King T P, Kochoumian L, Lu G
Rockefeller University, New York, NY 10021.
J Immunol. 1995 Jan 15;154(2):577-84.
White-face hornet venom allergen, Dol m 5.02, is a protein of 204 amino acid residues. This protein and its overlapping fragments, of 53-114 residues in size, containing the N-terminal, middle, and C-terminal regions of the molecule, can be expressed in high yield in bacteria by using the plasmid vector pQE12. Natural (n) and recombinant (r) Dol m 5.02s and the r-fragments are about equally immunogenic for IgG Ab response in BALB/c mice. n-Dol m 5.02 induces mainly murine IgG Abs specific for its discontinuous B cell epitopes and, to a lesser extent, Abs specific for its continuous epitopes. r-Dol m 5.02 and the r-fragments induce only Abs specific for continuous B cell epitopes that are common with those of the n-protein. Abs specific for the discontinuous epitopes show higher affinity than those specific for the continuous epitopes. r-Dol m 5.02 and the r-fragments are as efficient as n-Dol m 5.02 in inducing murine T cell responses specific for the n-protein. The differences in the immunogenicity of n- and r-proteins or r-peptide fragments for B and T cell responses are related to their conformations, inasmuch as only the n-protein is cross-linked by four disulfide bonds. These findings are relevant to the potential use of r-fragments as immunotherapeutic reagents in humans.
白面大黄蜂毒液过敏原Dol m 5.02是一种由204个氨基酸残基组成的蛋白质。该蛋白质及其大小为53 - 114个残基的重叠片段，包含分子的N端、中间和C端区域，可通过使用质粒载体pQE12在细菌中高产表达。天然（n）和重组（r）Dol m 5.02以及r片段在BALB/c小鼠中对IgG抗体反应的免疫原性大致相同。n - Dol m 5.02主要诱导针对其不连续B细胞表位的鼠IgG抗体，在较小程度上诱导针对其连续表位的抗体。r - Dol m 5.02和r片段仅诱导针对与n蛋白共有的连续B细胞表位的抗体。针对不连续表位的抗体比针对连续表位的抗体具有更高的亲和力。r - Dol m 5.02和r片段在诱导针对n蛋白的鼠T细胞反应方面与n - Dol m 5.02一样有效。n蛋白和r蛋白或r肽片段对B细胞和T细胞反应的免疫原性差异与其构象有关，因为只有n蛋白通过四个二硫键交联。这些发现与r片段作为人类免疫治疗试剂的潜在用途相关。
