Risk factors for hemorrhagic cystitis in pediatric allogeneic hematopoietic stem cell transplant recipients.

Background: Hemorrhagic cystitis (HC) results in significant morbidity among hematopoietic stem cell transplant (HSCT) recipients. Several potential causes for HC have been postulated, including viral infection, but definitive evidence is lacking, particularly in pediatric HSCT patients.

Methods: Ninety pediatric HSCT recipients were prospectively tested on a weekly basis for adenovirus (ADV) and BK virus (BKV) by quantitative real-time polymerase chain reaction in blood and urine samples.

Detection of respiratory viruses in asymptomatic children undergoing allogeneic hematopoietic cell transplantation.

Detection of respiratory viruses by molecular methods, in children without respiratory symptoms undergoing hematopoietic cell transplantation (HCT), has not been well described. A prospective study of 33 asymptomatic children detected respiratory viruses in 8 of 33 (24%) patients before HCT. Human rhinovirus (HRV) was detected in five patients, and human adenovirus (hADV) in three patients.

Isolation and characterization of an age-related antigen present on senescent human red blood cells.

Autologous membrane-bound IgG was isolated from a subpopulation of human red blood cells (RBC) with specific density greater than 1.110, by affinity chromatography of purified RBC membrane glycoprotein preparations using immobilized wheat germ agglutinin and immobilized anti-human immunoglobulin (Ig) as immunoabsorbents. The Ig-containing population thus obtained, when further separated by chromatography on Sephadex G-200 in the presence of chaotropic agents, yielded four peaks (Ia, Ib, II, and III).

Binding of immunoglobulin classes to subpopulations of human red blood cells separated by density-gradient centrifugation.

Human erythrocytes (RBC) from whole blood were separated according to their specific densities by centrifugation on a polyvinyl-pyrrolidine-coated colloidal silica matrix (Percoll) into four major subpopulations. By indirect immunofluorescence assay, the most dense RBC subpopulation, with specific density greater than 1.110 g/ml (3%-5% of total RBC), was positive for membrane-bound immunoglobulin; the remaining, less dense subpopulations were negative.

A comparison of the N-terminal amino acid sequences of early and late pools of anti-DNP and anti-DNP-p-aminobenzoylglutamate antibody light chains.

The N-terminal amino acid sequences of early and late pools of anti-DNP and anti-DNP-p-aminobenzoylglutamate (DNP-ABG) antibody light chains were quantitatively determined and compared. The amino acid composition at each locus of the N-terminal 20 amino acid residues of each light chain preparation was determined using automatic sequencing techniques coupled with high-pressure liquid chromatography and mass spectrometry. The sequence data obtained for the light chains corresponding to antibodies isolated early in the immune response (3-4 weeks) were essentially the same as those for light chains from antibodies isolated late in the response (12-14 weeks).

Antigens of human trophoblasts: a working hypothesis for their role in normal and abnormal pregnancies.

This report describes the preparation and characterization of antisera to human trophoblast membranes. Rabbit antisera were raised to trophoblast microvilli prepared by differential ultracentrifugation. Antibodies to serum proteins were removed by solid-phase immunoabsorption with normal human serum, and indirect immunofluorescence experiments with cryostat sections of human placentas showed that the absorbed anti-trophoblast sera reacted with trophoblasts as well as with stromal cells and endothelium of chorionic villi.

Immunological mechanism for spontaneous abortion in systemic lupus erythematosus.

The incidence of lymphocytotoxic antibodies in patients with systemic lupus erythematosus (S.L.E.

N-bromosuccinimide oxidation of anti-DNP and anti-DNP-p-aminobenzoylglutamate antibodies in the presence and absence of protecting hapten.

The reagent N-bromosuccinimide (NBS) has been employed to investigate the role of tryptophan in hapten binding in anti-DNP (H-1) and anti-DNP-p-aminobenzoylglutamate (DNP-ABG) (I-13) antibodies. In 0-1 M acetate (pH 4-0) buffer fifteen and sixteen moles of tryptophan in the anti-DNP and anti-DNP-ABG antibodies respectively were reactive toward NBS. The hapten DNP-lysine protected 1 tryptophan in antibody H-1 and three tryptophans in antibody I-13 from NBS modification.

A comparison of the isoelectrofocusing porperties of antibodies to DNP, DNP-glycylglycylglycine and DNP-p-aminobenzoylglutamate.

The charge of heterogeneity of antibodies to DNP-glycylglycylglycine (DNP-Gly-3) and DNP-p-aminobenzoylglutamate (DNP-pABG) has been investigated using preparative liquid isoelectric focusing techniques. The focusing profiles of the two antibody preparations are qualitatively similar, each containing four or five major peaks. The results are also similar to the profiles obtained earlier for DNP antibodies.

Purification of DNP specific antibodies using sepharose bound DNP-para-aminobenzoylglutamate.

An immunoabsorbent column employing a DNP derivative having restricted molecular freedom (dinitrophenylated-para-aminobenzoylglutamate--Sepharose 4B) was prepared in order to isolate and purify DNP specific antibodies in high yield. The antibodies were recovered from the immunoabsorbent column in yields from 80 to 90% using 3 M sodium thiocyanate, and these antibodies retained their immunologic activity. There appeared to be limited selection of antibodies with specific affinities as was noted in a parallel experiment comparing a second antibody purification procedure.

N-bromosuccinimide oxidation and amino acid content of isoelectrofocused fractions from a DNP antibody population.

The reagent -bromosuccinimide (NBS) has been employed to investigate the role of antibody tryptophan in the binding of 2,4-dinitrophenyl (DNP) ligands by isoelectric anti-DNP antibody components. NBS titration in 0.1 M acetate—8 M urea (pH 4) buffer established a direct relationship between the extinction coefficients, fluorescence yields, maximum quench values, and tryptophan contents of the isoelectric antibodies.

Binding studies on isoelectrofocused fractions from a DNP antibody population.

Preparative liquid isoelectric focusing has been used to isolate antibody fractions of restricted heterogeneity from typically heterogeneous anti-DNP antibody populations. The focusing distributions of anti-DNP antibody isolated from different rabbits and from different bleed periods (separated by 120 days) of the same rabbit were qualitatively very similar. All antibody preparations contained major components focusing in the pH ranges 7.

Affinity labelling of isoelectrofocused fractions from a DNP antibody preparation with the photoactive labels 2,4-dinitrophenyl-1-azide and 2,4-dinitrophenyl-epsilon-aminocaproyldiazoketone.

Isoelectric (I.E.F.

Antibodies to DNP, DNP-glycylglycylglycine and DNP-p-aminobenzoylglutamate.

The antibodies to a series of three dinitrophenyl (DNP) derivatives of increasing site-filling capacity (DNP; DNP—glycylglycylglycine and DNP—-aminobenzolylglutamate) have been isolated in order to assess the effect of the molecular dimensions of a haptenic group on the diversity of the immune response. A decrease in both the intensity and first appearance of precipitable antibody associated with an increase in molecular size of the hapten was observed. In a 5-week interval, the affinity constant for the homologous hapten of anti-DNP—BGG antibody increased approximately 100-fold.