In vivo study of targeted nanomedicine delivery into Langerhans cells by multiphoton laser scanning microscopy

Abstract

Epidermal Langerhans cells (LCs) function as professional antigen-presenting cells of the skin. We investigated the LC-targeting properties of a special mannose-moiety-coated pathogen-like synthetic nanomedicine DermaVir (DV), which is capable to express antigens to induce immune responses and kill HIV-infected cells. Our aim was to use multiphoton laser microscopy (MLM) in vivo in order to visualize the uptake of Alexa-labelled DV (AF546-DV) by LCs. Knock-in mice expressing enhanced green fluorescent protein (eGFP) under the control of the langerin gene (CD207) were used to visualize LCs. After 1 h, AF546-DV penetrated the epidermis and entered the eGFP-LCs. The AF546-DV signal was equally distributed inside the LCs. After 9 h, we observed AF546-DV signal accumulation that occurred mainly at the cell body. We demonstrated in live animals that LCs picked up and accumulated the nanoparticles in the cell body.

Handheld nonlinear microscope system comprising a 2 MHz repetition rate, mode-locked Yb-fiber laser for in vivo biomedical imaging

“…Red: fluorescence signal of AF546-labelled nanoparticles 1 hour after of the topical treatment by nanomedicine of the skin. One can observe that the Langerhans cells, which are a part of the immune system, accumulate the AF546-labelled nanoparticles [15]. Scale bar: 50 μm.…”

“…Epidermal Langerhans cells (LCs) function as professional antigen-presenting cells of the skin. In a recent experiment [15] we investigated the LC-targeting properties of a special mannose-moiety-coated pathogen-like synthetic nanomedicine DermaVir (DV), which is capable to express antigens to induce immune responses and kill HIV-infected cells. In order to visualize the uptake of Alexa-labelled DV (AF546-DV) by LCs in vivo, we used a commercial LSM 7MP scanning 2PEF microscope system of Carl Zeiss.…”

“…Among others, using SHG imaging of the collagen, one can investigate basal cell carcinoma [3] or follow the effects of obesity on dermal collagen alterations [14] for instance. For demonstration purposes in cosmetology and nanomedicine, we also used our two-channel 2PEF/SHG imaging system for in vivo visualization and for monitoring the uptake of Alexa Fluor 546 labelled nanomedicine by Langerhans cells [15] for a few hours.…”

Principles of nonlinear microscopy and application possibilities in dermatology

“…ábra) (14). Az epidermisz sejtjeinek kétfoton karakterisztikája, mint például a nagyság és az alak, fontos információként szolgálhatnak különbözô tumorok, gyulladásos betegségek diagnosztikájában, valamint in vivo farmakológiai vizsgála-tokban (15,16). Az utóbbi években számos publikáció jelent meg a melanóma és a nem pigmentált malignus bôrtu-morok kétfoton mikroszkópiás vizsgálatairól (17)(18)(19).…”

In vivosecond-harmonic generation andex vivocoherent anti-stokes raman scattering microscopy to study the effect of obesity to fibroblast cell function using an Yb-fiber laser-based CARS extension unit

“…An additional aim was to develop a system that allows us to continuously monitor these changes. For this reason, we set out to use nonlinear microscopy (Bognar et al, ; Kolonics et al, ) for detection of the dermal collagen and subcutaneous adipocytes.…”