Castilla J J, Sanchez-Moreno M, Mesa C, Osuna A
Instituto de Biotecnología, Facultad de Ciencias, Universidad de Granada, Spain.
Mol Cell Biochem. 1995 Jan 26;142(2):89-97. doi: 10.1007/BF00928929.
When Leishmania donovani promastigote forms, were cultured in TC-199 medium at 28 degrees C and subsequently incubated at 38 degrees C, they turned into aflagellate (amastigote-like) forms. A return of the incubation-culture temperature to 28 degrees C these amastigote-like forms to revert to promastigotes. The amastigotes obtained by heat-shock, were viable and retained antigenic capacity being recognized by the sera of naturally infected patients. These forms, remained also capable of multiplying inside the J-774A.1 macrophages. When the amastigote-like forms are kept in culture at 38 degrees C retained their rounded appearance and their biological characteristics for more than 3 months subculturing every 6 days. These amastigote-like forms, when used for subcultures at 28 degrees C, transformed into promastigotes capable of multiplying as flagellate forms. The amastigote-like forms obtained in vitro can be used in biochemical studies related to chemotherapy and immunology studies, as part of an effort to combat this parasite. The end-products of of glycolysis were studied in both the amastigote-like and promastigote forms of L. donovani, by proton magnetic resonance analysis of the culture media. Alanine, succinate, and acetate, were predominant, and to a lesser extent pyruvate, glycine and D-lactate. Our results suggest that both forms of Leishmania use different biochemical strategies to obtain their energy.
当杜氏利什曼原虫前鞭毛体在28℃的TC - 199培养基中培养，随后在38℃孵育时，它们会转变为无鞭毛体（类无鞭毛体）形式。将孵育培养温度恢复到28℃时，这些类无鞭毛体形式又会恢复为前鞭毛体。通过热休克获得的无鞭毛体是有活力的，并且保留了抗原能力，能被自然感染患者的血清识别。这些形式在J - 774A.1巨噬细胞内也能够增殖。当类无鞭毛体形式在38℃培养时，每6天传代一次，它们能保持圆形外观和生物学特性超过3个月。这些类无鞭毛体形式，当在28℃用于传代培养时，会转变为能够以前鞭毛体形式增殖的前鞭毛体。体外获得的类无鞭毛体形式可用于与化疗相关的生化研究和免疫学研究，作为对抗这种寄生虫努力的一部分。通过对培养基进行质子磁共振分析，研究了杜氏利什曼原虫类无鞭毛体和前鞭毛体形式的糖酵解终产物。丙氨酸、琥珀酸和乙酸占主导，丙酮酸、甘氨酸和D - 乳酸含量较少。我们的结果表明，利什曼原虫的两种形式使用不同的生化策略来获取能量。
