5-Amino-3-methyl-4-isoxazolecarboxylic acid hydrazide was used for the synthesis of a series of semicarbazides and thiosemicarbazides with immunotropic activity. In in vitro studies the compound showed no cytotoxic activity in murine cell lines and stimulated proliferation of murine lymphocytes isolated from peripheral lymphatic organs and murine peritoneal macrophages stimulated with mitogens [1]. The aim of this study was to determine the influence of 5-amino-3-methyl-4-isoxazolecarboxylic acid hydrazide on primary humoral immune response in sheep red blood cells (SRBC)-immunized mice. All experiments were carried out on Balb/c mice, 7-8 weeks of age, each weighing 18-20 g. The animals were immunized intraperitoneally (i.p.) with 0.2 ml of 10% SRBC suspension. The 5-amino-3-methyl-4-isoxazolecarboxylic acid hydrazide was administered  intraperitoneally (i.p.) four times at 24 h intervals at three different doses of 10, 1 and 0.1 mg/kg, before or after antigen stimulation. The measurements included number of plaque forming cells (PFC) in spleen and anti-SRBC haemagglutinin titre in the serum. The splenocytes producing hemolytic anti-SRBC antibodies (PFC) was determined by a local hemolysis technique in agar gel as described by Mishell and Dutton [2]. The total and 2-mercaptoethanol-(2-ME) resistant serum agglutination titres were defined by active haemagglutination test carried out on microplates [3]. The 5-amino-3-methyl-4-isoxazolecarboxylic acid hydrazide increased primary humoral immune response in SRBC-immunized mice, which resulted in an elevated number of splenocytes producing hemolytic anti-SRBC antibodies (PFC) and increased total and 2-mercaptoethanol-resistant serum hemagglutinins titers. Those modulating effects were dependent on the dose applied. Hydrazide administered before as well as after stimulation with antigen increased the number of PFC only at the lower doses- 1 and 0.1 mg/kg. On the other hand, four injections of the agent before immunization elevated the 2-ME resistant anti-SRBC antibody titers at all used doses. Total anti-SRBC haemagglutinin titre was increased only by the dose of 10 mg/kg of the hydrazide. Administration of the compound after priming changed only 2-ME resistant anti-SRBC antibody titre which was increased at the dose of 1 and 0.1 mg/kg.

References:

1. Drynda A., M. Mączyński, Ryng S., Obmińska-Mrukowicz B. In vitro immunomodulatory effects of 5-amino-3-methyl-4-isoxazolecarboxylic acid hydrazide on the cellular immune response. Immunopharmacol Immunotoxicol. 2014;36(2): 1. 2. Mishell R.J., Dutton R.W. Immunization of dissociated spleen cell cul50-7tures from normal mice. J Exp Med 1967; 126: 423-442. 3. Hudson L, Hay FC Practical Immunology. Blackwell Scientific Publications, Oxford, London, Edinburgh, Boston, Melbourne 1980;  447

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