Vibeke Ravn

Tissue distribution of histo‐blood group antigens.

The introduction of immunohistochemical techniques and monoclonal antibodies to specific carbohydrate epitopes has made it possible to study in detail the tissue distribution of histo‐blood group antigens and related carbohydrate structures. The present paper summarizes the available data concerning the histological distribution of histo‐blood group antigens and their precursor structures in normal human tissues. Studies performed have concentrated on carbohydrate antigens related to the ABO, Lewis, and TTn blood group systems, i.e. histo‐blood group antigens carried by type 1, 2, and 3 chain carrier carbohydrate chains. Histo‐blood group antigens are found in most epithelial tissues. Meanwhile, several factors influence the type, the amount, and the histological distribution of histo‐blood group antigens, i.e. the ABO, Lewis, and saliva‐secretor type of the individual, and the cell‐and tissue type. Oligosaccharides with blood‐group specificity are synthesized by the stepwise action of specific gene‐encoded glycosyltransferases. In general, this stepwise synthesis of histo‐blood group antigens correlates with cellular differentiation. The H and the Se genes both encode an α1–2fucosyltransferase, which is responsible for the synthesis of blood group antigen H from precursor disaccharides. A new model for the participation of the Se/H‐gene‐encoded glycosyl transferases in synthesis of terminal histo‐blood group antigens in human tissues is proposed; the type and degree of differentiation rather than the embryologic origin determines whether it is the H or the Se gene‐encoded transferases that influence expression of terminal histo‐blood group antigens in tissues.

Biochemical evaluation of endometrial function at the time of implantation

OBJECTIVE To review the literature on various endometrial factors assumed to be of importance to implantation and to evaluate their potential clinical value in the assessment of endometrial function at the time of implantation in infertile women in natural and stimulated cycles. DESIGN Literature review. RESULT(S) Cytokines such as leukemia inhibitory factor, colony-stimulating factor-1, and interleukin-1 have all been shown to play important roles in the cascade of events that leads to implantation. They participate in a synchronized cooperation between the endometrium and the preimplanting embryo under the influence of steroid hormones. The same applies to the integrin alpha(v)beta(3), glycodelin, and the polymorphic mucin 1. The usefulness of these factors to assess endometrial receptivity and to estimate the prognosis for pregnancy in natural and artificial cycles remains to be proven. CONCLUSION(S) The studies performed to date have mostly included only small groups of patients with a lack of fertile controls, and only a few prospective, controlled trials have been carried out. Therefore, definite conclusions about the clinical value of these factors in the assessment of endometrial function and prognosis for pregnancy after artificial reproductive therapy cannot be drawn at present. Further evaluation of their importance for and function during implantation is needed.

Reproducibility of Subjective Immunohistochemical Estrogen- and Progesterone Receptor Determination in Human Endometrium

To obtain a reliable scoring system for semi-quantitation of estrogen- (ER) and progesterone receptors (PgR) in human endometrial tissue, we investigated the reproducibility of subjective immunohistochemical ER and PgR determination. Specimens of frozen endometrial tissue were stained once (n = 129) or twice (n = 22) using the ER-ICA and PgR-ICA kits from Abbott. The semi-quantitative approach we used included subjective estimates of the overall staining intensity (I) and of the fraction of stained cells (%). Scoring was performed twice by the same observer and once by another observer (n = 87). Intra- and inter-observer agreement were evaluated using Kappa statistics. We found that more comprehensive scorings of I and % could not be agreed upon by the observers. Only simplified estimates of the fraction of cells stained and overall staining intensity were reproducible. Subjective estimates obtained by this method agreed with estimates obtained by counting (n = 38). Simplified H-scores, which were obtained by multiplication of the simplified estimates of % and I, were reproducible, too. In addition, semi-quantitation of ER and PgR by immunohistology was significantly correlated to quantitation by enzyme-immunoassay (n = 39). Thus, it was possible to reproducibly semi-quantitate ER and PgR only by employing a very simple immunohistochemical scoring of ER and PgR.

The distribution of type-2 chain histo-blood group antigens in normal cycling human endometrium

SummaryThe blood group ABO(H) determinants are major allogenic antigens in both erythrocytes and tissue of man. These antigens and related carbohydrates are markers of cellular maturation and differentiation in many epithelial tissues and have recently attracted great interest as tumor-associated antigens. Previous studies of endometrial tissues have indicated that glycosylation in this tissue may be related to hormonal stimulation. We have investigated the immunohistochemical distribution of type-2 chain histo-blood group-related carbohydrates in specimens of normal, cycling endometria obtained from hysterectomies on women with known ABO/Lewis erythrocyte type and saliva secretor status. N-acetyllactosamine and Lex were demonstrated to be uninfluenced by the genetic background. A and Aley antigens were exclusively demonstrated in endometria from blood group A individuals, while Ley was expressed in endometria from blood group 0 individuals mainly. The precursor N-acetyllactosamine as well as the terminal H, A, and ALey antigens were shown in only a few cells. In contrast, N-acetyllactosamine substituted by sialic acid and/or fucose residues (Lex, sialosyl-Lex, Ley) were demonstrated in epithelial cells of normal, cycling endometrium, but with both quantitative and qualitative differences in staining relating to the menstrual cycle, indicating that type-2 chain antigens are expressed under both genetic and hormonal influence in human cycling endometrium.

The distribution of type 1 chain ABH and related histo-blood group antigens in normal cycling human endometrium.

Summary:Cycling endometrial tissue was examined immunohistochemically for blood group antigens using a panel of monoclonal antibodies with specificity to type 1 chain carbohydrates. Staining was evaluated against the genetic background (ABO, Lewis, and saliva-secretor status) and the subphase of the menstrual cycle. Expression of type 1 chain antigens in most cases correlated with the genetic background; however, Lea and Leb antigens were in a few cases demonstrated in endometria with the erythrocyte Lea-b- phenotype and Leb antigen in erythrocyte Lea+b- endometria also. In addition, Leb antigen was preferentially expressed in endometria from blood group O individuals. Type I chain antigens were, in general, maximally demonstrated in the surface epithelium. Chain I A. H, ALeb, Lea, and disialosyl-Lea (dsLea) determinants were demonstrated only sporadically in functionalis and basalis glands of cycling endometria. Staining for most type I chain antigens showed variations that were related to the histomorphology (layer and menstrual phase), and monosialosyl-Lea (msLea) seemed to be a marker of secretory differentiation in the endometrium. Our results support the view that synthesis and expression of type 1 chain antigens in human endometrium is influenced by the genetic background and is modulated by the hormonal environment.

The use of polymerase chain reaction to detect metastatic cancer cells within lymph nodes in stage I cervical carcinoma.

The aim of the study was to determine whether human papillomavirus (HPV) in lymph nodes is a useful marker for the risk of recurrence in patients with HPV-related cervical cancer. The polymerase chain reaction and DNA-DNA hybridization techniques were used to examine 149 formalin-fixed, paraffin-embedded lymph nodes that had been resected from 24 patients undergoing radical hysterectomy for stage IB cervical carcinoma. The lymph nodes were examined for the HPV type, which in each case was found in the cervical tumor. Of 18 patients with histologically negative nodes, HPV DNA was found in a lymph node in two of 10 patients who later experienced a recurrence and in three of eight patients who were alive and well for > or = 5 years after surgery. In addition, HPV was detected in the lymph nodes of two of four patients with nodal metastases at the primary surgery; four of nine histologically positive lymph nodes in these patients contained HPV. It is concluded that detection of HPV in resected lymph nodes is probably not a useful means of identifying the cervical cancer patients who might benefit from adjuvant postoperative therapy.

Expression of type-2 histo-blood group carbohydrate antigens (Lex, Ley, and H) in normal and malignant human endometrium

Changes in expression of histo-blood group ABH and Lewis antigens are common alterations in carcinomas. Using immunohistochemistry, we have evaluated the expression of type-2 histo-blood group antigens in normal and malignant endometrial tissues in relation to genetic and hormonal factors. The Lex, sialosyl-Lex, and Ley antigens were inconstantly expressed in the normal endometrium. The expression was uninfluenced by the secretor status but was related to the ABO blood group status in Oestradiol (E2) stimulated endometria. Ley was expressed most frequently in proliferating endometria from blood group 0 individuals. Lex and Ley were maximally expressed in atrophic endometria, and Lex and Ley staining scores correlated inversely with serum levels of E2 in normal, non-secretory endometria. No correlation was found in adenomatous hyperplasias and endometrial carcinomas, which when compared with atrophic endometria, showed a loss of Lex and Ley and an increased H-carbohydrate expression at apical membranes. Carcinomas from non-secretors showed lower expression of Ley and H-antigens than carcinomas from secretors. Our findings suggest that the genetic and hormonal influence on glycosylation based on type-2 chain carbohydrates differ between normal and malignant endometrium. This difference is probably related to specific tumour-associated qualitative and quantitative changes in the fucosyltransferases.

Estrogen- and progesterone receptors in normal cycling endometrium as studied by end-point titration

A thorough knowledge of the normal physiological fluctuations in estrogen-(ER) and progesterone receptors (PgR) is essential to characterize the changes in ER and PgR in the abnormal endometrium. We investigated the distribution of ER and PgR in frozen human cycling endometrial tissue using the commercially available ER-and PgR-ICA kits. Two-fold end-point titration (EPT) of ER and PgR antibodies was implemented to semi-quantitate more accurately ER and PgR. Semiquantitation of ER and PgR using EPT was significantly correlated to results obtained using either simple scoring or enzyme-immunoassay (EIA) methods. ER and PgR staining fluctuated in relation to the menstrual cycle. In most subphases PgR exceeded ER in both epithelial and stromal cells. Highest levels of ER and PgR were demonstrated in the glands of the functionalis in mid-to-late proliferative phases, whereas both receptors were almost undetectable by immunohistology in the glands of mid-to-late secretory phases. Endometrial stromal cells had high and nearly constant EPT values for PgR, but low values for ER througout the menstrual cycle. EPT values for ER and PgR were generally higher in the basalis than in the functionalis but showed similar cyclic fluctuations. Our results further substantiate the view that the response to hormonal stimulation is cell-type specific, and suggest differences in steroid metabolism according to cell type and layer.

Inverted papilloma of upper urinary tract

Two new cases of inverted urothelial papilloma in the upper urinary tract are described and added to the 22 cases previously reported in the literature. In both cases inverted papilloma was localized beneath macroscopic normal surface, and in one of the cases the changes were found scattered widely in the upper urinary tract. The possible etiology and the symptomatology are discussed, and the need for follow-up of these patients is emphasized.

Thomsen-Friedenreich-related antigen in human endometrium

The Thomsen‐Friedenreich antigen (T‐antigen) Ga1β1‐3Ga1NAc, is a cryptic disaccharide structure on human erythrocytes. It is masked by sialic acid and uncovered by sialidases as neuraminidase. T‐antigen is a tumor‐associated antigen in epithelial tissues. This study reports preliminary immunohistochemical findings on the expression of the blood group related T‐antigen in human endometrium. Dewaxed sections of 10 proliferative‐, 6 interval‐, 14 secretory endometrium, and 16 endometrial carcinomas, either untreated or pretreated with neuraminidase, were subjected to an indirect immunoperoxidase technique using the monoclonal antibody 49H.8. The monoclonal antibody 49H.8 did not bind to normal cyclic endometrium, unless pretreated with neuraminidase. Neuraminidase treatment revealed staining of apical membranes of glandular‐ and surface epithelium in early or manifest secretory endometrium only. Thirteen of sixteen carcinomas stained without neuraminidase treatment, while all carcinomas stained after pretreatment with neuraminidase. Staining was more widespread than in untreated carcinomas, resembling that of secretory endometrium. Our results suggest: 1) That the sialylated 49H.8 antigen seems to be a marker of cellular differentation in the endometrium, 2) that the antigen defined by the monoclonal antibody 49H.8 might be a tumor‐associated antigen in endometrial epithelial tissue.