A single i.v. injection of a mAb 5-1-6 to rats was found to cause massive though transient proteinuria. This mAb 5-1-6, IgG1 was produced by immunization of BALB/c mice with collagenase-treated Wistar rat glomeruli and was highly organ and species specific. Immunoelectron microscopy using immunoperoxidase with the avidin-biotin complex and immunogold staining indicated mAb 5-1-6 to bind in vitro to the surface of glomerular epithelial foot processes, mainly to slit diaphragms. The recognized antigenic molecule was not susceptible to neuraminidase treatment and its Mr was about 51 kDa by immunoprecipitation. A one-shot i.v. injection of this mAb induced proteinuria in rats starting immediately, reaching the peak on day 8 (mean value of 150 mg/24 h), then gradually decreasing to normal level on day 18. The in vivo localization of administrated mAb 5-1-6 changed with time. Linear binding along glomerular capillary walls was observed 2 h after injection. However, 3 days later, it partially shifted to a fine granular pattern. The linear pattern disappeared and the size as well as intensity of the fluorescent granules decreased on day 12 to trace positive on day 18. Immunoelectron microscopy revealed the binding pattern of in vivo injected mAb 5-1-6 after 2 h to be similar to that in vitro. Three days later, injected mAb was observed within multivesicular bodies in glomerular epithelial cells as well as along the surface of foot processes and around slit diaphragms. Twelve days after injection, mAb along the surface of the foot processes and around slit diaphragms decreased but those in multivesicular bodies were observed more frequently. Rat IgG and C3 could not be detected throughout the period of observation. No histologic abnormalities were noted except for partial retraction of epithelial foot processes at the peak of proteinuria on day 8. This mAb thus provides a valuable means for examining the mechanism of proteinuria.

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