Immunospecific vesicle targeting facilitates microinjection into lymphocytes.

Area of Science:

• Biotechnology
• Cell Biology
• Immunology

Background:

• Polyethylene glycol (PEG)-mediated fusion is a common method for introducing substances into cells.
• Achieving efficient and specific cell microinjection remains a challenge, particularly for certain cell types like lymphocytes.
• Targeting strategies are crucial for directing cellular delivery systems to desired cell populations.

Purpose of the Study:

• To develop and evaluate an antibody-directed method for enhancing vesicle fusion and microinjection into specific cells.
• To demonstrate the efficacy of targeted delivery compared to non-targeted methods using polyethylene glycol (PEG).
• To establish a versatile technique for efficient cellular microinjection in various cell types.

Main Methods:

• Erythrocyte ghosts containing fluorescent bovine serum albumin were prepared.
• Targeting involved labeling cells with biotin-modified antibodies followed by incubation with avidin-coupled erythrocyte ghosts.
• The fusion and microinjection efficiency of targeted vesicles was assessed in murine spleen and thymus cells, and in lymphocyte, monocyte, and fibroblast cell lines.

Main Results:

• Antibody-directed targeting dramatically enhanced polyethylene glycol (PEG)-mediated fusion and microinjection.
• Targeted cell populations exhibited substantial levels of microinjection.
• Cells treated with PEG without targeting showed no significant microinjection.

Conclusions:

• Antibody-directed vesicle targeting is a highly effective strategy for enhancing cell microinjection.
• This method offers precise delivery to specific cell populations, including challenging cell types like lymphocytes.
• The technique improves injection efficiencies, making it valuable for targeted cellular delivery and when reagent use is limited.