IgD antibodies to DNP are specifically induced in ascitic fluid after intraperitoneal immunization

Abstract

The induction characteristics of IgD in serum and other body fluids are poorly understood. Here we report that Freund's complete adjuvant (FCA), when used in intraperitoneal (i.p.) immunization, resulted in significantly higher levels of IgD and specific IgD antibodies in ascitic fluid than in the serum of the same animals. This was in sharp contrast to the distribution of IgM and kappa light chain-bearing immunoglobulins (kappa Ig) between these compartments. Female rats were immunized subcutaneously (s.c.) on Day 0 with 1:1 FCA mixed with either 2,4 dinitrophenyl group A streptococcal vaccine (DNP-GASV), 2,4 dinitrophenyl ovalbumin (DNP-OVA), or saline. A week later (and weekly thereafter) they were boosted i.p. with these antigens mixed with FCA 1:9. Eleven of sixteen rats produced some ascites by Day 21, and 15/16 by Day 28. Total immunoglobulin and specific antibodies for the heavy-chain isotypes IgD and IgM and for kappa Ig were measured by ELISA. Between immunization groups no significant differences were observed in total immunoglobulin levels. Subcutaneous immunization with 1:1 FCA caused a significant two- to six-fold increase in serum IgD by Day 7 (P less than 0.01), while no change occurred with serum IgM or kappa Ig. Ascitic fluid collected on Day 21 had significantly high levels of IgD than that of serum IgD (P less than 0.01), with a mean level nine times that of the serum. In contrast, levels of total IgM, kappa Ig and total protein were significantly lower in ascitic fluid than in serum (all P less than 0.05). The high levels of IgD in ascitic fluid disappeared in Day 28 ascitic fluid, while IgM and kappa Ig levels were relatively unchanged. The ratio of ascitic fluid to serum IgD anti-DNP was 9.8 for the DNP-GASV-immunized group on Day 21, and 23.9 for the DNP-OVA-immunized group on day 28. The same ratios for IgM anti-DNP and kappa Ig anti-DNP were much lower, ranging from 0.4 to 2.3 and 0.3 to 3.3, respectively.