Cc. Chipev et al., CHARACTERIZATION OF AN IMMORTALIZED CELL-LINE FROM A PATIENT WITH EPIDERMOLYTIC HYPERKERATOSIS, Journal of investigative dermatology, 106(3), 1996, pp. 385-390
The most frequent mutation that causes the autosomal dominant skin dis
ease epidermolytic hyperkeratosis (EHK) is an arginine to histidine su
bstitution at position 10 in the 1A segment of the rod domain of kerat
in 10. As an initial step toward developing a strategy for treating EH
K, a cell line, EH18-1, was established after keratinocytes derived fr
om an EHK patient with this mutation were immortalized by a recombinan
t retrovirus encoding the E6 and E7 genes of human papillomavirus type
18. EH18-1 cells synthesize considerable amounts of keratin 10 mRNA a
nd protein when maintained in either submerged cultures or in organoty
pic cultures. When grown in organotypic culture, EH18-1 cells form mul
tiple layers and express keratin 10 and filaggrin predominantly in the
upper layers. Thus, the EH18-1 cell line exhibits several morphologic
al and biochemical markers of terminal epidermal differentiation. A se
miquantitative reverse transcriptase polymerase chain reaction assay f
or keratin 10 mRNA was developed to distinguish between expression of
the normal and the mutant alleles. The EH18-1 keratinocyte cell line w
ill be useful in developing protocols for gene therapy of EHK that may
be monitored by reverse transcriptase polymerase chain reaction of ei
ther allele.