Study On The General Pharmacology And Safety Evaluation Of BCG-CpG-DNA

OBJECTIVE: To study the pharmacology of BCG-CpG-DNA and evaluate its safety, for providing evidence of preclinical safety evaluation of BCG-CpG-DNA. METHODS: To reveal the pharmacology effect of BCG-CpG-DNA, we mainly investigated the proliferation effect of PMBC that can be stimulated by BCG-CpG-DNA and its influence of IL-6 and IL-12 originated from human PMBC to evaluate the immune stimulation; The change of total serum IgE in sensitization mice treated by BCG-CpG-DNA was observed to evaluate the immune regulation; the effects on expression of mice TLRs mRNA was determined to detect the immune targeted function.To reveal the safety evaluation of BCG-CpG-DNA ,we mainly studied of the change of total serum IgE in the mice which was treated with BCG-CpG-DNA more than twice, to evaluate the immune sensitization; Observation of the change of local injection skin and muscle, hematological, blood biochemical, major organ coefficients, and check for major immune organ on pathology after repeated administration with BCG-CpG-DNA to illustrate the Long-term toxicity in rabbit.RESULTS:1.Pharmacology research(1). Immune stimulation: BCG-CpG-DNA can stimulate the PMBC, and initialize its proliferation; BCG-CpG-DNA also can influence the IL-6 originated from PMBC, and stimulate PMBC to produce more IL-12 which play an important role in immune response as an cytokines, so we can obtain its immune stimulation.(2). Immune regulation: BCG-CpG-DNA treatment group has a less total serum IgE than control, and the difference was significant. That indicated BCG-CpG-DNA has activity on immune regulation.(3). Immune targeted: The effect of BCG-CpG-DNA was mainly regulated the expressions of mice TLR-7 and TLR-9, and produced more mRNA PCR product, so the immune targeted of it was identified.2. Safety evaluation(1). Sensitization in mice: BCG-CpG-DNA has no sensitization and would not qualify as an allergen to cause reaction.(2). Long-term toxicity in rabbit: There is no visual change in local muscle and skin; the immune cell counts in the blood were increased and no abnormalities in the other hematological indexes blood biochemical and major organ coefficients. The result of the pathology was that BCG-CpG-DNA caused inflammation in the local muscle and did not effect those main immune organ.CONCLUSION: BCG-CpG-DNA has pharmacology activity such as immune stimulation, regulation and targeted. Under the conditions of this experiment, we found that BCG-CpG-DNA has no sensitization in mice and no Long-term toxicity in rabbit. As stated previously, the BCG-CpG-DNA is an ideal tracer with wide application in vaccine as adjuvant.