Comparison of immature and mature bone marrow-derived dendritic cells by atomic force microscopy

Feiyue Xing,Guoliang Chen,Yu Yu,Jing Liu,Mingqian Hu,Jiongkun Wang

doi:10.1186/1556-276x-6-455

Abstract

A comparative study of immature and mature bone marrow-derived dendritic cells (BMDCs) was first performed through an atomic force microscope (AFM) to clarify differences of their nanostructure and adhesion force. AFM images revealed that the immature BMDCs treated by granulocyte macrophage-colony stimulating factor plus IL-4 mainly appeared round with smooth surface, whereas the mature BMDCs induced by lipopolysaccharide displayed an irregular shape with numerous pseudopodia or lamellapodia and ruffles on the cell membrane besides becoming larger, flatter, and longer. AFM quantitative analysis further showed that the surface roughness of the mature BMDCs greatly increased and that the adhesion force of them was fourfold more than that of the immature BMDCs. The nano-features of the mature BMDCs were supported by a high level of IL-12 produced from the mature BMDCs and high expression of MHC-II on the surface of them. These findings provide a new insight into the nanostructure of the immature and mature BMDCs.

Highlights

Dendritic cells (DCs) are the most potent specialized antigen-presenting cells, which bridge the innate and adaptive immune response, controlling both immunity and tolerance
Morphologic and functional characteristics of bone marrowderived dendritic cells (BMDCs) The bone marrow cells were cultured and induced in complete RPMI 1640 medium supplemented with a given dose of granulocyte macrophage-colony stimulating factor (GM-CSF) plus IL-4 for 6 days
MHC-II is one of activation molecules expressed on the surface of BMDCs, representing a phenotype of mature BMDCs

Summary

Introduction

Dendritic cells (DCs) are the most potent specialized antigen-presenting cells, which bridge the innate and adaptive immune response, controlling both immunity and tolerance. It is well known that DCs may be derived from bone marrow progenitors with two major developmental stages: immature and mature DCs [1]. The development of immature DCs can be induced with using cytokines, such as granulocyte macrophage-colony stimulating factor (GM-CSF) [2], FMS-like tyrosine kinase 3 (FLT3) [3], or cytokine cocktails containing GM-CSF +/-IL-4 [4] in vitro. After stimulation of lipopolysaccharide (LPS), poly I:C or thymic stromal lymphopoietin (TSLP), immature DCs can further differentiate into mature DCs, with increase of IL-12 and up-regulation of MHC-II, CD40, CD80, CD83, and CD86 molecules on the surface of DCs [5,6]. Force between both immature and mature bone marrowderived dendritic cells (BMDCs).

Materials and methods

Results and discussion

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