By Morris Reichin
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Immunol. (Paris) 135C, 11. , and Nisonoff, A. (1972). J. Exp. Med. 135, 579. D. (1983). Mol. Immunol. 20, 39. Tonegawa, S. (1983). Nature 302, 575. D. (1983). J. Exp. Med. 158, 1615. D. (1984). J. Immunol. 133, 3164. L. (1985). J. Immunol. 134, 1236. L. (1985). J. Immunol. J34' 2740. D. (1982). Science 216, 309. , and Gilbert, W. (1980). Methods Enzymol. 65, 499. , and Riblet, R. (1984). Eur. J. Immunol. 14, 922. , and Fougereau, M. EMBO J. 4, 1225. , and Marshak-Rothstein, A. (1982). Eur. J. Immunol.
These data (Figure 1) show that first round anti-micrococcal antibodies are clustered at basic pi's between pH 7 and 8. 4. Following a one-year rest, the thi^rd round immunization was given and the resulting clonotypes included all representatives of first and second round sera. Later, a fourth immunization with micrococcal vaccine elicited antibodies once again lacking the clonotypes representative of the first round Regulation of Antibody Clones in Rabbits 49 pattern and the fourth round sera contained substantial amounts of AAI antibodies.
New Zealand rabbits were purchased from a local breeder. 1 (γ1κ), were previously reported (9). 3 (γ1κ) specific for 2,4-dinitrophenyl (DNP) was described elsewhere (9). Preparation and characterization of anti-Id62 mAb 15C9 was already reported (12). Hybridoma antibody B10H2 was prepared (13) using the non-secreting Sp2/0 myeloma cell line as the fusion partner. Polyclonal anti-Id62 antibodies were pre- pared in rabbits (12). 15 (9,12). BALB/c polyclonal anti-Id62 antibodies were purified by affinity chromatography on a mAb 62 Sepharose4B immunoadsorbent from pooled sera of BALB/c mice repeatedly immunized with mAb 62 cross-linked to keyhole limpet hemocyanin (12).