Immunohistochemical properties of the endothelial cells in the human uterus during the menstrual cycle.

Expression of vascular cell adhesion molecule (VCAM) and endothelial leukocyte adhesion molecule (ELAM)-1 on human uterine endothelium was examined by immunohistochemistry. Endomyometrial blocks were obtained from 26 hysterectomy specimens (n = 6, menstrual; n = 6, follicular; n = 14, luteal). VCAM and ELAM-1 expression was compared to that of the classic endothelial markers Factor VIIIRA, CD31, CD34 and CD36, which were found to be present in samples taken from all stages of the cycle. Factor VIIIRA, CD31 and CD34 were expressed on most endothelium whereas CD36 was expressed primarily on that of small vessels. CD36 was also found to be strongly expressed on endometrial epithelium and no changes were apparent during the menstrual cycle. ELAM-1 expression was not detected in any of the tissues examined. VCAM expression varied during the cycle, appearing on endometrial endothelium in the mid- and late luteal phases, stages at which large granular lymphocytes migrate into endometrium. VCAM or a VCAM-like molecule may be involved in migration of large granular lymphocytes into endometrium.

Comparison between liver and serum concentrations of mannan binding protein.

AIMS: To investigate staining patterns for mannan binding protein (MBP) by immunocytochemistry in liver biopsy specimens from patients with various hepatic disorders; to measure the serum MBP concentration in the patients at the time of biopsy; and to compare these to define further the role of MBP in disease. METHODS: Fifty seven consecutive patients with a variety of types of liver disease were studied. Fresh liver biopsy specimens were immunostained with anti-MBP and graded for intensity of staining. Serum MBP concentrations were measured on samples obtained on the day of biopsy, as were a full range of liver blood tests. RESULTS: MBP was only detectable in liver biopsy specimens from patients with morphological evidence of liver disease. MBP was most prominent in the livers of patients with severe alcoholic liver disease; livers harbouring metastases or showing biliary disease had moderate concentrations. Patients with liver disease were more likely to have raised serum MBP concentrations, but there was no correlation between these values and those found in the biopsy specimens. There was also no significant correlation between either of these concentrations and liver blood test abnormalities. CONCLUSIONS: Patients with liver disease tend to have raised MBP concentrations in both the liver and serum, but the exact relation between the two is as yet undefined.

An immunohistologic study of endothelialization of uteroplacental vessels in human pregnancy--evidence that endothelium is focally disrupted by trophoblast in preeclampsia.

OBJECTIVE: Our objective was to study the endothelial status of the luminal lining of uteroplacental vessels in the human placental bed in normal and abnormal pregnancy in the third trimester. STUDY DESIGN: Six placental basal plates from uncomplicated pregnancies and five from pregnancies complicated by preeclampsia (n = 3), preeclampsia and a small-for-gestational-age infant (n = 1), and diabetes mellitus (n = 1) were accessioned from the archives because of documentation of their containing uteroplacental vessels. Five placental bed biopsy specimens with intraluminal endovascular trophoblast in the third trimester were also studied. Sections were subjected to immunohistochemical analysis with monoclonal and polyclonal antibodies labeling endothelium and trophoblast. RESULTS: In third-trimester normal uncomplicated pregnancies the uteroplacental arteries and veins were completely endothelialized with no disruption of the endothelium. In third-trimester abnormal pregnancies the uteroplacental veins were also completely endothelialized. However, intraluminal endovascular trophoblast was seen within the uteroplacental arteries in eight of the 10 complicated pregnancies; this finding was associated with disruption of the endothelium. CONCLUSION: In preeclampsia there is an aberrant wave of endovascular trophoblast migration in the third trimester, resulting in focal disruption of the endothelium. This may be responsible for the endothelial cell dysfunction thought to be of pathogenetic importance in preeclampsia.

Co-expression of markers for hepatitis delta and hepatitis B viruses in human liver.

Delta hepatitis (HDV) infection can only occur in the presence of hepatitis B (HBV) infection, as HDV requires a coat of HBV surface antigen (HBsAg) for assembly of complete virus. A number of studies have examined the variation of HBV markers in serum and liver during establishment of HDV infection, but none has systematically examined the relationship between the two viruses in individual hepatocytes. Liver biopsies from five patients with HDV/HBV infection were stained for HBsAg, HBV core antigen (HBcAg) and hepatitis D (delta) antigen (HDAg). Double immunostaining was performed with a combination of indirect immunoperoxidase and alkaline phosphatase/antialkaline phosphatase techniques. HDV and HBV antigens were expressed in all five liver biopsies. Co-localization of HBsAg was seen in up to 39% of HDAg positive cells, and HBcAg in up to 8% of HDAg positive cells. HBcAg was detectable in approximately 9% of HBsAg positive cells, and HBsAg in approximately 12% of HBcAg positive cells. HDV can replicate without HBV but ultimately requires HBV to produce complete virus and subsequently infect other cells. In this study the majority of HDV positive cells did not appear to contain HBV markers. This might suggest delta virus replication without assembly, or possibly sequential production/assembly of the virus.

Mannan-binding protein in human liver.

Mannan-binding protein (MBP) is a Ca(2+)-dependent lectin which was first described in 1978 in rabbit liver, and subsequently in serum and liver tissue from humans and a range of animal species. MBP structurally resembles C1q, and may act both as a focus for complement activation on the surface of microorganisms and as an opsonin in its own right. Low serum levels of serum MBP have been described in a group of children known to suffer from severe recurrent infections. MBP has also been reported to behave as an acute phase reactant. This preliminary study has investigated the localisation of MBP in human tissues using material obtained both at post mortem and from diagnostic liver biopsies. Using the IgG fraction of rabbit anti-human MBP, immunoperoxidase staining showed no evidence of significant MBP in a wide range of normal tissues, including liver taken both at post mortem and needle biopsy. However, there was a significant degree of staining for MBP in liver biopsies showing a variety of different pathologies, in particular severely damaged alcoholic livers, and those harbouring metastatic tumour. Moderate degrees of staining were also seen in liver biopsies from patients suffering from chronic biliary disease. It is uncertain whether this localisation of MBP in abnormal liver is an acute phase response, or represents a more fundamental link with liver disease. This question could be the focus for future studies.

JC70: a new monoclonal antibody that detects vascular endothelium associated antigen on routinely processed tissue sections.

A new monoclonal antibody, JC70, raised against a membrane preparation from a spleen affected by hairy cell leukaemia, recognises a membrane bound glycoprotein identical with that of the CD31 group of monoclonal antibodies. The antibody stains a fixation resistant epitope on endothelial cells in benign and malignant conditions in a wide variety of paraffin wax embedded tissue. JC70 stained malignant endothelial cells in 10 angiosarcomas with more consistency than monoclonal or polyclonal antibodies to factor VIII related antigen (FVIII-Rag). In four cases of Kaposi's sarcoma the antibody stained malignant endothelial cells but not spindle cells. It is concluded that antibody JC70 is of value for studying benign and malignant human vascular disorders in routinely processed tissue.

Kupffer cell number is normal, but their lysozyme content is reduced in alcoholic liver disease.

A quantitative immunohistochemical study of Kupffer cells in liver biopsies from alcoholics was carried out. Two markers were compared, lysozyme and an antimacrophage monoclonal antibody EMB/11. The results showed that there is no significant reduction in Kupffer cell numbers in acute and chronic alcoholic liver disease but that there is a reduction in the number of Kupffer cells staining positively for lysozyme. Thus, those clinical phenomena such as endotoxaemia in alcoholic liver disease are not due to reduction in Kupffer cell number but perhaps to a functional deficit in these cells.

Observations favouring Pneumocystis carinii pneumonia as a primary infection: a monoclonal antibody study on paraffin sections.

Pneumocystis carinii pneumonia is characteristic of immunodeficiency and the organism is probably acquired during early childhood. Since infection is only manifest in the lungs, it has been presumed that the organism lies dormant in these tissues following the primary infection. Conventional staining procedures have, however, failed in the absence of pneumonia to demonstrate consistently any forms of Pneumocystis carinii. To study this problem further, lung sections and hilar lymph nodes from immunodepressed adults with and without Pneumocystis carinii pneumonia as well as lung sections from presumed immunocompetent patients were examined for the cyst and trophozoite forms of Pneumocystis carinii using a monoclonal antibody. The organism was only identified in areas of pneumonia, and the source of the organism in these patients may therefore be a new infection with a different human subtype and not, as previously thought, reactivation of a primary infection.

Factor VIII related antigen positive macrophages and acquired immune deficiency syndrome (AIDS): a problem of antibody specificity.

A further histopathological complication of atypical mycobacterial infection in acquired immune deficiency syndrome (AIDS) is reported. Positive reactivity between mycobacterial antibodies within a polyclonal antiserum and mycobacteria within tissues resulted in false positivity with this reagent to factor VIII related antigen. This complication may be avoided either by prior testing of the antiserum, or by purification, or by the use of monoclonal antibodies. Histopathologists examining tissues from patients with AIDS and those with disseminated mycobacterial infections and using immunohistochemical techniques should be aware of this occurrence.

An immunohistological study of factor VIII related antigen and Kaposi's sarcoma using polyclonal and monoclonal antibodies.

Immunohistochemical techniques have been used to demonstrate Factor VIII related antigen (FVIIIRA) in endothelial cells and to study a variety of tumours including Kaposi's sarcoma. Conflicting reports on the presence of this antigen within the spindle cells of Kaposi's sarcoma have been made. These differences may be due to variations in techniques, in the immunoreagents used and in the method of fixation and handling of the specimen. A major factor may also be the lack of a uniform interpretation of positive peroxidase labelling. This study was confined to paraffin-embedded tissues and compared the results and distribution of FVIIIRA labelling in Kaposi's sarcoma using two commercially available polyclonal antibodies and three monoclonal antibodies. The effects of predigesting the sections with trypsin and protease were also evaluated. Positive labelling of tumour spindle cells was accepted only when it matched that seen in labelled endothelial cells. The polyclonal antibodies provided satisfactory localization of FVIIIRA and gave positive results more frequently than the monoclonal antibodies both with and without enzyme digestion. Endothelial labelling was evident in vessels both within and around the tumour but none was seen in the tumour spindle cells. Most previous studies gave no definition of positive labelling but spindle cells were reported as labelled. However, in those few reports where a definition was given the findings are analogous to those in this report, demonstrating the importance of defining positive labelling.