Kacskovics I, Sun J, Butler J E
Department of Microbiology, University of Iowa, Iowa City 52242.
J Immunol. 1994 Oct 15;153(8):3565-73.
We report the sequences of more than 40 partial and complete swine C gamma cDNAs obtained by PCR cloning of first strand cDNA, and from a cDNA expression library, all from a single animal. These seem to represent five IgG subclasses, that can be grouped into two clusters; one contains IgG1 and IgG3 and the other, IgG2a, IgG2b and IgG4. IgG2a and IgG2b differ by only three amino acids, but single strand conformational polymorphism analyses of PCR-amplified IgG2-specific segments in animals of different breeds, argues for their putative subclass status. Major subclass differences are found in the hinge and C gamma 3, but differences in upper hinge length, associated with segmental flexibility in the IgGs of other species, are absent. All subclasses have identical middle hinge segments that can accommodate three interheavy chain disulfide bridges. The putative swine IgG subclasses have their greatest similarity with those of the human except for the near absence of hinge region variation. Swine subclasses such as ruminant IgG2a, have a lower hinge deletion which in human IgG1, contains one of the motifs believed necessary for interaction with Fc gamma Rs. IgG1 was the most frequently encountered subclass cDNA (25 of 43) and the single swine-mouse hybridoma tested had a sequence identical to IgG1. Partial sequence analyses of genomic clones identified one clone identical with the IgG1/IgG3 subclass cluster, two identical to the IgG2b/IgG4 subclass group, and two identical to each other but different from any of the expressed sequences reported here. Genomic blots suggest that up to eight C gamma genes are present in the genome.
我们报告了通过对第一链cDNA进行PCR克隆以及从一个cDNA表达文库中获得的40多个部分和完整的猪Cγ cDNA序列,这些均来自同一只动物。这些序列似乎代表了五个IgG亚类,可分为两个簇;一个簇包含IgG1和IgG3,另一个簇包含IgG2a、IgG2b和IgG4。IgG2a和IgG2b仅相差三个氨基酸,但对不同品种动物中PCR扩增的IgG2特异性片段进行的单链构象多态性分析支持它们作为推定亚类的地位。主要亚类差异存在于铰链区和Cγ3,但不存在与其他物种IgG中片段灵活性相关的上铰链长度差异。所有亚类都有相同的中间铰链段,可容纳三个重链间二硫键。推定的猪IgG亚类与人类的IgG亚类最为相似,但铰链区几乎没有变异。猪的亚类如反刍动物的IgG2a,其铰链缺失较少,而在人类IgG1中,该区域包含一个被认为是与FcγR相互作用所必需的基序之一。IgG1是最常遇到的亚类cDNA(43个中有25个),并且测试的单个猪 - 小鼠杂交瘤的序列与IgG1相同。对基因组克隆的部分序列分析确定了一个与IgG1 / IgG3亚类簇相同的克隆,两个与IgG2b / IgG4亚类组相同的克隆,以及两个彼此相同但与本文报道的任何表达序列都不同的克隆。基因组印迹表明基因组中存在多达八个Cγ基因。