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

Quantificazione del leucocita Egress tramite vasi linfatici dalla pelle murina e tumori

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Quantifying Leukocyte Egress via Lymphatic Vessels from Murine Skin and Tumors

Quantificazione del leucocita Egress tramite vasi linfatici dalla pelle murina e tumori

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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

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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.

Qui, noi dimostrare i metodi per la quantificazione in vivo d'uscita del leucocita da ingenuo, infiammato e maligni della pelle murino. Eseguiamo un confronto testa a testa tra due modelli: transdermico FITC applicazione ed in situ fotoconversione. Inoltre, dimostriamo l'utilità di fotoconversione per servizi esterni del leucocita di rilevamento da tumori cutanei.

La fotoconversione fornisce un metodo in vivo tracciabile per l'analisi quantitativa dell'uscita dei leucociti per una visione meccanicistica della residenza dei leucociti e delle dinamiche di uscita da più siti tissutali in stati danneggiati. Bene, qui dimostriamo l'utilità del tracciamento dei leucociti dai tumori cutanei. L'applicazione di questo saggio ad altri tessuti e malattie è limitata solo dalla capacità di somministrare luce.

Il principale vantaggio di questa tecnica è che permette la quantificazione delle popolazioni di cellule immunitarie endogene, piuttosto che trasferite come uscita dai tessuti infiammati in vivo. Per indurre l'infiammazione, dopo aver confermato una mancanza di risposta al dito del piedi, utilizzare una micropipetta per somministrare 20 microlitri di ftalato di dibutile o DBP, al lato ventrale dell'pinna dell'orecchio di un topo transgenico Kaede anestetizzato. Dopo aver permesso al DBP di asciugarsi, tirare l'orecchio attraverso una fessura in un pezzo di foglio di alluminio e utilizzare nastro a doppia parte per fissare l'orecchio saldamente al foglio con il lato dorsale rivolto verso l'alto.

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