Isfahan University of Medical Sciences Journal of Isfahan Medical School 1027-7595 30 221 2013 02 19 Comparison of Protein Profile of Killed and Autoclaved Leishmania Major Using Polyacrylamide Gel Electrophoresis Comparison of Protein Profile of Killed and Autoclaved Leishmania Major Using Polyacrylamide Gel Electrophoresis 2460 2466 13991 FA Reza Arjmand PhD Student, Department of Parasitology and Mycology, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran Simindokht Soleimanifard PhD Student, Department of Parasitology and Mycology, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran Sedigheh Saberi PhD Student, Department of Parasitology and Mycology, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran Sepideh Tolouei Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran Ali Khamesipour Professor, Center for Research and Training in Skin Disease and Leprosy, Tehran University of Medical Sciences, Tehran, Iran Seyed Hossein Hejazi Associate Professor, Department of Parasitology and Mycology, School of Medicine AND Skin Disease and Leishmaniasis Research Center, Isfahan University of Medical Sciences, Isfahan, Iran Journal Article 2013 01 09 Background: Leishmania is a genus of trypanosomatid protozoa which causes a wide spectrum of diseases ranging from a self-healing cutaneous lesion to a vital visceral form of the disease. Although various experimental Leishmania vaccines have been prepared in different parts of the world, only a few of first generation vaccines have reached to human trials. Killed Leishmania major (KLM) and autoclaved Leishmania major (ALM) have been tested in human volunteers in Iran and other countries. We evaluated protein contents of KLM and ALM and fresh Leishmania major using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Methods: Leishmania major (MRHO/IR/75/ER) was isolated from infected BALB/c mice and cultured in Novy-MacNeal-Nicolle medium. It was then subcultured in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with fetal calf serum 20%. ALM and KLM were prepared by Razi Vaccine and Serum Institute of Iran. Electrophoresis of the two antigens and freshly cultured Leishmania major promastigotes was performed and their protein contents were compared using SDS-PAGE. Findings: The results of electrophoresis showed numerous bands from more than 100 kilodalton (kD) to less than 10 kD. KLM bands were found to be similar to freshly cultured intact Leishmania major (except for 57 and 24 kD bands). ALM contained two very close bands (71 kD) which were not seen in KLM or fresh Leishmania major. Conclusion: Identification and purification of protective immunogens in crude antigens and detection of their stability are essential in production of an effective vaccine. Keywords: Leishmania, Killed Leishmania major, autoclaved Leishmania major, Polyacrylamide gel electrophoresis Background: Leishmania is a genus of trypanosomatid protozoa which causes a wide spectrum of diseases ranging from a self-healing cutaneous lesion to a vital visceral form of the disease. Although various experimental Leishmania vaccines have been prepared in different parts of the world, only a few of first generation vaccines have reached to human trials. Killed Leishmania major (KLM) and autoclaved Leishmania major (ALM) have been tested in human volunteers in Iran and other countries. We evaluated protein contents of KLM and ALM and fresh Leishmania major using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Methods: Leishmania major (MRHO/IR/75/ER) was isolated from infected BALB/c mice and cultured in Novy-MacNeal-Nicolle medium. It was then subcultured in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with fetal calf serum 20%. ALM and KLM were prepared by Razi Vaccine and Serum Institute of Iran. Electrophoresis of the two antigens and freshly cultured Leishmania major promastigotes was performed and their protein contents were compared using SDS-PAGE. Findings: The results of electrophoresis showed numerous bands from more than 100 kilodalton (kD) to less than 10 kD. KLM bands were found to be similar to freshly cultured intact Leishmania major (except for 57 and 24 kD bands). ALM contained two very close bands (71 kD) which were not seen in KLM or fresh Leishmania major. Conclusion: Identification and purification of protective immunogens in crude antigens and detection of their stability are essential in production of an effective vaccine. Keywords: Leishmania, Killed Leishmania major, autoclaved Leishmania major, Polyacrylamide gel electrophoresis https://jims.mui.ac.ir/article_13991_7207cc49aa823ca90c015aacf4e9c161.pdf