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. 1999 Jun 7;189(11):1799–1814. doi: 10.1084/jem.189.11.1799

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AFCs in the spleen from ANA⁻ (4–6 wk) and ANA⁺ (10–12 wk) Tg⁻ MRL-lpr/lpr, VH3H9 MRL-lpr/lpr, and BALB/c mice. The number of (A) total Ig and (B) Igλ AFCs were determined by ex vivo ELISpot. The data are presented as the mean number of AFCs per 10⁴ cells ± the SD of triplicate wells. Total Ig⁺ and Igλ⁺ AFC numbers were calculated by normalizing for total B220⁺Ig⁺ and B220⁺Igλ⁺ numbers as determined by flow cytometry, respectively. Presented data are from a representative mouse of each genotype. Total number of mice from five experiments: n = 5 Tg⁻ BALB/c; n = 4 VH3H9 BALB/c; n = 4 ANA⁺ Tg⁻ and VH3H9 MRL-lpr/lpr; and n = 2 ANA⁻ Tg⁻ and VH3H9 MRL-lpr/lpr mice. Note that a higher number of Igλ⁺ AFCs than total Ig⁺ AFCs were detected for the ANA⁺ VH3H9 MRL-lpr/lpr mice in this assay. This difference is attributed to a sensitivity difference in the reagents used to detect total Ig versus Igλ. Therefore, although comparisons can be made for the relative number of AFCs of a given type (i.e., between Igλ⁺ numbers), they cannot be used to compare absolute frequencies between types (i.e., total Ig vs. Igλ).

AFCs in the spleen from ANA⁻ (4–6 wk) and ANA⁺ (10–12 wk) Tg⁻ MRL-lpr/lpr, VH3H9 MRL-lpr/lpr, and BALB/c mice. The number of (A) total Ig and (B) Igλ AFCs were determined by ex vivo ELISpot. The data are presented as the mean number of AFCs per 10⁴ cells ± the SD of triplicate wells. Total Ig⁺ and Igλ⁺ AFC numbers were calculated by normalizing for total B220⁺Ig⁺ and B220⁺Igλ⁺ numbers as determined by flow cytometry, respectively. Presented data are from a representative mouse of each genotype. Total number of mice from five experiments: n = 5 Tg⁻ BALB/c; n = 4 VH3H9 BALB/c; n = 4 ANA⁺ Tg⁻ and VH3H9 MRL-lpr/lpr; and n = 2 ANA⁻ Tg⁻ and VH3H9 MRL-lpr/lpr mice. Note that a higher number of Igλ⁺ AFCs than total Ig⁺ AFCs were detected for the ANA⁺ VH3H9 MRL-lpr/lpr mice in this assay. This difference is attributed to a sensitivity difference in the reagents used to detect total Ig versus Igλ. Therefore, although comparisons can be made for the relative number of AFCs of a given type (i.e., between Igλ⁺ numbers), they cannot be used to compare absolute frequencies between types (i.e., total Ig vs. Igλ).