www.IndianJournals.com Members Copy, Not for Commercial Sale Downloaded From IP - 124.247.209.58 on dated 30-Jan-2011 36 J. Immunol Immunopathol. 2009. 11(1): 36-41 Society for Immunology and Immunopathology IndianJournals.com Development of IL-2, IL-18 and IFN- γ gene based genetic adjuvants for use along with CSFV DNA vaccine in pigs Barkha Ratta 1 , Binita Nautiyal 2 , P V Ravindra 3 , Uttara Chaturvedi, Sudesh Palia, P K Subudhi, Rajiv Kumar, Sangeeta Tiwari 4 and A K Tiwari* *aktiwari63@yahoo.com, Molecular Biology Laboratory, Division of Animal Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122 (Uttar Pradesh), INDIA. 1 Immunochemistry Laboratory, Department of Biochemistry, Indian Veterinary Research Institute, Izatnagar-243122 (Uttar Pradesh), INDIA; 2, 4 Reader, Department of Animal Science, M J P Rohilkhand University, Bareilly, 243006 (Uttar Pradesh); 3 Department of Cardio-thoracic Surgery, Biological Science Division, University of Chicago, Chicago, Illinois-60637, U S A ABSTRACT Ratta B, Nautiyal B, Ravindra P V, Chaturvedi U, Palia S, Subudhi P K, Rajiv Kumar, Tiwari S and Tiwari A K. (2009). Development of IL-2, IL-18 and IFN-γ gene based genetic adjuvants for use along with CSFV DNA vaccine in pigs. J. Immunol Immunopathol. 11(1): 36-41. The cytokine genes when administered along with DNA vaccine may work as immunological adjuvant and enhance immune responses. Hence, the study was undertaken to clone and characterize the pig IFN-γ, IL-2 and IL-18 cytokine genes. The pig IFN-γ, IL-2 and IL-18 gene were amplified using specific primers by RT-PCR and cloned into pTZ57R/T cloning vector. The clone was confirmed by digestion analysis using KpnI and ApaI restriction enzymes. Further, the gene fragment was sub-cloned into eukaryotic expression vector pcDNA 3.1(+) and confirmed by restriction digestion as described above. These recombinant plasmids can be used as genetic adjuvants to DNA vaccine against CSFV. Keywords: RT-PCR, adjuvants, cytokines Abbreviations used: CSFV (Classical swine fever virus), CSF (Classical swine fever), IFN-γ (Interferon gamma), IL- 2 (Interleukin-2), IL-18 (Interleukin-18), RT-PCR (Reverse transcriptase-Polymerase chain reaction). INTRODUCTION Cytokines are prominent regulators of the immune response following infection or vaccination. Cytokines are proteins produced by cells of the immune system and act in autocrine, paracrine, or endocrine manners to control the type and extent of an immune response in various systemic responses. The effectiveness of cytokines in various vaccine adjuvants is well established. The use of cytokines as immunological adjuvants can enhance various types of immune responses when administered during the development of an immune response to a particular antigen (Dong et al., 1995). The adjuvant effects of cytokines were also observed in the case of genetic immunization. Plasmid vectors containing a CpG dinucleotide motif elicited much stronger humoral and cellular immune responses to the encoded antigens than did vectors which did not contain this sequence (Sato et al., 1996). The adjuvant activity of the CpG motif was closely related to its ability to elicit a rapid cytokine release from the transfected cells. Cytokines could influence the efficacy of DNA vaccination, when IL-2, IL-6, IL-7, or IL-12 was added following administration of DNA encoding a tumor- associated antigen, the number of established metastases was significantly reduced compared with that in mice treated with DNA only (Dong et al., 1995). Many approaches have been taken to improve the efficiency of DNA vaccine, such as incorporation of genes for cytokines and co-stimulatory molecules (Berzofsky et al., 2001).These cytokines and chemokines paves the path of leucocytes to the site of infection or injury, activating their antimicrobial functions and regulating the induction of the adaptive response to the pathogen. For some species, it has been previously described that co-administration of cytokine genes with DNA-vaccines increases and or modifies the immune response towards a Th1- or Th2-type reaction