Adhesion of mononuclear cells from multiple sclerosis patients to cerebral vessels in cryostat sections of normal human brain.

Leukocyte extravasation across the blood-brain barrier is a critical event in the pathogenesis of multiple sclerosis (MS). This complex multistep process includes the adhesion of leukocytes to the endothelial cells of the central nervous system microvasculature. To investigate this phenomenon in MS, we developed a modified version of the frozen-section assay. Peripheral blood mononuclear cells (PBM) from 26 MS patients, 26 healthy controls and 10 patients with other inflammatory non- neurological diseases (OIND) were co-incubated with cryostat sections of normal brain white matter, immunohistochemically labelled with anti-CD45 antibody and counterstained with Giemsa stain. CD45-positive PBM adherent to transected microvasculature were counted with an automated image analyzer. MS patients showed an increased number of vessel-bound PBM (48.8 +/- 36.4) with respect to healthy controls (27.4 +/- 20.7, P = 0.01) and OIND patients (22.6 +/- 7.8, P = 0.01). Significant differences were also obtained counting the number of vessel-bound PBM as a percent of total vascular cells between MS patients (12.7 +/- 7.2%) and healthy controls (6.9 +/- 5.4%, P = 0.002) or OIND patients (7.4 +/- 4.4%, P = 0.03). We confirm that PBM from MS patients show an increased potential of binding to cerebral vessels. The frozen-section assay provides a unique tool to study in situ the molecular interactions of leukocytes with brain vascular structures.

[Proliferative activity of stage I testicular neoplasms: evaluation by image analysis of immunoreactive MIB-1]. / L'attività proliferativa delle neoplasie del testicolo in stadio I: valutazione mediante analisi di immagine dell'immunoreattività MIB-1.

The proliferative activity has been evaluated in 55 cases of seminoma and 36 cases of non-seminomatous germ cell tumors of the testis (NSGCT) in clinical stage I from patients alive and well within 5 years, by a semi-quantitative image analysis method of determination of the percent MIB-1 positive nuclear areas, using a Leica Quantimet 500+ analyzer. Significant differences of growth fraction have been detected between classical seminomas and seminomas showing more than 3 mitoses/1HPF (anaplastic seminoma)-mean of MIB-1 positivity in classic seminoma 23.8%, versus 43.9% in anaplastic seminoma; confirming this value of cut-off in distinguishing two classes of cases with different growth fractions, but without any difference as far as prognosis is concerned. The non seminoma- germ cell tumours (NSGCT) in toto display a growth fraction significantly higher than that of seminomas (mean value of MIB-1 positivity: 26.3% in seminoma versus 40.4% in NSGCT), but absolute values of positivity as high as those obtained by others by subjective evaluation have not been observed. A higher growth fraction in NSGCT is positively correlated with an embryonal carcinoma component as high as 80% of the tumor mass and/or with vascular invasion, but not with different histotypes. The proliferative activity doesn't stratify subgroups with different prognosis within the two groups of germinal tumours of the testis (seminomas and NSGCT) in our cases of clinical stage I testicular tumours.

[Correlation of the histological and cytogenetic pictures in placental tissue from early abortion. Does immunohistochemistry have a role?]. / Correlazione tra aspetti istologici e quadri citogenetici nei tessuti placentari da aborto precoce. L'immunoistochimica ha un ruolo?

Ninety-four cases of early abortion have been studied. Five histological groups of lesion have been identified by routine histological techniques on abortion materials, group I corresponding to partial hydatidiform mole. Cytogenetic analyses have revealed chromosome anomalies in near 50% of cases with a prevalence of triploidies followed by trisomies and monosomies. Normal histological findings are more often associated with normal karyotypes and group I with abnormal karyotypes but a specific correlation between histological pattern and cytogenetic anomalies is lacking. Neither some histochemical reactions nor the well preserved immunohistochemical reactivities of beta-hCG, hPL, PLAP, AFP, cytokeratin, vimentin, desmin, factor VIII, CD 68, MIB1 (growth fraction), EGF-R, p53 and c-erbB-2 oncoproteins have disclosed specific chromosome anomalies. They have only allowed a better definition of histological groups. A simple histological evaluation, although extended to immunohistochemical reaction may not substitute the cytogenetic analyses, not even for purposes of preselection.

Scarff-bloom-richardson histoprognostic grading correlates with the immunohistochemical expressions of genomic alterations in infiltrating ductal carcinomas (nos) of the breast.

The Scarff-Bloom-Richardson (SBR) multiparametric histological grading has been correlated with the immunohistochemical expression of EGF-R, c-erbB-2 and p53 oncoproteins, with the growth fraction (Ki67 antibody) and with the receptor status (ER, PgR) in 365 infiltrating ductal carcinomas of the breast (IDC-NOS). Specimens of carcinomas after surgery were sectioned and a section of each lesion was formalin-fixed and paraffin-embedded, and stained by hematoxylin-eosin in order to classify and grade cases. Another section was liquid nitrogen frozen, cryostatcut and immunostained using monoclonal antibodies against EGF-R (455 and 528 clones), c-erbB-2 (3B5 clone), p53 (Pab 1801 clone) and Ki67 antigen. An ABC-peroxidase was used after incubation with biotinylated antimouse antibody. Colour was developed using a DAB solution. ER-ICA and PgR-ICA Kits (Abbott) served to detect the hormonal receptor status. A significant direct correlation between SBR and the immuno-histochemical markers (EGF-R, c-erbB-2, p53, Ki67 growth fraction) was found. An inverse relationship of grade to ER and a weaker one to PgR was evident. An increasing histological grade was found parallel with the progressive appearance of one, two or three immunohistochemical markers in the same tumour.

Relationships between epidermal growth factor receptor (EGF-R) and other predictors of prognosis in breast carcinomas. An immunohistochemical study.

The importance of epidermal growth factor receptor (EGF-R) as an immunohistochemical factor of prognosis has been investigated in 820 cases of breast carcinoma irrespective of subtyping. An immunohistochemical membrane positivity for EGF-R (Ab1, clone 455 and Ab2, clone 528, Oncogene Science Manhasset NY, USA, ABC-peroxidase method) has been observed in neoplastic cells of 131/820 breast carcinomas (15.9%); the percentage is lower than those of the majority of reported series, but the total number of cases is higher. A significant inverse relationship between EGF-R and estrogen/progesterone receptors has been found (ER-ICA, PgR-ICA, Abbott, PAP-method). Highly proliferating Ki67 positive (> = 20% stained nuclei-Dakopatts Denmark) and oncoprotein p53 (Pab 1801 clone, Oncogene Science) positive carcinomas are more frequently EGF-R positive (p < 0.001). No relationship exists between EGF-R expression, c-erbB-2 (3B5 clone, Oncogene Science) expression, tumor size and lymph node status. The detection of EGF-R may be a useful addition to other immunohistochemical prognosticators, but it must be related with clinical outcome in further studies.

Epidermal growth factor receptor immunohistochemistry in different histological types of infiltrating breast carcinoma.

AIMS: To determine the immunohistochemical expression of epidermal growth factor receptor (EGF-R) in high grade, intermediate, and low grade tumours. METHODS: Specimens from 931 breast carcinomas were partly formalin fixed and paraffin wax embedded, to classify cases, and partly frozen in liquid nitrogen, cryostat sectioned, and immunostained using two monoclonal antibodies from clone 455 and 528 to demonstrate EGF-R positive cells. An avidin-biotin complex and peroxidase method was used after incubation with biotinylated anti-mouse antibody; colour was developed using a diaiminobenzidine solution. RESULTS: Low grade carcinomas seldom expressed EGF-R (n = 3) compared with 106 high grade infiltrating ductal carcinomas: EGF-R positive cases were much less common in infiltrating lobular than in infiltrating ductal carcinoma. Medullary carcinomas did not differ from infiltrating ductal carcinomas. CONCLUSIONS: The very low incidence of EGF-R positive cases in the "special type" group of breast carcinomas with a good prognosis is in line with the absence of the homologous c-erbB-2 and p53 oncoproteins, and the rarity of highly proliferating and oestrogen/progesterone negative cases. EGF-R expression in infiltrating lobular carcinoma was in keeping with the intermediate behaviour of this kind of tumour. EGF-R expression in cases of pure medullary carcinoma is the same as that of high grade tumours.

Relationship between p53 expression and other prognostic factors in human breast carcinoma. An immunohistochemical study.

Among 843 cases of breast cancer, p53 oncoprotein was detected by the monoclonal antibody (MoAb) Pab-1801 in only 13%. Low-grade carcinomas (tubular, mucinous, papillary, and invasive cribriform types) did not express p53 protein, but it was observed in 4.2% of infiltrating lobular carcinomas (6 of 140 cases) and 50% of pure medullary carcinomas (5 of 10 cases). In intermediate-grade neoplasms, no correlation was seen between p53 status and other putative determinants of a poor prognosis. The latter included high tumor stage, lymph nodal involvement, high growth fraction (as determined by labeling with the MoAb Ki-67), negative results for estrogen receptor (ER) and progesterone receptor (PR) proteins, and amplification of the c-erbB-2 oncogene product in the neoplastic cells. Ninety-nine of 640 (15.5%) cases of high-grade, invasive, ductal breast carcinoma, however, showed an inverse relationship between expression of p53 protein and positive results for ER/PR proteins and a direct correlation with large tumor size, Ki-67-determined growth fraction, and amplification of c-erbB-2 oncopeptide. All of the latter associations were highly significant statistically. The authors conclude that mutant p53 protein may serve a prognostic role in a subset of cases of invasive ductal mammary carcinoma.