Immortalization of magnetically separated human lymphocytes by electrofusion.

Area of Science:

• Immunology
• Cell Biology

Background:

• Peripheral blood mononuclear cells (MNCs) are crucial for immune responses.
• Immortalization of lymphocytes is vital for monoclonal antibody production and cellular studies.
• Current methods for lymphocyte immortalization can be inefficient.

Purpose of the Study:

• To investigate the efficiency of electrofusion for immortalizing human peripheral blood lymphocytes.
• To determine the immortalization capacity of different lymphocyte subsets, including B cells and non-B cells.
• To compare the hybridoma formation rates across various cell populations.

Main Methods:

• Human peripheral blood mononuclear cells (MNCs) were isolated and separated using magnetic beads based on surface markers.
• Positive and negative cell fractions were subjected to electrofusion with the heteromyeloma line CB-Fu2.
• B cells were specifically isolated using anti-CD19 coated beads.
• Hybridoma clones were selected using HAT (hypoxanthine-aminopterin-thymidine) resistance.

Main Results:

• Successful immortalization of both positive and negative cell fractions from human peripheral blood lymphocytes was achieved.
• B cells, isolated via anti-CD19 beads, demonstrated high immortalization rates (10^-5 to 10^-4) even with limited cell numbers.
• Non-B cells, including T cells, also yielded HAT-resistant hybridoma clones, albeit at a lower fusion frequency.

Conclusions:

• Electrofusion is an effective method for immortalizing human peripheral blood lymphocytes.
• B cells are highly amenable to immortalization via electrofusion, enabling efficient hybridoma generation.
• The study demonstrates the potential for immortalizing non-B lymphocytes, broadening the scope of hybridoma technology.