Viral-specific T cells for Cytomegalovirus retinitis following hematopoietic stem cell transplantation: A success story.

Cytomegalovirus retinitis (CMVR) following hematopoietic stem cell transplantation (HCT) for a primary immunodeficiency is a rare but highly morbid condition with potential irreversible consequences despite optimal antiviral pharmacotherapy. Viral-specific T cells (VSTs) pose a promising and safe approach eradicating intractable viral disease. We describe the case of a 21-month-old male with Wiskott-Aldrich syndrome (WAS) and CMVR post HCT with sustained long-term virologic and clinical response after CMV-specific T-cell therapy. This case highlights the need to consider VST as an adjunct upfront strategy in refractory CMVR and for routine ophthalmologic screening and surveillance in high-risk patients post HCT.

Use of cardiac radiation therapy as bridging therapy to CAR-T for relapsed pediatric B-cell acute lymphoblastic leukemia.

The use of radiotherapy as bridging therapy to chimeric antigen receptor T-cell therapy (CAR-T) in pre-B acute lymphoblastic leukemia (B-ALL) has been minimally explored. Here, we present a boy with B-ALL who relapsed after allogeneic bone marrow transplant with disseminated disease, including significant symptomatic cardiovascular and gastrointestinal (GI) involvement. The cardiac and GI leukemic infiltrates were successfully treated with bridging radiation therapy (BRT) prior to CAR-T infusion. Using this approach, he successfully tolerated CAR-T with no evidence of disease or sequelae on 3-month follow-up. This is the first reported case of safe and effective delivery of cardiac BRT in B-ALL.

Immunomodulation with pomalidomide at early lymphocyte recovery after induction chemotherapy in newly diagnosed AML and high-risk MDS.

An immunosuppressive microenvironment promoting leukemia cell immune escape plays an important role in the pathogenesis of AML. Through its interaction with cereblon, a substrate receptor for the E3 ubiquitin ligase complex, pomalidomide leads to selective ubiquitination of transcription factors Aiolos and Ikaros thereby promoting immune modulation. In this phase I trial, 51 newly diagnosed non-favorable risk AML and high-risk MDS patients were enrolled and treated with AcDVP16 (cytarabine 667 mg/m2/day IV continuous infusion days 1-3, daunorubicin 45 mg/m2 IV days 1-3, etoposide 400 mg/m2 IV days 8-10) induction therapy followed by dose- and duration-escalation pomalidomide beginning at early lymphocyte recovery. Forty-three patients (AML: n = 39, MDS: n = 4) received pomalidomide. The maximum tolerated dose of pomalidomide was 4 mg for 21 consecutive days. The overall complete remission (CR + CRi) rate, median overall survival, and disease-free survival were 75%, 27.1 and 20.6 months, respectively. Subset analyses revealed 86% CR/CRi rate in AML patients with unfavorable-risk karyotype treated with pomalidomide. Pomalidomide significantly decreased Aiolos expression in both CD4+ and CD8+ peripheral blood and bone marrow T cells, promoted T cell differentiation, proliferation, and heightened their cytokine production. Finally, pomalidomide induced distinct gene expression changes in immune function-related ontologies in CD4+ and CD8+ T cells.

Connecting the Dots From Fever of Unknown Origin to Myelodysplastic Syndrome: GATA2 Haploinsufficiency.

Leukemia-predisposing conditions, such as GATA2 haploinsufficiency, are known for their high penetrance and expressivity profiles. These disorders pose a difficult diagnostic challenge to even the most experienced clinician when they first present. We describe the case of a 17-year-old male presenting with features of nontuberculous mycobacterial infection, pulmonary fibrinoid granulomatous vasculitis, and myelodysplasia in the setting of a pathogenic GATA2 frameshift mutation confirmed by next-generation sequencing. The broad differential for GATA2 haploinsufficiency requires prompt recognition of key clinical features and laboratory abnormalities towards directing diagnosis and guiding appropriate and perhaps life-saving therapy.

Signatures of CD8+ T cell dysfunction in AML patients and their reversibility with response to chemotherapy.

BACKGROUND: Our understanding of phenotypic and functional signatures of CD8+ T cell dysfunction in acute myeloid leukemia (AML) is limited. Deciphering these deranged T cell functional states and how they are impacted by induction chemotherapy is essential for incorporation of novel immune-based strategies to restore and maintain antileukemia immunity. METHODS: We utilized high-dimensional immunophenotyping, gene expression, and functional studies to characterize peripheral blood and bone marrow CD8+ T cells in 72 AML patients at diagnosis and after induction chemotherapy. RESULTS: Our data suggest that multiple aspects of deranged T cell function are operative in AML at diagnosis, with exhaustion and senescence being the dominant processes. Following treatment, the phenotypic and transcriptional profile of CD8+ T cells diverged between responders and nonresponders. Response to therapy correlated with upregulation of costimulatory, and downregulation of apoptotic and inhibitory, T cell signaling pathways, indicative of restoration of T cell function. In functional studies, AML blasts directly altered CD8+ T cell viability, expansion, co-signaling and senescence marker expression. This CD8+ T cell dysfunction was in part reversible upon PD-1 blockade or OX40 costimulation in vitro. CONCLUSION: Our findings highlight the uniqueness of AML in sculpting CD8+ T cell responses and the plasticity of their signatures upon chemotherapy response, providing a compelling rationale for integration of novel immunotherapies to augment antileukemia immunity. FUNDING: This work was supported by the Leukemia & Lymphoma Society grant no. 6449-13; NIH grants UM1-CA186691 and R01-HL110907-01; the American Society for Blood and Marrow Transplantation New Investigator Award/Gabrielle's Angel Foundation; the Vienna Fund for Innovative Cancer Research; and by fellowships from the Wenner-Gren Foundation and the Swedish Society for Medical Research.

Meniscal repair possibilities using bone morphogenetic protein-7.

This study analysed the influence of bone morphogenetic protein-7 (BMP-7) on cells and meniscal structure. The effect of treatment with BMP-7 was assessed in vitro and in vivo in lesions in the avascular area of the meniscus. Cells were extracted from the outer and inner part of eight menisci of four 2-year-old merino sheep. The menisci were digested with a collagenase mix, and meniscus cells of the synovium, vascular area and avascular area were extracted. The expression of genes for collagen (Col1 and Col2A), matrix metalloproteinases (MMP-2 and MMP-13) and aggrecan was analysed by real time quantitative polymerase chain reaction (qPCR) at baseline and after incubation with BMP-7. Eight sheep aged 2 years and weighing 35-40 kg were used for the in vivo study. Surgery was performed in both knees of every animal. Two holes were made in the avascular area of the medial meniscus of both knees and filled using Putty(®) (control groups) or OP-1 Putty(®), which comprises BMP-7 mixed with a cellulose putty carrier (experimental groups). Animals were sacrificed at 6, 12 and 25 weeks. Adding BMP-7 to vascular cells of the meniscus was associated with a 15-fold increase in Col2A expression and a 78-fold increase in BMP-7 expression. BMP-7 inhibited MMP-2 and MMP-13 expression. Adding BMP-7 to synovial cells inhibited the expression of Col1, doubled the expression of Col2A and reduced the expression of BMP-7; the expression of MMP-2 was inhibited, while that of MMP-13 was increased three-fold. Incubation of cells from the avascular region with BMP-7 was associated with a 2.4-fold increase in Col1 expression, and a 4.4-fold increase in Col2A expression compared with the control. The expression of MMP-2 and BMP-7 was inhibited. In the in vivo study, treatment of the holes in the avascular area of the meniscus with BMP-7 was associated with an important cell presence inside the holes and the appearance of fibrous tissue after 12 weeks; these features were not seen in the control groups. BMP-7 may be a suitable growth factor for stimulation of meniscal cell and collagen formation.

Cell study of the three areas of the meniscus: effect of growth factors in an experimental model in sheep.

Meniscus had two areas with different vascular supply. Cells of the two areas and the synovium were monolayer cultivated. We analyzed the expression of genes of Col1, Col 2A, MMP-2, MMP-13, and aggrecan in a baseline state and after incubation with VEGF, TGF-ß, FGF, and IGF. We found that the growth factors used produced a major increase in the MMP-13 in all three areas. In the vascular area, the stimulation of MMP-3 was produced by FGF, while in the synovial and avascular areas, it was caused by TGF-ß. MMP-2 was only stimulated in the synovial area by IGF. Col 2A was stimulated in the synovial area by VEGF, and in the avascular area by TGF-ß, FGF, and IGF, whereas Col 1 was stimulated in the avascular area by IGF, FGF, and VEGF. The vascular or avascular areas of the meniscus, behave differently in terms of repair, and their cells express different factors. The growth factors act in a different way in each meniscal area.

Efecto del almacenamiento de menisco a 4ºC sobre la morfología y la capacidad de respuesta celular a los factores TGF-ß1, IGF-1, aFGF, bFGF y BMP-7 en cultivo en monocapa / Effect of meniscus storage at 4ºC upon cell morphology and capacity to respond to growth factors TGF-ß1, IGF-1, aFGF, bFGF and BMP-7 in monolayer culture

Objetivo del trabajo: Estudiar la viabilidad del almacenamiento de menisco a 4ºC. Material y método: Se obtuvieron fragmentos de menisco procedentes de corderos sanos en condiciones de esterilidad. Como control, se utilizaron fragmentos de tejido digeridos inmediatamente después de la extracción del menisco, el resto se almacenaron a 4ºC sumergidos en medio de cultivo durante 15 y 30 días. Tras el almacenamiento, se realizó la digestión del material utilizando tripsina, colagenasa y dispasa y se cultivaron las células en monocapa. Se estudió la morfología celular al microscopio óptico y la respuesta a los factores de crecimiento TGF-ß1, IGF-1, aFGF, bFGF y BMP-7 mediante ensayos de proliferación. Resultados: La respuesta celular a bFGF e IGF-1 se incrementó tras los 15 días de tratamiento, para volver a descender tras los 30 días. La capacidad de respuesta tras el tratamiento con BMP-7 descendió a los 15 días y volvió a ser similar al control a los 30 días. Los factores aFGF y TGF-ß no modificaron la respuesta que se produjo en las células. Conclusiones: Tras el almacenamiento a 4ºC las células sufren cambios morfológicos y bioquímicos que las lleva a modificar su sensibilidad a algunos factores de crecimiento (AU)

Aim: To evaluate the viability of meniscus storage at 4ºC. Material and method: Meniscus fragments were obtained from healthy goats under sterile conditions. As control, use was made of tissue fragments digested immediately after meniscus harvesting; the rest were stored immersed at 4ºC in culture medium for 15 and 30 days. Following storage, the material was digested using trypsin, collagenase and dispase, with monolayer culture of the cells. Cell morphology was evaluated under the light microscope, and cell response to growth factors TGF-ß1, IGF-1, aFGF, bFGF and BMP-7 was assessed by proliferation tests. Results: The cell response to bFGF and IGF-1 increased after 15 days of treatment, followed by a decrease after 30 days.The response capacity after BMP-7 treatment decreased after 15 days and returned to control levels after 30 days. Factors aFGF and TGF-ß did not modify cell response. Conclusions: Following storage at 4ºC, the cells undergo morphological and biochemical changes that modify their sensitivity to certain growth factors (AU)

Accuracy of combined clinical findings and fine needle aspiration cytology for the diagnosis in palpable breast tumors

Objetivo. Un estudio prospectivo para evaluar el nivel de confianza y valor predictivo de la biopsia por aspiración con aguja fina fue llevado a cabo en el Servicio de Oncología, Hospital 20 de Noviembre del ISSSTE en la Ciudad de México. Material y método. Los casos clínicos que se presentaron de manera consecutiva de 1992 a 1994 con tumor mamario palpable, confirmado histológicamente, fueron incluidos en el estudio. Se realizó un aspirado por paciente y éstos fueron revisados por el mismo patólogo en todos los casos. Se determinaron sensibilidad, especificidad y valor predictivo de la prueba. La edad, características de los bordes tumorales, tamaño y movilidad del tumor fueron evaluados por análisis Bayesiano. Resultados. De 213 aspirados, 199 fueron adecuados para diagnóstico; 98 (46 por ciento) fueron diagnósticos de carcinoma, 13 fueron acelulares y uno sugestivo para carcinoma. Los diagnósticos citológicos acelulares fueron considerados como negativos y los sugestivos como positivos para propósitos de análisis. La edad promedio y el tamaño tumoral fueron: 46.6 años (rango 14-90) y 3.7 cm (rango 1-13), respectivamente. Se calcularon sensibilidad (.932), especificidad (.973) y valor predictivo de la prueba (96.9 por ciento). Observamos una alta probabilidad de resultados verdaderos positivos [P(D+/T+)] > 0.8 en pacientes entre los 40 y 60 años de edad, tumores con bordes irregulares, tamaño tumoral > 2 cm y en lesiones fijas. Conclusiones. La prueba tiene un alto nivel de confianza y en presencia de dos o más de los factores clínicos mencionados, se pueden tomar decisiones definitivas en el diseño del tratamiento sin necesidad de confirmación histológica