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Preparation and antitumor activity of nontoxic lipid A

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Summary

Highly refined, disaggregated endotoxic glycolipids (B5) from heptose-less (Re) mutant Salmonella typhimurium quantitatively converted to nontoxic (lethality for chick embryos) and nonpyrogenic (fever in rabbits) lipid A by treatment with boiling 0.1 N HCl (B5-HC1). Nontoxic B5-HCl, like toxic B5, caused regression of line-10 tumors and elimination of lymph node metastasis in 27 of 32 (84%) syngeneic strain 2 guinea pigs at a dosage of 150 μg. At this dosage, toxic B5 led to a cure in 54 of 67 (81%) tumor-bearing animals. All cured animals rejected a second line-10 tumor cell transplant. This activity depended on combining the toxic or nontoxic endotoxins with mycobacterial trehalose mycolate (P3) and an essentially nontoxic peptide-containing side-fraction (ACP) recovered during the isolation of B5. In contrast to toxic B5 or endotoxins in general, nontoxic B5-HCl did not cause endotoxic shock when combined with adjuvant dipeptide (MDP) and injected IV into guinea pigs. Chemical analysis showed that the phosphate content of nontoxic B5-HCl was about one-half that observed in toxic B5 or in toxic KDO-free lipid A, which was obtained by treating toxic B5 with sodium acetate at pH 4.5 at 100° C (B5-pH 4.5). The molar ratio of glucosamine: phosphorus: fatty acids was 2:1:4 for nontoxic B5-HCl and was 2:2:4 for toxic B5-pH 4.5. These results demonstrate that endotoxic extracts could be selectively detoxified while retaining antitumor properties. Thus, nontoxic B5-HCl may be a potential candidate for immunotherapy of human cancer.

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Authors and Affiliations

  1. Ribi Immuno Chem Research, Inc., P.O. Box 1409, 59840, Hamilton, Montana

    Edgar Ribi, John Cantrell, Steve Schwartzman & Reno Parker

  2. Rocky Mountain Laboratory, USPHS, National Institutes of Health, National Institute of Allergy and Infectious Diseases, 59840, Hamilton, Montana

    Edgar Ribi, Kenichi Amano, John Cantrell & Steve Schwartzman

  3. William S. Middleton Memorial Veterans Hospital, 53705, Madison, Wisconsin, USA

    Kuni Takayama

Authors
  1. Edgar Ribi
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  2. Kenichi Amano
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  3. John Cantrell
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  4. Steve Schwartzman
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  5. Reno Parker
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  6. Kuni Takayama
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Additional information

Abbreviations used in this paper: ACP, a nontoxic acetone-chloroform precipitated side-fraction of endotoxin that contains (an) ingredient(s) necessary for tumor regression of line-10 tumors in strain 2 guinea pigs; ReGl, endotoxic glycolipids from Re mutant gram-negative bacteria; ReGl-PCP, ReGl extracted with phenol-chloroform-petroleum ether (PCP); B5, refined endotoxin, free of phospholipids, divalent cations and disaggregated; B5-HCl, nontoxic lipid A prepared from B5 by treatment with hydrochloric acid; B5-pH 4.5, toxic lipid A prepared from B5 by treatment with sodium acetate at pH 4.5; lipid A, hydrochloric acid or sodium acetate hydrolysate of ReGl-PCP or B5; MDP, N-acetyl-muramyl-l-seryl-d-isoglutamine; KDO, keto-3-deoxyoctonate

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Ribi, E., Amano, K., Cantrell, J. et al. Preparation and antitumor activity of nontoxic lipid A. Cancer Immunol Immunother 12, 91–96 (1982). https://doi.org/10.1007/BF00205365

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  • DOI: https://doi.org/10.1007/BF00205365

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