Stable expression of a recombinant human antinucleosome antibody to investigate relationships between antibody sequence, binding properties, and pathogenicity

Table 1 Summary of results from two separate experiments implanting CHO cells into SCID mice

Implanted cells	Mean human IgG in sera (ng/ml) (no. of mice/group)	Terminal proteinuria (range for group)	Mean proteinuria (estimated^a g/l)	Mean day of death
Experiment 1
CHO-B3	<16 (3/5), 0 (2/5)	++/+++ to +++	2.4	20
CHO-B3(R27aS)	260 (2/5), <16 (3/5)	+ to ++	0.65	24
Nontransfected CHO only	0	Trace to trace/+	0.2	34^b,c
Pristane only	0	Trace/+	0.2	34^b
Experiment 2
CHO-B3	105 (4/5), NR (1/5)	++ to +++	2.4	26
CHO-B3(R27aS)	420 (5/5)	+ to ++/+++	0.99	39
CHO with empty vector	0	Trace to +/++	0.38	48
Pristane only	0	Trace/+	0.2	56^b

^aProteinuria was assessed using Albustix, which give a semiquantitative measure based on colour change. Based on values provided with the Albustix, in order to calculate the estimated proteinuria in g/l we assumed that trace = 0.1 g/l, trace/+ = 0.2 g/l, +/++ = 0.65 g/l, and ++/+++ = 2.0 g/l. The proteinuria data in both experiments were found to be normally distributed using the Kolmogorov–Smirnov test and compared using a one-way analysis of variance (ANOVA) followed by Bonferroni's multiple comparison test. In experiment 1, the ANOVA showed that there was a significant difference between the mean proteinuria values of all the groups (P < 0.0001). The Bonferroni post test indicated that there was a significant difference between mice implanted with CHO-B3 and CHO-B3(R27aS) (P < 0.001), nontransfected CHOdhfr^- cells (P < 0.001), and pristane only (P < 0.001). In experiment 2, the ANOVA showed that there was a significant difference between the mean proteinuria values of all the groups (P < 0.0003). The Bonferroni post test indicated that there was a significant difference between mice implanted with CHO-B3 and CHO-B3(R27aS) (P < 0.05), CHO cells containing the empty vector (P < 0.001), and pristane only (P < 0.01). ^bMice still healthy at the end of the experiment. ^cThese cells did not survive/grow in the mice. CHO, Chinese hamster ovary; NR, not recorded; SCID, severe combined immunodeficiency.

ISSN: 1478-6362