Zamora D O, Rosenbaum J T, Smith J R
Department of Ophthalmology, Casey Eye Institute, Oregon Health and Science University, Portland, OR 97239, USA.
Br J Ophthalmol. 2008 Jun;92(6):852-5. doi: 10.1136/bjo.2007.133314.
Toxoplasma gondii infection is a leading cause of posterior uveitis. Human retinal endothelial cells (HREC) are more susceptible to infection with T gondii tachyzoites than other subpopulations of endothelial cells. It is hypothesised that this phenomenon reflects differences in invasion efficiency.
YFP-expressing RH strain T gondii tachyzoites were added to confluent HREC or human dermal endothelial cells (HDEC) (MOI = 50:1). Tachyzoite invasion after 1 h was determined by microplate reading of fluorescence intensity or parasite counts obtained using image analysis software. Selected cultures were incubated for three subsequent days, at which time fluorescence intensity indicated intracellular tachyzoite proliferation.
HREC-tachyzoite cultures were more fluorescent than HDEC-tachyzoite cultures after 1 h (p = 0.020, paired t test, 3 experiments). Parasite counts also indicated that more tachyzoites invaded HREC than HDEC (p = 0.042, paired t test, 5 experiments). At 3 days, fluorescence intensity remained higher in HREC-tachyzoite cultures (p< or =0.002, t test, 3 experiments).
In culture, T gondii tachyzoites invade HREC with greater efficiency than they invade HDEC. This observation suggests that the relative susceptibility of HREC to infection may reflect a high efficiency of tachyzoite invasion which may be relevant to understanding how T gondii infects human retina.
弓形虫感染是后葡萄膜炎的主要病因。人视网膜内皮细胞(HREC)比其他内皮细胞亚群更容易受到弓形虫速殖子的感染。据推测,这种现象反映了侵袭效率的差异。
将表达黄色荧光蛋白(YFP)的RH株弓形虫速殖子加入汇合的HREC或人皮肤内皮细胞(HDEC)中(感染复数=50:1)。1小时后速殖子的侵袭情况通过酶标仪读取荧光强度或使用图像分析软件获得的寄生虫计数来确定。选择的培养物随后连续培养三天,此时荧光强度表明细胞内速殖子的增殖情况。
1小时后,HREC-速殖子培养物比HDEC-速殖子培养物荧光更强(p = 0.020,配对t检验,3次实验)。寄生虫计数也表明侵入HREC的速殖子比侵入HDEC的更多(p = 0.042,配对t检验,5次实验)。在第3天,HREC-速殖子培养物中的荧光强度仍然更高(p≤0.002,t检验,3次实验)。
在培养中,弓形虫速殖子侵入HREC的效率高于侵入HDEC。这一观察结果表明,HREC对感染的相对易感性可能反映了速殖子的高效侵袭,这可能与理解弓形虫如何感染人类视网膜有关。