Minimal size MIDGE vectors improve transgene expression in vivo.

Viral and plasmid vectors may cause immunological side-effects resulting from the expression of therapeutically unwanted genes and from CpG motifs contained in their sequence. A new vector type for minimalistic, immunological-defined gene expression (MIDGE) may overcome these problems. MIDGE is a minimal size gene transfer unit consisting of the expression cassette, including promotor, gene and RNA-stabilizing sequences, flanked by two short hairpin oligonucleotide sequences.

[Gene therapy in corneal diseases].

Gene therapy raised euphoric expectations in the past that have yet to be met and have even been lowered due to the absence of concrete clinical successes and the occurrence of some tragic incidents. In spite of the great future potential of gene therapy, numerous pilot studies in this field will probably be discontinued for a long time. However, despite these failures, diseases will continue to be the aim of gene transfer studies with experimental protocols using only temporary gene expression or those restricted to individual organs and thus requiring only a low dose of the vehicle (vector).

Ballistic CTLA4 and IL-4 gene transfer into the lower lid prolongs orthotopic corneal graft survival in mice.

Purpose: To explore outflow from the eye and to determine and modulate the influence of lymphatic drainage on corneal graft survival in mice.

Methods: Tracer experiments were conducted in BALB/c mice using the (99m)Tc colloidal albumin Nanocoll. Count rates were determined in the eyes, submandibular lymph nodes, spleen, liver and blood 24 h after subconjunctival, intracorneal, intracameral (anterior chamber), intravenous and subcutaneous lower-lid or upper-lid injections ( n=6 each).

Minimizing side effects of ballistic gene transfer into the murine corneal epithelium.

Background: The beneficial effect of modulating an allospecific immune response by ballistic IL-4 and CTLA4 gene transfer to deliver minimalistic immunologically defined gene expression (MIDGE) vectors into the corneal epithelium was demonstrated in corneal transplantation. However, side effects reduced graft survival in control animals after ballistic transfer without DNA.

Methods: An adapter was constructed for the gene gun apparatus to enlarge and keep constant the distance between the gun and the cornea.