de Swart R L, Kluten R M, Huizing C J, Vedder L J, Reijnders P J, Visser I K, UytdeHaag F G, Osterhaus A D
Seal Rehabilitation and Research Centre, Pieterburen, Netherlands.
Vet Immunol Immunopathol. 1993 Aug;37(3-4):217-30. doi: 10.1016/0165-2427(93)90195-a.
In vitro assays were developed for studies concerning the functioning of the immune system of the harbour seal (Phoca vitulina). Proliferative responses of peripheral blood mononuclear cells (PBMC) were measured after stimulation with different concentrations of the mitogens concanavalin A (Con A), pokeweed mitogen (PWM), phytohaemagglutinin (PHA) or lipopolysaccharide from Salmonella typhimurium (LPS). Con A and PWM induced strong proliferative responses, while PHA and LPS induced comparatively low proliferative responses. Responses of mitogen stimulated PBMC to recombinant human interleukin-2 (rhIL-2) and in vitro immunoglobulin production by mitogen stimulated PBMC were measured to discriminate between stimulation of T cells and B cells. It was found that Con A and PHA stimulate phocine T cells, PWM stimulates both T cells and B cells and LPS predominantly stimulates phocine B cells. Antigen-specific immune responses were measured after immunization of seals with an inactivated rabies vaccine and/or with tetanus toxoid. Antigen-specific proliferation of PBMC and the presence of antigen-specific antibody forming cells were demonstrated for both antigens in the PBMC of immunized animals. The responses measured in vitro correlated well with the development of specific serum antibody titers to these antigens.
已开发出体外试验用于研究斑海豹(Phoca vitulina)免疫系统的功能。在用不同浓度的促有丝分裂原刀豆球蛋白A(Con A)、商陆有丝分裂原(PWM)、植物血凝素(PHA)或鼠伤寒沙门氏菌脂多糖(LPS)刺激后,测量外周血单核细胞(PBMC)的增殖反应。Con A和PWM诱导强烈的增殖反应,而PHA和LPS诱导的增殖反应相对较低。测量促有丝分裂原刺激的PBMC对重组人白细胞介素-2(rhIL-2)的反应以及促有丝分裂原刺激的PBMC的体外免疫球蛋白产生,以区分T细胞和B细胞的刺激情况。结果发现,Con A和PHA刺激海豹T细胞,PWM刺激T细胞和B细胞,LPS主要刺激海豹B细胞。在用灭活狂犬病疫苗和/或破伤风类毒素免疫海豹后,测量抗原特异性免疫反应。在免疫动物的PBMC中,针对这两种抗原均证明了PBMC的抗原特异性增殖以及抗原特异性抗体形成细胞的存在。体外测量的反应与针对这些抗原的特异性血清抗体滴度的发展密切相关。