La glicoforina può essere dimostrata su biopsia osteomidollare fissata e decalcificata utilizzando le metodologie immunoistochimiche standard.

Antigene espresso dagli eritrociti e dai precursori eritroidi (dal pro-eritroblasto).

L'importanza del CD235a/Glicoforina come  marcatore immunofenotipico è limitata all'analisi del microambiente midollare in corso di emopatie (mielodisplasie, leucemie acute, malattie mieloproliferative croniche). La dimostrazione della componente eritroide con la glicoforina può essere occasionalmente utile in altri campi della patologia (ad es. nella dimostrazione di eritrociti nucleati nella patologia placentare, o nella dimostrazione di emopoiesi extramidollare (splenica, epatica).

Figura 1. Eritrociti nucleati in un villo placentare: glicoforina

Figura 2. E

Figura 3. Midollo con componente eritroide che appare displastica (eritroblasti multinucleati): glicoforina

Figura 4 . Midollo con marcata diminuzione della componente eritroide che appare displastica: glicoforina

Figura 5 . Completa assenza di precursori eritroidi  nucleati come evidenziato dalla  glicoforina in un caso di PRCA ("pure red aplasia" in pz con timoma). Solo rare emazie non-nucleate sono glicoforina positive.

J Histochem Cytochem. 1985 Nov;33(11):1103-9.

 Influence of fixation and decalcification on the immunohistochemical staining of cell-specific markers in paraffin-embedded human bone biopsies. 

Mullink H, Henzen-Logmans SC, Tadema TM, Mol JJ, Meijer CJ. 

    A number of fixation and decalcification procedures were evaluated to determine their suitability for immunohistochemistry on trephine samples of bone marrow after paraffin embedding. In particular, the immunoreactivity of antigens characteristic for various hematopoietic cell lines (immunoglobulin heavy and light chains for plasmacytoid cells; elastase for neutrophil myeloid cells; lysozyme, alpha-1-antitrypsin and alpha-1-antichymotrypsin for hystiocytic cells; leukocyte common antigen for lymphocytes; hemoglobin and glycophorin A for erythroid cells; Factor VIII-related antigen for thrombocytoid cells) as well as some antigens specific for epithelial tumors (CEA, 115D8, and keratin) were investigated. Fixation in a mercuric chloride-formaldehyde mixture followed by decalcification in acetic acid-formaldehyde-saline proved to be the best procedure for antigen preservation and retention of morphologic detail. Moreover, there is no need of trypsinization when using this procedure. The only exception was Factor VIII-related antigen in megakaryocytes, which was best demonstrated in trypsin-digested sections of formalin-fixed and acetic acid-decalcified biopsies.

Virchows Arch A Pathol Anat Histopathol. 1993;423(1):33-8. 

Myelodysplastic syndromes: immunohistochemical and morphometric evaluation of proliferative activity in erythropoiesis and endoreduplicative capacity of megakaryocytes. Thiele J, Hoffmann I, Bertsch HP, Fischer R. 

Institute of Pathology, University of Cologne, Germany. 

    An immunohistochemical and morphometric analysis was performed on bone marrow trephine biopsies in 40 patients with primary myelodysplastic syndromes (MDS) to evaluate the proliferative activity in erythropoiesis and the endoreduplicative capacity of megakaryocytes. Control groups included normal bone marrow and marrow from cases presenting with pernicious anaemia. Double-immunostaining was applied with a monoclonal antibody (PC10) directed against proliferating cell nuclear antigen (PCNA), followed by antibodies against glycophorin C (Ret40f) or platelet glycoprotein IIIa (Y2/51-CD61) for the identification of the erythroid and megakaryocytic cell lineage. Comparison with normal bone marrow showed a reduction of erythropoiesis accompanied by an increase in atypical (micro-) megakaryocytes. Erythroid precursors displayed significant enhancement of PCNA-immunostaining. Megakaryocytes showed no increase in the relative frequency of PC10-positive cells (PCNA-labelling index). In pernicious anaemia, predominance of macrocytic-megaloblastoid erythropoiesis was associated with a striking increase in PCNA-labelling. Cell kinetic studies in this disorder revealed an abnormal arrest, particularly in S-phase which generates an over-expression of PCNA. Similar conditions were believed to be present in MDS with secondary folate deficiency. This mechanism explains the relatively high rate of positively-reacting pro- and erythroblasts which is not invariably accompanied by an increase in cell proliferation. Determination of megakaryocyte size and PCNA-staining capacity resulted in a significant increase in PC10-positive cells among micromegakaryocytes. Our findings on this cell lineage are in keeping with the assumption of a block in endoreduplicative activity at higher ploidy levels, associated with an apparently not-deregulated endomitosis in small-sized megakaryocytes of lower ploidy stages.

Leukemia. 1993 Jun;7(6):838-47. 

Therapy-related myelodysplastic syndromes: FAB classification, bone marrow histology, and immunohistology in the prognostic assessment. 

Orazi A, Cattoretti G, Soligo D, Luksch R, Lambertenghi-Deliliers G. 

Division of Anatomic Pathology and Cytology, Istituto Nazionale per lo Studio e la Cura dei Tumori, Milano, Italy. 

    In this study we describe the morphologic and immunohistochemical evaluation of bone marrow biopsies from 14 patients with therapy-related myelodysplastic syndromes (t-MDS). We employed CD34, anti-HLA-Dr, anti-elastase, CD68, anti-glycophorin, CD61 monoclonal antibodies immunostaining, and enzyme histochemistry for chloroacetate esterase. Moreover, we used PC10, a MAb raised against the proliferating cell nuclear antigen, to study the proliferative capacity of these marrows. Our data suggest that diagnosis of refractory anemia with excess of blasts (versus chronic myelomonocytic leukemia), the abnormal localization of immature precursors, marrow fibrosis, and augmented CD34 expression in the bone marrow biopsy are ominous prognostic factors at a statistically significant level (p < 0.0005). A combined morpho-immunohistochemical analysis of bone marrow biopsy correctly classifies t-MDS cases according to the biologic and clinical aggressiveness.

J Clin Pathol. 1999 Dec;52(12):919-21. 

Immunohistochemical identification of erythroid precursors in paraffin embedded bone marrow sections: spectrin is a superior marker to glycophorin. 

Sadahira Y, Kanzaki A, Wada H, Yawata Y. 

Department of Pathology, Kawasaki Medical School, Kurashiki, Japan. sadapath@med.kawasaki-m.ac.jp AIM: 

    To investigate whether spectrin can be used as an immunohistochemical marker for erythroid precursors in routinely processed paraffin embedded bone marrow sections. METHODS: Bone marrow biopsies and clot sections were stained with rabbit antihuman erythrocyte spectrin antibodies, specific for erythroid cells as shown by western blotting and bone marrow smears, and compared to sections stained with antiglycophorin monoclonal antibodies (JC159 and Ret49f). RESULTS: Antispectrin antibodies resulted in diffuse cytoplasmic staining of early erythroblasts and membranous staining of late erythroblasts as well as erythrocytes. In haematopathological samples, immature erythroid cell clusters were clearly identified. In contrast, antiglycophorin monoclonal antibodies resulted in only membranous staining of late erythroblasts, and faint staining of early erythroblasts. CONCLUSIONS: Spectrin may be a superior marker to glycophorin for the identification of erythroid precursors in paraffin embedded sections.