Interstitial exclusion of positively and negatively charged IgG in rat skin and muscle

Abstract

Volume exclusion, i.e., the space not available for a specific probe, may be dependant on the probe charge. Therefore, interstitial exclusion was measured for positively and negatively charged immunoglobulin (IgG) in skin and muscle of rats by using a continuous infusion method (30). Steady-state concentration of125I-labeled IgG 1 (pI = 8.7) and131I- labeled IgG 4 (pI = 6.6) was maintained by infusion of tracer for 120–168 h with an implanted osmotic pump. At the end of the infusion period and before tissue sampling, the rat was anesthetized and nephrectomized, and51Cr-labeled EDTA was injected and allowed 4 h for equilibration to measure interstitial fluid volume (Vi). Interstitial fluid was isolated from skin and muscle by using nylon wicks implanted post mortem. The relative IgG available space was measured as the ratio between labeled IgG and51Cr-labeled EDTA wick fluid equivalent spaces, and relative excluded volume fraction (Ve/Vi) was calculated as 1 − Va/Vi. Ve/Viin hindlimb skin averaged 0.37 ± 0.05 (SE) and 0.65 ± 0.06 ( P < 0.01) for IgG 1 and 4, respectively, with corresponding figures of 0.24 ± 0.05 and 0.51 ± 0.04 ( P < 0.01) in hindlimb muscle ( n = 9 for both tissues). These experiments suggest that fixed negative charges, most likely glycosaminoglycans, influence distribution of macromolecules in the interstitium and therefore affect interstitial fluid balance.