Abstract
The saline extract from the roots of Phytolacca americana (pokeweed) possesses three biological properties; hemagglutinin, leukagglutinin, and mitogen. Fractionation and further purification on calcium phosphate column chromatography revealed that the biologically active substance was eluted in the front moving fraction with 0.05 M phosphate buffer pH 7.5. Analytical separation on polyacrylamide gels in disc electrophoresis yielded a single homogeneous band with an Rf value of 0.43 containing all three biological activities. This fraction had an ultraviolet absorption spectrum similar to PHA, was stable to both periodate and mercaptoethanol treatment and gave a single band in double diffusion and immunoelectrophoretic analysis against the antibody prepared to the crude PWM saline extract. Absorption studies with red cells or stroma revealed that the hemagglutinin could be selectively removed without significantly altering the mitogen, whereas absorption with leukocytes resulted in loss of both the mitogenic and leukagglutinating activities.
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- Börjeson J., Gardell S., Nordén A. A simple procedure for the determination of DNA synthesis in cultured lymphocytes. Scand J Haematol. 1966;3(2):158–164. doi: 10.1111/j.1600-0609.1966.tb01437.x. [DOI] [PubMed] [Google Scholar]
- CARSTAIRS K. The human small lymphocyte: its possible pluripotential quality. Lancet. 1962 Apr 21;1(7234):829–832. doi: 10.1016/s0140-6736(62)91841-x. [DOI] [PubMed] [Google Scholar]
- CHESSIN L. N., BRAMSON S., KUHNS W. J., HIRSCHHORN K. STUDIES ON THE A, B, O(H) BLOOD GROUPS ON HUMAN CELLS IN CULTURE. Blood. 1965 Jun;25:944–953. [PubMed] [Google Scholar]
- CROWLE A. J. A simplified agar electrophoretic method for use in antigen separation and serologic analysis. J Lab Clin Med. 1956 Oct;48(4):642–648. [PubMed] [Google Scholar]
- Chessin L. N., Börjeson J., Welsh P. D., Douglas S. D., Cooper H. L. Studies on human peripheral blood lymphocytes in vitro. II. Morphological and biochemical studies on the transformation of lymphocytes by pokeweed mitogen. J Exp Med. 1966 Nov 1;124(5):873–884. doi: 10.1084/jem.124.5.873. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FARNES P., BARKER B. E., BROWNHILL L. E., FANGER H. MITOGENIC ACTIVITY IN PHYTOLACCA AMERICANA (POKEWEED). Lancet. 1964 Nov 21;2(7369):1100–1101. doi: 10.1016/s0140-6736(64)92616-9. [DOI] [PubMed] [Google Scholar]
- GRABAR P., WILLIAMS C. A. Méthode permettant l'étude conjuguée des proprietés électrophorétiques et immunochimiques d'un mélange de protéines; application au sérum sanguin. Biochim Biophys Acta. 1953 Jan;10(1):193–194. doi: 10.1016/0006-3002(53)90233-9. [DOI] [PubMed] [Google Scholar]
- HIRSCHHORN K., KOLODNY R. L., HASHEM N., BACH F. Mitogenic action of phytohaemagglutinin. Lancet. 1963 Aug 10;2(7302):305–306. doi: 10.1016/s0140-6736(63)90213-7. [DOI] [PubMed] [Google Scholar]
- LING N. R., SPICER E., JAMES K., WILLIAMSON N. THE ACTIVATION OF HUMAN PERIPHERAL LYMPHOCYTES BY PRODUCTS OF STAPHYLOCOCCI. Br J Haematol. 1965 Jul;11:421–431. doi: 10.1111/j.1365-2141.1965.tb06604.x. [DOI] [PubMed] [Google Scholar]
- NOWELL P. C. Phytohemagglutinin: an initiator of mitosis in cultures of normal human leukocytes. Cancer Res. 1960 May;20:462–466. [PubMed] [Google Scholar]
- POLLEY M. J., ADINOLFI M., MOLLISON P. L. SEROLOGICAL CHARACTERISTICS OF ANTI-A RELATED TO TYPE OF ANTIBODY PROTEIN (7 S GAMMA OR 19 S GAMMA). Vox Sang. 1963 Jul-Aug;8:385–409. doi: 10.1111/j.1423-0410.1963.tb04159.x. [DOI] [PubMed] [Google Scholar]
- REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
- Reisfeld R. A., Small P. A., Jr Electrophoretic heterogeneity of polypeptide chains of specific antibodies. Science. 1966 May 27;152(3726):1253–1255. doi: 10.1126/science.152.3726.1253. [DOI] [PubMed] [Google Scholar]
- Sell S., Rowe D. S., Gell P. G. Studies on rabbit lymphocytes in vitro. 3. Proteins, RNA, and DNA synthesis by lymphocyte cultures after stimulation with phytohaemagglutinin, with staphylococcal filtrate, with antiallotype serum, and with heterologous antiserum to rabbit whole serum. J Exp Med. 1965 Oct 1;122(4):823–839. doi: 10.1084/jem.122.4.823. [DOI] [PMC free article] [PubMed] [Google Scholar]
- TANAKA Y., EPSTEIN L. B., BRECHER G., STOHLMAN F., Jr TRANSFORMATION OF LYMPHOCYTES IN CULTURES OF HUMAN PERIPHERAL BLOOD. Blood. 1963 Nov;22:614–629. [PubMed] [Google Scholar]
- VAN ROOD J. J., VAN LEEUWEN A. LEUKOCYTE GROUPING. A METHOD AND ITS APPLICATION. J Clin Invest. 1963 Sep;42:1382–1390. doi: 10.1172/JCI104822. [DOI] [PMC free article] [PubMed] [Google Scholar]
- YOFFEY J. M., WINTER G. C., OSMOND D. G., MEEK E. S. MORPHOLOGICAL STUDIES IN THE CULTURE OF HUMAN LEUCOCYTES WITH PHYTOHAEMAGGLUTININ. Br J Haematol. 1965 Jul;11:488–497. doi: 10.1111/j.1365-2141.1965.tb06613.x. [DOI] [PubMed] [Google Scholar]