All issues > Volume 35(7); 1992

Original Article

J Korean Pediatr Soc. 1992;35(7):902-908. Published online July 15, 1992.

In Vitro Immunization-IV: Antibody Response of Mouse Splenocytes Grown in a Mixture of Conditioned Media, Thymocytes, and Bone Marrow Cells

Dong Soo DS Kim¹, Geun Woong GW Noh¹

¹Department of Pediatrics, Yonsei University College of Medicine, Seoul, Korea

Abstract

We have previously reported several methods of in vitro immunization using different conditioned media, and the highest antibody titer was observed when a mixture of three conditioned media[PHA-stimulated adherent splenocytes conditioned medium (ASM-P), ASM-p-stimulated non-adherent splenocyte conditioned medium (NASM-A), ASM-P-stimulated thymocyte conditioned medium (TM-A)] was used. In this experiment, thymocytes and/or bone marrow cells was added instead of above conditioned media which were used in mKN in-vitro immunization methods. The added cells were mixed for free cell-to-cell contact and prohibited cell-to-cell contact. When cell-to-cell contact was prohibited, significant antibody titer could not be obtained, but when cells were mixed freely, antibody response was similar to that of mKN method using conditioned media. The same antigen was rechallenged to observe the secondary immune response which can be observed in the in-vivo system, but failed in all cases. Conclusively, cell-to-cell contact played an immportant role in immune responses and our conditioned media which were optimally induced according to immunologic circuit could replace cell-to-cell contact among splenocytes, thymocytes, and bone marrow cells.

Keywords :In-vitro immunization