Application of immune fluorescence technique to kidney biopsy – Mellors 1950s

Shankar Prasad Yadav
B.P. Koirala Institute of Health Sciences, Nepal

The use of immunofluorescence techniques to detect specific tissue antigens using fluorescein-labeled antibody was first described in the 1940s by Albert Hewett Coons and colleagues. The antibody, coupled with fluorescein (immunochemical reagent), reacts with tissue containing antigen and produces a light emission visible through a fluorescence microscope. Coons used fluorescein to detect pneumococcal antigen in Aschoff nodules, a pathognomonic marker of rheumatic fever (1,2).

It was not until the 1950s, however, that the use of this principle in kidney biopsy was demonstrated by R.C. Mellors. The technique was modified to localize the antibody in kidney tissue. In this landmark study (3), fifteen healthy rabbits were injected with bovine gamma globulin, while four rabbits were used as a control group: this led to the description of different patterns of acute glomerulonephritis among twelve of the experimental animals. In the second part of the experiment, antibody was prepared from the globulin fraction of chicken anti-serum and rat immunoglobulin, which was coupled with fluorescence thiocyanate to generate fluorochrome reagent. The application of this reagent to kidney tissue, with subsequent fluorescent microscopy, demonstrated that there was increased intensity of immunofluorescence in the glomerulus of affected rabbits in comparison to tubules, or unaffected, or control group. These findings helped to conceptualize the antigen-antibody reaction as central in the pathogenesis of human glomerulonephritis.

Current use of immunofluorescence in diagnosing glomerulonephritis including IgA nephropathy, C3 glomerulonephritis, lupus nephritis, or detection of C4d in humoral anti-graft reactions are based on the fundamental principles elucidated by Mellors more than 60 years ago.

References

  1. Coons AH, Creech H J & Jones R N. Immunological properties of an antibody containing a fluorescent group.  Soc. Expt. Biol. Med.47, 200–202 (1941).
  2. Coons AH., Creech HJ., Jones RN. & Berliner E. The demonstration of pneumococcal antigen in tissues by the use of fluorescent antibody.  Immunol.45, 159–170 (1942).
  3. Mellors RC, Arias-Stella J, Siegei M, & Pressman D. Analytic Pathology II. Histopathologic demonstration of glomerular·localizing antibodies in experimental glomerulonephritis. Am. J. Path., In Press, 1955.

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