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Intestinal microflora affects multi-herbal product JTX on the gene expression in the gut and liver.

These results show that vascular endothelial cells and perivascular nonparenchymal cells are activated after massive hemorrhage to produce inflammatory cytokines.

A serious drawback of tumor necrosis factor ␣ (TNF) as a clinical antitumor agent is that it also has hypotensive activity. To overcome this problem, derivatives of its sister cytokine lymphotoxin (TNF-␤ or LT) were prepared. One of them, mutein 2 (Mut2) has a deletion of amino acids 1-7 but contains substituted amino acids, Met-Phe-Pro at positions 8-10 of the mature human LT. This mutein has no hypotensive activity at the maximum dose (10 mg͞kg) tested on rats. In contrast, a much lower dose (1 mg͞kg) of TNF and LT caused a significant blood pressure drop. In vivo studies revealed that Mut2 was more effective than TNF or LT against MethA (a mouse tumor line) as judged by the therapeutic ratio [calculated as LD 50 (dose that kills 50% of the animals)͞ED 50 (dose that reduces the tumor size by 50%)]. With five other different mouse tumors and two different human tumors, Mut2 was also effective and the effectiveness was comparable or superior to that of TNF or LT. These results suggest the possibility that this derivative may be usable as a clinical antitumor agent without the serious side effects associated with TNF.Various attempts have been made to create muteins of tumor necrosis factor (TNF) and lymphotoxin (LT) to improve their therapeutic values (1). Results of these experiments suggest that up to nine NH 2 -terminal amino acids of TNF can be removed without losing cytotoxic activity in vitro (2). The replacement of acidic amino acids at the NH 2 -terminal region with basic amino acids resulted in a better (less toxic with high antitumor activity) TNF (3, 4). Replacement of the COOHterminal region amino acids (amino acids 141-146) with other amino acids often modified the in vitro cytotoxic activity of TNF. The addition of amino acids to TNF at the NH 2 -terminal position decreased a side effect of TNF (enhancement of metastasis) without diminishing its anticancer activity (5).On the other hand, relatively limited information is available for LT because efficient expression in Escherichia coli has become possible only recently (6). In vitro cytotoxic activity was maintained even after removal of the NH 2 -terminal amino acids residues from LT (7). In contrast, the removal of the COOH-terminal region of LT generally resulted in loss of the cytotoxic activity (8). To the best of our knowledge, LT muteins have never been tested in vivo for their antitumor or hypotensive activities.TNF is relatively ineffective against human cancers because its hypotensive activity prevents administration of doses that are high enough to cause tumor regression (9, 10). Systemic use of TNF causes hypotension in the experimental animals (11)(12)(13)(14). In this paper, we describe an LT mutein (mutein 2, Mut2) that lacks hypotensive effects but has high antitumor activity. The ratio hypotensive dose͞therapeutic dose for Mut2 was at least 9-fold higher than that of TNF suggesting the possible future clinical applicability of Mut2. MATERIALS AND METHODSExpression of TNF, LT, and Mut2 in E. coli. TNF and LT genes were clone...