Impact of Region-of-Interest Size on Immune Profiling Using Multiplex Immunofluorescence Tyramide Signal Amplification for Paraffin-Embedded Tumor Tissues.

INTRODUCTION: Representative regions of interest (ROIs) analysis from the whole slide images (WSI) are currently being used to study immune markers by multiplex immunofluorescence (mIF) and single immunohistochemistry (IHC). However, the amount of area needed to be analyzed to be representative of the entire tumor in a WSI has not been defined. METHODS: We labeled tumor-associated immune cells by mIF and single IHC in separate cohorts of non-small cell lung cancer (NSCLC) samples and we analyzed them as whole tumor area as well as using different number of ROIs to know how much area will be need to represent the entire tumor area. RESULTS: For mIF using the InForm software and ROI of 0.33 mm2 each, we observed that the cell density data from five randomly selected ROIs is enough to achieve, in 90% of our samples, more than 0.9 of Spearman correlation coefficient and for single IHC using ScanScope tool box from Aperio and ROIs of 1 mm2 each, we found that the correlation value of more than 0.9 was achieved using 5 ROIs in a similar cohort. Additionally, we also observed that each cell phenotype in mIF influence differently the correlation between the areas analyzed by the ROIs and the WSI. Tumor tissue with high intratumor epithelial and immune cells phenotype, quality, and spatial distribution heterogeneity need more area analyzed to represent better the whole tumor area. CONCLUSION: We found that at minimum 1.65 mm2 area is enough to represent the entire tumor areas in most of our NSCLC samples using mIF.

[Plasmablastic lymphoma: a case of rectal disease with bone marrow involvement in a HIV positive patient]. / Linfoma plasmablástico: un caso con enfermedad rectal y compromiso de médula ósea en un paciente VIH positivo.

Plasmablastic lymphoma is an aggressive form of lymphoma diffuse large B cell Lymphoma, initially described in HIV positive patients associated with lesions in the oral cavity. It is about 2% of NHL associated with HIV. This entity currently represents a challenge for the diagnosis and treatment, showing a poor long-term prognosis. This report describes a patient with VIH on HAART and CD4 count in 490 cells/ml associated with Plasmablastic lymphoma that involves rectum and bone marrow. The patient received 6 cycles of EPOCH regimen with complete response.

Linfoma plasmablástico: un caso con enfermedad rectal y compromiso de médula ósea en un paciente VIH positivo / Plasmablastic lymphoma: a case of rectal disease with bone marrow involvement in a HIV positive patient

El linfoma plasmablástico es una variante agresiva del linfoma difuso de células grandes B, descrito inicialmente en pacientes VIH positivos asociados a lesiones en cavidad oral. Este corresponde al 2% de LNH asociado a VIH. Esta entidad actualmente representa un reto para el diagnóstico y el tratamiento, presentando o un pobre pronóstico o. El presente reporte describe un paciente con HIV en tratamiento con TARGA, un recuento CD4 en 490 células/ml, asociado a un Linfoma plasmablástico con compromiso rectal y médula ósea. El paciente recibe el régimen EPOCH por 6 ciclos con respuesta completa...

Plasmablastic lymphoma is an aggressive form of lymphoma diffuse large B cell Lymphoma, initially described in HIV positive patients associated with lesions in the oral cavity. It is about 2% of NHL associated with HIV. This entity currently represents a challenge for the diagnosis and treatment, showing a poor long-term prognosis. This report describes a patient with VIH on HAART and CD4 count in 490 cells/ml associated with Plasmablastic lymphoma that involves rectum and bone marrow. The patient received 6 cycles of EPOCH regimen with complete response...