Monoclonal antibody T-199 directed against human medulloblastoma: characterization of a new antigenic system expressed on neuroectodermal tumors and natural killer cells.

A monoclonal antibody (mAb) (T-199) of IgG1 isotype was raised against medulloblastoma by immunizations of mice with the medulloblastoma cell line TE-671. Studies of the specificity of mAb T-199 on cell lines as well as fresh frozen sections of normal and malignant tissues revealed the antigen in high amounts on the cell surface of neuroectodermally derived tumors such as medulloblastoma, neuroblastoma, retinoblastoma, and astrocytoma. Some melanomas and a subgroup of rhabdomyosarcomas also expressed the antigen. In contrast, mesenchymal tumors, osteosarcomas, and Ewing's sarcomas did not bear the T-199 antigen. Reactivity of T-199 with normal tissues has not been found with few exceptions; in certain areas of the brain, especially in the cerebellum and part of the hypothalamus, in the adrenal glands, and in the pancreatic islet cells small amounts of antigen were detectable. Natural killer cells could also be demonstrated to express the T-199 antigen similar to the NKH-1 antigen. However, despite some striking similarities, the antigens or antigen epitopes recognized by mAbs T-199 and NKH-1 are not identical. Therefore, mAb T-199 seems to detect a unique differentiation antigen on neuroectodermal tumors, coexpressed in low amounts on normal neuroectodermally derived cells and natural killer cells. The pattern of reactivity and the biochemical properties of the T-199 antigen are different from other cell surface markers for neuroectodermal cells coexpressed on natural killer cells or T-cells (HNK-1, NKH-1, or Thy-1). Biochemical analysis of the T-199 antigen showed that it is a heat-labile protein.

Differential expression of cell surface antigens and glial fibrillary acidic protein in human astrocytoma subsets.

We have characterized five distinct cell surface antigens of human astrocytomas and correlated their expression with the expression of glial fibrillary acidic protein (GFAP) and four previously defined cell surface markers of astrocytomas. One of the newly studied antigens, A4, which was originally detected on rat central nervous system (but not peripheral nervous system) neurons, is expressed on GFAP+ human astrocytoma cells, but not on GFAP- astrocytomas or a wide range of other neuroectodermal, epithelial, and hematopoietic cells. Antigens F19 (Mr 140,000/90,000 glycoprotein) and F24 (Mr 90,000 glycoprotein) also show restricted distribution and are expressed on subsets of neuroectodermal and mesenchymal cells. Antigens G253 (Mr 95,000 glycoprotein) and S5 (Mr 120,000 glycoprotein) are more widely distributed on the cultured cell panel. The distribution of these antigens was determined on a series of 22 astrocytoma cell lines and in normal brain tissue and the results were compared with the distribution of 5 additional glial cell markers: GFAP and cell surface antigens A010 (Mr 110,000 glycoprotein); AJ8 (Mr 100,000 glycoprotein); LK26 (Mr 35,000 glycoprotein); and Thy-1. Distinct patterns of expression on cultured astrocytomas and in neural tissues were identified for all antigenic systems studied, and cell surface expression of antigen A4 was found to correlate closely with GFAP phenotype of cultured astrocytomas. The antigens described in this study provide new markers to study normal glial differentiation and to correlate the phenotypes and biological behavior of distinct subsets of astrocytomas.

Analysis of the fine specificities of 11 mouse monoclonal antibodies reactive with type 2 blood group determinants.

The specificity of 11 mouse monoclonal antibodies reacting selectively with type 2 blood group structures was analyzed in detail by studying their reactivities with a panel of standard glycolipids, glycolipids from erythrocytes and blood group glycoproteins. The antibodies reacted with monofucosyl type 2 H, difucosyl type 2 structures (Le gamma) or both; none of the antibodies reacted with type 1 (H, Lea, or Leb) structures. Only a small proportion of the antibodies were completely specific for either type 2H or Le gamma structures. None of the antibodies had identical patterns of reactivity and their specificities were individually distinct. Seven antibodies preferentially agglutinated O and A2 erythrocytes. Anti-Le gamma-specific antibodies, except mAb101, did not agglutinate erythrocytes or react with glycolipids from erythrocytes, indicating the absence of Le gamma structures in erythrocyte glycolipids. The ability of some antibodies to react with A erythrocytes was shown to be due to cross-reactivity of the antibodies with type 3 (repetitive) A structures. The study demonstrates that monoclonal anti-carbohydrate antibodies tend to react with a range of related, and even distantly related, structure in a pattern characteristic of each antibody and that very few antibodies have extremely restricted specificities.

Characterization of a continuous cell line (MHH-NB-11) derived from advanced neuroblastoma.

A continuous cell line was established from an explanted tumor biopsy obtained from a patient with advanced neuroblastoma, which showed no response to chemotherapy. This cell line MHH-NB-11 retained most properties of the original immature tumor, even after xenotransplantation into nude mice. The cell line consisted of small dense cells with scant cytoplasm and thin; long processes and expressed neuron-specific enolase and synaptophysin, but neither GFAP nor S-100 protein. Karyotyping showed karyograms with 49 to 54 chromosomes, with a modal at 52. Most cells had trisomy 2,7,8,20, but only few structural aberrations were observed. Two of four chromosomes 1 showed a rearrangement of the terminal 1p segment, and all cells had a long HSR on the long arm of one of the chromosomes 13. This region hybridized in situ with the N-myc probe pNB-1. N-myc was amplified 20-fold in this neuroblastoma cell line as determined in Southern blot analysis. This cell line should be a useful tool in vitro or as a xenograft model for neuroblastoma research.

Mouse monoclonal antibody with specificity for human interferon gamma.

We developed a monoclonal antibody (MAb) specific for human interferon gamma (HuIFN-gamma) by hybridizing cells from the NS-1 myeloma cell line with spleen lymphocytes from BALB/c mice immunized with partially purified HuIFN-gamma. Hybridoma culture supernatants were screened for neutralization of antiviral activity of HuIFN-gamma by the method determining the inhibition of nucleic acid synthesis assay (INAS), employing human fibroblasts infected with encephalomyocarditis virus (EMC). Clones exhibiting neutralization of antiviral activity of HuIFN-gamma were recloned, retested and an MAb with maximum neutralization activity was selected. This MAb was of IgM subclass and was specific for HuIFN-gamma. Antiviral activities either of human leukocyte-derived (HuIFN-alpha) or human fibroblast-derived interferon (HuIFN-beta) were not affected by this monoclonal antibody as determined by the INAS test. The specificity of the MAb for HuIFN-gamma was further confirmed by an indirect immunoprecipitation method, where monoclonal antibody-HuIFN-gamma complexes were immunoprecipitated with rabbit anti-mouse immunoglobulin and remaining IFN activity in the supernatants was determined by virus yield reduction assay. Ammonium sulfate precipitated preparations of this MAb were able to significantly increase (range of 230- to 1300-fold) the virus yield when compared with that obtained in the presence of IFN-gamma. SDS-PAGE analysis revealed that the MAb immunoprecipitates a molecule of Mr = 47 kD under nonreducing conditions. Under reducing conditions, two additional bands of Mr = 26 kD (major band) and Mr = 21 kD (minor band) were observed. A sepharose affinity column was constructed using this MAb and was able to retain approximately 60% of the partially purified HuIFN-gamma preparation applied. Significant amounts of HuIFN-gamma were eluted by increasing the ionic strength and decreasing the pH. HuIFN-alpha and HuIFN-beta were not retained by this column.

[Studies on the mechanism of action of D-penicillamine (DPA) on human lymphocytes (author's transl)]. / Untersuchungen zum Wirkungsmechanismus von D-Penicillamin (DPA) auf menschliche Lymphozyten.

From in vitro investigations we have evidence that the inhibition by D-penicillamine (DPA) of lymphocyte transformation is caused by an inhibitory effect on the cell membrane. This inhibition is reversible by washing the lymphocytes. Our results were confirmed by kinetic studies in lymphocyte cultures, using mitogens and DPA. The amount of DPA-molecules bound to lymphocytes was determined with about 2.5 x 10(6) molecules/lymphocyte.

Severe head injury in children: impact of risk factors on outcome.

BACKGROUND: Outcome after severe head injury has been shown in some studies to be more favorable in children than in adults. Mortality rates reported range between 20% and 40% for children. Only contradicting data are available regarding the impact of trauma modalities on long-term outcome, or the relative influence of head fractures, intracranial hemorrhages, and brain edema on survival or neurologic sequelae in children. METHODS: A retrospective study in a tertiary care facility of long-term outcome of children after severe head injury, and analysis of risk factors for poor outcome. All children up to 16 years of age with severe head injury (Glasgow Coma Scale [GCS] score < or = 8), which have been treated in the pediatric intensive care unit from 1977 until 1994 in a single institution. RESULTS: A total of 150 children with severe head injury (GCS score < or = 8) were treated, 92 of them (61.3%) had traffic-related injuries. The median age was 6.6 years (SD +/- 3.6). There were 96 boys (64%) and 54 girls (36%). Sixty-five children (43.3%) had skull fractures, 87 patients (58.0%) developed an intracranial hemorrhage, and 79 patients (52.7%) developed a diffuse brain swelling/edema visible in computed tomographic scans within 72 hours after trauma. Of 150 children treated, 33 died (22%). In most cases, death was related to the development of secondary brain edema. Fifty-nine children (39.3%) had severe neurologic impairments at the time of discharge. The most significant risk factors for adverse outcome, shown by multivariate analysis, were primary areflexia and secondary brain edema. The risk for development of brain edema and poor prognosis was well predicted by the GCS score. CONCLUSION: The overall death rate in this study of children with severe head injury was low (22%) compared with other studies. However, the incidence of severe neurologic impairment at discharge remained high. The major risks for death or neurologic impairment were primary areflexia and the development of secondary brain swelling/edema, indicated by a low GCS score.

Cell-surface antigens of human lung tumors detected by mouse monoclonal antibodies: definition of blood-group- and non-blood-group-related antigenic systems.

The pattern of antigen expression in human non-small-cell cancers of various histological subtypes has been studied. Mouse monoclonal antibodies (MAbs) were generated following immunizations with cell lines of squamous, adeno- and anaplastic large-cell carcinomas of the lung. Seven non-blood-group-related antigens were defined in addition to 5 antigens related to blood-group determinants. Detailed specificity was established with a large panel of cultured cell lines and normal and neoplastic tissues. MAb F-18 reacted in direct tests with the immunizing squamous lung carcinoma cell line, with 5 out of 5 choriocarcinoma cell lines, but with no other cell lines. No expression of F-18 antigen was observed in any normal or malignant tissue examined. The other 6 non-blood-group-reactive MAbs (F-7, F-8, F-11, F-15, F-16 and F-17) could be distinguished by their reactivity on a panel of cultured cells and tissues. One MAb in this group (F-17) reacted strongly with 19/35 lung tumor cell lines, 32/76 other tumor-derived cell lines, cultured normal kidney cells and fetal lung fibroblasts. This antibody did not react with any normal adult tissues examined, but did react with several cancer tissues including 1/17 lung tumors, 2/4 ovarian cancers and 1/5 colon tumors. Immunoprecipitation tests revealed that 5 of the antigens were glycoproteins: F-18 (Mr greater than 200,000), F-15 (Mr 44,000), F-16 (Mr 90,000), F-17 (Mr 95,000) and F-8 (Mr 95,000). Four MAbs detected Y blood-group antigen (Le(y)), only 2 of which were able to agglutinate O erythrocytes. Another antibody detected X blood-group antigen (Le(x)).

Granulocyte-macrophage colony stimulating factor does not improve neutrophil oxidative metabolism in a patient with variant X-linked chronic granulomatous disease.

Variant X-linked chronic granulomatous disease (CGD) is characterised by a decreased but still measurable respiratory burst and cytochrome b content of phagocytes resulting in a clinically milder form of the disease. We examined the in vivo effect of recombinant human granulocyte-macrophage colony stimulating factor (rh-GM-CSF) on the neutrophil functions of a patient treated for liver abscess. The number of white blood cells was markedly increased at the highest dose of GM-CSF injected (30 micrograms/kg per day). This was mainly due to a large increase in eosinophils and to a lesser extent in neutrophils. No change in the deficient neutrophil respiratory burst nitroblue tetrazolium (NBT)-reduction, superoxide (O2-)-production and cytochrome b content was observed during 6 weeks of therapy with increasing doses of GM-CSF. No significant clinical improvement of the liver abscess was observed during treatment with GM-CSF.

[Complex progressive palliation for univentricular circulation]. / Komplexe weiterführende Palliation univentrikulärer Kreislaufsysteme.

The type of palliative procedures (PP) for complex univentricular circulation (UC) changed significantly during the 80ies. Between 07/86 and 02/91 77 patients (pts) presented with UC to eventually undergo a modified Fontan-Kreutzer-type-operation (MFKTO). 33 pts had been previously palliated, 22 of whom were accepted for MFKTO as well as an additional 9 not palliated pts (group A: 31 pts; 20 PP). 11 previously palliated pts and 35 new pts required new palliations (group B: 46 pts; 89 PP), 34 of which resulted in 13 MFKTO. Prerequisites to be met for MFKTO are: undistorted PA-anatomy; absence of subaortic stenosis (SAS); Qp greater than 2.5 l/min/m2; PAP less than 20 mm Hg; Rp less than 3 Wood Units. All earlier PP in group A (31 BTS, 3 PAB, 2 PA-valvotomy/dil.) merely prolonged survival by only adjusting Qp. In group B PP comprise reconstruction of pulmonary arteries, aortic arch and isthmus as well as bypass or resection of SAS or atrial septectomy. Mortality for complex PP (group B) was 32.5% (15/46). 9 of these pts were neonates, 7 after modified Norwood procedures. In an attempt to plan a MFKTO for all pts presenting with UC (group B), complex PP was necessary. The high mortality is due to lesions in pts who would not have survived if only closed PP were applied.

Incidence, impact on survival, and risk factors for multi-organ system failure in children following liver transplantation.

Liver transplantation (LTx) in children currently offers long-term survival rates of more than 80%. Many causes for Tx failure have been identified. However, the incidence and impact of multi-organ system failure (MOSF) are, to date, unknown. Therefore, in this study the role of MOSF after LTx in children was investigated with regard to outcome. The data of 114 children (53 girls, 61 boys; median age 4.3 yr) after first LTx were evaluated retrospectively. The definition of MOSF, as used by Wilkinson et al. [Crit Care Med 1986: 14: 271-274], was modified with regard to age-adjusted values. The influence of MOSF on patient survival was investigated by Kaplan-Meier analysis and multivariate regression analysis. Thirty-one of 114 children with orthotopic LTx developed MOSF (involving two or more organs). In total, 18 children died (15.8%) during the hospitalization; 16 of these had MOSF. Mortality related to two-organ failure was 29.4% (n = 5), to three-organ failure 78% (n = 7), and to four-organ failure 80% (n = 4). The highest mortality rates were observed in children with central nervous system (CNS) and cardiovascular failure, leading to a decreased probability of survival of 0.40 (p < 0.0001). Multi-variate analysis showed that CNS and cardiovascular failure were the most important risk factors for survival (p < 0.0001 and 0.056, respectively). Respiratory and renal failure, in univariate analysis, were significant contributors to poor survival, but had no statistically significant influence on outcome in multivariate analysis. Bone marrow insufficiency was found to have no influence on survival in either analysis. In multivariate analysis, the risk of development of MOSF was significantly increased by high numbers of transfused units of fresh-frozen plasma (FFP), the absence of rejection episodes, or a high bilirubin level prior to Tx. Hence, MOSF is a major factor contributing to the death of children early after LTx. CNS and cardiovascular failure carried the highest risk for a poor outcome. Other risk factors associated with the development of MOSF were: numbers of transfused units of FFP, absence of rejection episodes, and a high pre-Tx bilirubin level.

[Nitric oxide in therapy of pulmonary hypertension after correction of congenital single atrium]. / Stickstoffmonoxid zur Therapie der pulmonalen Hypertonie nach Korrektur eines Atrium commune.

We report on a 19-month-old boy with congenital single atrium. Cardiac catheterization preceding the surgical repair revealed an elevated pulmonary artery pressure of 60/15 mmHg (mean pressure 40 mmHg). Pulmonary flow was 8.4 l/min.m2 and systemic flow was 5.5 l/min.m2. Pulmonary arteriolar resistance was elevated to 4.2 U.m2 with 64% left-right shunt and 25% right-left shunt. Arterial O2-saturation varied around 90%. After surgical repair (insertion of a Goretex patch), the patient required mechanical ventilation with 100% oxygen for adequate oxygenation. Cardiac catheterization was repeated on the first postoperative day. No residual shunts were found. The pulmonary artery pressure was 66/40 mmHg (mean pressure 50 mmHg), systemic arterial pressure was 85/62 mmHg (mean pressure 68 mmHg). Cardiac index was 2.8 l/min.m2, pulmonary vascular resistance was 12 U.m2. After administration of prostacyclin a significant decrease of pulmonary artery pressure was observed, but without changing the ratio between pulmonary and systemic pressure. The AaDO2 varied between 400 and 580 mmHg and the oxygenation-index (PaO2/FiO2) was less than 1.0. In this situation, an attempt with inhaled nitric oxide (NO) was performed. After adding 20 ppm NO to the inspired gas, the AaDO2 decreased significantly from 580 to 270 mmHg and the oxygenation-index (OI) rose from 0.9 to 1.5. The inspired fraction of oxygen could be reduced quickly to 60%. During the next days, the concentration of NO was reduced stepwise to 1 ppm. Finally, the AaDO2 was within the normal range (25-65 mmHg) and the OI rose to a level about 4.0. The FiO2 could be reduced to 30% and nitric oxide therapy could be stopped and the child could be extubated.

[Treatment of acute lymphoblastic leukemia in childhood and adolescence: results of the multicenter therapy study ALL-BFM 81]. / Die Behandlung der akuten lymphoblastischen Leukämie im Kindes- und Jugendalter: Ergebnisse der multizentrischen Therapiestudie ALL-BFM 81.

In therapy study ALL-BFM 81 633 previously untreated patients with acute lymphoblastic leukemia (ALL) less than 18 years of age have been recruited from April 1, 1981 to September 30, 1983 and treated in 37 institutions throughout West-Germany and Austria. Here only therapy results of 611 patients with non-B-ALL are presented. Patients with ALL of B-type are described elsewhere. In this fourth consecutive trial of the BFM study group three major questions have been asked: 1. Is it possible to assess the individual risk for relapse more accurately by the use of a risk factor rather than by the risk score which was the discriminator in studies ALL-BFM 76 and ALL-BFM 79? Does this risk factor discriminate more precisely patients at the highest risk for relapse? Offers more intensive risk-adapted therapy to this patient group a better chance for disease-free survival? 2. In patients at a standard risk for relapse with a risk factor below 1.2--approximately 60% of patients with non-B-ALL--can radiotherapy for prevention of CNS disease effectively be replaced by chemotherapy (intermediate dose Methotrexate)? 3. It is possible to reduce duration of maintenance therapy by 6 months to a total duration of 18 months with no unfavorable effect? To assess the radiation problem in standard risk patients and to evaluate the importance of duration of maintenance therapy two randomisations have been utilized. After a median duration of study ALL-BFM 81 of 4 1/2 years and 3 1/4 years after the study had been closed (date of evaluation January 1, 87) the answers are as follows: 1. For the majority of patients risk-adapted therapy had a curative effect. The probability for event-free survival (EFS) in standard risk patients in slightly above 70%, in medium risk patients 67%. In high-risk patients risk-adapted therapy did not improve prognosis, the EFS being still in the order of 50%. A good assessment of the individual risk for relapse is possible by the newly introduced risk factor. This principle is superior to the risk score used in former studies ALL-BFM 76 and ALL-BFM 79 because a low risk group (risk factor below 0.8) could be identified including approximately 25% of all patients with non-B ALL. Selection, quality, and timing of therapy elements remain the decisive prognostic factors, however. 2. Standard risk patients with a risk factor below 0.8 can effectively be protected for CNS relapse by treatment with intermediate dose Methotrexate.(ABSTRACT TRUNCATED AT 400 WORDS)