MURINE T-CELL RESPONSE TO NATIVE AND RECOMBINANT PROTEIN ANTIGENS OF RICKETTSIA-TSUTSUGAMUSHI
MURINE T-CELL RESPONSE TO NATIVE AND RECOMBINANT PROTEIN ANTIGENS OF RICKETTSIA-TSUTSUGAMUSHI
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DOI：
10.1128/iai.61.5.1674-1681.1993
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发表时间：
1993-05-01
影响因子：
3.1
通讯作者：
OAKS, EV
中科院分区：
文献类型：
--
作者：
HICKMAN, CJ;STOVER, CK;OAKS, EV
使用具有TH 1特征的多克隆T细胞系来评估小鼠对天然和重组恙虫病立克次体抗原的细胞免疫应答。该T细胞系的增殖被观察到响应于许多天然抗原组分，这表明鼠T辅助细胞应答针对多种恙虫病抗原，没有明显的抗原免疫优势。重组R.恙虫病抗原使得鉴定刺激多克隆T细胞系的47-kDa恙虫病抗原（Sta 47）成为可能。编码56-、58-和110-kDa抗原（分别为Sta 56、Sta 58和Sta 110）的重组克隆不能诱导该T细胞系的增殖。对编码Sta 47蛋白的克隆立克次体插入片段的DNA序列分析显示存在4个开放阅读框，可能编码47、30、18和13 kDa的蛋白。对十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离和洗脱的重组HB 101（pRTS47B4.3）裂解物级分的分析表明，含有47-kDa蛋白质的级分以及含有小于18 kDa蛋白质的级分是刺激性的。从Sta 47抗原序列衍生的选择的合成两亲性肽鉴定了一种20个氨基酸的肽，其在T细胞增殖中比类似长度的对照疟疾肽增加10倍。47-kDa抗原的识别与TH 1特性的T细胞系暗示这种蛋白质作为一个潜在的重要性，在保护研究和未来的疫苗开发。
A polyclonal T-cell line with TH1 characteristics was used to assess the murine cellular immune response to native and recombinant Rickettsia tsutsugamushi antigens. Proliferation of this T-cell line was observed in response to numerous native antigen fractions, which indicates that the murine T-helper-cell response is directed at multiple scrub typhus antigens with no apparent antigenic immunodominance. Subsequent analysis of recombinant R. tsutsugamushi antigens made it possible to identify a 47-kDa scrub typhus antigen (Sta47) that was stimulatory for the polyclonal T-cell line. Recombinant clones encoding 56-, 58-, and 110-kDa antigens (Sta56, Sta58, and Sta110, respectively) were unable to induce proliferation of this T-cell line. DNA sequence analysis of the cloned rickettsial insert encoding the Sta47 protein revealed the presence of four open reading frames potentially encoding proteins of 47, 30, 18, and 13 kDa. Analysis of sodium dodecyl sulfate-polyacryl-amide gel electrophoresis-separated and eluted fractions of lysates from the recombinant HB101(pRTS47B4.3) demonstrated that the fractions containing the 47-kDa protein as well as those containing proteins less than 18 kDa were stimulatory. Selected synthetic amphipathic peptides derived from the Sta47 antigen sequence identified a 20-amino-acid peptide that gave a 10-fold increase in T-cell proliferation over a control malarial peptide of similar length. Recognition of the 47-kDa antigen by a T-cell line with TH1 characteristics implicates this protein as one of potential importance in protection studies and future vaccine development.