Improved Titer and Gene Transfer by Lentiviral Vectors Using Novel, Small ß-Globin Locus Control Region Elements.

ß-globin lentiviral vectors (ß-LV) have faced challenges in clinical translation for gene therapy of sickle cell disease (SCD) due to low titer and sub-optimal gene transfer to hematopoietic stem and progenitor cells (HSPCs). To overcome the challenge of preserving efficacious expression while increasing vector performance, we used published genomic and epigenomic data available through ENCODE to redefine enhancer element boundaries of the ß-globin locus control region (LCR) to construct novel ENCODE core sequences. These novel LCR elements were used to design a ß-LV of reduced proviral length, termed CoreGA-AS3-FB, produced at higher titers and possessing superior gene transfer to HSPCs when compared to the full-length parental ß-LV at equal MOI. At low vector copy number, vectors containing the ENCODE core sequences were capable of reversing the sickle phenotype in a mouse model of SCD. These studies provide a ß-LV that will be beneficial for gene therapy of SCD by significantly reducing the cost of vector production and extending the vector supply.

Reacción transfusional hemolítica retardada por aloanticuerpo anti-E / Reação transfusional hemolítica tardia por aloanticorpo anti-E / Delayed hemolytic transfusion reaction by anti-E alloantibody

Se presentó el caso de una señora de 47 años, grupo sanguíneo O Rh D positivo, G4P2A2, sin antecedente transfusional, que requería intervención quirúrgica por fibroma uterino sangrante. Las pruebas pretransfusionales resultaron compatibles, transfundiéndole dos concentrados de hematíes, sin presentar manifestaciones clínicas; trascurridos siete días, presentó una crisis hemolítica, con caída brusca de los parámetros hematológicos, palidez cutáneo-mucosa e íctero. Los estudios inmunohematológicos fueron positivos; la muestra de sangre de la enferma fue incompatible con un hijo, esposo y los hematíes transfundidos. Seis meses después se identificó un aloanticuerpo anti-E (es aquél anticuerpo que se produce como resultado de la exposición de un organismo a antígenos extraños) (128 diluciones) en la paciente. Se concluye que la paciente se aloinmunizó durante sus embarazos con un anticuerpo anti-E, donde la transfusión sanguínea fue el estímulo antigénico causante de la reacción transfusional hemolítica tardía(AU)

We present the case of a 47-year-old woman, blood group O Rh D positive, G4P2A2, with no transfusion history, which required surgical intervention due to bleeding uterine fibroma. The pretransfusional tests were compatible, transfusing two concentrates of red blood cells, without presenting clinical manifestations; after seven days, he presented a haemolytic crisis, with abrupt fall in hematological parameters, cutaneous-mucosal pallor and icterus. The immunohematological studies were positive; the patient's blood sample was incompatible with a son, husband and the transfused red blood cells. Six months later, an anti-E alloantibody (128 dilutions) was identified in the patient. It is concluded that the patient was alloimmunized during her pregnancies with an anti-E antibody, where the blood transfusion was the antigenic stimulus causing the late hemolytic transfusion reaction. Apresentamos o caso de uma mulher de 47 anos, grupo sanguíneo O Rh D positivo, G4P2A2, sem histórico transfusional, que necessitou de intervenção cirúrgica devido ao sangramento do fibroma uterino. Os exames pré-transfusionais foram compatíveis, transfundindo dois concentrados de hemácias, sem apresentar manifestações clínicas; Após sete dias, apresentou crise hemolítica, com queda abrupta nos parâmetros hematológicos, palidez cutâneo-mucosa e icterícia. Os estudos imuno-hematológicos foram positivos; a amostra de sangue do paciente era incompatível com um filho, marido e hemácias transfundidas. Seis meses depois, um aloanticorpo anti-E (128 diluições) foi identificado no paciente. Conclui-se que a paciente foi aloimunizada durante a gestação com anticorpo anti-E, sendo a hemotransfusão o estímulo antigênico causador da reação transfusional hemolítica tardia(AU)

Apresentamos o caso de uma mulher de 47 anos, grupo sanguíneo O Rh D positivo, G4P2A2, sem histórico transfusional, que necessitou de intervenção cirúrgica devido ao sangramento do fibroma uterino. Os exames pré-transfusionais foram compatíveis, transfundindo dois concentrados de hemácias, sem apresentar manifestações clínicas; Após sete dias, apresentou crise hemolítica, com queda abrupta nos parâmetros hematológicos, palidez cutâneo-mucosa e icterícia. Os estudos imuno-hematológicos foram positivos; a amostra de sangue do paciente era incompatível com um filho, marido e hemácias transfundidas. Seis meses depois, um aloanticorpo anti-E (128 diluições) foi identificado no paciente. Conclui-se que a paciente foi aloimunizada durante a gestação com anticorpo anti-E, sendo a hemotransfusão o estímulo antigênico causador da reação transfusional hemolítica tardia(AU)

Propagating Humanized BLT Mice for the Study of Human Immunology and Immunotherapy.

The humanized bone marrow-liver-thymus (BLT) mouse model harbors a nearly complete human immune system, therefore providing a powerful tool to study human immunology and immunotherapy. However, its application is greatly limited by the restricted supply of human CD34+ hematopoietic stem cells and fetal thymus tissues that are needed to generate these mice. The restriction is especially significant for the study of human immune systems with special genetic traits, such as certain human leukocyte antigen (HLA) haplotypes or monogene deficiencies. To circumvent this critical limitation, we have developed a method to quickly propagate established BLT mice. Through secondary transfer of bone marrow cells and human thymus implants from BLT mice into NSG (NOD/SCID/IL-2Rγ-/-) recipient mice, we were able to expand one primary BLT mouse into a colony of 4-5 proBLT (propagated BLT) mice in 6-8 weeks. These proBLT mice reconstituted human immune cells, including T cells, at levels comparable to those of their primary BLT donor mouse. They also faithfully inherited the human immune cell genetic traits from their donor BLT mouse, such as the HLA-A2 haplotype that is of special interest for studying HLA-A2-restricted human T cell immunotherapies. Moreover, an EGFP reporter gene engineered into the human immune system was stably passed from BLT to proBLT mice, making proBLT mice suitable for studying human immune cell gene therapy. This method provides an opportunity to overcome a critical hurdle to utilizing the BLT humanized mouse model and enables its more widespread use as a valuable preclinical research tool.

Manejo de antisépticos y desinfectantes en el servicio de cirugía del Instituto Gastroenterológico Boliviano Japonés: 2004 / manage of antiseptic and disinfectant in the service of surgery of the Institute Japanese Bolivian Gastroenterológico: 2004

El tema de este trabajo de investigación trata de los diferentes antisépticos y desinfectantes más utilizados hasta nuestros días, tomando en cuenta la clasificación, mecanismo de acción y principalmente el manejo de estas sustancias químicas para garantizar su efectividad beneficiando de esta manera al Servicio de Cirugía del Instituto Gastroenterológico Boliviano Japonés. Mediante la observación y una encuesta dirigida al personal profesional de enfermería, se obtuvieron como resultados criterios diversos: desconocimiento y la no existencia de protocolos específicamente elaborados en cuanto a los antisépticos y desinfectantes. Estas coclusiones motivaron la inquetud de plantear una propueta de intervención que consiste en la elaboración de un protocolo que será un instrumento práctico y útil que unifique criterios para el manejo de cada antiséptico y desinfectante. Con el apoyo del marco teórico se describen parámetros más sobresalientes como ser: sinónimos, nivel de acción, categoría, actividad microbiana, uso, dilución, propiedades, efectos adversos, conservación, precauciones, ventajas, desventajas y presentación.