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Antibody Repertoire Development in Fetal and Neonatal Piglets. I. Four V_H Genes Account for 80 Percent of V_H Usage During 84 Days of Fetal Life^{1 ,2}

J. Sun^*, C. Hayward^*, R. Shinde^*, R. Christenson, S. P. Ford and J. E. Butler^3,*

^* Department of Microbiology and Interdisciplinary Immunology Program, University of Iowa, Iowa City, IA 52242; Roman L. Hruska Agricultural Research Service, U.S. Meat Animal Research Center, U.S. Department of Agriculture, Clay Center, NE 68933; and Department of Animal Science, Iowa State University, Ames, IA 50011

VDJ rearrangement and V_H gene usage during fetal development in 35 outbred piglets was examined by PCR amplification of VDJs; VDJs were subsequently characterized by hybridization with V_H-specific gene probes and by sequencing. VDJ rearrangement was first seen in the fetal liver on day 30 of a 114-day gestation. Four V_H genes (V_HA, V_HB, V_HC, and V_HE) accounted for ~80% of all V_H gene usage regardless of gestational age, choice of piglet, or lymphoid tissue tested; D_HA and D_HB were used in >90% of the fetal VDJs examined. Evidence of somatic hypermutation during fetal development was not found. The proportion of the four prominent fetal V_H genes did not differ significantly between cDNA and DNA, suggesting the absence of selective B cell differentiation. A comparison of recombination signal sequences, flanking sequences, and framework sequences of these fetal genes with other germline V_H genes of swine offered no clue as to their selective usage. N-region additions were prominent on day 40 but not on day 30, suggesting that the onset of terminal deoxynucleotidyltransferase activity occurs after 30 days of fetal development. These collective findings indicate that the preimmune, "natural Ab" repertoire of the fetal piglet is largely restricted to the use of four nonpolymorphic and nonmutated V_H genes and two nonmutated D_H segments. This suggests that the preimmune repertoire of swine is either highly restricted or almost entirely determined by junctional diversity in complementarity-determining region-3.

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