Fernandes A P, Herrera E C, Mayrink W, Gazzinelli R T, Liu W Y, de Costa C A, Tavares C A, Melo M N, Michalick M S, Gentz R, Nascimento E
Departamento de Parasitologia, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Rev Inst Med Trop Sao Paulo. 1997 Mar-Apr;39(2):71-8. doi: 10.1590/s0036-46651997000200002.
In the search for Leishmania recombinant antigens that can be used as a vaccine against American Cutaneous Leishmaniasis, we identified a Leishmania (Leishmania) amazonensis recombinant protein of 33 kD (Larp33) which is recognized by antibodies and peripheral blood leukocytes (PBL) from subjects vaccinated with Leishvacin, Larp33 was expressed in Escherichia coli after cloning of a 2.2 kb Sau3 digested genomic fragment of L. (L.) amazonensis into the pDS56-6 His vector. Immunoblotting analysis indicated that Larp33 corresponds to an approximately 40-kD native protein expressed in promastigotes of L. (L.) amazonensis and L. (Viannia) braziliensis. Northern blots of total RNA also demonstrated that the gene coding for this protein is expressed in promastigotes of the major lineages of Leishmania causing American Cutaneous Leishmaniasis. Larp33 induced partial protection in susceptible mouse strains (BALB/c and C57BL/10) against L. (L.) amazonensis after vaccination using Bacille Calmette-Guerin (BCG) as adjuvant. In vitro stimulation of splenocytes from BALB/c protected mice with Larp33 elicited the secretion of IL-2 and IFN-gamma, suggesting that a Th1 cell-mediated protective response is associated with the resistance observed in these mice. As revealed by its immunogenic and antigenic properties, this novel recombinant antigen is a suitable candidate to compose a vaccine against cutaneous leishmaniasis.
在寻找可用于预防美洲皮肤利什曼病疫苗的利什曼原虫重组抗原的过程中，我们鉴定出一种33 kD的亚马逊利什曼原虫（Leishmania (Leishmania) amazonensis）重组蛋白（Larp33），接种Leishvacin疫苗的受试者的抗体和外周血白细胞（PBL）可识别该蛋白。将亚马逊利什曼原虫2.2 kb Sau3酶切基因组片段克隆到pDS56-6 His载体后，Larp33在大肠杆菌中表达。免疫印迹分析表明，Larp33对应于亚马逊利什曼原虫和巴西利什曼原虫（Leishmania (Viannia) braziliensis）前鞭毛体中表达的一种约40-kD的天然蛋白。总RNA的Northern印迹也表明，编码该蛋白的基因在引起美洲皮肤利什曼病的主要利什曼原虫谱系的前鞭毛体中表达。使用卡介苗（BCG）作为佐剂接种后，Larp33在易感小鼠品系（BALB/c和C57BL/10）中诱导了对亚马逊利什曼原虫的部分保护作用。用Larp33体外刺激BALB/c保护小鼠的脾细胞可诱导IL-2和IFN-γ的分泌，这表明Th1细胞介导的保护性反应与这些小鼠中观察到的抗性有关。从其免疫原性和抗原性特性来看，这种新型重组抗原是组成抗皮肤利什曼病疫苗的合适候选物。
