A complex glyconutritional (GN) mixture of mono-, di-and polysaccharides was investigated to assess its capacity to protect two different types of rodent cells, rat hepatocytes and mouse splenocytes, from depletion of glutathione by a sulfhydryl-reactive mycotoxin, patulin, or by coxsackievirus B3 (CVB3) infection, respectively. Rat hepatocytes were treated with the GN mixture in vitro or received carrier medium only prior to treatment with patulin. When treated with the GN mixture prior to patulin exposure hepatocytes demonstrated protection against depletion of intracellular reduced glutathione (GSH). Cells treated with the GN for up to 15 hours prior to patulin exposure showed no increase in protection of GSH above that demonstrated by cells treated for 3 hours. Mice were infected with CVB3 and one treatment group was injected intraperitoneally with the GN once a week. Animals were splenectomized each month over a ten month treatment for analysis of spleen monocytic cells. Splenocytes from mice treated with the GN mixture did not show the virally-associated depletion of intracellular GSH or damage to pancreatic acini observed in CVB3 inoculated but non-GN-treated mice. Animals from which spleen cells were taken for analysis showed no decrease in anti-CVB3 antibodies and no decrease in viral titers to accompany or explain the normal levels of intracellular GSH. These data strongly suggest that a complex mixture of exogenous saccharides exerts a protective effect on liver cells in vitro in that the cells are protected from chemically initiated depletion of intracellular GSH, and on spleen cells in vivo in that the cells are protected against a CVB3-initiated decrease in intracellular GSH and increase in pancreatic acini damage.

Challenge of adolescent male CD-1 mice with a coxsackievirus B3 (CVB3) strain (CVB3 m ) induces mild to severe destruction of pancreatic acinar cells, but causes no deaths and does not induce hyperglycemia. A weekly parenteral (intraperitoneal) administration of a glyconutritional mixture (Ambrotose® to virus-challenged mice was assessed to determine if there were any benefits to recovery over an eight month period. Virus-challenged mice showed a significant weight loss over the initial five weeks of the experiment, but injection of Ambrotose® to similar virus-challenged mice restored the total body weight to levels found in normal mice. Normal mice given Ambrotose® exhibited a small weight gain. Mice given Ambrotose® showed reduced severity of pancreatitis, as evidenced by significant reductions in percentages of pancreatic acinar cells destroyed and proportion of sections of pancreata with destroyed acinar cells, compared to virus control-mice not injected with Ambrotose®. Statistical analyses of the extent of acinar cell pathology in all virus-challenged mice showed that Ambrotose® contributed significantly to recovery of the acinar cell population in virus-inoculated mice. Anti-viral antibody titers were not affected by Ambrotose® injections. One potential mechanism to explain the benefits derived from Ambrotose® injections came from studies of antioxidant levels of glutathione in splenic macrophages/monocytes. Whereas CVB3 challenge of mice reduced glutathione levels in the latter cells, Ambrotose® injections to virus-challenged mice restored glutathione levels to those found in normal mice. In summary, most but not all mice derived benefits from Ambrotose® injections, i.e. a reduction in pathology in the pancreas and restored levels of the antioxidant glutathione in macrophages/monocytes. Higher doses of Ambrotose® could provide greater benefits for more mice, a study for the future.

An extract from the parenchyma of Aloe barbadensis Miller shown to contain long chain polydispersed beta (1,4)-linked mannan polymers with random O-acetyl groups (acemannan, Carrisyn) was found to initiate the phagocyte production of monokines that supported antibody dependent cellular cytotoxicity and stimulated blastogenesis in thymocytes. Acemannan, in both enriched and highly purified forms, was administered intraperitoneally to female CFW mice into which murine sarcoma cells had been subcutaneously implanted. The rapidly growing, highly malignant and invasive sarcoma grew in 100% of implanted control animals, resulting in mortality in 20 to 46 days, dependent on the number of cells implanted. Approximately 40% of animals treated with acemannan at the time of tumor cell implantation (1.5 x 10(6) cells) survived. Tumors in acemannan-treated animals exhibited vascular congestion, edema, polymorphonuclear leukocyte infiltration, and central necrosing foci with hemorrhage and peripheral fibrosis. The data indicate that in vivo treatment of peritoneal macrophages stimulates the macrophage production of monokines, including interleukin-1 and tumor necrosis factor. The data further indicate that sarcomas in animals treated i.p. with acemannan at the time of tumor cell implantation were infiltrated by immune system cells, became necrotic, and regressed. The combined data suggest that acemannan-stimulated synthesis of monokines resulted in the initiation of immune attack, necrosis, and regression of implanted sarcomas in mice.

Acemannan (ACE-M), a beta-(1,4)-linked acetylated mannan, was evaluated for in vitro activity against human immunodeficiency virus type 1 (HIV-1). Castanospermine (CAS), deoxymannojirimycin (DMN), swainsonine (SWS), azidothymidine (AZT), and dideoxythymidine (DDC) were tested in parallel as control compounds. In vitro antiviral efficacy of ACE-M was evaluated in a variety of cell lines including human peripheral mononuclear, CEM-SS1 and MT-2(2) cells. The virus strain, number of infectious units per cell, and target cell line were important factors in determining the degree of inhibition of viral cytopathic effect in the presence of ACE-M and other control compounds tested. Maximum inhibitory effect was observed in CEM-SS cells infected with the RFII strain of HIV-1. This inhibitory effect was determined to be concentration-dependent. Assay design included primary screening to measure cell viabilities of infected target cells in the presence and absence of test compounds. When tested on HIV-1/RFII-infected CEM-SS cells, the 50% inhibitory effect of CAS (IC50 = 28), an inhibitor of alpha-glucosidase I, was determined to be similar to that observed for ACE-M (IC50 = 45). However, DMN and SWS, inhibitors of mannosidase I and II, tested in parallel to CAS and ACE-M, exhibited no IC50 values. Antiviral potential of ACE-M as an inhibitor of syncytia formation was also explored using CEM-SS cells. Suppression of syncytia formation was observed at an ACE-M concentration of 31.25 micrograms/ml, and complete inhibition was observed at 62.5 micrograms/ml. In addition, HIV-1 RNA levels were studied to establish the antiviral potential of ACE-M in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)