Hybridoma autoantibodies to erythrocytes from NZB mice and the induction of hemolytic anemia (1).
New zealand black (NZB) mice spontaneously elicit autoantibodies to erythrocytes and develop autoimmune hemolytic anemia. There are two possible autoantigens: autoantigen X on the erythrocyte surface and cryptic surface antigen, HB. Autoantibodies against these two antigens play roles pathogenetically in the autoimmune hemolytic anemia of NZB mice.
Spleen cells from non-immunized NZB mice were fused with P3-X63-Ag8.653 cells. The antibodies were screened using C57BL/6 erythrocytes by a binding assay.
Two IgM monoclonal antibodies (mAbs) were developed from 26 positive cells in 3936 hybridoma-positive wells. B13H4C8 has 22C and B13H2E12 has 4C as each optimal hemagglutination temperature.
These mAbs could be absorbed with mouse erythrocytes from strains, NZB, NZW and C57BL/6.
Aliquots of 10^6 to 10^7 cells of hybridoma cells were implanted intraperitoneally (i.p.) into 4-month-old BALB/c female mice, previously given i.p. 0.5ml of pristane (2,6,10,14-tet- ramethylpentadecane) 7 days prior to the implantation. Although B13H4C8 or B13H2EI2 hybridoma cell lines are semi-allogeneic in BALB/c mice,they grew in BALB/c mice and induced symptoms of hemolytic anemia. All the hybridoma recipients died within 14 days after the implantation. The hematocrit values on day 7 decreased to 12.3 +/- 0.6 in B13H4C8 recipients and to 26.6 +/- 3.4% in B13H2E12 recipients, compared to 47.3 +/- 0.6% in control groups.
On day 7, serum hemagglutination titers were negative in control mice, 1:320 to 1:2560 in B13H4C8 recipients and 1:10240 or more in B13H2E12 recipients.
B13H4C8 induced more severe disease than B13H2E12. This may be due to the difference in the optimal reaction temperature.
