The pre- and postpartum maternal serum anti-D concentrations of 28 women with severe Rh(D) immunization who received high-dose intravenous immunoglobulin treatment has been determined. In all cases, including 1 in which the newborn was D negative, a sharp increment in the anti-D titer was observed after delivery. The specific immunoglobulin concentration rose to levels ranging from 4.7 to 204.0 µg/ml and, in 20% of the patients, increments of fifty times or greater were observed. Human monoclonal antibodies (hmAb) have been produced from Epstein-Barr virus-transformed lymphoblastoid B cell lines derived from 1 of these naturally hyperimmunized patients whose serum contained an anti-D-category D^VI antibody. Four anti-D-secreting cell lines (97.E3.39.214, 44.E4.R1.257, E7.R1.126.83.115 and E11V. 117.63; hereafter referred to as 214, 257, 115 and 63) have been established and maintained in continuous culture for periods ranging from several months to 3 years, without loss of antibody production capacity. Antibodies 115 and 214 recognize all D^u samples tested at the same level as the polyclonal positive control. Antibodies 63 and 257 show a significantly lower reaction strength with some of the D^u samples. Studies with D category cells showed that the D^VI category was recognized only by hmAb 214. The reactivity pattern of this antibody is that of an anti-epD4, although the reaction strength varied greatly with different D^IVa cell samples. Results obtained with hmAb 257 and 115 using papain-treated D category cells suggest that booth react as anti-epD6/7. Antibody 63 reacts as anti-epD1, although it shows only weak reactivity with the single sample of D^VII cells tested. A mixture of hmAb 214 and 115 should thus be capable of identifying most weak and partial D erythrocytes by an appropriate method.

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