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Title: Effects of OK-432 on murine bone marrow and the production of natural killer cells

Abstract

The streptococcal preparation, OK-432, which augments anti-tumor responses in humans and mice, has been shown to be a potent immunomodulator. Among its effects is a pronounced augmentation of natural killer (NK) activity. The hypothesis that OK-432 alters the rates of production and maturation of NK cells in the bone marrow was tested. Studies to determine the kinetic parameters of NK cell production in normal C57BL/6J mice using tritiated thymidine, /sup 3/H-TdR, as a DNA marker are described. We are now extending those studies to determine the effect of OK-432 on the bone marrow and on the production of NK cells in the marrow. Initial observations are reported which indicate that OK-432 has profound effects on the cellularity and mitotic activity of the bone marrow, and in particular, on cells with the characteristics of natural killer cells within the marrow. 17 refs., 3 figs., 4 tabs.

Authors:
;
Publication Date:
Research Org.:
Washington Univ., Seattle (USA). Dept. of Biological Structure
OSTI Identifier:
5072424
Report Number(s):
DOE/EV/10270-T7
ON: DE87002046
DOE Contract Number:
AT06-79EV10270
Resource Type:
Technical Report
Resource Relation:
Other Information: Microfiche only, copy does not permit paper copy reproduction
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; BONE MARROW CELLS; CELL DIFFERENTIATION; ENDOTOXINS; BIOLOGICAL FUNCTIONS; LEUKOCYTES; ANTINEOPLASTIC DRUGS; MICE; TRACER TECHNIQUES; TRITIUM COMPOUNDS; ANIMAL CELLS; ANIMALS; ANTIGENS; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CONNECTIVE TISSUE CELLS; DRUGS; FUNCTIONS; ISOTOPE APPLICATIONS; LABELLED COMPOUNDS; MAMMALS; MATERIALS; RODENTS; SOMATIC CELLS; TOXIC MATERIALS; TOXINS; VERTEBRATES; 551001* - Physiological Systems- Tracer Techniques; 550301 - Cytology- Tracer Techniques

Citation Formats

Pollack, S.B., and Rosse, C.. Effects of OK-432 on murine bone marrow and the production of natural killer cells. United States: N. p., 1985. Web.
Pollack, S.B., & Rosse, C.. Effects of OK-432 on murine bone marrow and the production of natural killer cells. United States.
Pollack, S.B., and Rosse, C.. Tue . "Effects of OK-432 on murine bone marrow and the production of natural killer cells". United States. doi:.
@article{osti_5072424,
title = {Effects of OK-432 on murine bone marrow and the production of natural killer cells},
author = {Pollack, S.B. and Rosse, C.},
abstractNote = {The streptococcal preparation, OK-432, which augments anti-tumor responses in humans and mice, has been shown to be a potent immunomodulator. Among its effects is a pronounced augmentation of natural killer (NK) activity. The hypothesis that OK-432 alters the rates of production and maturation of NK cells in the bone marrow was tested. Studies to determine the kinetic parameters of NK cell production in normal C57BL/6J mice using tritiated thymidine, /sup 3/H-TdR, as a DNA marker are described. We are now extending those studies to determine the effect of OK-432 on the bone marrow and on the production of NK cells in the marrow. Initial observations are reported which indicate that OK-432 has profound effects on the cellularity and mitotic activity of the bone marrow, and in particular, on cells with the characteristics of natural killer cells within the marrow. 17 refs., 3 figs., 4 tabs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Jan 01 00:00:00 EST 1985},
month = {Tue Jan 01 00:00:00 EST 1985}
}

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  • In vitro effects of OK-432 (polysaccharide extract of Streptococcus haemolyticus) on irradiated mouse bone marrow cells are examined. Bone marrow cells of BDF1 mouse (1 [times] 10[sup 6] cells/ml) were incubated with alpha medium, 2% fetal calf serum and OK-432 in a CO[sub 2] incubator at 37[degrees]C for 24, 48 and 72 h, respectively. After centrifugation, each supernatant was collected and used for conditioned medium in a CFU-GM assay: Changes in CFU-GM as a function of incubation time and OK-432 dose were examined; changes of CFU-GM according to various doses of OK-432 were examined in two mouse strains, BDF[sub 1]more » and BALB/c mouse; changes in the protective effect of OK-432 in terms of CFU-GM as a function of administration timing of OK-432 in relation to irradiation. As a radiation source, [sup 137]Cs at a dose rate of 500 cGy/min was used. The CFU-GM decreased with the incubation time when OK-432 was not administered, while it significantly increased with incubation time when OK-432 was added at 0.5 and 1.0 KE/ml at 48-72 h of incubation. The former showed marked increase at 48-72 h of incubation. CFU-GM of BDF[sub 1] mouse was always higher than that of BALB/c mouse for any dose of OK-432. CFU-GM per femur according to the timing of administration of OK-432 from 24 h before to 24 h after irradiation showed 10299 [+-] 2300 (24 h before), 10783 [+-] 2463 (3 h before), 10045 [+-] 1501 (immediately after), 8504 [+-] 1188 (3 h after), 4898 [+-] 1212 (6 h after), 1214 [+-] 736 (12 h after) and 181 [+-] 113 (24 h after irradiation), respectively. OK-432 stimulates cultures mouse bone marrow cells to produce GM-CSF in vitro by direct contact action. This direct stimulating action of OK-432 on GM-CSF production of bone marrow cells can be kept from 24 h before to at least 3 h after irradiation. 6 refs., 4 figs.« less
  • Human bone marrow (BM) cells, depleted of nylon wool-adherent cells, T cells, and natural killer (NK) cells, were cultured in medium containing recombinant interleukin 2 (rIL2). After 21 or 24 days in culture, numerous lymphoid cells with multiple azurophilic granules and a morphology similar to large granular lymphocytes (LGL) were found. Two-color analysis of surface phenotype showed many of these cells to be NKH1-positive and a limited number of cells had other NK markers such as CD16, CD2, or CD8. The CD3 antigen was not coexpressed with NKH1. The cultured BM cells were cytotoxic for K562, Daudi, and Raji cellmore » lines. The NKH1+, CD2-, CD3-, CD16- cells were sorted and, in addition to having the LGL morphology, were found to be cytotoxic for K562 cells (NK (K562)). The generation of NK(K562) activity was significantly suppressed by 5-bromodeoxyuridine plus ultraviolet light treatment, indicating that DNA synthesis is required. These experiments suggest that the described culture conditions allow differentiation of progenitor cells, into immature, but functionally active, NK cells.« less
  • Spleen cells from irradiated, bone marrow-reconstituted mice were tested for their ability to mediate antibody-dependent cellular cytotoxicity against P815 target (ADCC-P815), ADCC against sheep red blood cells (ADCC-SRBC), and natural killer (NK) activity judged as YAC-1 lysis at different times after bone marrow reconstitution. Donor-derived ADCC-P815 effectors were found to appear in the spleens 10-12 days after bone marrow reconstitution simultaneously with the appearance of donor-derived NK cells. NK cells recently derived from bone marrow are known to express the Thy-1 antigen; the phenotype of the ''early'' ADCC-P815 effectors was found to be the same as that of NK cells,more » i.e., Thy-1+, asialo-GM1+. These data suggest that ADCC-P815 effector cells belong to the NK cell population. ADCC-SRBC, in contrast to ADCC-P815 and NK activity, was already high on Day 7 after bone marrow reconstitution. However, it was mediated partly by recipient-derived effectors. ADCC-SRBC effectors were characterized to be different from ADCC-P815 effectors.« less
  • Previous data generated by Murphy and McDaniel indicate that normal murine nylon wool nonadherent splenic cells, with the characteristics of natural killer (NK) cells, effectively inhibit the in vitro growth of Cryptococcus neoformans, a yeast-like pathogen. Nylon wood nonadherent cells from spleens of 7-8 week old mice were further fractionated on discontinuous Percoll gradients. The enrichment of NK cells in Percoll fractions 1 and 2 was confirmed by morphological examination, immunofluorescent staining, and by assessing the cytolytic activity of each Percoll cell fraction against YAC-1 targets in the 4 h /sup 51/Cr release assay. Cells isolated from each Percoll fractionmore » were tested for growth inhibitory activity against C neoformans, using an in vitro 18 h growth inhibition assay. The results showed that NK cell enrichment was concomitant with the enrichment of anti-cryptococcal activity the Percoll fractions 1 and 2. An immunolabeling method combined with scanning electron microscopy was used to demonstrate that the effector cells attached to C. neoformans were asialo GM/sub 1/ positive and, therefore, had NK cell characteristics. NK cells have Fc receptors on their surfaces , and are capable of antibody-dependent cell-mediated cytotoxicity (ADCC) against IgG-coated target cells. The author examined the effects of the IgG fraction of rabbit anti-cryptococcal antibody on the NK cell-mediated growth inhibition of C. neoformans. The data indicated that the effector cells involved in antibody-dependent growth inhibition of cryptococci are either NK cells or copurify and coexist in the same population with NK cells.« less
  • The authors previously reported that interleukin 1 (IL-1) administration 20 hr before irradiation protects mice from lethal effects of radiation. The recovery of total nucleated bone-marrow cells and of hematopoietic progenitor cells was enhanced in IL-1 treated, as compared to untreated, irradiated mice. This suggests that IL-1 administration may affect the cells in the bone marrow of normal mice. Intraperitoneal administration of recombinant IL-1 resulted in bone marrow cell enlargement and increased cycling of these enlarged cells. In addition, mice to proliferate in response to granulocyte macrophage-colony-stimulating factor (GM-CSF) in cell suspension cultures was enhanced. The authors have also previouslymore » shown that IL-1 induces the appearance of high titers of CSF in the serum. Consequently, hematopoietic growth factors that are generated at local sites following IL-1 administration may mediate the observed cell cycling effect.« less