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Updated: Jan 20, 2026

Collection and Processing of Lymph Nodes from Large Animals for RNA Analysis: Preparing for Lymph Node Transcriptomic Studies of Large Animal Species

Collection and Processing of Lymph Nodes from Large Animals for RNA Analysis: Preparing for Lymph Node Transcriptomic Studies of Large Animal Species

Published on: May 19, 2018

Integración del perfil de citometría de flujo y el análisis de mutaciones genéticas de ganglios linfáticos intactos

Qun-Yi Peng¹, Hong-Yuan Chen¹, Yan Zheng¹

¹Department of Hematology, Quanzhou First Hospital of Affiliated to Fujian Medical University Quanzhou 362000, Fujian, China.

American journal of translational research

|January 19, 2026

Insights

La integración de la citometría de flujo multiparamétrica (FCM) con el análisis de mutaciones genéticas mejora la clasificación del linfoma. Este enfoque combinado mejora la precisión diagnóstica de los linfomas de células B y de células T/NK utilizando muestras de ganglios linfáticos intactos.

Área de la Ciencia:

Hematología; Oncología; Inmunología

Sus antecedentes:

La clasificación precisa del linfoma es crucial para un tratamiento eficaz.
Los métodos de diagnóstico tradicionales pueden ser limitados para diferenciar ciertos subtipos de linfoma.
La citometría de flujo multiparamétrica (FCM) y el análisis de mutaciones genéticas ofrecen información diagnóstica complementaria.

Objetivo del estudio:

Evaluar la utilidad diagnóstica combinada de FCM, histopatología y análisis de mutaciones genéticas para la clasificación del linfoma.
Evaluar la concordancia entre FCM e histopatología en el diagnóstico del linfoma.
Explorar el papel de mutaciones genéticas específicas y la expresión de proteínas asociadas en el refinamiento del diagnóstico del linfoma.

Principales métodos:

Análisis retrospectivo de 109 muestras de ganglios linfáticos (LN) intactos de pacientes con linfadenopatía.
Se utilizó FCM multicolor, histopatología y secuenciación de próxima generación (NGS) dirigida a 62 genes relacionados con el linfoma.
Se correlacionó la inmunofenotipificación por FCM y los perfiles de mutación con los diagnósticos patológicos.

Principales resultados:

La FCM demostró una alta concordancia con la histopatología en la detección del linfoma.
Los perfiles distintos de tamaño celular (FSC) y la expresión de PD-1 en las células T diferenciaron subtipos de linfoma.
Se identificaron mutaciones genéticas específicas (por ejemplo, IGHV, BCL2, EZH2, BCL6) en diferentes subtipos de linfoma.
La FCM guiada por mutaciones confirmó la expresión de proteínas, lo que ayudó en el diagnóstico.
Los linfomas de células T/NK exhibieron expresión de antígenos aberrante y clonación TRBC1 restringida.

Conclusiones:

La FCM en LN intactos es una herramienta diagnóstica fiable con una fuerte concordancia patológica.
La integración de datos de mutaciones genéticas con FCM proporciona una estrategia de diagnóstico multiparamétrica potente.
Este enfoque integrado refina la clasificación del linfoma más allá de la inmunofenotipificación estándar, mejorando la precisión diagnóstica.

Read full analysis

Abstracto

Objective:

To evaluate the diagnostic use of integrating multi-parameter flow cytometry (FCM), histopathology, and gene mutation analysis for lymphoma classification using intact lymph node (LN) samples.

Methods:

Intact LN samples from 109 patients with lymphadenopathy were retrospectively analyzed by pathology, multi-color FCM, and next-generation sequencing (NGS) targeting 62 lymphoma-related genes.

Results:

FCM immunophenotyping showed high concordance with pathology (56/56 lymphoma cases detected by FCM were pathologically confirmed). PD-1 on T cells was significantly elevated in B-cell lymphoma (BCL), especially diffuse large BCL (DLBCL) (P<0.05). All BCLs exhibited monotypic intracellular cKappa or cLambda expression. Significant differences in cell size (FSC) were observed: Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma (CLL/SLL) (mean FSC: 89.42±6.01) and Follicular Lymphoma cells (FL; 93.88±4.94) were smaller than normal B-cells (102.09±11.58), while DLBCL cells (121.84±9.17) were larger (all P<0.05). Subtypes showed distinct mutation profiles, including IGHV (9/11) in CLL/SLL; BCL2 (5/7) and EZH2 (4/7) in FL; and BCL6 (5/13) in DLBCL. Mutation-guided FCM confirmed BCL2 protein expression in two FL cases and BCL6 in one DLBCL case. T/NK-cell lymphomas showed aberrant antigen expression and restricted TRBC1 clonality (1.17%±1.61% or >96.4%) outside the normal polyclonal range (36.60%±7.21%).

Conclusion:

FCM on intact LNs is a robust tool with high pathologic concordance. Integrating genetic mutation data with FCM provides a powerful, multi-parameter strategy. This approach moves beyond standard immunophenotyping to include mutation-associated antigens, thereby refining lymphoma classification and enhancing diagnostic accuracy.

Palabras clave:

Perfiles de expresión de antígenos PD-1 TRBC1 mutación genética inmunofenotipo ganglio linfático

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