Background/Aims: The lectin phytohemagglutinin is a mitogen for intestinal epithelial cells in vivo. The mechanisms of action are unknown and were therefore analyzed in vitro. Methods: Human (Intestine-407) and rat (IEC-6; IEC-18) intestinal epithelial cell lines were stimulated with phytohemagglutinin. Proliferation was assayed by 3H-thymidine incorporation, activation of mitogen-activated protein kinase (MAPK) by Western blotting, and induction of c-fos mRNA expression by semiquantitative polymerase chain reaction. Control experiments were performed with phenyl-N-acetyl-α-D-galactosaminide or the tyrosine kinase inhibitor tyrphostin A25. Results: Phytohemagglutinin (0.1 µg/ml) significantly stimulated proliferation in all three cell lines after 48–72 h. MAPK activation was detected after 15–30 min, and an induction of c-fos mRNA expression after 15– 30 min of stimulation. Mitogenic effects were blocked by preincubation with phenyl-N-acetyl-α-D-galactosaminide or tyrphostin A25. Conclusion: Phytohemagglutinin stimulated proliferation, MAPK activation and induction of c-fos mRNA expression. The lectin may contribute to intestinal mucosal growth and regeneration thereby preventing gut atrophy.

Journal Section:
Original Paper: Intestinal Disorders
Keywords:
Gut growth, Mitogen-activated protein kinase, Intestinal epithelial cell lines, Phytohemagglutinin
1.
Leavitt RT, Felsted RL, Bachur NR: Biological and biochemical properties of phaseolus vulgaris isolectins. J Biol Chem 1977;252:2967–2971.
[PubMed]
2.
Pusztai A, Ewen SWB, Grant G, Pneumans WJ, Van Damme EJM, Rubio L, Bardocz S: Relationship between survival and binding of plant lectins during small intestinal passage and their effectiveness as growth factors. Digestion 1990;46:308–316.
[PubMed]
3.
King TP, Pusztai A, Clarke EMW: Immunocytochemical localization of ingested kidney bean (Phaseolus vulgaris) lectins in rat gut. Histochem J 1980;12:201–208.
[PubMed]
4.
Banwell JG, Howard R, Kabir I, Adrian TE, Diamond RH, Abromowsky C: Small intestinal growth caused by feeding red kidney bean phytohaemagglutinin lectin to rats. Gastroenterology 1993;104 1669–1677.
5.
Herzig KH, Bardocz S, Grant G, Nustedt R, Fölsch UR, Pusztai A: Red kidney bean lectin is a potent cholecystokinin releasing stimulus in the rat inducing pancreatic growth. Gut 1997;41:333–338.
[PubMed]
6.
Bardocz S, Grant G, Duguid TJ, Brown DS, Sakhri M, Pusztai A, Pryme IF, Mayer D, Wayss K: Phytohaemagglutinin in the diet induced growth of the gut and modified some organ weights in mice. Med Sci Res 1994;22:101–103.
7.
Bardocz S, Grant G, Ewen SWB, Duguid TJ, Brown DS, Englyst K, Pusztai A: Reversible effect of phytohaemagglutinin on the growth and metabolism of rat gastrointestinal tract. Gut 1995;37:353–360.
[PubMed]
8.
Oliveira JT, Pusztai A, Grant G: Changes in organs and tissues induced by feeding of purified kidney bean (Phaseolus vulgaris) lectins. Nutr Res 1988;8:943–947.
9.
Greer F, Brewe A, Pusztai A: Effect of kidney bean (Phaseolus vulgaris) on tissue weight and composition and some metabolic functions of rats. Br J Nutr 1985;54:95–103.
[PubMed]
10.
Hendriks HG, Kik MJ, Koninkx JF, van den Ingh TS: Binding of kidney bean (Phaseolus vulgaris) isolectins to differentiated human colon carcinoma CaCo-2 cells and their effect on cellular metabolism. Gut 1991;32:196–201.
[PubMed]
11.
Bardocz S, Brown DS, Grant G, Pusztai A, Stewart JC, Palmer RM: Effect of the β-adrenoceptor agonist clenbuterol and phytohaemagglutinin on growth, protein synthesis and polyamine metabolism of tissues of the rat. Br J Pharmacol 1992;106:476–482.
[PubMed]
12.
Jordinson M, Goodlad RA, Brynes A, Bliss P, Ghatei MA, Bloom SR, Fitzgerald A, Grant G, Bardocz S, Pusztai A, Pignatelli M, Calam J: Gastrointestinal response to a panel of lectins in rats maintained on total parenteral nutrition. Am J Physiol 1999;276:G1235-G1242.
[PubMed]
13.
Banwell JG, Boldt DH, Meyers J, Weber FL Jr: Phytohemagglutinin derived from red kidney bean (phaseolus vulgaris): A cause for intestinal malabsorption associated with bacterial overgrowth in the rat. Gastroenterology 1983;84:506–515.
[PubMed]
14.
Banwell JG, Abramowsky CR, Weber F, Howard R, Boldt DH: Phytohemagglutinin-induced diarrheal disease. Dig Dis Sci 1984;29:921–929.
[PubMed]
15.
Weinman MD, Allan CH, Trier JS, Hagen SJ: Repair of microvilli in the rat small intestine after damage with lectins contained in the red kidney bean. Gastroenterology 1989;97:1193–1204.
[PubMed]
16.
Kiss R, Camby I, Duckworth C, De Decker R, Salmon I, Pasteels JL, Danguy A, Yeaton P: In vitro influence of phaseolus vulgaris, griffonia simplicifolia, concavalin A, wheat germ, and peanut agglutinin on HCT-15, LoVo, and SW 837 human colorectal cancer cell growth. Gut 1997;40:253–261.
[PubMed]
17.
Koninkx JF, Hendriks HG, van Rossum JM, van den Ingh TS, Mouwen JM: Interaction of legume lectins with the cellular metabolism of differentiated Caco-2 cells. Gastroenterology 1992;102:1516–1523.
[PubMed]
18.
Koninkx JF, Brown DS, Kok W, Hendriks HG, Pusztai A, Bardocz S: Polyamine metabolism of enterocyte-like Caco-2 cells after exposure to Phaseolus vulgaris lectin. Gut 1996;38:47–52.
[PubMed]
19.
Pusztai A, Watt WB: Isolectins of Phaseolus vulgaris. A comprehensive study of fractionation. Biochim Biophys Acta 1974;365:57–71.
[PubMed]
20.
Pusztai A, Grant G, Brown DS, Ewen SWB, Bardocz S: Phaseolus vulgaris lectin induces growth and increases the polyamine content of rat small intestine in vivo. Med Sci Res 1988;16:1283–1284.
21.
Pryme IF, Pusztai A, Bardocz S, Ewen SW: The induction of gut hyperplasia by phytohaemagglutinin in the diet and limitation of tumour growth. Histol Histopathol 1998;13:575–583.
[PubMed]
22.
Pryme IF, Bardocz S, Pusztai A, Ewen SW: The growth of an established murine non-Hodgkin lymphoma tumour is limited by switching to a phytohaemagglutinin-containing diet. Cancer Lett 1999;146:87–91.
[PubMed]
23.
Hussain MA: A biological function for glucagon-like peptide-2. Eur J Endocrinol 1998;139:265–267.
[PubMed]
24.
Grant BG, Dorward PM, Buchan WC, Armour JC, Pusztai A: Consumption of diets containing raw soya beans (Glycine max), kidney beans (Phaseolus vulgaris), cowpeas (Vigna unguiculata) or lupin seeds (Lupinus angustifolius) by rats for up to 700 days: Effects on body consumption and organ weights. Br J Nutr 1995;73:17–29.
[PubMed]
25.
Kik MJL, Koninkx JFJG, van den Muysenberg A, Hendriksen F: Pathological effects of phaseolus vulgaris isolectins on pig jejunal mucosa in organ culture. Gut 1991;32:886–892.
[PubMed]
26.
Lorenzsonn V, Olsen WA: In vivo responses of rat intestinal epithelium to intraluminal dietary lectins. Gastroenterology 1982;82:838–848.
[PubMed]
27.
Ovelgonne JH, Koninkx JF, Pusztai A, Bardocz S, Kok W, Ewen SW, Hendriks HG, van Dijk JE: Decreased levels of heat shock proteins in gut epithelial cells after exposure to plant lectins. Gut 2000;46:679–687.
[PubMed]
28.
Seger R, Krebs EG: The MAPK signaling cascade. FASEB J 1995;9:726–735.
[PubMed]
29.
Leaman DW, Pisharody S, Flickinger TW, Commane MA, Schlessinger J, Kerr IM, Levy DE, Stark GR: Roles of JAKs in activation of STATs and stimulation of c-fos gene expression by epidermal growth factor. Mol Cell Biol 1996;16:369–375.
[PubMed]
30.
Dieckgraefe BK, Weems DM, Santoro SA, Alpers DH: ERK and p38 MAP kinase pathways are mediators of intestinal epithelial wound-induced signal transduction. Biochem Biophys Res Commun 1997;233:389–394.
[PubMed]
31.
Li YQ, Hii CS, Der CJ, Ferrante A: Direct evidence that ERK regulates the production/secretion of interleukin-2 in PHA/PMA-stimulated T lymphocytes. Immunology 1999;96:524–528.
[PubMed]
32.
Pahlavani MA, Harris MD, Richardson A: Activation of p21ras/MAPK signal transduction molecules decreases with age in mitogen-stimulated T cells from rats. Cell Immunol 1998;185:39–48.
[PubMed]
33.
Li YQ, Hii CS, Costabile M, Goh D, Der CJ, Ferrante A: Regulation of lymphotoxin production by the p21ras-raf-MEK-ERK cascade in PHA/PMA-stimulated Jurkat cells. J Immunol 1999;162:3316–3320.
[PubMed]
34.
Goldstein IJ, Poretz RD: Isolation, physiochemical characterization and carbohydrate-binding specifity of lectins; in Liener IE, Sharon N, Goldstein IJ (eds): The Lectins. Properties, Functions, and Applications in Biology and Medicine. London, Academic Press, 1986, pp 33–247.
35.
Borberg H, Woodruff J, Hirschhorn R, Gesner B, Miescher PA, Silber R: Phytohemagglutinin: Inhibition of the agglutinating activity by N-acetyl-D-galactosamine. Science 1966;154:1019–1020.
[PubMed]
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