Immunostimulatory compositions and methods of use thereof

Abstract

Immunostimulatory compositions and methods of use are described to either enhance or diminish the immune stimulation effects of a honey or honey isolate by recognition of the presence of type II arabinogalactan compounds and utilising this knowledge to tailor the concentration of such compounds thereby adjusting the immune stimulation effects.

1 . A honey isolate containing type II arabinogalactan, wherein the average molecular weight of the arabinogalactan is 40,000 to 1,100,000 and wherein the ratio of arabinose to galactose ranges from 0.1 to 2.0 parts arabinose to 1.0 parts galactose. 
     
     
         2 . (canceled) 
     
     
         3 . The honey isolate as claimed in  claim 1  wherein the type II arabinogalactan has a structure including: 
       
         
           
           
               
               
           
         
       
       Where n=5 to 25 and R 1 = 
       
         
           
           
               
               
           
         
       
       Where R 2 = 
       
         
           
           
               
               
           
         
       
       Where R 3 = 
       
         
           
           
               
               
           
         
       
     
     
         4 . The honey isolate as claimed in  claim 1 , wherein the isolate contains at least 5 μg/g type II arabinogalactan. 
     
     
         5 . The honey isolate as claimed in  claim 1  wherein the isolate contains at least 10 μg/g type II arabinogalactan. 
     
     
         6 . A honey analogue containing an isolate as claimed in  claim 1 . 
     
     
         7 . (canceled) 
     
     
         8 . A composition formulated for topical application to a body area of a patient in need thereof containing:
 (a) an isolate as claimed in  claim 1 ; or   (b) a honey analogue as claimed in  claim 6 ; or   (c) combinations of the above.   
     
     
         9 . The composition as claimed in  claim 8  wherein the composition is selected from: a dressing, a cream, an ointment, a gel, and combinations thereof. 
     
     
         10 . The composition as claimed in  claim 8  wherein the type II AG is an arabinogalactan protein (AGP). 
     
     
         11 . The composition as claimed in  claim 8  wherein a non-honey based AG containing compound or isolate is added to the composition. 
     
     
         12 . A method of stimulating the immune system of a patient in need thereof by topical application of the composition as claimed in  claim 8  to a body area of the patient. 
     
     
         13 . The method as claimed in  claim 12  wherein the composition is formulated for application to a recalcitrant topical body area on a patient. 
     
     
         14 . (canceled) 
     
     
         15 . A method of minimising the immune-stimulatory effects of a honey based topical composition by the step of removing type II arabinogalactan (AG) from the honey in the composition. 
     
     
         16 . The method as claimed in  claim 15  wherein the composition has less than 5 μg/g AG compounds. 
     
     
         17 . The method as claimed in  claim 15  wherein the composition has less than 1 μg/g AG compounds. 
     
     
         18 . The method as claimed in  claim 15  wherein the composition is formulated for application to a sensitive topical body area on a patient. 
     
     
         19 . The method as claimed in  claim 15  wherein the honey based topical composition still provides anti-microbial effects associated with honey. 
     
     
         20 - 22 . (canceled) 
     
     
         23 . A method of treatment of a recalcitrant skin condition on a topical body area on a subject in need thereof by:
 a. application of a composition as claimed in  claim 8  initially; and, as skin healing progresses,   b. applying a honey or honey analogue based composition with type II AG compounds removed from the honey or analogue.   
     
     
         24 . The method as claimed in  claim 23  wherein the composition of part b. has less than 5 μg/g AG compounds. 
     
     
         25 . The method as claimed in  claim 24  wherein the composition has less than 1 μg/g AG compounds. 
     
     
         26 - 28 . (canceled) 
     
     
         29 . A method of treatment of a sensitive skin condition on a topical body area on a subject in need thereof by:
 a. applying a honey or honey analogue based composition with type II AG compounds removed from the honey; and as skin healing progresses,   b. application of a composition as claimed in  claim 8 .   
     
     
         30 . The method as claimed in  claim 29  wherein the composition of part a. has less than 5 μg/g AG compounds. 
     
     
         31 . The method as claimed in  claim 29  wherein the composition of part a. has less than 1 μg/g AG compounds. 
     
     
         32 - 34 . (canceled) 
     
     
         35 . A method of producing a honey based immune-stimulatory composition by the step of:
 (a) testing a series of honey batches for the presence of type II AG compounds;   (b) selecting and blending together honeys based on the test results to form a honey blend with type II AG levels greater than 5 μg/g.   
     
     
         36 . The method as claimed in  claim 35  wherein the honeys selected and blended result in a honey blend with type II AG levels greater than 10 μg/g. 
     
     
         37 . The method as claimed in  claim 35  where in the honey includes kanuka honey. 
     
     
         38 . The method as claimed in  claim 35  wherein the honey tested is produced from a plant or plants selected and/or bred to maximise the level of type II AG compounds in the plant nectar from which the honey is produced. 
     
     
         39 . The method as claimed in  claim 38  wherein the plant nectar is selected from the genus:  Leptospermum, Kunzea, Weinmannia, Knightia, Metrosideros, Fagus, Trifolium,  Myrtaceae, and combinations thereof. 
     
     
         40 . The method as claimed in  claim 35  wherein the composition further includes non-honey based arabinogalactans. 
     
     
         41 . A method of producing a honey based composition that minimises immune-stimulatory effects by:
 (a) testing a series of honey batches for the presence of type II AG compounds;   (b) selecting and blending together honeys based on the test results sufficient to leave less than 5 μg/g type II AG compounds in the honey blend.   
     
     
         42 . The method as claimed in  claim 41  wherein the honey tested is produced from a plant or plants selected and/or bred to minimise the level of type II AG compounds in the plant nectar from which the honey is produced. 
     
     
         43 . The method as claimed in  claim 42  wherein the plant nectar is selected from the genus:  Leptospermum, Kunzea, Weinmannia, Knightia, Metrosideros, Fagus, Trifolium,  Myrtaceae, and combinations thereof. 
     
     
         44 . The method as claimed in  claim 35  wherein the type II AG compound is AGP. 
     
     
         45 . A method of determining the immunostimulatory properties of a honey or honey analogue by testing the content of type II AG in the honey or honey analogue and wherein:
 a. if the amount of type II AG in the honey or honey analogue is greater than approximately 5 μg/g, a measurable immunostimulatory response is predicted; and,   b. if the amount of type II AG in the honey or honey analogue is less than approximately 5 μg/g, a measurable immunostimulatory response is not predicted.   
     
     
         46 . The method as claimed in  claim 45  wherein the amount in step a. is greater than 10 μg/g. 
     
     
         47 . The method as claimed in  claim 45  wherein the amount in step b. is less than 1 μg/g. 
     
     
         48 . The method as claimed in  claim 45  wherein an immunostimulatory topical formulation is produced from the honey or honey analogue if the measured type II AG is greater than 5 μg/g. 
     
     
         49 . The method as claimed in  claim 45  wherein a non-immune stimulatory topical formulation is produced from the honey or honey analogue if the measured type II AG is less than 5 μg/g. 
     
     
         50 . The method as claimed in  claim 35  wherein testing is completed utilising reactions that bind type II AG compounds with specific antibodies. 
     
     
         51 . The method as claimed in  claim 35  wherein testing is completed using ELISA analysis. 
     
     
         52 . A method of producing an isolate as claimed in  claim 1  by obtaining a honey and then processing the honey by steps selected from: filtration, ultra-filtration, reverse osmosis, solvent extraction, precipitation, or combinations thereof and collecting a high molecular weight isolate from the processing step. 
     
     
         53 . The method as claimed in  claim 52  wherein the honey is processed through a 10 kDa filter and the high molecular isolate post filtration is collected.