Preferential expression of variable region heavy chain gene segments by predominant 2,4-dinitrophenyl-specific BALB/c neonatal antibody clonotypes.

Abstract

The B-cell repertoire in neonatal mice contains predominant clonotypes that are reproducibly expressed at particular times after birth. We have isolated and sequenced heavy and light chain cDNA clones from three 2,4-dinitrophenyl-specific neonatal hybridomas. Two of these hybridomas (TF2-36 and TF5-139) express idiotypes (Ids) that predominate during the first days after birth, and the third hybridoma (TF2-76) expresses Although the adult primary B-cell repertoire ofBALB/c mice exceeds 107 conotypes (1, 2), the antibody repertoire at birth is limited to 104-105 different specificities (3, 4). During the first weeks after birth, this repertoire undergoes a reproducible and patterned expansion (3-9). Thus, in the first 3 days of neonatal life, all BALB/c neonates share three predominant 2,4-dinitrophenyl (DNP)-specific and three predominant 2,4,6-trinitrophenyl (TNP)-specific clonotypes (3). The same mice do not express predominant clonotypes for fluorescein (3), phosphocholine (6), and either a(1-*3) or a(1--6) dextran determinants (7,8). Phosphocholine-specific clones reproducibly appear several days after DNP-or TNP-specific clones in both fetal liver (9) and neonatal spleen (6). Two weeks after birth, the antibody repertoire, which contains 106 specificities, is still more restricted than the adult repertoire (5).Recently, several investigators have attempted to correlate this patterned acquisition of the antibody repertoire with the preferential rearrangement of specific variable (V) region gene segments (10, 11). Murine germ-line heavy (H) chain V region gene segments (VH) have been grouped by sequence homologies into eight families (12, 13). VH recombinant strains have been used to obtain the order of these VH families, whose names relate to cell lines, on chromosome 12 as: 5' (VH 3609, VH 36-60, VH J606, VH X24)-VH J558-VH S107-VH Q52-VH 7183-D-JH 3' (14). The relative order ofthe set of VH gene families 5' to the J558 VH family cannot be mapped with the existing recombinant strains. The potential correlation between sequential rearrangement of V segments and sequential expression of antibody specificities is based on the observation that the rearrangement of VH families in fetal liver pre-B-cells is not random (10, 11). Cells with a rearranged member of the 7183 VH family have been the most frequently found among Abelson murine leukemia virus (Ab-MuLV)-transformed pre-B-cells and among fetal liver pre-B-cell hybridomas. This is especially striking because the 7183 VH family represents only z10% of all VH genes (13). One member of the 7183 VH family, VH 81X, rearranges at a particularly high frequency. VH 81X, one of the most JHproximal VH regions, is rearranged in almost half of AbMuLV-transformed fetal liver pre-B-cells (10).In order to evaluate the contribution of such a developmentally ordered rearrangement of V segments to the repertoire of antigen-responsive B cells, we have examined the V genes utilized by three hybridomas derived by fusion of DNP-hemocyanin-stimulated neonatal spleen cell...

Comparison of the fetal and adult functional B cell repertoires by analysis of VH gene family expression.

“…Furthermore, an important observation was the major contribution of VH 36-60 expression to the DNP response . This was most pronounced in the fetal response and is consistent with our previous results indicating that predominant idiotypes 36 (52) and 460 (53)(54)(55), both associated with VH 36-60 (35,55,56), were produced in the DNP-specific fetal response (56) . In addition, the 460 idiotype is also a predominant idiotype found in the adult DNP response (53)(54)(55).…”

“…The results suggest that the anti-DNP response, in general, is very heterogeneous and that member(s) of all of the VH gene families tested can potentially code for anti-DNP antibodies in both the fetus and adult. The heterogeneous nature of the anti-DNP response and the ability of anti-DNP antibodies to be encoded by more than one VH gene family is consistent with other reports (35,(52)(53)(54)(55) .…”

“…Hood, Brodeur, Riblet, and Riley and subcloned into pT7/T3-18 (Bethesda Research Laboratories, Gaithersburg, MD) so that radiolabeled single-stranded RNA probes could be prepared (30,33). The probes used were PV 3621 (36-60), pV14RI (J606), pVJ558 (J558), PVS107 (S107), pVQ52 (Q52), pVSAPC-15 (7183), and pVX24 (X24) and have been described elsewhere (11,34,35). The recombinant plasmids were linearized with the appropriate restriction enzyme and radioactive RNA probes were generated using T3 or T7 polymerase (Bethesda Research Laboratories) and ["S]UTP (New England Nuclear, Boston, MA) (36).…”

Two murine natural polyreactive autoantibodies are encoded by nonmutated germ-line genes.

“…In fact, the same difference between the E7 VH and VH 1210.7 is found in all previously described mAbs expressing a VH homologous to VH 1210.7. Thus, two mAbs have exactly the same VH sequence (22,23), and two mAbs have exactly the same 5' untranslated region and leader peptide sequences as E7 (22). Similar findings were also made for D23 VH (26,29).…”

Programmed Development of the Antibody Repertoire

“…Interestingly, antigens which stimulate responses in fetal B cells, such as DNP, elicit antibodies that frequently utilize members of the VH 7183 gene family (RILEY et al 1986) or of the V H 36-60 family (TEALE and KEARNEY 1986), which has not yet been fully evaluated in terms of either its chromosomal location or the time at which its elements typically rearrange. Antibodies that utilize late-rearranging VH elements (J558, J606) are frequently induced by antigens Based on adoptive transfer studies and the data presented in Fig.…”