Bernier J, De Guise S, Martineau D, Béland P, Beaudet M, Fournier M
INRS-Institut Armand Frappier, Centre Santé humaine, Pointe-Claire, Quebec, Canada.
Dev Comp Immunol. 2000 Sep-Oct;24(6-7):653-62. doi: 10.1016/s0145-305x(99)00086-5.
In an effort to gain knowledge on immune functions in beluga whales, Delphinapterus leucas, we have used two physical methods for the purification of T lymphocytes of spleen cells. Isolation by sheep red blood cells (SRBC) rosetting and by adherence on nylon wool columns were tested. SRBC-rosetting gave unreliable results in obtaining purified T cells. Therefore, the purification of T cells was done using nylon wool columns. Less than 3% of the IgM(+) B cells remained in effluent populations. In the later population, 45% gave positive staining with mouse anti-human CD4 allowing us to verify functionality of the cells. The study of calcium mobilization and tyrosine kinase activation, mediated by CD4 cross-linking permitted verification of the functionality of cells. We also showed that upon activation with mitogens, beluga T cells upregulate the density of MHC class II molecules on their surfaces. CD4 cross-linking with a specific antibody inhibited the proliferation response. Overall, the activation of beluga whales lymphocytes did not differ markedly from what is known in other species. This study can help in the groundwork for functional investigation of the beluga whale's immune system.
为了了解白鲸（Delphinapterus leucas）的免疫功能，我们采用了两种物理方法来纯化脾细胞中的T淋巴细胞。对通过绵羊红细胞（SRBC）花环试验分离以及通过尼龙毛柱黏附进行了测试。SRBC花环试验在获得纯化T细胞方面给出了不可靠的结果。因此，使用尼龙毛柱进行T细胞的纯化。流出群体中残留的IgM(+) B细胞不到3%。在后续群体中，45%的细胞用小鼠抗人CD4呈阳性染色，这使我们能够验证细胞的功能。对由CD4交联介导的钙动员和酪氨酸激酶激活的研究证实了细胞的功能。我们还表明，在用有丝分裂原激活后，白鲸T细胞会上调其表面MHC II类分子的密度。用特异性抗体进行CD4交联会抑制增殖反应。总体而言，白鲸淋巴细胞的激活与其他物种已知的情况没有明显差异。这项研究有助于为白鲸免疫系统的功能研究奠定基础。
