BCG is an immune modulatory that inducing humoral and cellular immune responses during a number of infections including cutaneous leishmaniasis. Killed Leishmania Vaccine (KLV) has been applied for its immunogenicity in hosts. This study was carried out between 2010 and 2011 at the Department of Parasitology, Pasteur Institute of Iran. In the present study, alterations of Nitric Oxide (NO) levels, total content of essential trace elements (Cu, Zn) and liver enzymes (SGOT, SGPT) were investigated in Balb/c mice infected with Leishmania major. Results showed that BCG induced NO in liver; Zn increased in KLV/BCG and KLV test groups and Cu decreased in KLV test group, but increased in BCG test group. Moreover to decreasing SGOT level in KLV/BCG test group, SGPT decreased in BCG test group. Our findings showed that serum trace element concentrations were altered in CL infection, probably in accordance with host defense mechanisms. It is indicated that application of KLV/BCG as a combined vaccine/adjuvant could be indicated as a potent inducer of immuno-biochemical characters during infection with Leishmaniasis. In conclusion, some dependency is observed between KLV/BCG and NO levels, Cu, Zn concentrations and SGOT, SGPT production. The mechanism of KLV/BCG action may be associated more on BCG rather than KLV.