Monoclonal antibody to an alloantigenic determinant onβ2-microglobulin (β2M) of the mouse

✍ Scribed by Michael Chorney; Fung-Win Shen; James Michaelson; Edward A. Boyse

• Publisher: Springer-Verlag
• Year: 1982
• Tongue: English
• Weight: 158 KB
• Volume: 16
• Category: Article
• ISSN: 0093-7711
• DOI: 10.1007/bf00364446

✦ Synopsis

Polymorphism of mouse fi2M (B2m locus) has been indicated by strain differences observed in SDS-PAGE (Michaelson et al. 1980, Robinson et al. 198 la) and IEF (Goding and Walker 1980), corresponding to a single amino acid substitution (Gates et al. 1981). The B2m locus has been mapped in the region of H-3 on chromosome 2 (Michaelson 1981, Goding 1981, Robinson et al. 1981b).

Tli6 immunization giving rise to the monoclonal anti-fi2M alloantibody, provisionally called mc-/~2M-B :clone 23-IgG2a(~c), described in this report, was SJL/J anti-B10.S spleen and thymus. The mc-fl2M-B reagent used was pooled serum and ascites from pristane-primed (BALB/c x SJL/J)F t mice carrying clone 23 hybridoma in passage. In the cytotoxicity assay with guinea pig serum as complement (rabbit complement gave a somewhat higher titer but a lower proportion of lysed cells), the strain distribution shown in Table 1 accords without exception to the strain distribution of Ly-ml 1 given by Tada and co-workers (1980); the Ly-ml 1 phenotype of the BXD-6 RI strain was originally reported to be Ly-ml 1 negative, apparently because of an error in identification of the tested mice, and is now known to be Ly-mll positive (U. H~immerling, personal communication). Also, the cytotoxicity activity is in agreement with fl2M-B distinguished by SDS-PAGE as described (Michaelson 1981). The proportions of positive cells was > 95~o for LNC, ~ 85~ for spleen, -40~ for bone marrow, and ~ 10~ for thymus, roughly conforming to the comparative sensitivities of these four populations to H-2 antibody in cytotoxicity assays, to a titer of 1 : 640. The bands precipitated by the mc-fi2M from labeled spleen cells are shown in Figures 1 and2.