Milich D R, Hughes J L, McLachlan A, Thornton G B, Moriarty A
Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, CA 92037.
Proc Natl Acad Sci U S A. 1988 Mar;85(5):1610-4. doi: 10.1073/pnas.85.5.1610.
Previous studies located T-cell recognition of the nucleocapsid of the hepatitis B virus (HBcAg) to residues 120-140 in mice bearing the H-2s or H-2b haplotypes. Herein, we demonstrate that B10.S (H-2s) and B10 (H-2b) H-2 congenic strains recognize distinct T-cell sites within the p120-140 (a synthetic peptide corresponding to residues 120-140 of HBcAg) sequence defined by p120-131 and p129-140, respectively. Peptide p120-131 stimulates B10.S HBcAg-primed T cells, and reciprocally p120-131-primed T cells recognize HBcAg. Similarly, the p129-140 sequence is a T-cell recognition site relevant to the native HBcAg in the B10 strain. It is also shown that these 12-residue peptides efficiently prime T-helper cells, which are capable of eliciting antibody production to HBcAg in vivo. These observations prompted us to examine the ability of the HBcAg-specific p120-140 sequence to function as a T-cell carrier moiety as a component of a totally synthetic hepatitis B vaccine. For this purpose a synthetic B-cell epitope from the pre-S(2) region (p133-140) of the viral envelope was chosen because this sequence represents a dominant antibody-binding site of the envelope. Immunization of B10.S and B10 strains with the synthetic composite peptide c120-140-(133-140) elicited anti-peptide antibody production, which was crossreactive with the native viral envelope. Furthermore, c120-140-(133-140) immunization primed p120-131-specific T cells in the B10.S strain and p129-140-specific T cells in the B10 strain, which recognized HBcAg and provided T-helper cell function for anti-envelope antibody production in vivo. These results demonstrate the feasibility of constructing complex synthetic immunogens that represent multiple proteins of a pathogen and are capable of engaging both T and B cells relevant to the native antigens.
先前的研究在携带H-2s或H-2b单倍型的小鼠中确定了乙型肝炎病毒核衣壳（HBcAg）的T细胞识别位点位于第120 - 140位氨基酸残基处。在此，我们证明B10.S（H-2s）和B10（H-2b）H-2同源品系分别识别由p120 - 131和p129 - 140定义的p120 - 140（与HBcAg第120 - 140位氨基酸残基对应的合成肽）序列内不同的T细胞位点。肽p120 - 131刺激经HBcAg致敏的B10.S T细胞，反之，经p120 - 131致敏的T细胞识别HBcAg。同样，p129 - 140序列是B10品系中与天然HBcAg相关的T细胞识别位点。还表明这些12个氨基酸残基的肽能有效地启动T辅助细胞，这些T辅助细胞能够在体内引发针对HBcAg的抗体产生。这些观察结果促使我们研究HBcAg特异性p120 - 140序列作为完全合成的乙型肝炎疫苗的一个组成部分作为T细胞载体部分发挥作用的能力。为此，选择了来自病毒包膜前S(2)区（p133 - 140）的一个合成B细胞表位，因为该序列代表包膜的一个主要抗体结合位点。用合成复合肽c120 - 140-(133 - 140)免疫B10.S和B10品系可引发抗肽抗体产生，该抗体与天然病毒包膜具有交叉反应性。此外，c120 - 140-(133 - 140)免疫在B10.S品系中启动了p120 - 131特异性T细胞，在B10品系中启动了p129 - 140特异性T细胞，这些T细胞识别HBcAg并在体内为抗包膜抗体产生提供T辅助细胞功能。这些结果证明了构建代表病原体多种蛋白质且能够与天然抗原相关的T细胞和B细胞相互作用的复杂合成免疫原的可行性。
