Separation and purification of lymphocyte chemotactic factor (LCF) and interleukin 2 produced by human peripheral blood mononuclear cells - PubMed (original) (raw)

Separation and purification of lymphocyte chemotactic factor (LCF) and interleukin 2 produced by human peripheral blood mononuclear cells

J W Potter et al. Cell Immunol. 1987 Mar.

Abstract

Two T-cell chemotactic factors, lymphocyte chemotactic factor (LCF) and interleukin 2 (IL-2), were separated and characterized from culture supernatants of concanavalin A-stimulated human peripheral blood mononuclear cells. LCF was purified approximately 7800-fold to homogeneity from culture supernatant using gel filtration and high-performance liquid chromatography (HPLC). LCF was found to be distinct from both IL-2 and interleukin-1. Sephadex G-100 gel filtration of crude supernatants from concanavalin A-stimulated mononuclear cells showed two molecular weight regions of T lymphocyte chemotactic activity. A 10,000- to 25,000-Da region contained both IL-2 and LCF and a 45,000- to 75,000-Da region contained only a high molecular weight form of LCF. Both high and low molecular weight species of LCF eluted with 40-44% acetonitrile from a reversed-phase C18 HPLC column. IL-2 present only in the low molecular weight region eluted from the C18 column with 65-75% acetonitrile. The migration of T lymphocytes to IL-2 was totally inhibited by anti-interleukin 2 receptor antibody while the response of T cells to LCF was unaffected. LCF eluting off the C18 column was purified to homogeneity by two subsequent cycles of gel filtration HPLC. The resultant protein showed a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis corresponding to a molecular weight of 10,500. The data presented here demonstrate that IL-2 and LCF are distinct lymphocyte chemotactic factors and although they are not readily separable from crude supernatants by molecular sieve chromatography, they can easily be distinguished by reversed-phase HPLC.

PubMed Disclaimer

Cited by

Chemoattractant activity of IL-2 for human lymphocytes: a requirement for the IL-2 receptor beta-chain.
Wilkinson PC, Newman I. Wilkinson PC, et al. Immunology. 1994 May;82(1):134-9. Immunology. 1994. PMID: 8045589 Free PMC article.
Chemoattraction of human blood T lymphocytes by interleukin-15.
Wilkinson PC, Liew FY. Wilkinson PC, et al. J Exp Med. 1995 Mar 1;181(3):1255-9. doi: 10.1084/jem.181.3.1255. J Exp Med. 1995. PMID: 7869044 Free PMC article.
Recombinant IL-4 and IFN-gamma activate locomotor capacity in human B lymphocytes.
Wilkinson PC, Islam LN. Wilkinson PC, et al. Immunology. 1989 Jun;67(2):237-43. Immunology. 1989. PMID: 2502496 Free PMC article.
Production of interleukin-8 by human dermal fibroblasts and keratinocytes in response to interleukin-1 or tumour necrosis factor.
Larsen CG, Anderson AO, Oppenheim JJ, Matsushima K. Larsen CG, et al. Immunology. 1989 Sep;68(1):31-6. Immunology. 1989. PMID: 2478449 Free PMC article.

Separation and purification of lymphocyte chemotactic factor (LCF) and interleukin 2 produced by human peripheral blood mononuclear cells - PubMed (original) (raw)