PMID- 10482009
OWN - NLM
STAT- MEDLINE
DCOM- 19991014
LR  - 20190905
IS  - 0340-3696 (Print)
IS  - 0340-3696 (Linking)
VI  - 291
IP  - 7-8
DP  - 1999 Jul-Aug
TI  - In vivo introduction of the interleukin 6 gene into human keratinocytes: 
      induction of epidermal proliferation by the fully spliced form of interleukin 6, 
      but not by the alternatively spliced form.
PG  - 400-4
AB  - The achievement of keratinocyte gene therapy in clinical practice requires 
      fundamental experiments using human keratinocytes or skin. We have recently 
      demonstrated that the in vivo introduction of the interleukin 6 (IL-6) gene into 
      rat keratinocytes induces epidermal proliferation and lymphocyte infiltration 
      into the skin. In this study, we first amplified the human IL-6 cDNA from 
      oligo-dT-primed keratinocyte cDNA and then detected the fully spliced (FS) form 
      and the alternatively spliced (AS) form of IL-6 cDNA. Sequence analysis showed 
      that the AS form, which was composed of the IL-6 coding region with all of exon 
      II deleted except for the first guanine, was identical to that reported to be 
      present in lymphocytes. We constructed the expression vectors phIL6 of the FS 
      form and phIL6S of the AS form. We transplanted human skin onto nude rats and 
      introduced phIL6 and phIL6S into the human keratinocytes using the naked DNA 
      method. Keratinocytes prepared 24 h after introduction from the areas treated 
      with them were examined by reverse transcriptase (RT)-PCR and enzyme linked 
      immunosorbent assay (ELISA). RT-PCR showed that the amounts of FS IL-6 mRNA and 
      AS IL-6 mRNA were similar, whereas the ELISA showed that the amount of FS IL-6 
      peptide was four times that of the AS IL-6 peptide. Histological examination 48 h 
      after introduction showed that the FS form had induced epidermal proliferation, 
      whereas the AS form had not. The epidermal thickening without lymphocyte 
      infiltration induce by the FS form indicates that keratinocyte proliferation is 
      caused by a direct effect of overexpressed IL-6, and not by a secondary effect of 
      infiltrating lymphocytes. This is the first report of the introduction of a human 
      gene into human keratinocytes to produce a biologically active transgenic gene 
      product in human skin using the naked DNA method.
FAU - Sato, M
AU  - Sato M
AD  - Department of Dermatology, Hirosaki University School of Medicine, Japan.
FAU - Sawamura, D
AU  - Sawamura D
FAU - Ina, S
AU  - Ina S
FAU - Yaguchi, T
AU  - Yaguchi T
FAU - Hanada, K
AU  - Hanada K
FAU - Hashimoto, I
AU  - Hashimoto I
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - Arch Dermatol Res
JT  - Archives of dermatological research
JID - 8000462
RN  - 0 (DNA, Complementary)
RN  - 0 (DNA, Recombinant)
RN  - 0 (Interleukin-6)
RN  - 0 (Proliferating Cell Nuclear Antigen)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - *Alternative Splicing
MH  - Animals
MH  - Cell Division/physiology
MH  - DNA, Complementary/genetics
MH  - *DNA, Recombinant
MH  - Enzyme-Linked Immunosorbent Assay
MH  - *Epidermal Cells
MH  - *Gene Transfer Techniques
MH  - Humans
MH  - Interleukin-6/*genetics/metabolism/physiology
MH  - Keratinocytes/cytology/metabolism/*physiology
MH  - Proliferating Cell Nuclear Antigen/metabolism
MH  - RNA, Messenger/metabolism
MH  - Rats
MH  - Rats, Nude
MH  - Reverse Transcriptase Polymerase Chain Reaction
EDAT- 1999/09/11 00:00
MHDA- 1999/09/11 00:01
CRDT- 1999/09/11 00:00
PHST- 1999/09/11 00:00 [pubmed]
PHST- 1999/09/11 00:01 [medline]
PHST- 1999/09/11 00:00 [entrez]
AID - 10.1007/s004030050429 [doi]
PST - ppublish
SO  - Arch Dermatol Res. 1999 Jul-Aug;291(7-8):400-4. doi: 10.1007/s004030050429.