RESUMEN
BACKGROUND AND OBJECTIVES: The isolation of neutrophils and subsequent detection of anti-human neutrophil antigens (HNA) antibodies are crucial in clinical medicine for the diagnosis of autoimmune neutropenia, neonatal alloimmune neutropenia (NAIN) and transfusion-related acute lung injury (TRALI). This study reports two cases of maternal anti-Fc-gamma-receptor-IIIb (FcγRIIIb) isoimmunization without NAIN symptoms and compares the efficiency of immunomagnetic negative selection (IMNS) with traditional dextran/Ficoll for neutrophil isolation in HNA serological assays. MATERIALS AND METHODS: Investigating two cases of maternal anti-FcγRIIIb isoimmunization, neutrophils from three donors were isolated from 8 mL of whole blood using IMNS and dextran/Ficoll. Serological assays included the granulocyte agglutination and immunofluorescence test, monoclonal antibody immobilization of granulocyte antigens and the LABScreen Multi (One Lambda). IMNS and dextran/Ficoll were compared in terms of cell yield, viability, time, cost and purity. RESULTS: Maternal anti-FcγRIIIb isoantibodies with FCGR3B gene deletion were detected in both cases. Newborns and fathers exhibited specific gene combinations: FCGR3B*02/FCGR3B*02 (Case 1) and FCGR3B*02/FCGR3B*03 (Case 2). IMNS outperformed dextran/Ficoll, yielding four times more neutrophils (average neutrophil counts: 18.5 × 103/µL vs. 4.5 × 103/µL), efficiently removing non-neutrophil cells and reducing processing time (30-40 min vs. 70-90 min), although it incurred a higher cost (2.7 times). CONCLUSION: Two cases of maternal anti-FcγRIIIb isoantibodies, unrelated to NAIN, were identified. Although neutropenia has not been described in these cases, we emphasize the importance of identifying asymptomatic cases with the potential for severe neutropenia. Additionally, IMNS is introduced as a rapid, high-yield, high-purity neutrophil isolation technique, beneficial for serological assays detecting anti-HNA antibodies.
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Isoanticuerpos , Neutrófilos , Receptores de IgG , Humanos , Neutrófilos/inmunología , Femenino , Receptores de IgG/inmunología , Isoanticuerpos/inmunología , Isoanticuerpos/sangre , Recién Nacido , Proteínas Ligadas a GPI/inmunología , Masculino , Separación Inmunomagnética/métodos , Adulto , Embarazo , Neutropenia/inmunología , Neutropenia/sangreRESUMEN
Neutrophils are essential to control several fungal infections. These cells are commonly known for their pro-inflammatory activities. However, some studies have demonstrated the anti-inflammatory properties of neutrophils during certain infectious diseases, culminating in the inhibition of T cell proliferation. Chromoblastomycosis (CBM) is a deep and progressive mycosis that affects thousands of people worldwide. Although neutrophil infiltrates are observed in the lesion histopathology, the fungus can overtake the immune system response and destroy the host-infected tissue. The present study demonstrated that neutropenic animals had an increase in the IL-6 production in the spleen and liver, followed by a lower fungal burden in these organs up to 14 days of infection. Neutropenic animals also showed a lower F. pedrosoi-specific antibody production 14-days post infection and higher T-cell proliferation in the in vitro experiments after stimulation with F. pedrosoi-purified proteins. Taken together, our results suggest that the presence of regulatory neutrophils in the mouse model of F. pedrosoi infection could act favoring the spread of the fungus and the chronicity of the infection. These findings shed light on the CBM treatment, which might target neutrophil polarization as a new therapy approach to treat CBM lesions.
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Anticuerpos/efectos adversos , Antígenos Ly/inmunología , Cromoblastomicosis/inmunología , Fonsecaea/patogenicidad , Neutropenia/inmunología , Neutrófilos/metabolismo , Linfocitos T/metabolismo , Animales , Polaridad Celular , Proliferación Celular , Cromoblastomicosis/complicaciones , Modelos Animales de Enfermedad , Fonsecaea/inmunología , Humanos , Interleucina-6/metabolismo , Hígado/inmunología , Activación de Linfocitos , Ratones , Neutropenia/inducido químicamente , Bazo/inmunologíaRESUMEN
BACKGROUND: Neonatal alloimmune neutropenia results from maternal alloimmunization to human neutrophil antigens. The alloantibodies involved in neonatal alloimmune neutropenia are against human neutrophil antigens HNA-1a, HNA-1b, HNA-1c, HNA-1d, HNA-2, HNA-3a, HNA-4a, HNA-4b, and HNA-5a; however, to date, antibodies specific to HNA-3b have not been reported. STUDY DESIGN AND METHODS: Blood samples from 10,000 unselected neonates were analyzed, resulting in the selection of 88 neutropenic newborns (neutrophil count <1.5 × 109 /L) from 83 mothers (three pairs of twins and one triplet). HNA-3 genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism to identify the cases of maternal-fetal HNA-3 incompatibility. Serologic studies for detecting maternal HNA-3 alloantibodies were performed with the granulocyte agglutination test, the white blood cell immunofluorescence test, and a LABScreen Multi-HNA Kit. RESULTS: Genotyping studies identified 13 of 88 (14.8%) instances of maternal-fetal HNA-3 incompatibility, with all mothers typed as HNA-3a/a and neonates typed as HNA-3a/b. Serologic studies revealed that five of 13 (38.5%) mothers carried anti-HNA-3b plus human leukocyte antigen antibodies and that three of 13 (23.1%) mothers had anti-HNA-3b without human leukocyte antigen antibodies. CONCLUSION: Here, we report the first three cases of neonatal alloimmune neutropenia associated with HNA-3b antibodies resulting in a neonatal alloimmune neutropenia incidence of one in 3333 live births.
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Enfermedades del Recién Nacido/inmunología , Isoanticuerpos/inmunología , Isoantígenos/inmunología , Neutropenia/inmunología , Incompatibilidad de Grupos Sanguíneos/etiología , Genotipo , Humanos , Lactante , Recién Nacido , Enfermedades del Recién Nacido/etiología , Isoanticuerpos/efectos adversos , Neutropenia/etiologíaRESUMEN
Abstract Introduction Invasive mold disease is an important complication of patients with hematologic malignancies, and is associated with high mortality. A diagnostic-driven approach has been an alternative to the classical empiric antifungal therapy. In the present study we tested an algorithm that incorporated risk stratification using the D-index, serial serum galactomannan and computed tomographic-scan to guide the decision to start antifungal therapy in neutropenic patients. Patients and methods Between May 2010 and August 2012, patients with acute leukemia in induction remission were prospectively monitored from day 1 of chemotherapy until discharge or death with the D-index and galactomannan. Patients were stratified in low, intermediate and high risk according to the D-index and an extensive workup for invasive mold disease was performed in case of positive galactomannan (≥0.5), persistent fever, or the appearance of clinical manifestations suggestive of invasive mold disease. Results Among 29 patients, 6 (21%), 11 (38%), and 12 (41%) were classified as high, intermediate, and low risk, respectively. Workup for invasive mold disease was undertaken in 67%, 73% and 58% (p = 0.77) of patients in each risk category, respectively, and antifungal therapy was given to 67%, 54.5%, and 17% (p = 0.07). Proven or probable invasive mold disease was diagnosed in 67%, 45.5%, and in none (p = 0.007) of high, intermediate, and low risk patients, respectively. All patients survived. Conclusion A risk stratification using D-index was a useful instrument to be incorporated in invasive mold disease diagnostic approach, resulting in a more comprehensive antifungal treatment strategy, and to guide an earlier start of treatment in afebrile patients under very high risk.
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Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Anciano , Adulto Joven , Aspergilosis/tratamiento farmacológico , Algoritmos , Fusariosis/tratamiento farmacológico , Mananos/sangre , Antifúngicos/uso terapéutico , Neutropenia/inmunología , Aspergilosis/diagnóstico , Aspergilosis/inmunología , Síndromes Mielodisplásicos/inmunología , Síndromes Mielodisplásicos/microbiología , Leucemia Mieloide Aguda/inmunología , Leucemia Mieloide Aguda/microbiología , Tomografía Computarizada por Rayos X , Estudios Prospectivos , Sensibilidad y Especificidad , Medición de Riesgo , Fusariosis/diagnóstico , Fusariosis/inmunología , Mananos/inmunología , Neutropenia/microbiologíaRESUMEN
INTRODUCTION: Invasive mold disease is an important complication of patients with hematologic malignancies, and is associated with high mortality. A diagnostic-driven approach has been an alternative to the classical empiric antifungal therapy. In the present study we tested an algorithm that incorporated risk stratification using the D-index, serial serum galactomannan and computed tomographic-scan to guide the decision to start antifungal therapy in neutropenic patients. PATIENTS AND METHODS: Between May 2010 and August 2012, patients with acute leukemia in induction remission were prospectively monitored from day 1 of chemotherapy until discharge or death with the D-index and galactomannan. Patients were stratified in low, intermediate and high risk according to the D-index and an extensive workup for invasive mold disease was performed in case of positive galactomannan (≥0.5), persistent fever, or the appearance of clinical manifestations suggestive of invasive mold disease. RESULTS: Among 29 patients, 6 (21%), 11 (38%), and 12 (41%) were classified as high, intermediate, and low risk, respectively. Workup for invasive mold disease was undertaken in 67%, 73% and 58% (p=0.77) of patients in each risk category, respectively, and antifungal therapy was given to 67%, 54.5%, and 17% (p=0.07). Proven or probable invasive mold disease was diagnosed in 67%, 45.5%, and in none (p=0.007) of high, intermediate, and low risk patients, respectively. All patients survived. CONCLUSION: A risk stratification using D-index was a useful instrument to be incorporated in invasive mold disease diagnostic approach, resulting in a more comprehensive antifungal treatment strategy, and to guide an earlier start of treatment in afebrile patients under very high risk.
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Algoritmos , Antifúngicos/uso terapéutico , Aspergilosis/tratamiento farmacológico , Fusariosis/tratamiento farmacológico , Mananos/sangre , Neutropenia/inmunología , Adulto , Anciano , Aspergilosis/diagnóstico , Aspergilosis/inmunología , Femenino , Fusariosis/diagnóstico , Fusariosis/inmunología , Galactosa/análogos & derivados , Humanos , Leucemia Mieloide Aguda/inmunología , Leucemia Mieloide Aguda/microbiología , Masculino , Mananos/inmunología , Persona de Mediana Edad , Síndromes Mielodisplásicos/inmunología , Síndromes Mielodisplásicos/microbiología , Neutropenia/microbiología , Estudios Prospectivos , Medición de Riesgo , Sensibilidad y Especificidad , Tomografía Computarizada por Rayos X , Adulto JovenRESUMEN
Therapeutic nonequivalence of generic antibiotics may lead to treatment failure and enrichment of resistance. However, there has been no demonstration that an equivalent generic displays the same resistance selection profile as the innovator drug. We aimed to test this hypothesis with five generic versions of ciprofloxacin by assessing their pharmaceutical equivalence with microbiological assays and their efficacy against Pseudomonas aeruginosa PAO1 in the neutropenic murine thigh infection model. One equivalent generic was selected for analysis by high-pressure liquid chromatography-tandem mass spectrometry (LC-MS/MS), to confirm chemical identity, and resistance selection experiments in a hollow-fiber (HF) system simulating two clinical dosing regimens. Total and resistant populations were measured, and the MICs of the resistant cells with and without an efflux pump inhibitor were determined. LC-MS/MS found no differences between products, and the innovator and the generic selected resistance with the same magnitude and mechanism after 7 days of treatment in the HF system, supporting the fact that a generic with demonstrated equivalence in vivo is also equivalent regarding resistance selection.
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Antibacterianos/farmacocinética , Ciprofloxacina/farmacocinética , Medicamentos Genéricos/uso terapéutico , Infecciones por Pseudomonas/tratamiento farmacológico , Pseudomonas aeruginosa/efectos de los fármacos , Animales , Antibacterianos/uso terapéutico , Ciprofloxacina/uso terapéutico , Ratones , Pruebas de Sensibilidad Microbiana , Neutropenia/inmunología , Equivalencia TerapéuticaRESUMEN
BACKGROUND: Interleukin-1 receptor antagonist polymorphism (ILRN) 2 (ILRN*2) has been associated with a poor outcome in septic patients because of an elevated production of anti-inflammatory cytokines. In >70% of patients, morbidity and mortality in childhood acute lymphoblastic leukemia is caused by infections. The aim of this study was to determine the association between this polymorphism and the frequency of septic shock from the time of diagnosis until completion of treatment. METHODS: This cohort study was conducted in 57 consecutive children with acute lymphoblastic leukemia. At the end of follow-up, children were stratified according to their IL1RN polymorphism (ILRN*1/ILRN*2), evaluating the impact of genotype on the severity of febrile neutropenic events during their treatment. RESULTS: Overall survival was 80% at 55 months after treatment. The average number of febrile neutropenic events in this cohort was 2.82 per patient. Genotype distribution was 50.9% for homozygote IL-1RN*1, 38.6% for heterozygote ILRN*1/ILRN*2 and 10.5% for homozygote IL-1RN*2. The risk of presenting septic shock for homozygote IL1RN*2/IL1RN*2 and heterozygote ILRN*1/ILRN*2 patients was significantly greater (odds ratio, 45; P = 0.001) adjusted for age, gender, risk of leukemia and presence of pathogenic bacteria. Genotype IL-1RN*2 is associated with the risk of development of septic shock in children with acute lymphoblastic leukemia. Further research in larger population-based studies is needed to replicate these findings. CONCLUSIONS: This information would allow us to identify more predictive factors in this group of acute lymphoblastic leukemia patients in whom this information is lacking to establish an earlier and more aggressive approach.
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Proteína Antagonista del Receptor de Interleucina 1/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Choque Séptico/genética , Adolescente , Distribución de Chi-Cuadrado , Niño , Preescolar , Femenino , Fiebre/genética , Fiebre/inmunología , Estudios de Seguimiento , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Lactante , Proteína Antagonista del Receptor de Interleucina 1/inmunología , Modelos Logísticos , Masculino , Neutropenia/genética , Neutropenia/inmunología , Leucemia-Linfoma Linfoblástico de Células Precursoras/inmunología , Choque Séptico/inmunología , Estadísticas no ParamétricasRESUMEN
BACKGROUND: Febrile neutropenia (FN) and infection-related mortality are major problems for children with cancer in low-income countries. Identifying predictors for adverse outcome of FN in low-income countries permits targeted interventions. We describe the nature and predictors of microbiologically documented infection (MDI) and mortality of FN in children with cancer in El Salvador. METHODS: We examined Salvadoran pediatric oncology patients admitted with FN over a 1-year period. Data were collected prospectively. Demographic, treatment, and admission-related variables were examined as predictors of outcomes. RESULTS: Hundred six FN episodes among 85 patients were included. Twenty-three of 106 episodes (22%) were microbiologically documented; 13 of 106 episodes (12%) resulted in death. Gram-positive and gram-negative organisms were isolated in 14 of 23 and 11 of 23 specimens; polymicrobial infections were common (11 of 23 episodes of MDI). Older age decreased the MDI risk [odds ratio (OR) per year=0.87, 95% confidence interval (CI), 0.75-0.99; P=0.04] while increasing number of days since the last chemotherapy increased the risk (OR=1.03 per day, 95% CI, 1.01-1.04; P=0.002). Pneumonia diagnosed either clinically (OR=6.6, 95% CI, 1.8-30.0; P=0.005) or radiographically (OR=5.5, 95% CI, 1.7-18.1; P=0.005) was the only predictor of mortality. CONCLUSIONS: In El Salvador, polymicrobial infections were common. Pneumonia at admission identified children with FN at high risk of death; these children may benefit from targeted interventions.
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Fiebre/mortalidad , Infecciones por Bacterias Gramnegativas/mortalidad , Infecciones por Bacterias Grampositivas/mortalidad , Neoplasias/mortalidad , Neutropenia/mortalidad , Adolescente , Niño , Preescolar , El Salvador/epidemiología , Femenino , Fiebre/inmunología , Fiebre/microbiología , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Grampositivas/inmunología , Humanos , Lactante , Masculino , Neoplasias/inmunología , Neutropenia/inmunología , Neumonía Bacteriana/inmunología , Neumonía Bacteriana/mortalidad , Valor Predictivo de las Pruebas , Prevalencia , Factores de RiesgoRESUMEN
Las enfermedades autoinmunes son trastornos secundarios a una des regulación del sistema inmune; éste pierde la capacidad de autotolerancia, y produce un daño crónico, continuo y progresivo. Son patologías muy diversas y pueden llegar a afectar hasta el 5 por ciento de la población. Pueden ser clasificadas en sistémicas u órgano-especificas, dependiendo si el antígeno reconocido es exclusivo del tejido target. A este grupo de enfermedades corresponden las citopenias autoinmunes, condiciones que se caracterizan por disminución del recuento de glóbulos rojos, plaquetas, leucocitos o la mezcla de ellos. Si bien los mecanismos patogénicos que participan en producción de cada una de ellas son similares, cada la tiene características que la hacen particularmente interesante. De esta manera tenemos que la anemia hemolítica autoinmune (AHAI) en un síndrome clínico que se produce debido a la disminución de glóbulos rojos secundaria a una destrucción exagerada de ellos, mediada por la alteración de la respuesta inmune, en la que los antígenos de la membrana de estas células son reconocidos como extraños por nuestro sistema inmune. La clasificación de la AHA/ está dada por la temperatura óptima de unión entre el anticuerpo y la membrana del glóbulo rojo; de esta forma, tenemos tres grandes grupos: por Anticuerpos calientes (WAHA), por Anticuerpos fríos (CAHA) y Mixtas (mediadas por anticuerpos calientes y frios). El diagnóstico de AHAl se confirma con la demostración de anticuerpos y/o proteínas del complemento en la superficie de los eritrocitos; entre ellos encontramos: Coombs directo y Coombs indirecto, test de Coombs recto anti-lgM , búsqueda de C3b en la membrana del glóbulo rojo y finalmente la prueba cualitativa de Donath-Landsteine Las trombopenias autoinmunes corresponden a un grupo de desórdenes que se caracterizan por tener un recuento plaquetario bajo 150.000, mediado por una alteración del sistema inmune. (3.14) La trombocitopenia autoinmune puede ser...
Autoimmune diseases are disorders secondary to a deregulation of the immune system, which loses the capacity for self-tolerance, resulting in chronic, continuous and progressive damage. These pathologies are very diverse and affect up to 5 percent of the population. They can be classified into systemic or organ-specific, depending on whether the recognized antigen is unique to the target tissue. Autoimmune cytopenias belong to this group of diseases and are characterized by a decreased count of red blood cells, platelets, leukocytes, or a mix of these. While the pathogenic mechanisms involved in the production of each are similar, each has features that make it particularly interesting. We have autoimmune hemolytic anemia (AlHA), which is a clinical syndrome that occurs due to a decreased number of red blood cells secondary to an exaggerated destruction of the same, mediated by an alteration of the immune response, in which the membrane antigens of these cells are recognized as foreign by our immune system. The classification of AIHA is given by the optimum bond temperature between the antibody and red blood cell membrane. There are 3 main groups: warm antibodies(WAHA), Cold antibodies (CAHA) and mixed (mediated by hot and cold antibodies). AIHA diagnosis is confirmed with the demonstration of antibodies and/or complement proteins on the surface of red blood cells, among these are the direct and indirect Coombs test, direct anti-IgM Coombs test, the search for C3b in the membrane of the red cell and, finally, the qualitative Donath-Landsteiner test. Autoimmune thrombocytopenia isassociated with a group of disorders characterized by a platelet count under 150,000, mediated by an alteration of the immune system.(3-14) Autoimmune thrombocytopenia can be classified as primary or secondary, and these, in turn, into acute and chronic, depending on whether they last 6 months or more, respectively. Antibodies that react against the glycoproteins of the plate...
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Humanos , Anemia Hemolítica Autoinmune/inmunología , Neutropenia/inmunología , Púrpura Trombocitopénica Idiopática/inmunología , Anticuerpos , Autoinmunidad , Enfermedades Autoinmunes/inmunología , Granulocitos , Hemoglobinuria Paroxística/inmunología , Sistema Inmunológico , Receptores Fc , Linfocitos TRESUMEN
Auto or alloantibodies reactive with neutrophils define immune neutropenia. Alloimmune neonatal neutropenia is caused by maternal sensitization to paternal neutrophil antigens, resulting in IgG antibodies that are transferred to the fetus through the placenta. We present the studies in 4 children from 3 families with neutropenia of unknown origin (two of them were brothers). They were evaluated by flow cytometry in parallel with leukoagglutination. Reference values were established for serum reactive IgG in healthy volunteers for three dilutions (1/2, 1/5 and 1/20), both for the autologous reaction (serum and cells of the same individual) and for the heterologous reaction (serum and cells of different individuals). Results were expressed by an index defined by the quotient of the mean fluorescence intensity of the patient's serum divided by that of the reference serum. Serum reactive/agglutinant factors and circulating immune complexes were evaluated in patients and parents serum. Neutrophil specific phenotypes were determined for HNA-1a, HNA-1b and HNA-2a. Reactive IgG/agglutinant factors were found in 4 children. Two maternal sera were reactive against paternal and/or children neutrophils. Circulating immune complexes were detected in 2/4 children sera and were negative in 3/3 maternal sera. Maternal/children incompatibility was detected in the four cases. The three mothers had the same phenotype: homozygous NA1/NA1, NB1+.
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Anticuerpos Anticitoplasma de Neutrófilos/inmunología , Inmunoglobulina G/sangre , Neutropenia/inmunología , Neutrófilos/inmunología , Aglutinación/inmunología , Anticuerpos Anticitoplasma de Neutrófilos/sangre , Eosinófilos/metabolismo , Femenino , Citometría de Flujo/métodos , Factor Estimulante de Colonias de Granulocitos , Humanos , Lactante , Recuento de Leucocitos , Masculino , Neutropenia/sangre , Neutrófilos/metabolismo , Fenotipo , Embarazo , Proteínas Recombinantes , Valores de ReferenciaRESUMEN
A neutropenia febril(NF) é uma complicação muito freqüente nos pacientes com neoplasias hematológicas submetidos à quimioterapia. Determinar a prevalência e a sensibilidade aos antimicrobianos dos germes isolados de hemoculturas(HMC) de pacientes hematológicos com NF internados no HCPA. Foram analisadas, retrospectivamente, todas as HMC solicitadas para pacientes com NF(contagem total de neutrófilos<500/uL) internados na Unidade de Hematologia do HCPA entre fev/2003 e fev/2005. Considerou-se febre uma temperatura axilar>38,5oC ou duas medidas>38oC em 24 horas. Comparou-se o perfil de sensibilidade das infecções em pacientes com NF em relação às demais unidades de internação clínica do hospital. Foram solicitadas no período em estudo 2389 hemoculturas para 178 pacientes. Destas, 719 (30,1 por cento) foram positivas sendo que este percentual subiu para 38,7 por cento quando consideradas apenas as hemoculturas coletas sem a vigência de antibiótico. A análise dos germes isolados revelou que a maioria das infecções foi causada por Bacilos Gram-negativos(53,8 por cento) seguido pelos Cocos Gram-positivos(31,2 por cento). Os microrganismos isolados com maior freqüência foram: E. coli(14,6 por cento), S. aureus(13,8 por cento), K. pneumoniae(12,2 por cento), Streptococcus sp.(8,5 por cento), Pseudomonas sp. (7,9 por cento), Staphylococcus coagulase-negativo (7,3 por cento). A sensibilidade dos Bacilos Gram-negativos aos diferentes antimicrobianos foi: Cefepime(63 por cento), Amicacina(65 por cento), Ciprofloxacin(63 por cento), Piperacilina/Tazobactam(71 por cento), Ceftazidima(63 por cento), Meropenem(97 por cento). A sensibilidade dos Cocos Gram-positivos foi: Oxacilina(30 por cento), Levofloxacin(38 por cento), Vancomicina(100 por cento). Comparativamente ao perfil de sensibilidade das unidades de internação clínica adulta não-hematológicas constatou-se diferença estatisticamente significante(p<0,05) entre a sensibilidade dos Bacilos Gram-negativos ao Cefepime...
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Humanos , Masculino , Femenino , Neutropenia , Neutropenia/epidemiología , Neutropenia/inmunología , Protocolos Clínicos , HematologíaRESUMEN
Bothrops atrox crude venom injected intraperitoneal (i.p.) into BALB/c mice induced local afflux of inflammatory cells, one neutrophil-rich peak after 6h and another macrophage-rich peak after 48 h. A similar pattern of local cell afflux plus edema, Delta lesions of some skeletal muscle cells, and hemorrhage were observed in mice intramuscular (i.m.) injected with the venom. Measurement of serum cytokines in neutrophil-depleted (by anti-mouse rat monoclonal antibody (mAb) RB6-8C5) and non-depleted BALB/c mice was performed by ELISA. With the exception of IL-1beta (78 pg/ml), higher levels of IL-6 (1348 pg/ml), MIP-1beta (437 pg/ml) and MIP-2 (904 pg/ml) were observed in neutrophil-depleted mice, in comparison to the values found in non-neutrophil depleted mice: IL-1beta (437 pg/ml), IL-6 (750 pg/ml), MIP-1beta (165 pg/ml) and MIP-2 (90 pg/ml). TNF-alpha was not detected. NO was detected (18 microM) 24h after venom injection in neutrophil-depleted mice. RT-PCR using representative primers detected expression of mRNA in cells from BALB/c mice injected with B. atrox venom: (a) for IL-1beta, IL-6, inducible nitric oxide synthase (iNOS), CXCR2, MIP-2 and RANTES in cells from mice that were neutrophil-depleted or not; (b) for CCR1, CCR5 and MIP-1beta in cells from neutrophil-depleted mice; (c) for MIP-1alpha in cells from non-neutrophil-depleted mice; (d) TNF-alpha and TGF-beta were not detected in either of the mice. These results indicate that neutrophils play a role in regulating the production of some cytokines and chemokines as well as locally expressed or liberated iNOS/NO in tissues injected with B. atrox crude venom.
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Quimiocinas/biosíntesis , Venenos de Crotálidos/administración & dosificación , Neutrófilos/inmunología , Neutrófilos/metabolismo , Óxido Nítrico Sintasa/biosíntesis , Óxido Nítrico/biosíntesis , Animales , Anticuerpos Monoclonales/administración & dosificación , Bothrops , Línea Celular , Movimiento Celular/inmunología , Quimiocinas/genética , Venenos de Crotálidos/toxicidad , Ratones , Ratones Endogámicos BALB C , Neutropenia/enzimología , Neutropenia/inmunología , Neutropenia/metabolismo , Neutrófilos/enzimología , Neutrófilos/patología , Óxido Nítrico Sintasa/genética , Ratas , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/inmunologíaRESUMEN
La neutropenia inmune se diagnostica por la presencia de auto o aloanticuerpos reactivos con losneutrófilos. La neutropenia aloinmune neonatal es consecuencia de la sensibilización materna alos antígenos específicos de los neutrófilos paternos que afectan al neonato al atravesar la barrera placentaria. Se presentan 4 casos de niños, 2 de ellos hermanos consanguíneos con doble vínculo. Se estudiaron los sueros de los pacientes y sus padres. Por citometría de flujo se establecen los valores de referencia de la IgG sérica reactiva con los neutrófilos en voluntarios sanos, para 3 diluciones (1/2, 1/5 y 1/20) en reacción autóloga(suero y células de un mismo individuo) y heteróloga (suero y células de diferentes individuos). Los resultadosse expresan por un índice definido como el cociente entre la mediana de la intensidad de fluorescencia media del suero incógnita y la de un suero utilizado como referencia. Por leucoaglutinación se evaluó la dilucióndel suero 1/20. Se determinó el nivel de complejos inmunes circulantes. Se determinó el fenotipo, para los epitopes HNA-1a, HNA-1b y HNA-2a. En los 4 niños se encontró IgG reactiva y/o factores aglutinantes; 2/3 sueros maternos fueron reactivos con los neutrófilos del cónyuge y de los hijos. Los complejos inmunes circulantes fueron positivos en 2/4 sueros negativos en 3/3 sueros maternos. Se encontró incompatibilidad materno-infantil en los 4 casos. Las 3 madres tenían igual fenotipo: homocigotos NA1/NA1, NB1+. En síntesis, se presenta el hallazgo de 4 casos con neutropenia inmune: 3/4 auto-inmune, 1/3 se asocia a complejos inmunes circulantes y 1/4 con neutropenia neonatal aloinmune (AU)
Auto or alloantibodies reactive with neutrophils define immune neutropenia. Alloimmune neonatal neutropenia is caused by maternal sensitization to paternal neutrophil antigens, resulting in IgG antibodies that are transferred to the fetus through the placenta. We present the studies in 4 children from 3 families with neutropenia of unknown origin (two of them were brothers). Theywere evaluated by flow cytometry in parallel with leukoagglutination. Reference values were established forserum reactive IgG in healthy volunteers for three dilutions (1/2, 1/5 and 1/20), both for the autologous reaction (serum and cells of the same individual) and for the heterologous reaction (serum and cells of differentindividuals). Results were expressed by an index defined by the quotient of the mean fluorescence intensityof the patientãs serum divided by that of the reference serum. Serum reactive/agglutinant factors and circulating immune complexes were evaluated in patients and parents serum. Neutrophil specific phenotypes weredetermined for HNA-1a, HNA-1b and HNA-2a. Reactive IgG/agglutinant factors were found in 4 children. Twomaternal sera were reactive against paternal and/or children neutrophils. Circulating immune complexes weredetected in 2/4 children sera and were negative in 3/3 maternal sera. Maternal/children incompatibility wasdetected in the four cases. The three mothers had the same phenotype: homozygous NA1/NA1, NB1+ (AU)
Asunto(s)
Humanos , Masculino , Femenino , Embarazo , Lactante , Anticuerpos Anticitoplasma de Neutrófilos/inmunología , Citometría de Flujo/métodos , Inmunoglobulina G/sangre , Neutropenia/inmunología , Neutrófilos/inmunología , Aglutinación/inmunología , Anticuerpos Anticitoplasma de Neutrófilos/sangre , Eosinófilos/metabolismo , Factor Estimulante de Colonias de Granulocitos , Recuento de Leucocitos , Neutropenia/sangre , Neutrófilos/metabolismo , Fenotipo , Valores de ReferenciaRESUMEN
La neutropenia inmune se diagnostica por la presencia de auto o aloanticuerpos reactivos con losneutrófilos. La neutropenia aloinmune neonatal es consecuencia de la sensibilización materna alos antígenos específicos de los neutrófilos paternos que afectan al neonato al atravesar la barrera placentaria. Se presentan 4 casos de niños, 2 de ellos hermanos consanguíneos con doble vínculo. Se estudiaron los sueros de los pacientes y sus padres. Por citometría de flujo se establecen los valores de referencia de la IgG sérica reactiva con los neutrófilos en voluntarios sanos, para 3 diluciones (1/2, 1/5 y 1/20) en reacción autóloga(suero y células de un mismo individuo) y heteróloga (suero y células de diferentes individuos). Los resultadosse expresan por un índice definido como el cociente entre la mediana de la intensidad de fluorescencia media del suero incógnita y la de un suero utilizado como referencia. Por leucoaglutinación se evaluó la dilucióndel suero 1/20. Se determinó el nivel de complejos inmunes circulantes. Se determinó el fenotipo, para los epitopes HNA-1a, HNA-1b y HNA-2a. En los 4 niños se encontró IgG reactiva y/o factores aglutinantes; 2/3 sueros maternos fueron reactivos con los neutrófilos del cónyuge y de los hijos. Los complejos inmunes circulantes fueron positivos en 2/4 sueros negativos en 3/3 sueros maternos. Se encontró incompatibilidad materno-infantil en los 4 casos. Las 3 madres tenían igual fenotipo: homocigotos NA1/NA1, NB1+. En síntesis, se presenta el hallazgo de 4 casos con neutropenia inmune: 3/4 auto-inmune, 1/3 se asocia a complejos inmunes circulantes y 1/4 con neutropenia neonatal aloinmune
Auto or alloantibodies reactive with neutrophils define immune neutropenia. Alloimmune neonatal neutropenia is caused by maternal sensitization to paternal neutrophil antigens, resulting in IgG antibodies that are transferred to the fetus through the placenta. We present the studies in 4 children from 3 families with neutropenia of unknown origin (two of them were brothers). Theywere evaluated by flow cytometry in parallel with leukoagglutination. Reference values were established forserum reactive IgG in healthy volunteers for three dilutions (1/2, 1/5 and 1/20), both for the autologous reaction (serum and cells of the same individual) and for the heterologous reaction (serum and cells of differentindividuals). Results were expressed by an index defined by the quotient of the mean fluorescence intensityof the patients serum divided by that of the reference serum. Serum reactive/agglutinant factors and circulating immune complexes were evaluated in patients and parents serum. Neutrophil specific phenotypes weredetermined for HNA-1a, HNA-1b and HNA-2a. Reactive IgG/agglutinant factors were found in 4 children. Twomaternal sera were reactive against paternal and/or children neutrophils. Circulating immune complexes weredetected in 2/4 children sera and were negative in 3/3 maternal sera. Maternal/children incompatibility wasdetected in the four cases. The three mothers had the same phenotype: homozygous NA1/NA1, NB1+
Asunto(s)
Humanos , Masculino , Femenino , Embarazo , Lactante , Anticuerpos Anticitoplasma de Neutrófilos/inmunología , Citometría de Flujo/métodos , Inmunoglobulina G/sangre , Neutropenia/inmunología , Neutrófilos/inmunología , Aglutinación/inmunología , Anticuerpos Anticitoplasma de Neutrófilos/sangre , Eosinófilos/metabolismo , Recuento de Leucocitos , Neutropenia/sangre , Neutrófilos/metabolismo , Fenotipo , Valores de ReferenciaRESUMEN
La neutropenia inmune se diagnostica por la presencia de auto o aloanticuerpos reactivos con losneutrófilos. La neutropenia aloinmune neonatal es consecuencia de la sensibilización materna alos antígenos específicos de los neutrófilos paternos que afectan al neonato al atravesar la barrera placentaria. Se presentan 4 casos de niños, 2 de ellos hermanos consanguíneos con doble vínculo. Se estudiaron los sueros de los pacientes y sus padres. Por citometría de flujo se establecen los valores de referencia de la IgG sérica reactiva con los neutrófilos en voluntarios sanos, para 3 diluciones (1/2, 1/5 y 1/20) en reacción autóloga(suero y células de un mismo individuo) y heteróloga (suero y células de diferentes individuos). Los resultadosse expresan por un índice definido como el cociente entre la mediana de la intensidad de fluorescencia media del suero incógnita y la de un suero utilizado como referencia. Por leucoaglutinación se evaluó la dilucióndel suero 1/20. Se determinó el nivel de complejos inmunes circulantes. Se determinó el fenotipo, para los epitopes HNA-1a, HNA-1b y HNA-2a. En los 4 niños se encontró IgG reactiva y/o factores aglutinantes; 2/3 sueros maternos fueron reactivos con los neutrófilos del cónyuge y de los hijos. Los complejos inmunes circulantes fueron positivos en 2/4 sueros negativos en 3/3 sueros maternos. Se encontró incompatibilidad materno-infantil en los 4 casos. Las 3 madres tenían igual fenotipo: homocigotos NA1/NA1, NB1+. En síntesis, se presenta el hallazgo de 4 casos con neutropenia inmune: 3/4 auto-inmune, 1/3 se asocia a complejos inmunes circulantes y 1/4 con neutropenia neonatal aloinmune (AU)
Auto or alloantibodies reactive with neutrophils define immune neutropenia. Alloimmune neonatal neutropenia is caused by maternal sensitization to paternal neutrophil antigens, resulting in IgG antibodies that are transferred to the fetus through the placenta. We present the studies in 4 children from 3 families with neutropenia of unknown origin (two of them were brothers). Theywere evaluated by flow cytometry in parallel with leukoagglutination. Reference values were established forserum reactive IgG in healthy volunteers for three dilutions (1/2, 1/5 and 1/20), both for the autologous reaction (serum and cells of the same individual) and for the heterologous reaction (serum and cells of differentindividuals). Results were expressed by an index defined by the quotient of the mean fluorescence intensityof the patientãs serum divided by that of the reference serum. Serum reactive/agglutinant factors and circulating immune complexes were evaluated in patients and parents serum. Neutrophil specific phenotypes weredetermined for HNA-1a, HNA-1b and HNA-2a. Reactive IgG/agglutinant factors were found in 4 children. Twomaternal sera were reactive against paternal and/or children neutrophils. Circulating immune complexes weredetected in 2/4 children sera and were negative in 3/3 maternal sera. Maternal/children incompatibility wasdetected in the four cases. The three mothers had the same phenotype: homozygous NA1/NA1, NB1+ (AU)
Asunto(s)
Humanos , Masculino , Femenino , Embarazo , Lactante , Anticuerpos Anticitoplasma de Neutrófilos/inmunología , Citometría de Flujo/métodos , Inmunoglobulina G/sangre , Neutropenia/inmunología , Neutrófilos/inmunología , Aglutinación/inmunología , Anticuerpos Anticitoplasma de Neutrófilos/sangre , Eosinófilos/metabolismo , Factor Estimulante de Colonias de Granulocitos , Recuento de Leucocitos , Neutropenia/sangre , Neutrófilos/metabolismo , Fenotipo , Valores de ReferenciaRESUMEN
Chronic idiopathic neutropenia of adults (CINA) is a granulocytic disorder characterised by the "unexplained" decrease in the number of circulating neutrophils. Serum inflammatory cytokines and chemokines are increased in CINA. In addition, cytokines gene polymorphisms are associated with increased levels of respective products and related with inflammatory diseases. The aim of the present study was to investigate the association of polymorphisms of IL1B-511C/T and +3953C/T, IL1RN intron 2, IL6-174G/C and TNF-308G/A genes with CINA. We analysed 29 CINA and controls by polymerase chain reaction and restriction fragment length polymorphism. Statistical analyses were performed using chi2 test, and the Hardy-Weinberg equilibrium (HWE) was investigated. All alleles analysed were in HWE in both populations. Similar frequencies of IL1B-511C/T, IL1B+3953C/T, IL1RN, IL6-174G/C and TNF-308G/A genotypes were observed in CINA and controls. These results suggest that cytokine polymorphisms associated with control of gene expression and protein levels were not associated with occurrence of CINA and were not responsible for the increased cytokine in CINA patients.
Asunto(s)
Interleucina-1/genética , Interleucina-6/genética , Neutropenia/genética , Polimorfismo Genético , Polimorfismo de Nucleótido Simple , Factor de Necrosis Tumoral alfa/genética , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Familia de Multigenes , Neutropenia/inmunologíaRESUMEN
The incidence of systemic fungal infections increased during the last two decades. Rare fungi, such as Mucor, Fusarium and Paecilomyces, are emerging as causes of systemic fungal infections in immunocompromised hosts. There are reports of cutaneous infections, endophthalmitis, keratitis, sinusitis, neuropathy and fungemia in immunocompromised and immunocompetent adult patients. We report a 5 years old neutropenic patient with acute myeloid leukemia treated with multiple courses of chemotherapy, with a fungemia caused by Paecilomyces lilacinus (PL). His initial clinical course was characterized by fever, skin lesions, respiratory distress and shock. Blood and bone marrow cultures were positive. The patient was treated with amphotericin B and itraconazole with a good clinical response.
Asunto(s)
Huésped Inmunocomprometido/inmunología , Micosis/microbiología , Neutropenia/complicaciones , Infecciones Oportunistas/microbiología , Paecilomyces , Enfermedad Aguda , Preescolar , Humanos , Leucemia Mieloide/tratamiento farmacológico , Leucemia Mieloide/inmunología , Leucemia Mieloide/microbiología , Masculino , Micosis/inmunología , Neutropenia/inmunología , Neutropenia/microbiología , Infecciones Oportunistas/inmunologíaRESUMEN
To study the role of neutrophils in the innate resistance to Entamoeba histolytica intestinal infection in mice, animals were treated with anti-neutrophil monoclonal antibodies prior to intracecal parasite inoculation and the resulting lesions were compared with normal mice that had been equally infected. In contrast to our previous finding that neutrophils are critical in eliminating E. histolytica infection in the liver, we show here that neutrophils are not absolutely required to eliminate E. histolytica infection from the intestine. Although the neutrophils are not critical for resolution of the E. histolytica infection, neutrophils do appear to provide some measure of protection as the intestinal amoeba burden was higher at early timepoints after infection in the neutropenic animals. In addition, we found that while both the normal and the neutrophil-depleted mice developed ulcerative lesions in the colon, the neutropenic mice had an increased frequency of granulomas that formed around the amoeba. Thus, our findings appear to be the first evidence showing that granulomatous inflammation can occur after intestinal infection in mice using axenically cultured amoeba.
Asunto(s)
Disentería Amebiana/inmunología , Entamoeba histolytica/inmunología , Granuloma/inmunología , Intestinos/patología , Neutrófilos/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Ciego/parasitología , Disentería Amebiana/patología , Entamoeba histolytica/fisiología , Femenino , Granuloma/parasitología , Granuloma/patología , Inmunoglobulina G/inmunología , Intestinos/inmunología , Intestinos/parasitología , Masculino , Ratones , Ratones Endogámicos BALB C , Neutropenia/inmunología , Neutropenia/parasitología , Neutropenia/patología , Úlcera/inmunología , Úlcera/parasitología , Úlcera/patologíaRESUMEN
PURPOSE: To identify clinical and laboratory parameters present at the time of a first evaluation that could help predict which children with cancer, fever, and neutropenia were at high risk or low risk for an invasive bacterial infection. PATIENTS AND METHODS: Over a 17-month period, all children with cancer, fever, and neutropenia admitted to five hospitals in Santiago, Chile, were enrolled onto a prospective protocol. Associations between admission parameters and risk for invasive bacterial infection were assessed by univariate and logistic regression analyses. RESULTS: A total of 447 febrile neutropenic episodes occurred in 257 children. Five parameters were statistically independent risk factors for an invasive bacterial infection. Ranked by order of significance, they were as follows: C-reactive protein levels of 90 mg/L or higher (relative risk [RR], 4.2; 95% confidence interval [CI], 3.6 to 4.8); presence of hypotension (RR, 2.7; 95% CI, 2.3 to 3.2); relapse of leukemia as cancer type (RR, 1.8, 95% CI, 1.7 to 2.3); platelet count less than or equal to 50,000/mm(3) (RR, 1.7; 95% CI, 1.4 to 2.2); and recent (< or = 7 days) chemotherapy (RR, 1.3; 95% CI, 1.1 to 1.6). Other previously postulated risk factors (magnitude of fever, monocyte count) were not independent risk factors in this study population. CONCLUSION: In a large population of children, common clinical and laboratory admission parameters were identified that can help predict the risk for an invasive bacterial infection. These results encourage the possibility of a more selective management strategy for these children.