Subba Raju B V, Singh R, Sreenivas G, Singh S, Salotra P
Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi-110 029, India.
Parasitology. 2008 Jan;135(Pt 1):23-32. doi: 10.1017/S0031182007003484. Epub 2007 Aug 28.
Post-kala-azar dermal leishmaniasis (PKDL) is an unusual dermatosis that develops as a sequel in 5-15% of cured cases of kala-azar (KA) after months or years of treatment in India. Molecular differences are reported to exist between the KA and PKDL isolates which may underlie the diversity in clinical manifestations of the disease. Here, arbitrary primed-PCR (AP-PCR) has been used for genetic fingerprinting of parasite isolates from dermal lesions of PKDL patients (n=14) and compared with bone-marrow derived parasites from KA patients (n=3). All isolates showed an identical AP-PCR pattern with 4 arbitrary primers. Further, AP-PCR was exploited to identify the stage regulated genes of the parasite. Six polymorphic fragments were identified in PKDL in comparison with KA isolates, and were subjected to Northern blot analysis. Five polymorphic fragments represented transcribed sequences; 4 out of 5 drew differential expression in pro- and amastigote stages, although the expression was comparable between PKDL and KA isolates. The study led to the identification of genes, which exhibit stage-regulated expression in Leishmania donovani derived from PKDL or KA patients, including a putative phosphodiesterase, DEAD box RNA helicase, iron superoxide dismutase b (fesodb) and a hypothetical protein. Demonstration of transcripts of DEAD box RNA helicase in PKDL and KA diseased tissues implicates its role in disease pathogenesis.
黑热病后皮肤利什曼病(PKDL)是一种罕见的皮肤病,在印度,5%至15%的治愈黑热病(KA)患者在治疗数月或数年之后会继发该病。据报道,KA和PKDL分离株之间存在分子差异,这可能是该疾病临床表现多样性的基础。在此,任意引物PCR(AP-PCR)已用于对PKDL患者(n = 14)皮肤病变中的寄生虫分离株进行基因指纹分析,并与KA患者(n = 3)骨髓来源的寄生虫进行比较。所有分离株用4种任意引物均显示出相同的AP-PCR模式。此外,利用AP-PCR鉴定寄生虫的阶段调控基因。与KA分离株相比,在PKDL中鉴定出6个多态性片段,并对其进行Northern印迹分析。5个多态性片段代表转录序列;其中4个在前鞭毛体和无鞭毛体阶段表达存在差异,尽管PKDL和KA分离株之间的表达相当。该研究鉴定出了在源自PKDL或KA患者的杜氏利什曼原虫中表现出阶段调控表达的基因,包括一个假定的磷酸二酯酶、DEAD盒RNA解旋酶、铁超氧化物歧化酶b(fesodb)和一个假定蛋白。在PKDL和KA患病组织中证实DEAD盒RNA解旋酶的转录本表明其在疾病发病机制中的作用。
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