more detail. METHODS: α-Gal-specific IgE was determined by ImmunoCAP. IgE reactivity to meat extract and bovine gamma globulin (BGG) was assessed by immunoblotting and ELISA, respectively. In some experiments, sera were pre-incubated with α-gal or protein G to deplete IgG Ab. α-Gal-specific IgG1-4 Ab in individuals with and without meat allergy were assessed by ELISA. RESULTS: In immunoblots, BGG was the most frequently recognized meat protein. Binding of IgE and IgG to BGG was confirmed by ELISA and completely abolished after pre-incubation with α-gal. Neither the depletion of autologous α-gal-specific IgG Ab nor the addition of α-gal-specific IgG Ab from nonallergic individuals changed the IgE recognition of BGG of meat-allergic patients. Meat-allergic patients showed significantly higher α-gal-specific IgG1 and IgG3 Ab than nonallergic individuals, whereas the latter showed significantly higher levels of α-gal-specific IgG4 Ab. CONCLUSION: Patients with delayed meat allergy display IgE and IgG Ab that selectively recognize the α-gal epitope on BGG. Their enhanced α-gal-specific IgE levels are accompanied by high levels of α-gal-specific IgG1 devoid of IgE-blocking activity. This subclass distribution is atypical for food allergies and distinct from natural α-gal IgG responses in nonallergic individuals.

Sustained effect of SQ-standardized grass allergy immunotherapy tablet on rhinoconjunctivitis quality of life.

BACKGROUND: The prevalence of allergic rhinoconjunctivitis has increased significantly over the past decades with grass pollen being a common trigger. The impact of allergy on patient's quality of life is substantial. AIM: To investigate the sustained effect on quality of life during the grass pollen season 1 year after 3 years of treatment with the SQ-standardized grass allergy immunotherapy tablet (AIT), Graza (Phleum pratense 75,000 SQ-T/2800 BAU; ALK, Denmark). METHODS: The trial was a randomized, parallel-group, double-blind, placebo-controlled trial in adult subjects with a history of moderate-severe grass pollen induced rhinoconjunctivitis inadequately controlled by symptomatic medications. Subjects received 3 years of grass AIT (n = 157) or placebo (n = 126), followed by 1 year of follow-up. Quality of life assessments were based on the standardized rhinoconjunctivitis quality of life questionnaire (RQLQ(S)); completed weekly during the entire grass pollen season. RESULTS: During follow-up, the overall RQLQ(S) score for the entire grass pollen season was significantly improved in the active group (relative difference to placebo: 23%, P = 0.004). The improvement was higher during the peak pollen season (28%, P = 0.001). The treatment effect of grass AIT during the follow-up year and the previous three treatment years was similar. Improvements were found in all seven RQLQ(S) domains. The RQLQ(S) as a function of the weekly average pollen counts showed a clear separation between the treatment groups (P < 0.001). CONCLUSION: In subjects inadequately controlled by symptomatic medications, grass AIT provided sustained and clinically relevant improvements in rhinoconjunctivitis quality of life compared to placebo. The effect increased with increasing grass pollen exposure.

CD34-positive cell proportions in peripheral blood correlate with colony-forming capacity.

Blood samples were examined from 25 children with malignancies during hematopoietic recovery following chemotherapy-induced aplasia and from 9 children undergoing tonsillectomy. The proportion of CD34-positive peripheral blood mononuclear cells (PBMNC) evaluated by flow cytometry was compared with the number of colonies (granulocyte-macrophage colony-forming units, CFU-GM; mixed-lineage colony-forming units, CFU-GEMM; and erythroid burst-forming units, BFU-E) grown in methylcellulose medium within 2 weeks. A mean of 1387 myeloid colonies (495-4480) per 10(5) PBMNC seeded developed from 13 samples with detectable CD34 populations (between 0.9% and 5.6%), whereas only 152 (9-386, p = 0.002) and 65 (12-137, p = 0.005) colonies were formed from 12 patient and from 9 control samples in which the percentage of CD34-positive cells was too low for analysis. Linear regression analysis revealed that CD34 positivity correlates with colony-forming capacity (p = 0.0008, r = 0.782). Flow cytometric evaluation of the CD34 proportions can thus predict the in vitro colony-forming capacity of peripheral blood prior to leukapheresis.

First experience with prognostic factors in unselected neuroblastoma patients. The Austrian Neuroblastoma 87 Study.

Between January 1987 and December 1993, 117 patients were registered in the Austrian A-NB87 study. The male/female ratio was 1.18, with 50 patients below the age of 1 year at diagnosis. Patients were assigned to stage according to the result of primary surgery in localised disease. Age, ferritin and neuron specific enolase were used in addition in stage III disease for risk-adapted treatment. Adrenal or pelvic primary tumour sites were mainly associated (81%) with advanced disease. The median observation time of the study is 3.5 years. The overall survival at 3 years was excellent in low stage disease and IVs patients, i.e. 100% for stage I and IIA (20 patients), 92% in stage IVs (13 patients), 81% in stage IIIA (18 patients) and 69% in stage IIB (8 patients). Stage IV (38 patients) showed a survival rate of 51%, whereas stage IIB (10 patients) had the worst outcome in this study, i.e. 20%, due to treatment-related toxicity. Significant unfavourable prognostic factors were neuron specific enolase (NSE) > 100 ng/ml, ferritin > 300 micrograms/ml and amplified MYCN. This study achieved a better survival rate in stage IV patients and a subgroup of stage III in comparison to our previous study (Pädiatrie und Pädologie 1986, 21, 269) and gives the basis to further reduce treatment intensity in low-risk disease based on biological factors. However, prognosis for high-risk cases was still poor in spite of a very aggressive treatment concept.

Myeloid progenitor cells (CFU-GM) predict engraftment kinetics in autologous transplantation in children.

In autologous stem cell transplantation the duration of aplasia may be particularly long. We studied 27 children with a median age of 5 years (1-16), who received 31 autologous grafts, seven of them receiving both bone marrow and peripheral stem cells. Five were transplanted with blood derived stem cells only. The median number of nucleated cells (NC) grafted was 1.04 x 10(8)/kg body weight (range 0.09-6.72). The median number of granulocyte-macrophage progenitors (CFU-GM) transfused was 0.66 x 10(4)/kg body weight (range 0-11.8). The numbers of NC correlated with the CFU-GM numbers (p less than 0.001), and were inversely correlated with time to recover 1 x 10(9)/l leukocytes and 0.5 x 10(9)/l granulocytes (p less than 0.01), as well as with the appearance of reticulocytes (p less than 0.05). Moreover, the logarithm of the CFU-GM numbers transfused was linearly correlated with the time to recover 1 x 10(9)/l leukocytes and 0.5 x 10(9)/l granulocytes (p less than 0.001), respectively. There also existed an inverse correlation with the first appearance of reticulocytes in the peripheral blood (p less than 0.01). No correlation could be detected with the duration of platelet transfusion dependence. Based on our findings a prediction of time to recover leukocytes, granulocytes and reticulocytes is feasible before grafting with autologous bone marrow and peripheral stem cells.

The median rise of leucocytes predicts the optimal days for leukapheresis in children.

22 consecutive patients with a median age of 11 years (range 3-23) underwent 103 leucaphereses after a chemotherapy induced aplasia. They suffered from various solid tumors and hematological malignancies. The stem cell yield of 69 aphereses which were done during the 8 days following the first platelet rise, was median 1.02 x 10(4) myeloid committed stem cells (CFU-GM)/kg/apheresis (range 0.02-13.3), whereas only 0.27 x 10(4) CFU-GM/kg/apheresis (range 0.01-1.11) were obtained in 34 "random" collections (p less than .05). The CFU-GM yield of 69 "well timed" collections depended on the absolute number of circulating leucocytes (p = .002) and mononuclear cells (p less than .001). The median daily increment of leucocytes was a reliable predictor of the stem cell yield (p = .002) and could be used to select the optimal days for leucaphereses during the well timed period. By careful timing alone high stem cell numbers could be collected in various malignancies without using exogenous growth factors.

The median daily increment of leukocytes during hematopoietic recovery reflects the myeloid progenitor cell yield during leukapheresis in children.

Twenty-two consecutive patients with a median age of 11 years (range 3-23) underwent 103 leukaphereses for collection of peripheral stem cells. They suffered from various solid tumors and hematologic malignancies. Our aim was to collect at least 2.5 x 10(4) myeloid committed stem cells (CFU-GM) per kg body weight. This stem cell number has reliably resulted in rapid and sustained hematopoietic reconstitution after autografting in our institution. Peripheral stem cell mobilization was induced by myelosuppressive therapy alone. If the stem cell collections were started at the first platelet rise and were continued during the following 8 days, the stem cell yield of one procedure was high - median 1.02 x 10(4) CFU-GM/kg body weight (range 0.02-13.3). In 69 such 'well-timed' collections, a median of only 4 leukaphereses (range 2-6) were needed to collect high numbers of CFU-GM. The yield from leukaphereses carried out at any other time was unsatisfactory. In 34 'random' collections, a median of only 0.27 x 10(4) CFU-GM/kg body weight (range 0.01-1.11) was obtained in one procedure. In 69 well-timed collections the CFU-GM yield depended on the absolute number of circulating leukocytes (r = 0.369, p = 0.002) and mononuclear cells (r = 0.476, p less than 0.001). The median daily increment of leukocytes to the day of apheresis proved to be an excellent parameter for the prediction of the yield and could be used to select the best days for leukaphereses (r = 0.403, p = 0.002). These findings may help to make the collection of blood-derived stem cells more efficient in pediatric patients.

Unusually long survival after autografting in second partial remission of translocation t(4;11) acute infant leukemia.

About 70% of children with acute lymphoblastic leukemia may be cured by conventional chemotherapy. The prognosis is considerably worse in infant leukemia with a translocation t(4;11). We report an infant with a diagnosis of cytochemically undifferentiated acute hybrid leukemia (pre pre B-ALL coexpressing one myelomonocytic marker) and t(4;11). Initial clinical presentation and the course of the disease were typical for t(4;11) acute leukemia. After an early hematologic relapse intensive chemotherapy resulted only in a second partial remission 7 months after initial diagnosis. Subsequent bone marrow transplantation with 16 mg/kg busulfan and 200 mg/kg cyclophosphamide followed by the infusion of autologous purged bone marrow resulted in a continuous second remission which has lasted 46 months so far.

Amphotericin B serum levels in pediatric bone marrow transplant recipients.

We studied amphotericin B (AMB) serum levels (n = 590) in 41 pediatric patients, who underwent allogeneic (21) or autologous (20) bone marrow transplantation (BMT). All patients received AMB orally as part of a total gut decontamination; 30/41 patients (73%) had AMB i.v. either for prophylaxis or therapy of fungal infections. Rapid initial dose escalation of AMB and the infusion over 1 h only were well tolerated by the children. Serum level monitoring allowed AMB long-term treatment safely to be administered in children suffering from transplantation-related complications (veno-occlusive disease of the liver, graft-versus-host disease of the liver). An h.p.l.c. method was used for monitoring AMB serum trough levels to avoid levels exceeding 2 mg/l. One lethal fungal infection was observed in 41 pediatric BMT recipients (2.4%). Rapidly increasing doses of AMB at start of therapy and drug monitoring by h.p.l.c. might help to reduce fungal mortality and renal toxicity by a dose sparing effect in BMT recipients.

Is treatment intensification by adding etoposide and carboplatin to fractionated total body irradiation and melphalan acceptable in children with solid tumors with respect to toxicity?

Conventional chemotherapy results in high mortality rates in patients with solid tumors involving the bones or the bone marrow. High dose melphalan (MEL) with or without total body irradiation followed by bone marrow transplantation (BMT) has prolonged survival, but curative potential has remained disappointing. In order to improve survival 20 children with generalized or relapsed solid tumors (neuroblastoma, peripheral neuroectodermal tumor, Ewing's sarcoma, rhabdomyosarcoma) underwent autologous (n = 16) or allogeneic (n = 4) BMT. The myeloablative regimen consisted of 12 Gy fractionated total body irradiation (FTBI) and high dose MEL. In 12 of these patients this regimen was intensified by giving 60 mg/kg etoposide (1800 mg/m2 VP), and 1.5 g/m2 carboplatin (CBDCA) was added in seven of these 12 patients. The intensification of FTBI and MEL by adding VP and CBDCA was followed by acceptable toxicity. Acute liver toxicity in 15/20 patients (75%) and acute renal toxicity in 17/20 patients (85%) did not exceed WHO grade 1. The use of the conditioning regimen FTBI-MEL-VP-CBDCA during first chemotherapy response is a promising approach in treatment of children suffering from generalized solid tumors.

Treatment of infant leukemia with busulfan, cyclophosphamide +/- etoposide and bone marrow transplantation.

Infants with acute leukemia have a poor chance of being cured by conventional chemotherapy. We therefore treated cases of infant leukemia with high dose chemotherapy followed by bone marrow transplantation (BMT). Six suffered from acute leukemia and one from refractory anemia with excess of blasts (RAEB-t). The conditioning regimen consisted of busulfan (BU) and cyclophosphamide (CY), and was intensified by adding etoposide (VP) in four cases. At the time of BMT the children were 4, 5, 12, 13, 13, 14, and 20 months old. Three children were autografted, three received HLA-identical marrow from a sibling donor, and one child received matched unrelated donor marrow. All five children who were grafted in complete (CR) or partial remission (PR) are alive and well in CR 7, 13, 24, 37, and 46 months after allogeneic (two patients) or autologous (three patients) BMT, and 13, 17, 29, 42, and 53 months after initial diagnosis. The child with RAEB-t and the one transplanted in second chemotherapy-resistant relapse of acute non-lymphoblastic leukemia relapsed at 7 and 17 months respectively. The chemotherapy regimen was well tolerated. BU-CY-VP is a promising alternative treatment to regimens including total body irradiation for very young children suffering from acute leukemia.

Organ toxicity and quality of life after allogeneic bone marrow transplantation in pediatric patients: a single centre retrospective analysis.

One hundred and fifty five pediatric patients underwent allogeneic bone marrow transplantation between 1980 and 1996 in the St Anna Children's Hospital in Vienna with an overall survival of 52.3% (81 patients). Seventy-three patients with a minimum observation time of 1 year (1-13 years, median: 4.6) were analyzed retrospectively for chronic GVHD, organ toxicity (WHO score), growth and pubertal development. Chronic GVHD was diagnosed in 20 patients (27.3%), being extensive in 17 cases. Maximum organ toxicity was WHO III in two patients (3%) and WHO II in 11 patients (15%) 1 year after BMT and WHO III in one patient (2%) and WHO II in five patients (11%) 3 years after BMT. Impaired growth and pubertal development were detected in more than 30% 3 years after BMT. As all patients presented with a Karnofsky or Lansky score of more than 80%, they were asked to complete a questionnaire comprising 12 questions concerning physical state of health and psychosocial state of health. Restricted contacts were classified as imposing a severe handicap by six patients (8%), restriction in mobility and 'normal life activities' by three patients (4%) and two patients classified themselves as severely physically handicapped. Most patients (75%) reported no physical or psychical impairment.

Stem cell procurement and transplantation in paediatric patients.

Harvesting of peripheral blood stem cells (PBSC) with cell separators has become a safe procedure used in many centers. However, when PBSC harvesting in small children is considered, problems due to the small volume, loss of protein and red cell trapping arise. Priming the tubing of the cell separator with either albumin or red cells is necessary in children with a blood volume of less than 1500 mL. Ten children aged from 0.8 to 5 years underwent PBSC-apheresis without major problems despite low blood volume. Forty-one children with various malignancies had a total of 189 PBSC-aphereses with a median of 5 phereses for an individual patient. Harvesting of PBSC was started after recovering from neutropenia following scheduled chemotherapy. An average of 3.7 x 10(4)/kg CFU-GM per child was harvested. Sustained engraftment after myeloablative chemotherapy could be achieved when the dose of the retransfused PBSCs was greater than 3 x 10(4)/kg. Long-lasting complete remission was observed in 5 out of 25 patients.

Enhanced production of IL4 but not of IFN gamma and IL 10 by peripheral blood mononuclear cells from atopic children in response to CD2 plus CD28 stimulation.

Peripheral blood mononuclear cells from 16 children with atopic disease (range of IgE levels: 33 - 2892 kU/l) and 12 age matched controls were stimulated either with mAbs specific for CD3, CD2, CD3 plus CD28, CD2 plus CD28, with Tetanus Toxoid, SEA, or PHA plus PMA and their cell proliferation was determined. In addition, their cytokine production (IL2, IL4, IL10, IFN gamma) following selected stimuli was measured. We found that the cells from atopics proliferated significantly better in response to CD2 stimulation than control cells, with no difference in response to CD3 or SEA stimulation. Furthermore, cells from atopics produced significantly higher amounts of IL4 than cells from controls, a difference most pronounced following CD2 plus CD28 stimulation. No differential production was found for IL10 and IFN gamma. We conclude that in atopic children with moderately elevated IgE a hyperreactivity of the CD2 pathway of stimulation and a clear elevation of IL4 but not of IL10 or IFN gamma production can be demonstrated.

CD14(dim)/CD16(bright) monocytes in hemophagocytic lymphohistiocytosis.

Hemophagocytic lymphohistiocytosis (HLH) is an extremely rare and highly lethal chronic inflammatory disease, which is mediated by proinflammatory cytokines. In the peripheral blood of a boy suffering from HLH, a chronic expansion of CD14(dim)/CD16(bright) inflammatory monocytes was detected. Compared with CD14(bright) monocytes, their immunophenotype correlated with more mature monocytic cells differentiating to macrophages: they showed lower expression of CD11b, CD64 and CD35. Such CD14(dim)/CD16(bright) monocytes produce the inflammatory cytokines IL-1beta, IL-6 and TNF-alpha. They fit in well with the pathophysiological concept of HLH as an inflammatory state of lymphocytes and of the monocyte/macrophage system. In the presented patient the percentage of these circulating inflammatory monocytes decreased over time during clinical response to immunosuppressive therapy. This finding may indicate that CD14(dim)/CD16(bright) monocytes represented the degree of inflammation in this extremely rare and highly lethal disease.

Limited tolerance of intensified conditioning regimens in children receiving methotrexate/cyclosporin A for graft-versus-host disease prophylaxis.

Twenty-one patients with a median age of 9 years (0.5-19) underwent intensified myeloblative therapy: 1800 mg/m2 etoposide (VP) was added to 120 mg/kg cyclophosphamide (CY) and 12 Gy fractionated total body irradiation (FTBI) or 12-16 mg/kg busulfan (BU) for treatment of acute lymphoblastic leukemia (11 patients), acute myeloid leukemia (8 patients), non-Hodgkin's lymphoma (1 patient), or myelodysplastic syndrome (1 patient). Severe liver toxicity occurred in 5 of 7 children (71%) receiving short-term methotrexate (MTX) and additional cyclosporin A (CSA) for prophylaxis of graft-versus-host disease (GVHD). Three of them died of subsequent acute renal failure on days 8, 13, and 34. In contrast, acute severe organ toxicity occurred in only 1 of 14 children (7%) receiving the same intensified regimens who were autografted (7 pts) or received MTX alone for GVHD prophylaxis (7 pts). These observations suggest that GVHD prophylaxis with MTX and CSA may adversely influence the tolerance of intensified antileukemic regimens in children.

Kinking and stenosis of the carotid artery associated with homolateral ischaemic brain infarction in a patient treated with cyclosporin A.

UNLABELLED: We present a 3-year-old patient with stenotic kinking of the left internal carotid artery (ICA) who developed an ischaemic infarction of the left brain hemisphere followed by severe neurological sequelae after a prolonged generalized seizure. At time of the seizure the boy was in biological remission of a nephrotic syndrome and received prednisolone and cyclosporin A (CsA) treatment. The haemodynamic consequences of inborn kinking of the ICA is discussed controversely in the literature. The presented case shows that stenotic kinking of the ICA may significantly impair the blood flow towards the homolateral hemisphere and therefore may result in an ischaemic infarction. The influence of CsA on seizure activity is discussed. CONCLUSION: This case provides clinical and radiological evidence supporting an association between stenotic kinking of the carotid artery and homolateral hemispheric brain infarction.

[Therapy of urinary tract infection]. / Die Therapie des Harnwegsinfektes.

The choice of therapy in urinary tract infections depends on the infection site, complications, previous history, and recrudescence. Acute uncomplicated urinary tract infections which do not respond to general measures as increased fluid intake will be cured with antibiotics for 3 days. If the compliance of the patient is sufficient a single shot therapy may be applied. Infections of the upper urinary tract with typical signs such as fever, nausea and increase of the so-called acute phase proteins have to be treated with antibiotics for 7 to 14 days. When treating the first episode of urinary tract infection the choice of antibiotics is unimportant. Trimethoprim and quinolones are standard. In recurrent infections urine cultures and susceptibility testing are mandatory. Patients with recurrent urinary tract infections should receive a long-term suppressive therapy with low-dose trimethoprim or a quinolone. Patients with urinary catheters must not receive any long-term therapy.

High-dose melphalan, etoposide +/- carboplatin (MEC) combined with 12-gray fractionated total-body irradiation in children with generalized solid tumors.

Long-term disease-free survival is poor in patients with primary generalized or relapsed solid tumors. High-dose chemoradiotherapy with stem cell rescue improved survival, but more effective protocols are needed. From January 1988 to November 1988, we treated 7 patients (median age, 9 years; range, 3-23 years) with an intensified treatment program. They received 12-Gy fractionated, total-body irradiation (FTBI). High-dose chemotherapy (MEC) consisted of melphalan (120-140 mg/m2 Mel) and etoposide (40-60 mg/kg VP-16) with or without carboplatin (1.5 g/m2 CBDCA). Although we combined 12-Gy FTBI with Mel, VP-16, +/- CBDCA in doses used previously for high-dose single-agent chemotherapy, the extramedullary toxicity of FTBI with ME(C) was tolerable. Two of the four patients who were grafted without delay after good initial chemotherapy response are still alive in continued complete remission 30 and 33 months, respectively, after initial diagnosis. Early application of FTBI and ME(C) during first chemotherapy response might improve outcome in patients with primarily generalized solid tumors.

[Liposomal amphotericin B in the treatment of candida infection in a 3-month-old infant]. / Liposomales Amphotericin B zur Behandlung einer Candidasepsis bei einem drei Monate alten Säugling.

Amphotericin B (Amph-B) is the treatment of choice for systemic fungal infections. The application of high doses is limited due to the high incidence of acute side effects (fever, chills) and organ toxicity (reduction in glomerular filtration rate, renal tubular damage). Amph-B was applied without success in a three months old infant, who suffered from systemic candidiasis. After a change to high doses (3-5 mg/kg/day) of liposomal Amph-B (Amph-lip) rapid improvement of the patient's condition occurred and the pulmonary lesions disappeared during six weeks of treatment (total dose 185.5 mg/kg). Acute side effects or renal function abnormalities did not occur. The reported case indicates that the application of high doses of Amph-lip is an alternative to conventional Amph-B in treatment of systemic fungal infections.