Table II.

F4/80+, slow-sedimenting (3–4.5 mm/h), LPS-stimulated spleen cells stimulated in the presence of OX2:Fc provide optimal inhibition of CTL and type 1 cytokine production in vitroa

LPS-Stimulated Cells AddedOX2:Fc AddedPercent Lysis 51Cr Targets (50:1, E:T)Cytokines in Culture (pg/ml)
IL-2IFN-γ
None31 ± 4.2910 ± 1601060 ± 150
None+17 ± 3.2b460 ± 70b510 ± 90
3–4.5 mm/h:Unfx30 ± 4.1915 ± 1501040 ± 140
3–4.5 mm/h:Unfx+6.9 ± 2.3**240 ± 55**230 ± 45**
3–4.5 mm/h:F4/80+28 ± 5.3935 ± 1601050 ± 160
3–4.5 mm/hr F4/80++3.0 ± 1.7**90 ± 30**105 ± 35**
3–4.5 mm/h:F4/8027 ± 4.8915 ± 150990 ± 145
3–4.5 mm/h:F4/80+16 ± 3.2b380 ± 50b390 ± 40b
>6 mm/h14 ± 3.7b310 ± 80b390 ± 85b
>6 mm/h+8.6 ± 2.9b270 ± 75b230 ± 55b
  • a As for Table I, 5 × 106 responder C3H spleen cells pooled from three 8-wk donors were cultured in triplicate with 2.5 × 106 mitomycin-c-treated C57BL/6 DC in the presence or absence of 1.5 × 106 (or 3 × 105 for the F4/80+ pool) cells from velocity-sedimented, LPS-stimulated, T-depleted spleen cells. Cultures received additional OX2:Fc protein as shown. Further separation of small cells into F4/80+ and F4/80 populations used an anti-PE column and PE-anti-F4/80 (see Materials and Methods). Total recovery was 80%, and recovery of the F4/80+ population (10% of the 3–4.5 mm/h pool) was 70%.

  • b , p < 0.05, compared with control cultures in first row (ANOVA followed by pair-wise t test)

  • c ∗, p < 0.05, compared with all other ∗ groups (pair-wise t test).