Epidemiologic Profile of Patients Transplanted With Hematopoietic Stem Cells in a Reference Service in the State of Rio Grande do Norte, Brazil.

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) consists of the intravenous infusion of healthy hematopoietic stem cells to restore the medullary and immunologic function of patients affected by a series of hematologic, oncologic, immunologic, malignant and nonmalignant inherited or acquired diseases, with the possibility of cure or increase of disease-free survival. OBJECTIVE: To characterize the epidemiologic profile and the cases of death of patients who underwent HSCT. METHODS: This is a cohort quantitative study, nested with a retrospective, descriptive, and analytical study of a hospital-based cohort that included the patients who underwent HSCT at a referral service in the state of Rio Grande do Norte, a region of northeastern Brazil. RESULTS: There was a slight male prevalence (52.94%), the age of the patients ranged from 2 to 73 years old, 18.38% were brown, 47.06% were married, 15.07% were students, 78.31% had a diagnosis of multiple myeloma, 93.38% developed gastrointestinal toxicities, all patients received chemotherapeutic treatment, 54.78% had allogeneic HSCT, and the cause of the most recorded deaths was septic shock (48.19%). CONCLUSIONS: This study showed relevant scientific evidence on the clinical and epidemiologic profile of patients who underwent HSCT. In general, sociodemographic data are similar to national and international research results.

Solid versus liquid phase assays in detection of insulin antibodies. Influence of iodination site on labelled insulin binding.

On type 1 newly diagnosed and on insulin treated diabetic patients, anti-insulin autoantibodies (IAA) and antibodies (IA) having the same specificity are respectively induced. Such immune response may be evaluated either by radiobinding assay (RBA) or enzyme-linked immunosorbent assay (ELISA). Both methodologies have been compared at previous International Workshops, which pointed out discrepancies in results. In this work, IAA/IA prevalence was assessed by displacement RBA and ELISA, in normal subjects, type 2 (treated with hypoglycaemic agents), insulin treated and newly diagnosed type 1 diabetic patients. Results showed a lack of RBA-ELISA agreement. An attempt was then made to determine whether such results were, at least in part, attributable to iodination site in Tyr-A14. For this purpose parallel RBA assays were carried out by using radiolabelled insulin at A14 and A19 Tyr residues. Control sera and samples from insulin treated and type 1 newly diagnosed diabetic patients were tested. Our results suggest that labelling position is not involved in artifactual binding of tracers, at least as a systematic phenomenon. In the majority of cases the variability in RBA-ELISA signal ratios are best explained in terms of differences in the basic principles operating in both methods instead of artifacts due to tracer preparation.

A case of allergy to human insulin associated with high IgG/IgE ratio for specific antibodies.

A 37-year-old woman with a previous history of allergic rhinitis and bronchial asthma presented local and generalized allergic reactions to bovine, porcine and human insulins. She developed ketoacidosis, high insulin requirements and transient hypoglycemic episodes. Several desensitizing schedules were applied in order to induce tolerance to human insulin therapy. In addition, the following parameters of the humoral immune response were measured in different serum samples taken within 13 days after one of the anaphylactic episodes: insulin antibodies (binding range = 29.4-47.8%; cutoff = 2.6%), total IgE (range = 500-850 IU/ml; normal values = 3.7-269 IU/ml), specific IgE (range = 0.42-0.83 PRU/ml; class 2) and subclasses of specific IgG (IgG1 = 97.2 SDS; IgG2 = 41.1 SDS; IgG3 = 9.9 SDS; IgG4 = 0.3 SDS, on day 1). A binding capacity of 31.8 IU insulin/l obtained by Scatchard analysis was in agreement with episodes of elevated insulin requirements and hypoglycemia. A high anti-insulin IgG/IgE ratio, along with high levels of specific IgG1 antibodies, suggested that the latter antibodies could be involved in the development of anaphylactic episodes.

Improvement in soil and sorghum health following the application of polyacrylate polymers to a Cd-contaminated soil.

Contamination of soils with cadmium (Cd) is a serious global issue due to its high mobility and toxicity. We investigated the application of insoluble polyacrylate polymers to improve soil and plant health. Sorghum was grown in a Cd-contaminated sandy soil. Polyacrylate polymers at 0.2% (w/w) were added to half of the soil. Control soil without plants was also included in the experiment. Growth of sorghum was stimulated in the polymer-amended soil. The concentration of Cd in the shoots, and the activities of catalase and ascorbate peroxidase decreased in plants from polymer-amended soil compared with unamended control. The amount of CaCl(2)-extractable Cd in the polymer-amended soil was 55% of that in the unamended soil. The Cd extracted in sorghum shoots was 0.19 mg per plant grown on soil without polymer and 0.41 mg per plant grown on polymer-amended soil. The total amount of Cd removed from each pot corresponded to 1.5 and more than 6% of soil CaCl(2)-extractable Cd in unamended and polymer-amended soil, respectively. The activities of soil acid phosphatase, beta-glucosidase, urease, protease and cellulase were greatest in polymer-amended soil with sorghum. In conclusion, the application of polyacrylate polymers to reduce the bioavailable Cd pool seems a promising method to enhance productivity and health of plants grown on Cd-contaminated soils.

Analysis of the contribution of CTL epitopes in the immunobiology of morbillivirus infection.

In Balb/c (H-2d) mice, the nucleoprotein (NP) of measles virus (MV) induces a MHC class I restricted-CTL response to a single 9-amino-acid epitope (aa 281--289). This L(d)-restricted epitope is also present in the NP of the closely related canine distemper virus (CDV). To investigate whether this epitope is immunologically effective when it is present within the primary sequence of a nonviral protein, we have incorporated the 281--289 motif into the human CD36 protein. When cells are infected with vaccinia virus (VV) recombinants expressing this protein, CD36NP, the MV epitope is correctly processed and the cells are lysed by MVNP-specific CTLs. In vivo, VV-CD36NP induced CTLs which protected mice from a lethal dose of CDV, but did not block virus replication. The MVNP contains four other potential L(d)-restricted motifs. To investigate if these could be utilized in the absence of the dominant epitope, a mutant NP was produced in which one of the anchor residues in the aa 281--289 motif was mutated. Cells infected with a VV recombinant expressing this protein (VV-NP F289S) were only poorly lysed by MVNP-specific CTLs. Similarly, immunization of Balb/c mice with VV-NP F289S induced a lower level of CTL activity compared to the VV-NP, but the activity was now directed to three other epitopes. When mice were vaccinated with VV-NP F289S they were only partially protected from a lethal CDV challenge. The significance of these results for MV vaccine development is discussed.

The ectodomain of measles virus envelope glycoprotein does not gain access to the cytosol and MHC class I presentation pathway following virus-cell fusion.

To unravel the intracellular fate of measles virus (MV) haemagglutinin (H) following fusion of the virus envelope with the cell membrane, its presentation by MHC molecules to T cells was explored. After MV infection, murine cells expressing CD46 were lysed by MHC class I-restricted CD8 CTLs specific for the ectodomain of H. In contrast, when sensitized with UV-inactivated MV, they were not lysed by these effectors, but were recognized by H-specific and class II-restricted CD4 CTLs. Thus, after MV binding and fusion, H becomes associated with plasma membrane and its ectodomain can reach the endosomal MHC-II but not the cytosolic MHC-I antigen presentation pathway. From these data and a reappraisal of previous reports, it appears that the ectodomains of both MV haemagglutinin fusion proteins, having undergone the fusion step, are not translocated into the cytosol and end up in the endosomes.

Measles virus DNA vaccination: antibody isotype is determined by the method of immunization and by the nature of both the antigen and the coimmunized antigen.

Plasmids encoding the measles virus hemagglutinin (HA) and nucleoprotein (NP) proteins inoculated into the skin of BALB/c mice by the gene gun method induced both humoral and cytotoxic lymphocyte class I-restricted immune responses. Although intramuscular immunization induces the immunoglobulin G2a (IgG2a) antibody isotype for both antigens, with gene gun immunization, the NP still generated mainly IgG2a and the major isotype induced by the HA was IgG1. Interestingly, gene gun coimmunization of HA and NP plasmids resulted in a dominant IgG1 HA response and the switching of antibodies generated against the NP to the IgG1 isotype.

Formaldehyde inactivation of measles virus abolishes CD46-dependent presentation of nucleoprotein to murine class I-restricted CTLs but not to class II-restricted helper T cells.

To induce an MHC-restricted specific CTL or Th response, an antigen must be delivered into the appropriate cellular compartment. We explored the role of CD46 in the presentation of measles virus (MV) nucleoprotein (NP) to murine NP-specific and MHC Class I-restricted polyclonal CTLs and the effect of inactivating MV by uv or formaldehyde. CD46(-)- and CD46(+)-transfected murine cells were used as target cells. After MV infection, only the targets which expressed CD46 were lysed by NP-specific class I-restricted CTLs. When MV was uv-inactivated, NP presentation by MHC class I molecules was retained but could be blocked by fusion inhibitors which block virus cell entry. When MV was inactivated with formaldehyde, NP was no longer presented by MHC class I molecules, although it was still presented by MHC class II molecules to a NP-specific class II-restricted T cell hybridoma. These data show that MV binding to the CD46 molecule is a prerequisite for virus-to-cell fusion and that cytosolic delivery of NP is necessary for presentation by class I molecules. Moreover, formaldehyde inactivation of virus induces the loss of class I-restricted presentation of NP due to selective abrogation of fusion and cytosolic delivery of NP.

Detection of autoantibodies against hGH in sera of idiopathic hypopituitary children.

Using reliable displacement radiobinding assay (RBA) and ELISA, the existence of anti-human growth hormone autoantibodies (hGHAA) was confirmed in idiopathic hypopituitary patients with growth impairment. Six of 35 hypopituitary patients (17.1%) and 1/85 (1.2%) control children proved positive for hGHAA by RBA (>control mean + 3 SD). IgG isotype-hGHAA by ELISAIgG (> control mean + 3 SD) were positive for 6/34 (17.7%) and 3/85 (3.5% hypopituitary and control children, respectively. Due to an asymmetry to the right of the ELISAIgG distribution, an alternative cutoff based on a nonparametric method was obtained, and positive results for hypopituitary children increased to 10/34 (29.4%). Three of 34 hypopituitary patients but no control children were positive for hGHAA of IgM isotype. The hGHAA were detected in children with or without perinatal problems. These autoantibodies may represent markers of a major autoimmune process involving a portion of the anterior pituitary and may contribute to the development of hypopituitarism in over 15% of hypopituitary children.

Immunization with plasmid DNA encoding for the measles virus hemagglutinin and nucleoprotein leads to humoral and cell-mediated immunity.

We have evaluated the DNA vaccination strategy for measles virus (MV) hemagglutinin (HA) and nucleoprotein (NP) genes. Plasmids encoding either the MV, HA, or NP proteins inoculated intramuscularly into Balb/c mice induced both humoral and CTL class I restricted responses. Antibody responses were not increased by multiple inoculations. The major antibody isotype induced by both the HA and NP was IgG2a consistent with a Th1 response. In contrast, immunization with a plasmid which directed the synthesis of a partially secreted form of HA gave mainly IgG1 antibodies. When the amount of DNA was reduced for the HA plasmid (1 or 10 microg/animal), although the antibody was not induced, a CTL response was observed.

Cellular and humoural autoimmunity markers in type 2 (non-insulin-dependent) diabetic patients with secondary drug failure.

In some cases patients with Type 2 (non-insulin-dependent) diabetes mellitus fail to respond to treatment with oral hypoglycaemic agents. These patients may respond in the same way as Type 1 (insulin-dependent) diabetic patients. Cellular immune aggression (defined as the capacity of peripheral mononuclear cells to inhibit stimulated insulin secretion by dispersed rat islet cells), insulin autoantibodies, C-peptide response and HLA antigens were determined in 31 Type 2 diabetic patients with secondary failure to oral hypoglycaemic agents and in 22 control subjects. Nine (29.03%) of the 31 Type 2 diabetic patients showed positive cellular immune aggression (2 SD below control group) and 22 (70.97%) presented no cellular immune aggression. There was a relationship between positive cellular immune aggression and each of the following parameters: age, body mass index and microangiopathy. No correlation was found between positive cellular immune aggression and glycaemia, HbA1, blood lipids or atherosclerosis. Patients with positive cellular immune aggression showed a significantly lower glucagon-stimulated C-peptide response vs those with no cellular immune aggression. Within a sub-group of patients who had never been treated with insulin, insulin autoantibodies were present in four of six patients with positive cellular immune aggression. DR2 antigen was found with decreased frequency in patients whereas no DR3/DR4 heterozygotes were observed. Our data support the hypothesis that a group of Type 2 diabetic patients with secondary failure to oral hypoglycaemic agents presented autoimmunity towards pancreatic Beta cells.

Heterologous humoral immune response in patients treated with human growth hormone from different sources.

The existence of homologous anti-human growth hormone (anti-hGH) and heterologous anti-bovine growth hormone (anti-bGH) humoral immune responses in hypopituitary patients under hGH therapy has been reported previously. In order to study the influence of the hormone source, both responses were compared by radiobinding assays performed with [125I]hGH or [125I]bGH as tracers. Fifty-seven hypopituitary patients treated with extractive hGH, recombinant methionyl hGH or authentic recombinant hGH were studied. A very low incidence of heterologous antibodies was found in patients under recombinant hGH therapy, contrary to the high incidence observed in patients treated with extractive hGH preparations. In addition, immunochemical studies performed with a synthetic peptide (hGH 44-128) indicated that this peptide exhibited, in the anti-bGH/[125]bGH radioimmunoassay system, higher reactivity than the native hGH, suggesting that such a fragment resembled an altered conformation of the hormone. The high heterologous response elicited only by the extractive hGH along with the behaviour of the hGH 44-128 fragment supports the fact that the extraction and purification procedures in extractive preparations may alter slightly the structure of the hGH molecule and trigger a heterologous immune response.

Presença dos antígenos C (rh') e/ou E (rh") em doadores de sangue Rh negativos analisados no Centro de Hematologia e Hemoterapia do Rio Grande do Norte - Hemonorte / Presence of the C (rh') and/or R (rh\")antigenes blood donors Rh negatives analysed at the Centro de Hematologia e Hemoterapia do Rio Grande do Norte - Hemonorte

Foram analisados o fator Rh de 32.455 doadores de sangue no Centro de Hamatologia e Hemoterapia do Rio grande do Norte – HEMONORTE e a freqüência dos ant´genos C (rh´) e E (rh”) de 3.3193 doadores Rh negativos, no período de janeiro de 1993 a maio de 1995, onde 2.966 (92,9 por cento) são do sexo masculino e 227 (7,1 por cento) são do sexo feminino. Para a demonstração dos dados, foram elaboradas tabelas com a freqüência do fator Rh, onde 90,16 por cento são fator Rh Positivos e 9,84 por cento são fator Rh negativos. Quanto a presença ou ausência dos antígenos C (rh’) e E (rh”), 172 (5,39 por cento) apresentam antígeno C (rh’), enquanto 77 (2,40 por cento) possuem o antígeno E (rh”), ficando 2.944 (92,21 por cento) dos doadores Rh Negativos sem a presença dos referidos Rh antígenos.

Los anticuerpos anti-insulina: origen, detección y relevancia clínica / Insulin antibodies: origin, detection and clinical relevance

Los anticuerpos anti-insulina aparecen en el paciente diabético como consecuencia del tratamiento con insulina exógena o bien espontáneamente en algunos de ellos con diagnóstico reciente que no han sido tratados con insulina (autoanticuerpos), como indicadores de una alteración inmune humoral. La magnitud de la respuesta inmune depende de factores tales como el tipo y especie de la preparación utilizada en la insulinoterapia y factores constitutivos propios del paciente. Los autoanticuerpos son marcadores de autoinmunidad y su presencia indica una tendencia del portador para desarrollar una diabetes, participando probablemente en el mecanismo de alteración insular. Los anticuerpos generados en respuesta a la insulina modifican la biodisponibilidad de la hormona, pudiendo incrementar el requerimiento diario de insulina, ocasionalmente ser responsable de algunos casos de hipoglucemias y participar en el mecanismo de produción de complicaciones tardías de la diabetes. La importancia de su detección ha generado gran espectativa y se han estandarizado las condiciones para su determinación