Regulation of key molecules of immunological synapse by T11TS immunotherapy abrogates Cryptococcus neoformans infection in rats.

Cryptococcus neoformans infects and disseminates in hosts with diminished T cell responses. The immunomodulator T11TS (T11 target structure) had profound potential in glioma as well as C. neoformans infected model for disease amelioration. It is been established by our group that T11TS potentiates Calcineurin-NFAT pathway in T cells of C. neoformans infected rats. We investigated the upstream Immunological Synapse (IS) molecules that are vital for the foundation of initial signals for downstream signaling, differentiation and proliferation in T cells. Improved RANTES level in the T11TS treated groups suggests potential recruitment of T cells. Down-regulation of TCRαß, CD3ζ, CD2, CD45 and CD28 molecules by cryptococcus were boosted after T11TS therapy. Heightened expression of inhibitory molecule CTLA-4 in cryptococcosis was dampened by T11TS. The decline of MHC I, MHC II and CD80 expression on macrophages by C. neoformans were enhanced by T11TS. The dampening of positive regulators and upsurge of negative regulators of the IS during cryptococcosis was reversed with T11TS therapy resulting in enhanced clearance of fungus from the lungs as envisaged by our histological studies. This preclinical study with T11TS opens a new prospect for potential immunotherapeutic intervention against the devastating C. neoformans infection with positive aspect for the long-term solution and a safer immunotherapeutic regimen.

Indian freshwater edible snail Bellamya bengalensis lipid extract prevents T cell mediated hypersensitivity and inhibits LPS induced macrophage activation.

ETHNOPHARMACOLOGICAL RELEVANCE: Soup prepared from the foot of fresh water edible snail, Bellamya bengalensis, is traditionally consumed by the tribes of Jharkhand against rheumatism like bone and joint inflammation. As rheumatism has underlying involvement of cell mediated hypersensitivity, in vivo delayed-type hypersensitivity (DTH) model and in vitro LPS-induced macrophage signaling were studied to delineate the mechanism by which Bellamya bengalensis exerts its ethnomedicinal function. Since the whole meat is consumed, the lipid of Bellamya bengalensis (BBL) was hypothesized to be the active part. METHODS AND MATERIALS: BBL isolated from the foot part of this species, was characterized and given by gavage daily (10mg BBL/kg; 20mg BBL/kg) to mice for 3 weeks prior to initiating development of DTH. Effects of DTH induced changes in paw diameter, serum nitric oxide (NO), serum tumor necrosis factor (TNF)-α level, CINC1 level, splenic CD4(+)/CD8(+) cell ratios, and level of splenic Treg cells were then compared with values in untreated control mice. In vitro effect of BBL on LPS-stimulated macrophage, the immune cell that is active in DTH, was assessed by NF-kB p65 nuclear translocation, reactive oxygen species (ROS), TNFα, and NO production. RESULTS: BBL was characterized, and its supplementation in situ led to significant decrease in paw edema, tissue myeloperoxidase activity, NO level, serum TNFα level and CINC 1 level as well as decrease in splenic CD4(+)/CD8(+) ratios and increase in level of Treg cells. BBL was shown to inhibit ROS, NO, and TNFα production along with NF-kB p65 nuclear translocation in LPS stimulated macrophage. CONCLUSION: Bellamya bengalensis, traditionally used against diseases with underlying etiology of cell mediated immunity as in rheumatism, which acts through inhibition of overexpressed cell mediated immunity. The factor exerting this activity probably is the oleic acid and cyclopropane fatty acid rich lipid, isolated after the ethnomedicinal clue, from the foot of this species.

Prevention of arthritis markers in experimental animal and inflammation signalling in macrophage by Karanjin isolated from Pongamia pinnata seed extract.

Karanjin, the furanoflavonoid reported to possess gastroprotective and anti-diabetic properties, was investigated against experimental arthritis and its molecular signalling in inflammation was explored in macrophages. Karanjin was isolated from hexane extract of Pongamia pinnata seeds and was evaluated on arthritis markers in adjuvant induced arthritis model (AIA) in two doses (per oral; 10 mg/kg/day and 20 mg/kg/day). Karanjin dose dependently reduced collagen and cartilage breakdown markers viz. urinary hydroxyproline and glucosamine, respectively, serum lysosomal enzymes responsible for articular cartilage damage, and major proinflammatory cytokine TNFα, secreted by macrophages involved in articular inflammation and destruction. Karanjin also prevented joint damage as evidenced from arthritis score, radiographic and histopathological analysis. To delineate the molecular target of Karanjin, in vitro study on LPS induced macrophages were performed at calibrated non toxic doses (4 µg/mL and 6 µg/mL). Karanjin reduced TNFα production and also showed potent inhibitory effect on nitric oxide and reactive oxygen species production which is generally induced by TNFα from activated macrophages. NF-κB, the key regulator of TNFα signalling during inflammation was significantly suppressed by Karanjin. Our study for the first time highlights the anti-inflammatory role of Karanjin in experimental arthritis model as well as on macrophage signalling, thereby depicting its probable mechanism of action.

Suppression of NF-κB p65 nuclear translocation and tumor necrosis factor-α by Pongamia pinnata seed extract in adjuvant-induced arthritis.

Pongamia pinnata is a plant known for its therapeutic usage in Indian traditional medicine. Despite the controversy regarding toxic flavonoid and erucic acid content, the seed of this plant is consumed in tribal medicine and its oil is used in Ayurveda to treat psoriasis and arthritis. This study explored the potential anti-arthritic effects of a P. pinnata seed (hexane) extract (PSE) at non-lethal doses in an adjuvant-induced arthritic rat model; possible mechanisms of any observed effects were also explored. After establishing the lethal doses arising from oral exposure to the extract, the material was administered per os daily at two doses (0.3 g/kg/day; 0.5 g/kg/day) to arthritic rats. Other rats received indomethacin or vehicle (control). Treatments were performed for a total of 14 days. One day after the final exposure, the rats were euthanized to permit harvest of various cells, blood, and tissues for analyses. Paw diameter and tissue myeloperoxidase activity in the paws were evaluated as indices for edema and neutrophil infiltration into the tissue. The severity of arthritis in the experimental rats was assessed via measures of urinary hydroxyproline (HP) and glucosamine, and of serum pro-inflammatory TNFα and anti-inflammatory IL-10. The extent of NF-κB p65 nuclear translocation in peritoneal macrophages harvested from naïve rats and then treated in vitro was also assessed. The results indicated that exposure to PSE significantly decreased paw diameter, tissue myeloperoxidase level, and levels of urinary HP and glucosamine, as well as of serum TNFα and IL-10 in adjuvant-injected (arthritic) rats. In vitro PSE treatment also resulted in a marked inhibition of NF-κB p65 nuclear translocation in primary cultures of peritoneal macrophages. Thus, PSE appears to be able to prevent experimental arthritis, in part, by helping to maintain the balance between pro- and anti-inflammatory cytokines and by inhibiting NF-κB activation.

Antioxidant content and activity of the Indian fresh-water pearl mussel in the prevention of arthritis in an experimental animal model.

The flesh of the Indian fresh-water mussel, Lamellidens marginalis (LM; Lamarck, 1819), is the byproduct of pearl culture and a cheap protein source. The present study investigated the antioxidant content of this ethnomedicinally cited species to outline its importance in food security and disease prevention. LM was found to be rich in polyphenol antioxidants with good correlation with its reducing capacity. LM also showed a significant free-radical-scavenging activity, H(2)O(2)-scavenging activity and Fe-chelating activity. To study the effect of this dietary antioxidant against oxidative stress, we took inflammatory arthritis as a model. LM-treated arthritis rats showed a higher antioxidant defence system with elevated superoxide dismutase, total thiol, glutathione S transferase, glutathione peroxidase, total antioxidant status and catalase concentration of haemolysate. Oxidative stress markers like serum thiobarbituric acid-reacting substances, methyl glyoxal, NO and total oxidant status levels were decreased in LM-treated arthritis rats. Hence, the dietary antioxidants of LM were found to be effective in the prevention of oxidative stress in inflammatory arthritis. In conclusion, LM, the cash-crop byproduct, provides a rare opportunity for income and nutrition, not only by providing cheap and available energy, protein and dietary factors, but also by providing antioxidants effective against chronic inflammatory disease.

Anticancer potential of animal venoms and toxins.

Anticancer drug development from natural resources are ventured throughout the world. Animal venoms and toxins a potential bio resource and a therapeutic tool were known to man for centuries through folk and traditional knowledge. The biodiversity of venoms and toxins made it a unique source of leads and structural templates from which new therapeutic agents may be developed. Venoms of several animal species (snake, scorpion, toad, frog etc) and their active components (protein and non protein toxins, peptides, enzymes, etc) have shown therapeutic potential against cancer. In the present review, the anticancer potential of venoms and toxins from snakes, scorpions, toads and frogs has been discussed. Some of these molecules are in the clinical trials and may find their way towards anticancer drug development in the near future. The implications of combination therapy of natural products in cancer have been discussed.