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B Lymphocyte Selection and Age-Related Changes in V_H Gene Usage in Mutant Alicia Rabbits¹

Department of Microbiology and Immunology, Loyola University of Chicago, Maywood, IL 60153

Young Alicia rabbits use V_Ha-negative genes, V_Hx and V_Hy, in most VDJ genes, and their serum Ig is V_Ha negative. However, as Alicia rabbits age, V_Ha2 allotype Ig is produced at high levels. We investigated which V_H gene segments are used in the VDJ genes of a2 Ig-secreting hybridomas and of a2 Ig⁺ B cells from adult Alicia rabbits. We found that 21 of the 25 VDJ genes used the a2-encoding genes, V_H4 or V_H7; the other four VDJ genes used four unknown V_H gene segments. Because V_H4 and V_H7 are rarely found in VDJ genes of normal or young Alicia rabbits, we investigated the timing of rearrangement of these genes in Alicia rabbits. During fetal development, V_H4 was used in 60–80% of nonproductively rearranged VDJ genes, and V_Hx and V_Hy together were used in 10–26%. These data indicate that during B lymphopoiesis V_H4 is preferentially rearranged. However, the percentage of productive V_Hx- and V_Hy-utilizing VDJ genes increased from 38% at day 21 of gestation to 89% at birth (gestation day 31), whereas the percentage of V_H4-utilizing VDJ genes remained at 15%. These data suggest that during fetal development, either V_H4-utilizing B-lineage cells are selectively eliminated, or B cells with V_Hx- and V_Hy-utilizing VDJ genes are selectively expanded, or both. The accumulation of peripheral V_H4-utilizing a2 B cells with age indicates that these B cells might be selectively expanded in the periphery. We discuss the possible selection mechanisms that regulate V_H gene segment usage in rabbit B cells during lymphopoiesis and in the periphery.

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