Beck M A, Tracy S, Coller B A, Chapman N M, Hufnagel G, Johnson J E, Lomonossoff G
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198.
Virology. 1992 Jan;186(1):238-46. doi: 10.1016/0042-6822(92)90078-4.
An in vitro murine T cell proliferation assay was used to determine whether an antigenic epitope(s) recognized by enterovirus-immune T cells is held in common between plant comoviruses and human enteroviruses. Splenocytes isolated from C3H/HeJ mice infected with coxsackievirus B3 (CVB3) proliferated in vitro not only against a variety of enterovirus (CVB2, CVB3, CVB6, CVA16, PV1) antigens, but against comovirus (CPMV, BPMV) antigens as well. Splenocytes from mice inoculated with bean pod mottle virus (BPMV) also proliferated in response to comoviral and enteroviral antigens in vitro. However, if the viral inocula were highly purified prior to inoculation, then the splenocyte response was generated only against the group used to inoculate, suggesting that the epitope shared between the comoviruses and the enteroviruses resided in the nonstructural region. B (nonstructural) and M (structural) genomic segments of CPMV were translated in rabbit reticulocyte lysates and used as in vitro antigens. Splenocytes from mice inoculated with live CVB3 proliferated in response to the B-RNA-encoded but not the M-RNA-encoded polypeptides, confirming the nonstructural coding region location of the common epitope. Comparison of predicted amino acid sequences in the nonstructural coding regions of the comoviruses and picornaviruses suggested a potentially immunogenic linear epitope in protein 2C. The consensus peptide LEEKGI was synthezized and shown to be immunogenic for both BPMV- and CVB3-immune splenocytes.
采用体外小鼠T细胞增殖试验来确定肠道病毒免疫T细胞识别的抗原表位是否在植物豇豆花叶病毒和人肠道病毒之间存在共性。从感染柯萨奇病毒B3（CVB3）的C3H/HeJ小鼠中分离出的脾细胞，在体外不仅能针对多种肠道病毒（CVB2、CVB3、CVB6、CVA16、PV1）抗原增殖，还能针对豇豆花叶病毒（CPMV、BPMV）抗原增殖。接种菜豆荚斑驳病毒（BPMV）的小鼠的脾细胞在体外也能对豇豆花叶病毒和肠道病毒抗原产生增殖反应。然而，如果在接种前对病毒接种物进行高度纯化，那么脾细胞反应仅针对用于接种的病毒组产生，这表明豇豆花叶病毒和肠道病毒之间共有的表位位于非结构区域。CPMV的B（非结构）和M（结构）基因组片段在兔网织红细胞裂解物中进行翻译，并用作体外抗原。接种活CVB3的小鼠的脾细胞对B-RNA编码的多肽而非M-RNA编码的多肽产生增殖反应，证实了共同表位位于非结构编码区。对豇豆花叶病毒和小RNA病毒非结构编码区预测氨基酸序列的比较表明，在蛋白2C中存在一个潜在的免疫原性线性表位。合成了共有肽LEEKGI，并证明其对BPMV免疫和CVB3免疫的脾细胞均具有免疫原性。
