Acellular pro-inflammatory compositions, process for making same and methods of using same

A process for making an acellular pro-inflammatory preparation and decreasing a ratio of endogenous regulatory T (Treg) cell concentration to endogenous pro-inflammatory T cell concentration in a subject, said process comprising:

• (i) contacting a first leukocyte with a second leukocyte in vitro under conditions to allow a pro-inflammatory allo-recognition to provide a conditioned preparation, wherein the first leukocyte is allogeneic to the second leukocyte;
• (ii) separating whole first leukocyte cells and whole second leukocyte cells from the conditioned preparation to obtain an acellular conditioned preparation;
• (iii) selecting miRNA components having an individual average molecular weight of less than about 10 kDa from the acellular conditioned preparation under conditions to inhibit RNA degradation and to maintain a relative abundance of each of the miRNA components so as to obtain a composition enriched in acellular pro-inflammatory components;
• (iv) formulating the composition of step (iii), under conditions to inhibit RNA degradation and to maintain a relative abundance of each of the miRNA components, in the acellular pro-inflammatory preparation for administration to the subject, and
• (v) administering a therapeutically effective amount of the acellular pro-inflammatory preparation to the subject,
• wherein the acellular pro-inflammatory preparation, when administered to the subject, decreases the ratio of endogenous regulatory T (Treg) cell concentration to endogenous pro-inflammatory T cell concentration in the subject.