CROSS-REACTIVITY OF HUMAN-IGG ANTI-F(AB'), ANTIBODY WITH DNA AND OTHER NUCLEAR ANTIGENS

Objective. To characterize immunologic specificity and possible antiid iotype activity of IgG anti-F(ab')(2) in normal subjects as well as in patients with active and inactive systemic lupus erythematosus (SLE). Methods. IgG anti-F(ab')(2) and anti-double-stranded DNA (anti-dsDNA) were affinity isolated from immunoadsorption columns of F(ab')(2) and dsDNA linked to Sepharose 4B. Affinity-purified IgG anti-F(ab')(2) (A PAF) and affinity-isolated IgG anti-dsDNA (APAD) were tested by enzyme -linked immunosorbent assay (ELISA) for other cross-reacting specifici ties including anti-Sm, anti-Sm/RNP, and anti- Crithidia binding. Anti -DNA specificity of APAF and APAD was assayed by S1 nuclease treatment of heat-denatured DNA. Rabbit antiidiotypic antisera were prepared by immunization with APAF and APAD from normal subjects and SLE patients and absorption with insolubilized human Cohn fraction II (Fr II). V-L and V-H regions of 5 monoclonal IgM antibodies with anti-F(ab')(2)/an ti-DNA specificity generated by Epstein-Barr virus B cell stimulation were sequenced by polymerase chain reaction and characterized for V-H and V-L subgroup. APAF and APAD were also examined by high-resolution electron microscopy for possible ring forms indicative of antiidiotypi c V-region interactions. Results. APAF from normal subjects, represent ing 0.08-0.18% of serum IgG, showed striking relative concentrations o f both anti-F(ab')(2) and anti-DNA, as well as anti-Sm and anti-Sm/RNP ELISA reactivity. Both APAF and APAD reacting with F(ab')(2) or dsDNA on the ELISA plate could be cross-inhibited by F(ab')(2) or DNA in so lution. Anti-DNA reactivity in normal APAF and APAD was much more sens itive to S1 nuclease treatment than similar fractions from SLE patient s. Neither APAF nor APAD from controls produced positive antinuclear i mmunofluorescence or positive Crithidia staining, whereas these were s trongly positive using SLE APAF and APAD. Absorbed rabbit antisera aga inst normal or SLE APAF and APAD showed strong ELISA reactivity agains t both APAF and APAD, but no residual reactivity with normal Fr II. V- L and V-H sequencing of monoclonal human IgM antibodies showing both a nti-F(ab')(2) and anti-DNA reactivity showed relative V(H)3, V kappa 1 or V(H)1, V kappa 3 restriction. No evidence of ring forms or V-regio n ''kissing'' dimers was obtained when normal or SLE APAD or APAF was examined by high-resolution electron microscopy. Conclusion. IgG anti- F(ab')(2) in both normal subjects and SLE patients represents a polyre active Ig subfraction with concomitant anti-DNA, anti-Sm, and anti-Sm/ RNP specificities. Anti-DNA reactivity in SLE is qualitatively differe nt from that in normal APAD and APAF since normal APAD and APAF anti-D NA is much more sensitive to S1 nuclease digestion of denatured dsDNA. APAF and APAD share distinct V-region antigens which may be related t o prominent V(H)3 or V(H)1 antigenic components. No evidence for in vi vo complexing of anti-DNA and anti-F(ab')(2) as ring forms or antiidio type-IgG complexes was observed during ultrastructural studies. In bot h normal individuals and SLE patients, APAF may represent a small poly reactive IgG subfraction which also contains antinuclear and anti-DNA specificities.