Optimization of immunocytochemical P-glycoprotein assessment in multidrug-resistant plasma cell myeloma using three antibodies

T. Grogan, W. Dalton, J. Rybski, Catherine S Perry, P. Meltzer, L. Richter, M. Gleason, J. Pindur, A. Cline, R. Scheper, T. Tsuruo, S. Salmon

Research output: Contribution to journalArticle

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Abstract

Seeking to optimize the immunocytochemical assay of P-glycoprotein, a 170-kilodalton (P-170) molecule associated with multidrug resistance, we experimented with a variety of antibodies (JSB-1, C219, and MRK-16), fixation conditions, and titers using both human myeloma cell lines and clinical myeloma specimens. Under optimized conditions, using all three antibodies and the cell lines as standards and controls, the ICC method proved sensitive, specific, reliable, rapid, and within the realm of every day hospital laboratory expertise. The 3 anti-P-glycoprotein antibodies revealed different reactivities with P-170. Both C219 and JSB1 were optimized by fixation in cold acetone. With MRK-16 optimal results were obtained on unfixed or formalin fixed specimens. Under optimal fixation and titering conditions, low level (DOX 4) detection was possible. Given that the three antibodies differ in reactivity and recognize different P-170 epitopes, it follows that using the antibodies in a small panel in a useful strategy in increasing the likelihood of detecting true P-glycoprotein expression by the immunocytochemical method. In dilution experiments, the immunocytochemical method was as sensitive as RNase protection asay and more sensitive than Western blot detection. Immunocytochemistry coupled to computer-assisted single-cell densitometry, showed a strong correlation (R= 0.98) between cellular P-170 density and in vitro resistance to doxorubicin. Multidrug-resistant specific probes for RNA expression and Western blot assays confirmed the specificity of P-170 expression in both cell lines and clinical samples. Thus, a small panel of antibodies, under optimized immunocytochemical conditions, appears to have potential as a rapid, sensitive, clinically useful assay for multidrug resistance in myeloma.

Original languageEnglish (US)
Pages (from-to)815-824
Number of pages10
JournalLaboratory Investigation
Volume63
Issue number6
StatePublished - 1990

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P-Glycoprotein
Multiple Myeloma
Antibodies
Multiple Drug Resistance
Cell Line
Western Blotting
RNA Probes
Hospital Laboratories
Densitometry
Ribonucleases
Acetone
Doxorubicin
Formaldehyde
Epitopes
Immunohistochemistry

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Optimization of immunocytochemical P-glycoprotein assessment in multidrug-resistant plasma cell myeloma using three antibodies. / Grogan, T.; Dalton, W.; Rybski, J.; Perry, Catherine S; Meltzer, P.; Richter, L.; Gleason, M.; Pindur, J.; Cline, A.; Scheper, R.; Tsuruo, T.; Salmon, S.

In: Laboratory Investigation, Vol. 63, No. 6, 1990, p. 815-824.

Research output: Contribution to journalArticle

Grogan, T, Dalton, W, Rybski, J, Perry, CS, Meltzer, P, Richter, L, Gleason, M, Pindur, J, Cline, A, Scheper, R, Tsuruo, T & Salmon, S 1990, 'Optimization of immunocytochemical P-glycoprotein assessment in multidrug-resistant plasma cell myeloma using three antibodies', Laboratory Investigation, vol. 63, no. 6, pp. 815-824.
Grogan, T. ; Dalton, W. ; Rybski, J. ; Perry, Catherine S ; Meltzer, P. ; Richter, L. ; Gleason, M. ; Pindur, J. ; Cline, A. ; Scheper, R. ; Tsuruo, T. ; Salmon, S. / Optimization of immunocytochemical P-glycoprotein assessment in multidrug-resistant plasma cell myeloma using three antibodies. In: Laboratory Investigation. 1990 ; Vol. 63, No. 6. pp. 815-824.
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AU - Grogan, T.

AU - Dalton, W.

AU - Rybski, J.

AU - Perry, Catherine S

AU - Meltzer, P.

AU - Richter, L.

AU - Gleason, M.

AU - Pindur, J.

AU - Cline, A.

AU - Scheper, R.

AU - Tsuruo, T.

AU - Salmon, S.

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