Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors

Maria M. Steele; Madeline J. Churchill; Alec P. Breazeale; Ryan S. Lane; Nicholas A. Nelson; Amanda W. Lund

doi:10.3791/58704

Cuantificación de leucocitos salida vía los recipientes linfáticos de la piel murina y tumores

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors

Cuantificación de leucocitos salida vía los recipientes linfáticos de la piel murina y tumores

Full Text

9,244 Views

08:39 min

January 7, 2019

DOI: 10.3791/58704-v

Maria M. Steele¹, Madeline J. Churchill², Alec P. Breazeale¹, Ryan S. Lane¹, Nicholas A. Nelson¹, Amanda W. Lund^1,2,3,4

¹Department of Cell, Developmental, & Cancer Biology,Oregon Health and Science University, ²Department of Molecular Microbiology & Immunology,Oregon Health and Science University, ³Department of Dermatology,Oregon Health and Science University, ⁴Knight Cancer Institute,Oregon Health and Science University

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study investigates the in vivo quantification of leukocyte egress from various skin conditions, including naive, inflamed, and malignant murine skin. The authors compare two photoconversion techniques to track leukocyte movement and examine their utility in understanding immune dynamics in cutaneous tumors.

Key Study Components

Research Area

Immunology
Cancer biology
Leukocyte dynamics

Background

Leukocyte egress is crucial in inflammation and cancer.
Understanding leukocyte behavior can provide insights into disease progression.
Existing techniques may not effectively quantify endogenous immune cell populations.

Methods Used

In vivo photoconversion techniques for leukocyte tracking.
Use of Kaede transgenic mice.
Flow cytometry for analysis of leukocyte populations.

Main Results

Photoconversion efficiently tracked leukocyte egress from tumors.
Increased leukocyte infiltration was observed following violet light exposure.
Photoconversion efficiency varied significantly with tumor type and size.

Conclusions

This study establishes effective methodologies for quantifying leukocyte behavior in vivo.
Findings enhance understanding of immune responses in skin inflammation and cancer.

Frequently Asked Questions

What is photoconversion?

Photoconversion is a technique that allows tracking of specific leukocyte populations by converting fluorescent proteins in vivo using light exposure.

Why focus on leukocyte egress?

Leukocyte egress is crucial for studying immune responses during inflammation and tumor progression.

What models were compared in the study?

The study compared transdermal FITC application and in situ photoconversion methods.

What organism was used in the experiments?

Kaede transgenic mice were used as the biological model for this study.

How was leukocyte infiltration measured?

Leukocyte infiltration was quantified using flow cytometry of the harvested tissues post-photoconversion.

What implications do the study findings have?

The findings have implications for understanding immune cell dynamics in disease, which can inform therapeutic strategies.

Can these methods be used for other tissues?

Yes, the techniques can potentially be applied to other tissue types, contingent on the ability to administer light for photoconversion.

Aquí, demostramos los métodos de cuantificación en vivo de la salida de leucocitos de ingenuo, inflamado y mala piel murina. Realizamos una comparación directa de dos modelos: photoconversion de aplicación y en situ FITC transdérmica. Además, demostramos la utilidad de photoconversion para el seguimiento de salida de leucocitos de tumores cutáneos.

La fotoconversión proporciona un método in vivo rastreable para el análisis cuantitativo de la salida de leucocitos para una visión mecanicista de la residencia de leucocitos y la dinámica de salida de múltiples sitios de tejido en estados enfermos. Bueno, aquí demostramos la utilidad del seguimiento de leucocitos de tumores cutáneos. La aplicación de este ensayo a otros tejidos y enfermedades está limitada sólo por la capacidad de administrar la luz.

La principal ventaja de esta técnica es que permite la cuantificación de poblaciones endógenas, en lugar de células inmunitarias transferidas como una salida de tejidos inflamados in vivo. Para inducir la inflamación, después de confirmar la falta de respuesta al pellizco del dedo del otro, utilice una micropipeta para administrar 20 microlitros de ftalato de dibutilo o DBP, en el lado ventral del pinna del oído de un ratón transgénico Kaede anestesiado. Después de permitir que el DBP se seque, tire de la oreja a través de una hendidura en un pedazo de papel de aluminio y utilice cinta adhesiva de doble cara para fijar la oreja planamente a la lámina con el lado dorsal hacia arriba.

View the full transcript and gain access to thousands of scientific videos