ABSTRACT
Trichosanthin (TCS) is a potent allergen to mice. According to our previous experiments, it could bring out the IgE response to ovabumin (OVA) if TCS was given one day before OVA immunization, while OVA alone could not induce IgE to it. In this work, the kinetics of interleukin 4(IL-4) and interferon gamma(IFN-gamma) gene expression in the mesenteric lymph node (MLN) of TCS-immunized mice was investigated using a semi-quantitative RT-PCR method. It indicated that TCS induced significant IL-4 gene expression and the peaks of IL4 gene expression were on day one after TCS immunization in both primary and secondary response. In contrast, the IFN-gamma gene expression was suppressed. Furthermore, the IL-4 gene expression in the secondary response was lower than that in the primary response. Thus the presence of IgE memory B cells were studied. Results showed that the amount of mature IgE mRNA arose significantly and rapidly one day after TCS restimulation, while in the MLN of the mice primed 30 days before and without boost, it was almost as the same amount of the unimmunized control. These findings suggest the existence of the IgE memory B cells in the mice after the primary TCS immunization.
Subject(s)
Gene Expression , Interferon-gamma/genetics , Interleukin-4/genetics , Linolenic Acids/immunology , Animals , B-Lymphocytes/immunology , Female , Immunization , Immunoglobulin E/genetics , Kinetics , Mice , Mice, Inbred C57BLABSTRACT
The covalently linked antigenic complex, bovine serum albumin-alpha-linolenic acid, was used to immunize Balb/c mice against the hapten. Hybridization between splenocytes and the myeloma cell line, P 3 X63 Ag 8,651, resulted in stable clones synthesizing monoclonal antibodies (Mab) that were subsequently purified and characterized. Four Mab (A, B, C, D) were retained and their specificities studied by ELISA. Antibody D only recognized 18-carbon fatty acids having a cis,cis,-cis-1,4,7 unsaturated system in the omega-3 position: it was specific for alpha-linolenic acid. B recognized all fatty acids containing the structure cis,cis,cis-1,4,7-octatriene. A and C recognized polyunsaturated fatty acids with a degree of unsaturation superior to two double bonds.
